Treatment of complex regional pain syndrome with extracellular vesicle compositions

Abstract

A method for treating a target complex regional pain syndrome is disclosed, which involves administering a therapeutic MSC secretome product prepared by a method comprising the step of culturing bone marrow-derived MSCs under conditions including an oxygen pressure of less than 5% and a culture medium with a pH of less than 7.

Description

[Technical Field]

[0001] cross reference This application claims the benefits of U.S. Provisional Patent Application No. 63 / 507,207, filed on 9 June 2023, which is incorporated herein by reference in whole. [Background technology]

[0002] Complex regional pain syndrome (CRPS) is a severely painful physical disorder that can occur in the limbs after major trauma or surgery, or can develop spontaneously. CRPS is a multi-system disorder involving elements distinct from the autonomic nervous system, but inflammation is the primary driving force behind the overall symptoms. Currently, there are no treatments proven effective for patients with CRPS identified through clinical trials. Safe and effective management of CRPS is needed. [Overview of the Initiative]

[0003] This specification discloses a method for treating a subject with complex regional pain syndrome (CRPS). One aspect of this disclosure is a method for treating a subject with complex regional pain syndrome (CRPS) requiring treatment, comprising the step of administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition comprising extracellular vesicles, wherein at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63 + CD9 - CD81 -This is a method. In some embodiments, the subject's visual analog pain scale (VAS) score decreases by at least 50% after administration compared to before administration. Also disclosed herein is a method for treating complex regional pain syndrome (CRPS) in a subject requiring treatment of CRPS, comprising the step of administering a composition to the subject comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles, wherein the subject's visual analog pain scale (VAS) score decreases by at least 50% after administration compared to before administration. In some embodiments, at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63 + CD9 - CD81 -In some embodiments, the minimum pre-administration VAS score in the affected limb of the subject is greater than 6. In some embodiments, the subject's Brief Pain Scale (BPI) score decreases after administration compared to before administration. In some embodiments, the subject's BPI score decreases by at least 10%, at least 20%, at least 30%, or more than 30% after administration compared to before administration. In some embodiments, the subject's BPI pain interference score decreases by at least 10%, at least 20%, at least 30%, or more than 30% after administration compared to before administration. In some embodiments, the subject's range of motion score increases by at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 95%, or at least 100% after administration compared to before administration. In some embodiments, the subject's Jamar grip strength test score increases after administration compared to before administration. In some embodiments, the increase in the Jamar grip strength test score is in the upper limb. In some embodiments, the subject's grip strength is equalized after administration. In some embodiments, the subject's grip strength increases by at least 5 kg, at least 10 kg, at least 15 kg, or at least 20 kg after administration compared to before administration. In some embodiments, the subject's grip strength increases by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, or at least 210% after administration compared to before administration. In some embodiments, the subject's grip strength increases in the affected limb. In some embodiments, the subject's grip strength increases in the control limb. In some embodiments, the subject experiences improvement in one or more of the following after administration compared to before administration: range of motion of the limbs, range of motion of the joints, changes in spontaneous movement, edema, allodynia, and paresthesia. In some embodiments, the subject's Short Form 36 Health SurveyQuestionnaire (SF-36) scores improve after administration compared to before administration. In some embodiments, the improvement in SF-36 scores includes one or more of the following: improvement in pain, improvement in energy levels, improvement in social functioning, improvement in physical functioning, a positive trend in emotional health, and improvement in overall health. In some embodiments, the subject suffers from chronic, intractable pain in one or more upper or lower limbs. In some embodiments, the subject does not experience any increase or change in pain during the 30 days prior to administration. In some embodiments, the subject experiences a decrease in pain onset, a decrease in swelling, and / or a decrease in burning sensation in the affected limb after administration compared to before administration. In some embodiments, the administration improves hair and / or nail growth in the subject's affected limb. In some embodiments, the therapeutic MSC secretome composition includes the following proteins: ferritin, NUP85, LAMP2, GPR115, serpin Fl, OPN, PAI-1, DAPP1, cathepsin B, semaphorin 6C, PDGF R-alpha, soltirin, serpin B6, Dkk-3, thrombomodulin, PF4, MIF, periostin, furin, TIMP-1, decorin, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, lumican, TIMP-2, SLITRK5, FAP, artemin, DPPII, cIAP-1, pentraxin 3, visfatin, neprilysin, albumin, galectin-1, UNC5H3, IL-20 R-beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midoquin, Calreticulin, Osteoactivin, Regmine, TAZ, Cathepsin L, RBP4, Serpine A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin-3, SIRP alpha, and Syndecane-4 (one or more), as well as: Ferritin, IGFBP-4, IL-1, R6, GSTM1, NUP85, LAMP2, Meprin A, IL-1F10, bIG-H3, GPR115, TGFbl, Ephrin-A4, CD109, Serpine Fl, IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF R-Alpha, Sortiline, Serpine B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, Thrombomodulin, Endoglycan, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periostin, Fulin, TIMP-1, PAPP-A, Decorin, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin Alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin D1, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Parcephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R Beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNFRI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matryptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nidogen-1, CD2, Kallikrein 1, TSLP R, LAMP1, TROY, VCAM-1, Siglec-11, S100A1, PAR1, Thyroid peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF -1 Alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Nuturin, Clement-2, EMMPRIN, Activin RIB, Neuroligin 2, Epilegulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A, NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Paparin-2, Syndecane-3, Jagged1, AKR1C4, Olfactomedine-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin Alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p2'7, Siglec-6, Dectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, C A125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Regmine, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 Beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR Alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Newdecine, ILT4, uPAR, Axl, WIF-1, IL-7 R-Alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Kokurin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL-17 RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, P D-1, Serpine A4, ADAM23, NOV, Galectin-2, Neurexin 3 Beta, TLR3, Sirtuin 2, Numb, IL-28 R Alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR Alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-Selectin, Hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-1RA, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpine A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP(TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-Alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirre13, FLRT1, Galectin-3, CXCL16, JAM-B,DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R Alpha 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurograinin, HE4, IL-23 R, Galectin-7, GALNT3, GITR L, CD14, R-Spondin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospongin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulisin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin-3, ARSB, LIF R Alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP Alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 Alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF-AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP-2, PU.1, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-Gamma R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2,Further comprising at least one protein selected from resistin, HVEM, ENPP-7, syndecane-4, IL-2 Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, galectin-8, CA4, cystatin EM, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7, or IL-13. In some embodiments, the extracellular vesicles are the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106 a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a- 5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-1 36-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-1 46a, hsa-miR-146a-5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir -16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1'7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR- 193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p,hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, h sa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-m ir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3 p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30d- 5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-342-3p, hsa-miR-345-5p, hsa-miR-34a-5p , hsa-miR-361-5p, hsa-miR-376a-3p, hsa-miR-376c-3p, hsa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486 -1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p, or hsa-miR-99b-5p. In some embodiments, the composition is obtained by culturing bone marrow-derived MSCs under the following conditions: (a) an oxygen tension of less than 5%, and (ii) a culture medium with a pH of less than 7.The steps of producing an MSC-conditioned medium, (b) collecting the MSC-conditioned medium, and (c) formulating the MSC-conditioned medium to produce a therapeutic MSC secretome composition, wherein the therapeutic MSC secretome composition is produced by the steps of including proteins and extracellular vesicles produced by bone marrow-derived MSCs in step (a). In some embodiments, the culture medium is serum-free. In some embodiments, the glucose concentration of the culture medium is less than 4.5 g / L. In some embodiments, the subject is human. In some embodiments, the bone marrow-derived MSCs are derived from human bone marrow. In some embodiments, the step of administration includes intravenous administration. In some embodiments, the dosage of the therapeutic MSC secretome composition administered to the subject is a cell equivalent dosage of 0.7 to 7 million cells per kg. In some embodiments, the therapeutic MSC secretome composition contains 4×10, 10 ~10×10 10 cells per ml. In some embodiments, the therapeutic MSC secretome composition contains 5×10 11 ~1.5×10 12 extracellular vesicles. In some embodiments, the composition is administered monthly over a period of 2 months or more, or once every 1 month, 2 months, or 3 months or more. In some embodiments, the composition is administered twice. In some embodiments, the composition is administered at 4-day intervals.

[0004] One aspect of the present disclosure is a method for preparing a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition for treating complex regional pain syndrome (CRPS) in subjects requiring treatment of CRPS, the method comprising: (a) culturing bone marrow-derived MSCs in a culture medium under the following conditions: (i) oxygen tension less than 5%, and (ii) pH less than 7 to produce an MSC-conditioned medium; (b) collecting the MSC-conditioned medium; and (c) formulating the MSC-conditioned medium to produce a therapeutic MSC secretome composition, wherein the therapeutic MSC secretome composition comprises proteins and extracellular vesicles produced by the bone marrow-derived MSCs in step (a). In some embodiments, the culture medium is serum-free. In some embodiments, the glucose concentration of the culture medium is less than 4.5 g / L. In some embodiments, at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63 + CD9 - CD81 -In some embodiments, the bone marrow-derived MSCs are derived from human bone marrow. In some embodiments, the administration step includes intravenous administration. In some embodiments, the therapeutic MSC secretome composition includes the following proteins: ferritin, NUP85, LAMP2, GPR115, serpin Fl, OPN, PAI-1, DAPP1, cathepsin B, semaphorin 6C, PDGF R-alpha, soltirin, serpin B6, Dkk-3, thrombomodulin, PF4, MIF, periostin, furin, TIMP-1, decorin, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, lumican, TIMP-2, SLITRK5, FAP, artemin, DPPII, cIAP-1, pentraxin 3, visfatin, neprilysin, albumin, galectin-1, UNC5H3, IL-20 R-beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midoquin, Calreticulin, Osteoactivin, Regmine, TAZ, Cathepsin L, RBP4, Serpine A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin-3, SIRP alpha, and Syndecane-4 (one or more), as well as: Ferritin, IGFBP-4, IL-1, R6, GSTM1, NUP85, LAMP2, Meprin A, IL-1 F10, bIG-H3, GPR115, TGFbl, Ephrin-A4, CD109, Serpine Fl, IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF R-Alpha, Sortiline, Serpine B6, Dkk-3, CNTF, TSP-1, GM-CSFRa, Thrombomodulin, Endoglycan, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periostin, Fulin, TIMP-1, PAPP-A, Decorin, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin Alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin D1, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Parcephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R Beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matryptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAKR, Nidogen-1, CD2, Kallikrein-1, TSLP R, LAMP1, TROY, VCAM-1, Siglec-11, S100A1, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R Beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF -1 Alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Nuturin, Clement-2, EMMPRIN, Activin RIB, Neuroligin 2, Epilegulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A, NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Paparin-2, Syndecane-3, Jagged1, AKR1C4, Olfactomedine-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin Alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p2'7, Siglec-6, Dectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, C A125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Regmine, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 Beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR Alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Newdecine, ILT4, uPAR, Axl, WIF-1, IL-7 R-Alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Kokurin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL-17 RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGFR, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, Serpine A4, ADAM23, NOV, Galectin-2, Neurexin 3 Beta, TLR3, Sirtuin 2, Numb, IL-28 R Alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-IRA, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpine A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP(TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirre13, FLRT1, Galectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R-alpha 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurograinin, HE4, IL-2 3 R, Galectin-7, GALNT3, GITR L, CD14, R-Spongin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospongin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulisin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin-3, ARSB, LIF R Alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP alfa, uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 alfa, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF-AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP-2, PU.1, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-Gamma-R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein-11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 Rg, MICA, dopadecarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, galectin-8, CA4, cystatin EM, FUT8, B7-H3,The extracellular vesicle further comprises at least one protein selected from GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7, or IL-13. In some embodiments, the extracellular vesicle comprises the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p, hsa-m ir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, h sa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a-5p , hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1'7 -5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p , hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir- 21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p,Comprising one or more of hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-342-3p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-376a-3p, hsa-miR-376c-3p, hsa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p, or hsa-miR-99b-5p.,

[0005] One aspect of this disclosure is the use of a composition produced by any one of the methods described herein in the treatment of complex regional pain syndrome (CRPS) in a subject requiring treatment for CRPS. In some embodiments, the subject is human. In some embodiments, the subject's visual analog pain scale (VAS) score decreases by at least 50% after administration compared to before administration. In some embodiments, the subject's minimum pre-administration VAS score in the affected limb is greater than 6. In some embodiments, the subject's brief pain assessment (BPI) score decreases after administration compared to before administration. In some embodiments, the subject's BPI score decreases by at least 10%, at least 20%, at least 30%, or more than 30%, after administration compared to before administration. In some embodiments, the subject's BPI pain interference score decreases by at least 10%, at least 20%, at least 30%, or more than 30%, after administration compared to before administration. In some embodiments, the range of motion assessment score of the subject increases by at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 95%, or at least 100% after administration compared to before administration. In some embodiments, the Jamar grip strength test score of the subject increases after administration compared to before administration. In some embodiments, the increase in the Jamar grip strength test score is in the upper limb. In some embodiments, the grip strength of the subject is equalized after administration. In some embodiments, the grip strength of the subject increases by at least 5 kg, at least 10 kg, at least 15 kg, or at least 20 kg after administration compared to before administration. In some embodiments, the grip strength of the subject increases by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, or at least 210% after administration compared to before administration. In some embodiments, the grip strength of the subject increases in the affected limb. In some embodiments, the grip strength of the subject increases in the control limb. In some embodiments, subjects experience improvement in one or more of the following after administration compared to before administration: range of motion of the limbs, range of motion of the joints, changes in spontaneous movement, edema, allodynia, and paresthesia.In some embodiments, the Short Form 36 Health Survey Questionnaire (SF-36) scores of the subject improve after administration compared to before administration. In some embodiments, the improvement in the SF-36 questionnaire scores includes one or more of improvement in pain, improvement in energy level, improvement in social function, improvement in physical function, a positive trend in emotional health, and improvement in general health. In some embodiments, the subject suffers from chronic refractory pain in one or more upper or lower limbs. In some embodiments, the subject does not experience an increase or change in pain during the 30 days prior to administration. In some embodiments, the subject experiences a sudden decrease in pain, a decrease in swelling, and / or a decrease in the feeling of burning in the affected limb after administration compared to before administration. In some embodiments, administration improves hair and / or nail growth in the affected limb of the subject. In some embodiments, the dosage of the therapeutic MSC secretome composition administered to the subject is a cell equivalent dosage of 0.7 to 7 million cells per kg. In some embodiments, the therapeutic MSC secretome composition contains 4×10 10 ~10×10 10 cells per ml. In some embodiments, the therapeutic MSC secretome composition contains 5×10 11 ~1.5×10 12 extracellular vesicles. In some embodiments, the composition is administered monthly over a period of 2 months or more, or once every 1 month, 2 months, or 3 months or more. In some embodiments, the composition is administered twice. In some embodiments, the composition is administered at 4-day intervals.

[0006] One aspect of the present disclosure is the use of a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles in the treatment of complex regional pain syndrome (CRPS) in a subject requiring treatment of CRPS, wherein at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63 + CD9 - CD81 -This is the use. In some embodiments, the composition is administered intravenously to the subject. In some embodiments, the subject's Visual Analog Pain Scale (VAS) score decreases by at least 50% after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition. In some embodiments, the minimum VAS score in the affected limb of the subject before administration of the therapeutic MSC secretome composition is greater than 6. In some embodiments, the subject's Brief Pain Response Inventory (BPI) score decreases after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

[0007] Another aspect of this disclosure is the use of a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles in the treatment of complex regional pain syndrome (CRPS) in subjects requiring treatment of CRPS, wherein the subject's visual analog pain scale (VAS) score decreases by at least 50% after administration of the therapeutic MSC secretome composition compared to before administration. In some embodiments, the minimum VAS score in the affected limb of the subject before administration of the therapeutic MSC secretome composition is greater than 6. In some embodiments, the subject's brief pain assessment (BPI) score decreases after administration of the therapeutic MSC secretome composition compared to before administration. In some embodiments, at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63 + CD9 - CD81 -In some embodiments, the therapeutic MSC secretome composition includes the following proteins: ferritin, NUP85, LAMP2, GPR115, serpin Fl, OPN, PAI-1, DAPP1, cathepsin B, semaphorin 6C, PDGF R-alpha, soltirin, serpin B6, Dkk-3, thrombomodulin, PF4, MIF, periostin, furin, TIMP-1, decorin, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, lumican, TIMP-2, SLITRK5, FAP, artemin, DPPII, cIAP-1, pentraxin 3, visfatin, neprilysin, albumin, galectin-1, UNC5H3, IL-20 R-beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midoquin, Calreticulin, Osteoactivin, Regmine, TAZ, Cathepsin L, RBP4, Serpine A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin-3, SIRP alpha, and Syndecane-4 (one or more), as well as: Ferritin, IGFBP-4, IL-1, R6, GSTM1, NUP85, LAMP2, Meprin A, IL-1 F10, bIG-H3, GPR115, TGFbl, Ephrin-A4, CD109, Serpine Fl, IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF R-Alpha, Sortiline, Serpine B6, Dkk-3, CNTF, TSP-1, GM-CSFRa, Thrombomodulin, Endoglycan, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periostin, Fulin, TIMP-1, PAPP-A, Decorin, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin Alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin D1, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Parcephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R Beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matryptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAKR, Nidogen-1, CD2, Kallikrein-1, TSLP R, LAMP1, TROY, VCAM-1, Siglec-11, S100A1, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R Beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF -1 Alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Nuturin, Clement-2, EMMPRIN, Activin RIB, Neuroligin 2, Epilegulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A, NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Paparin-2, Syndecane-3, Jagged1, AKR1C4, Olfactomedine-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin Alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p2'7, Siglec-6, Dectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, C A125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Regmine, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 Beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR Alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Newdecine, ILT4, uPAR, Axl, WIF-1, IL-7 R-Alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Kokurin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL-17 RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGFR, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, Serpine A4, ADAM23, NOV, Galectin-2, Neurexin 3 Beta, TLR3, Sirtuin 2, Numb, IL-28 R Alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-IRA, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpine A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP(TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirre13, FLRT1, Galectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R-alpha 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurograinin, HE4, IL-23 R, Galectin-7, GALNT3, GITR L, CD14, R-Spongin 2, CK19, Cardiotrophin-1, TREML1, HAPLIN1, CD27, ANG4, Siglec-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospongin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulisin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin-3, ARSB, LIF R Alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP Alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 Alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF-AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP-2, PU.1, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-Gamma-R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein-11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 The present invention further comprises at least one protein selected from Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, galectin-8, CA4, cystatin EM, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7, or IL-13. In some embodiments,Extracellular vesicles contain the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a-5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1’7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19’7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a-5phsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-m iR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30 d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa- miR-342-3p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-376a-3p, hsa-mi R-376c-3p, hsa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-48 6-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, h Includes one or more of the following: sa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p, or hsa-miR-99b-5p. In some embodiments, the composition is produced by the steps of (a) culturing bone marrow-derived MSCs in a culture medium with a pH of less than 7 under the following conditions: (i) an oxygen tension of less than 5% and (ii) a pH of less than 7 to produce an MSC-conditioned medium; (b) collecting the MSC-conditioned medium; and (c) formulating the MSC-conditioned medium to produce a therapeutic MSC secretome composition, wherein the therapeutic MSC secretome composition comprises proteins and extracellular vesicles produced by the bone marrow-derived MSCs in step (a). In some embodiments, the culture medium is serum-free. In some embodiments, the glucose concentration of the culture medium is less than 4.5 g / L. In some embodiments, the target BPI score is at least 10%, at least 20%, at least 30% after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.or decreases by more than 30%. In some embodiments, the subject's BPI pain interference score decreases by at least 10%, at least 20%, at least 30%, or more than 30% after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition. In some embodiments, the subject's range of motion assessment score increases by at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 95%, or at least 100% after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition. In some embodiments, the subject's Jamar grip strength test score increases after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition. In some embodiments, the increase in the Jamar grip strength test score is in the upper limbs. In some embodiments, the subject's grip strength is equalized after administration of the therapeutic MSC secretome composition. In some embodiments, the subject's grip strength increases by at least 5 kg, at least 10 kg, at least 15 kg, or at least 20 kg after administration of the therapeutic MSC secretome composition compared to before administration. In some embodiments, the subject's grip strength increases by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, or at least 210% after administration of the therapeutic MSC secretome composition compared to before administration. In some embodiments, the subject's grip strength increases in the affected limb. In some embodiments, the subject's grip strength increases in the control limb. In some embodiments, in the subject, one or more of the following improve after administration of the therapeutic MSC secretome composition compared to before administration: range of motion of the limbs, range of motion of the joints, changes in spontaneous movement, edema, allodynia, and paresthesia. In some embodiments, the Short Form 36 Health Survey Questionnaire (SF-36) score in question was compared to the score before administration of the therapeutic MSC secretome composition.Improvement occurs after administration of the therapeutic MSC secretome composition. In some embodiments, improvement in SF-36 questionnaire scores includes one or more of the following: improvement in pain, improvement in energy levels, improvement in social functioning, improvement in physical functioning, positive trend in emotional health, and improvement in overall health. In some embodiments, the subject suffers from chronic refractory pain in one or more upper or lower limbs. In some embodiments, the subject does not experience any increase or change in pain during the 30 days prior to administration of the therapeutic MSC secretome composition. In some embodiments, the subject experiences a reduction in pain onset, a reduction in swelling, and / or a reduction in burning sensation in the affected limb after administration of the therapeutic MSC secretome composition compared to before administration. In some embodiments, administration of the therapeutic MSC secretome composition improves hair and / or nail growth in the subject's affected limb.

[0008] Reference The patents, publications, and non-patent documents cited herein are each incorporated herein by reference in whole, as if each were each incorporated by reference individually. If any publication or patent or patent application incorporated by reference conflicts with any disclosure contained herein, this specification is intended to supersede and / or take precedence over any such conflicting material. [Brief explanation of the drawing]

[0009] The features of this disclosure are specifically stated in the appended claims. The features and advantages of this disclosure will be better understood by referring to the following detailed description, which specifies exemplary embodiments in which the principles of this disclosure are utilized, and to the appended drawings (also referred to herein as "Figure" and "FIG.").

[0010] [Figure 1]This is a box plot of the mean visual analog scale (VAS) pain assessment results over a 6-month trial period. The box identifies the 25th and 75th percentiles with an internal line identifying the median and a "+" indicating the mean. Upper and lower whisker lines indicate the maximum and minimum data values, respectively. Individual data points are represented as single dots (·) (*p<0.05, **p<0.01). [Figure 2] This plot shows the results of the simplified pain index (A for pain interference, B for pain severity) assessment over a 6-month trial period. The plot and p-value markers are shown in Figure 1. [Figure 3] This plot shows the results of the SF-36 assessment over a 6-month trial period. The categories are A) Pain, B) Energy / Fatigue, C) Social Functioning, D) Physical Functioning, E) Physical Limitation, F) Emotional Limitation, G) Emotional Health, and H) General Health. The plots and p-value markers are shown in Figure 1. [Figure 4] This figure shows the results of range of motion evaluations over a 6-month trial period as a bar graph. [Figure 5] This graph quantifies the results of the Jamar grip strength meter test evaluation over a 6-month trial period and shows them in bar graph format. [Modes for carrying out the invention]

[0011] I. Definition As used herein and in the appended claims, the singular forms “a,” “an,” and “the” refer to multiple objects unless otherwise specified. Also note that the term “or” is generally used to mean “and / or” unless otherwise specified. As used herein, the terms “and / or,” “any combination thereof,” and their grammatical equivalents are interchangeable. These terms can convey that any combination is specifically intended. For illustrative purposes only, the following phrases “A, B, and / or C” or “A, B, C, or any combination thereof” may mean A individually, B individually, C individually, A and B, B and C, A and C, and A, B, and C. The term “or” may be used conjunctively or disjunctively unless the context specifically refers to a disjunctive use.

[0012] The terms “about” or “approximately” can mean within an acceptable margin of error for a particular value, which may depend in part on how that value is measured or determined, for example, on the limitations of the measurement system. For example, “about” could mean within or above one standard deviation. Alternatively, “about” could mean a range of up to 20%, up to 10%, up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term could mean being within one order of magnitude, up to five times, or up to two times the value. Where a particular value is described in this application and claims, unless otherwise specified, the term “about” should be assumed to mean within an acceptable margin of error for that particular value.

[0013] Throughout this disclosure, numerical features are presented in range form. It should be understood that this range form is for convenience and brevity only and should not be interpreted as an inflexible limitation on the scope of all embodiments. Therefore, unless otherwise explicitly indicated in the context, range descriptions should be interpreted as disclosing possible subranges as well as individual numerical values ​​within that range to a lower limit of one-tenth of a unit. For example, a range description such as 1–6 should be considered to disclose not only subranges such as 1–3, 1–4, 1–5, 2–4, 2–6, 3–6, as well as individual values ​​within that range, e.g., 1.1, 2, 2.3, 5, and 5.9. This applies regardless of the width of the range. These intervening upper and lower limits may independently be included within smaller ranges and, furthermore, are subject to any specifically excluded limitations within the ranges contained and expressed in this disclosure. If a specified range includes one or both of the limit values, the range excluding one or both of those limit values ​​is also included in this disclosure unless the context explicitly indicates otherwise.

[0014] As used herein and in the claims, the words “comprising” (and any form such as “comprise” or “comprises”), “having” (and any form such as “have” or “has”), “including” (and any form such as “includes” or “include”), or “containing” (and any form such as “contains” or “contain”) are inclusive or open-ended and do not preclude any additional elements or method steps not enumerated. All embodiments discussed herein can be implemented with respect to any method or composition of the Disclosure, and vice versa. Furthermore, the compositions of the Disclosure can be used to achieve the methods of the Disclosure.

[0015] References to “some embodiments,” “an embodiment,” “one embodiment,” or “other embodiments” in this specification mean that certain features, structures, or characteristics described in relation to that embodiment are included in at least some embodiments of this disclosure, but not necessarily in all embodiments. To facilitate understanding of this disclosure, several terms and phrases are defined below.

[0016] To provide a complete understanding of the various embodiments, certain specific details are described herein. However, those skilled in the art will understand that this disclosure can be implemented without these details. In other instances, well-known techniques or methods are neither shown nor described in detail to avoid unnecessarily obscuring the description of the embodiments. Unless otherwise required in context, throughout this specification and the following claims, the word “comprise,” and variations such as “comprises” and “comprising,” should be interpreted as having an open and comprehensive meaning, i.e., “including, but not limited.” Furthermore, the headings provided herein are for convenience only and do not imply any interpretation of the scope or meaning of the claimed embodiments.

[0017] Similar or equivalent methods and materials described herein may be used in the implementation or testing of this disclosure, although preferred methods and materials are described below.

[0018] II. Therapeutic composition In addition to the components used to prepare the compositions disclosed herein, the compositions themselves used in the methods disclosed herein are disclosed. These and other materials are disclosed herein, and where combinations, subsets, interactions, groups, etc., of these materials are disclosed, specific references to each of the various individual combinations, collective combinations, and permutations of these compounds may not be expressly disclosed, and should be understood that each is specifically intended and described herein. For example, where a particular MSC secretome (including, but not limited to, MSC exosomes (with or without growth factors) referred herein as extracellular vesicle isolation products (EVIPs)) is disclosed and discussed, and several modifications that can be made to several molecules containing the MSC secretome are discussed, specifically intended are all possible combinations and permutations of the MSC secretome, and modifications that are possible unless specifically indicated in the opposite sense. Therefore, if molecular classes A, B, and C, as well as molecular classes D, E, and F, and example molecular combination AD are disclosed, even if each is not individually enumerated, it is construed individually and collectively that combinations AE, AF, BD, BE, BF, CD, CE, and CF are disclosed. Similarly, any subset or combination thereof is also disclosed. For example, the subgroups AE, BF, and CE are considered to be disclosed. This concept applies to all aspects of this application, including but not limited to steps in a method for preparing and using the compositions of this disclosure. Therefore, if there are various additional steps that can be performed, it is understood that each of these additional steps can be performed by any particular embodiment or combination of embodiments of the method of this disclosure.

[0019] The primary nutritional characteristics of MSCs are the secretion of growth factors and exosomes that induce cell proliferation and angiogenesis. Exosomes express mitotic proteins such as transforming growth factor-alpha (TGF-α), TGF-β, hepatocyte growth factor (HGF), epidermal growth factor (EGF), basic fibroblast growth factor (FGF-2), and insulin-1-like growth factor-1 (IGF-1). These promote the division of fibroblasts, epithelial cells, and endothelial cells. Vascular endothelial growth factor (VEGF), IGF-1, EGF, and angiopoietin-1 are released to recruit endothelial cells and initiate angiogenesis. MSCs support this process through paracrine mechanisms and regulate the regenerative environment through anti-inflammatory and immunomodulatory mechanisms. In response to inflammatory molecules such as interleukin-1 (IL-1), IL-6, IL-2, IL-12, tumor necrosis factor-α (TNF-α), and interferon-gamma (INF-γ), MSCs secrete an array of growth factors and anti-inflammatory proteins with complex feedback mechanisms among many types of immune cells. Key immunomodulatory cytokines include prostaglandin 2, TGF-131, HGF, SDF-1, nitrous oxide, indoleaniline 2,3-dioxygenase, IL-4, IL-10, IL-1 receptor antagonists, and soluble tumor necrosis factor-α receptors. MSCs interfere with the proliferation and function of many inflammatory immune cells, including T cells, natural killer cells, B cells, monocytes, macrophages, and dendritic cells. While MSCs can modulate T cell activity across species, these mechanisms are not identical across mammalian species.

[0020] A characteristic feature of a chronic inflammatory environment is a persistent imbalance in helper T cell and macrophage types. MSC exosomes indirectly promote the transition from TH1 to TH2 cells by reducing INF-γ and increasing IL-4 and IL-10. Restored TH1 / TH2 balance has been shown to improve tissue regeneration in cartilage, muscle, and other soft tissue injuries, alleviate symptoms of autoimmune diseases, and exert anti-diabetic effects. Similarly, reduced INF-γ and increased IL-4 secretion promote the shift of macrophages from M1 (pro-inflammatory, anti-angiogenic, and tissue growth inhibitory) to M2 (anti-inflammatory, pro-remodeling, and tissue healing) types, which are necessary for the healing and regeneration of the skeleton, muscle, and nerves.

[0021] This specification discloses complex compositions of extracellular vesicles containing secreted biomolecules (proteins, lipids, and ribonucleic acids) and / or biomolecules derived from mesenchymal cells. In one embodiment of this specification, a composition is disclosed comprising a therapeutically effective amount of MSC secretome (e.g., a composition comprising MSC growth factors, MSC exosomes, MSC extracts, and / or extracellular vesicles) and one or more biomolecules (e.g., peptides, polypeptides, proteins, siRNAs, shRNAs, and / or microRNAs (miRNAs)).

[0022] In some embodiments, the therapeutic composition includes the following proteins: ferritin, NUP85, LAMP2, GPR115, serpine Fl, OPN, PAI-1, DAPP1, cathepsin B, semaphorin 6C, PDGF R-alpha, soltirin, serpine B6, Dkk-3, thrombomodulin, PF4, MIF, periostin, furin, TIMP-1, decorin, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, lumican, TIMP-2, SLITRK5, FAP, artemin, DPPII, cIAP-1, pentraxin 3, visfatin, neprilysin, albumin, galectin-1, UNC5H3, IL-20 R-beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midoquin, Calreticulin, Osteoactivin, Regmine, TAZ, Cathepsin L, RBP4, Serpine A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin-3, SIRP alpha, and Syndecane-4 (one or more), as well as: Ferritin, IGFBP-4, IL-1, R6, GSTM1, NUP85, LAMP2, Meprin A, IL-1 F10, bIG-H3, GPR115, TGFbl, Ephrin-A4, CD109, Serpine Fl, IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF R-Alpha, Sortiline, Serpine B6, Dkk-3, CNTF, TSP-1, GM-CSFRa, Thrombomodulin, Endoglycan, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periostin, Fulin, TIMP-1, PAPP-A, Decorin, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin Alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin D1, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Parcephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R Beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matryptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAKR, Nidogen-1, CD2, Kallikrein-1, TSLP R, LAMP1, TROY, VCAM-1, Siglec-11, S100A1, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R Beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF -1 Alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Nuturin, Clement-2, EMMPRIN, Activin RIB, Neuroligin 2, Epilegulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A, NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Paparin-2, Syndecane-3, Jagged1, AKR1C4, Olfactomedine-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin Alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p2'7, Siglec-6, Dectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, C A125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Regmine, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 Beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR Alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Newdecine, ILT4, uPAR, Axl, WIF-1, IL-7 R-Alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Kokurin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL-17 RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGFR, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, Serpine A4, ADAM23, NOV, Galectin-2, Neurexin 3 Beta, TLR3, Sirtuin 2, Numb, IL-28 R Alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-IRA, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpine A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP(TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirre13, FLRT1, Galectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R-alpha 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurograinin, HE4, IL-23 R, Galectin-7, GALNT3, GITR L, CD14, R-Spongin 2, CK19, Cardiotrophin-1, TREML1, HAPLIN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospongin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulisin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin-3, ARSB, LIF R Alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP Alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 Alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, M er, TREM-2, Flt-3L, CDS, IL-6, CD229, insulin, syntaxin 6, GRO, Bcl-w, lipocalin-2, PDGF-AA, IL-2 Ra, angiogenin, LYVE-1, CD4, RAGE, CDNF, brevican, NAP-2, PU.1, EDAR, ADAMTS13, kynureninase, PTH1R, IFN-gamma-R1, CrkL, B7-1, PARC, draxin, VE-cadherin, procalcitonin, SOX15, kallikrein-11, BCMA, dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, resistin, HVEM, ENPP-7, syndecane-4, IL-2 Rg, MICA, Dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, Glycoprotein V, Semaphorin 4G, IL-12p40, PSA-Total, IL-15, MAP1D, Clq, TNF4, Dtk, Endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, Galectin-8, CA4, Cystatin EM, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S10 Any combination of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 or more of these proteins, or all of these proteins.

[0023] Extracellular vesicles (EVs) are small, membrane-bound spheres containing proteins and RNA (of which exosomes are a subset). Exosomes are small-diameter (e.g., 20–150 nm) lipid bilayer vesicles secreted by cells to enable paracrine signaling. Other EV populations originate directly from the plasma membrane or are formed during apoptosis (apoptotic bodies). This specification discloses compositions containing therapeutically effective amounts of MSC secretomes (e.g., compositions comprising MSC growth factors, MSC exosomes, MSC extracts, and / or extracellular vesicles). In some embodiments, the therapeutic composition includes the following nucleic acids: HSA-LET-7A-5P, HSA-LET-7B-5P, HSA-LET-7C-5P, HSA-LET-7D-3P, HSA-LET-7E-5P, HSA-LET-7G-5P, HSA-LET-7I, HSA-LET-7I-5P, HSA-MIR-100-5P, HSA-MIR-103a-3P, HSA-MIR-106a-5 p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hs a-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, h sa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-m iR-146a-5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-m ir-16-2, hsa-miR-16-5p, hsa-miR-1'7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-m iR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p,hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p hsa-mir-25 hsa-miR-25-3p hsa-miR-26a-5p hsa-miR-27a-3p hsa-mir-27b hsa-miR-27b-3p hsa-miR-29a-3p hsa-miR-29c-3p hsa-miR-30a- 5p、hsa-miR-30a-5p、hsa-miR-30b-5p、hsa-miR-30c-5p、hsa-mir-30d、hs a-miR-30d-5p、hsa-mir-30e、hsa-miR-30e-5p、hsa-miR-31-3p、hsa-miR-3 1-5p, hsa-miR-320a, hsa-miR-342-3p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-376a-3p, hsa-miR-376c-3p, hsa-miR-423-3p hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a- 5p、およびhsa-miR-99b-5pのうち1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、 17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、40、45、50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 or more in any combination, or containing all of these nucleic acids.

[0024] Exemplary microRNA contents may include human miRNA sequences hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7g-5p, hsa-let-7i-5p, hsa-miR-214-3p, and hsa-miR-27a-3p, all of which have binding sites in TMPRSS2 mRNA.

[0025] In some embodiments, the therapeutic composition is CD63 + CD9 - CD81 - The composition contains extracellular vesicles having the phenotype of CD63. In some embodiments, at least 70, 75, 80, 85, 90, 91, 92, 93, 94, or 95% of the extracellular vesicles in the therapeutic composition are CD63. + CD9 - CD81 - In some embodiments, at least 50, 60, 70, 80, 85, 90, 91, 92, 93, 94, or 95% of the extracellular vesicles in the therapeutic composition are CD9 - In some embodiments, at least 50, 60, 70, 80, 85, 90, 91, 92, 93, 94, or 95% of the extracellular vesicles in the therapeutic composition are CD81. - That is the case.

[0026] In some embodiments, MSCs cultured to produce therapeutic compositions have the ability to undergo tricellular differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes. In some embodiments, MSCs are positive for CD73, CD105, CD166, and CD90, and negative for CD14, CD31, CD34, and CD45.

[0027] The MSC secretome is understood and intended herein to include exosomes and growth factors. The growth factors and exosomes may be allogeneic or autologous. The growth factors and exosomes may originate from any cell in the human body, such as ectoderm cells, endoderm cells, or mesoderm cells. For example, the MSC secretome may include mesenchymal stem cell (MSC)-derived growth factors, MSC-derived exosomes, or both MSC-derived growth factors and exosomes. In some embodiments, the method further includes the step of adding at least one additive together with the exosomes and growth factors. Specifically, MSCs under appropriate wound healing conditions may produce suitable therapeutic agents, such as exosomes and growth factors, that can provide treatment for inflammatory lung diseases. In one embodiment herein, the MSC secretome composition comprises prostaglandin E2 (PGE2), transforming growth factor 131 (TGF-I31), hepatocyte growth factor (HGF), stromal cell-derived factor-1 (SDF-1), nitrate oxide, indoleamine 2,3-dioxygenase, interleukin-4 (IL-4), IL-6, interleukin-10 (IL-10), IL-1 receptor antagonist and soluble TMF-α receptor, insulin-like growth factor, fibroblast growth factor (FGF) 1-23 (Patent A composition is disclosed further comprising FGF1 and FGF2), bone morphogenetic proteins (BMPs) 1-15, epidermal growth factor (EGF), transforming growth factor-α (TGF-α) macrophage-stimulating protein (MSP), platelet-derived growth factor (PLGF), vascular endothelial growth factor (VEGF), macrophage colony-stimulating factor (M-CSF), insulin, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF estrogen), and / or thyroid hormones.

[0028] Embodiments of therapeutic compositions described herein may include proteins and microRNAs, some of which may be embedded in or surrounded by lipid membranes to form vesicles ranging in size from approximately 20 nm to approximately 200 nm. The number of vesicles in the composition may be between approximately 1 million and 100 billion per mL when suspended, or between approximately 10 million and 1 trillion when formulated as a lyophilized powder.

[0029] A. Delivery of pharmaceutical carriers / medicines The therapeutic compositions described herein may be administered in vivo in a pharmaceutically acceptable carrier. “pharmaceutically acceptable” means a material that is not biologically or otherwise undesirable; that is, the material can be administered to a subject together with a nucleic acid or vector without causing any undesirable biological effects or adverse interactions with any other components of the pharmaceutical composition containing the material. The carrier is, of course, selected to minimize any degradation of the active ingredient and minimize any adverse side effects in the subject, as is well known to those skilled in the art. The composition may be administered orally, parenterally (e.g., intravenously), by intramuscular injection, by intraperitoneal injection, transdermally, extracorporeally, or topically, including by intranasal administration or inhalation. As used herein, “intranasal administration” means delivery of the composition to the nose and / or nasal passages through one or both nostrils, and may include delivery by spray or droplet mechanisms, or by aerosolization of the nucleic acid or vector.

[0030] The composition can be administered by inhalation via the nose or mouth, for example, by delivery using a spray or droplet mechanism such as a metered-dose inhaler, dry powder inhaler, nebulizer, or vaporization device. Delivery can also be directly to any area of ​​the respiratory system (e.g., the lungs) via intubation. The exact amount of composition required varies from subject to subject, depending on the species, age, weight, and general condition of the subject, the severity of the disorder being treated, and the mode of administration.

[0031] Parenteral administration of a composition, when used, is characterized by injection as a whole. Injectable preparations can be prepared conventionally as a liquid solution or suspension, as a solid suitable for solution of a suspension in a liquid before injection, or as an emulsion. Parenteral administration may involve the use of a sustained-release or continuous-release system such that a constant dose is maintained.

[0032] Suitable carriers and their formulations are described in Remington: The Science and Practice of Pharmacy (19th edition) ed. ARGennaro, Mack Publishing Company, Easton, PA 1995. Typically, the formulation is made isotonic by using an appropriate amount of a pharmaceutically acceptable salt in the formulation. Examples of pharmaceutically acceptable carriers include, but are not limited to, physiological saline, Ringer's solution, and dextrose solution. The pH of the solution is preferably about 5 to about 8, more preferably about 7 to about 7.5. Furthermore, the carrier comprises a sustained-release preparation such as a semipermeable matrix of a solid hydrophobic polymer containing the antibody, the matrix being in the form of a molded article, e.g., a film, liposomes, or microparticles. It will be obvious to those skilled in the art that the specific carrier may more preferably depend, for example, on the route of administration and the concentration of the composition being administered.

[0033] Pharmaceutical carriers are known to those skilled in the art. These are, most typically, standard carriers for drug administration to humans, including solutions such as sterile water, physiological saline, and buffered solutions of physiological pH. The composition may be administered intramuscularly or subcutaneously. Other compounds will be administered according to standard procedures used by those skilled in the art.

[0034] The pharmaceutical composition may contain, in addition to the selected molecule, a carrier, a thickener, a diluent, a buffer, a preservative, a surfactant, and the like. The pharmaceutical composition may also contain one or more active ingredients such as an antibacterial agent, an anti-inflammatory agent, or an anesthetic.

[0035] The pharmaceutical composition may be administered in several ways, depending on whether topical or systemic treatment is desired and the area to be treated. Administration may be topical (including ophthalmic, vaginal, rectal, and intranasal administration), oral, inhalation, or parenteral administration, such as intravenous infusion, subcutaneous injection, intraperitoneal injection, or intramuscular injection. The antibodies of this disclosure may be administered intravenously, intraperitoneally, intramuscularly, subcutaneously, intracavitarially, or transdermally.

[0036] Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcohol solutions / aqueous solutions, emulsions, or suspensions, including saline and buffering media. Parenteral vehicles include sodium chloride solutions, ringer's dextrose, dextrose, and sodium chloride, lactated Ringer's solution, or fixative oils. Intravenous vehicles include fluids and nutritional supplements, electrolyte supplements (such as those based on ringer's dextrose), etc. Preservatives and other additives may also be present, such as antibacterial agents, antioxidants, chelating agents, and inert gases. Preparations for topical administration may include ointments, lotions, creams, gels, drops, suppositories, sprays, solutions, and powders. Conventional pharmaceutical carriers, aqueous bases, powder bases, or oily bases, thickeners, etc., may be essential or desirable.

[0037] Compositions for oral administration may include powders or granules, suspensions or solutions in water or a non-aqueous medium, capsules, sachets, or tablets. Thickeners, flavoring agents, diluents, emulsifiers, dispersing agents, or binders may also be desirable.

[0038] Possibly, portions of this composition may be administered as pharmaceutically acceptable acid addition salts or base addition salts formed by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, and phosphoric acid, as well as organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, and fumaric acid, or by reaction with inorganic bases such as sodium hydroxide, ammonium hydroxide, and potassium hydroxide, as well as organic bases such as mono-, di-, trialkyl and arylamines and substituted ethanolamines.

[0039] B. Therapeutic use Effective dosages and schedules for administering this composition can be determined empirically, and making such determinations is within the scope of the art. The dosage range for administering this composition should be large enough to produce the desired effect in which the symptoms of the disorder are affected. The dosage should not be large enough to cause adverse side effects such as unwanted cross-reactions or anaphylactic reactions. In general, the dosage can be determined by those skilled in the art, depending on the patient's age, condition, sex, and the severity of the disease, the route of administration, or whether other drugs are included in the regimen. The dosage may be adjusted by the individual physician if any contraindications exist. The dosage may vary and may be administered once or multiple times daily for one or several days, and / or once or multiple times monthly for two months or more, or once every one, two, or three months or more. Guidelines can be found in the literature on appropriate dosages for a given class of medicinal products.

[0040] C. Mesenchymal stem cells and therapeutic secretome compositions The therapeutic compositions disclosed herein utilize MSC secretomes and / or growth factors derived from mesenchymal stem cells (MSCs). In one embodiment herein, MSC secretome compositions (including, but not limited to, compositions comprising MSC growth factors, MSC exosomes, MSC extracts, and / or extracellular vesicles) are disclosed. These therapeutic compositions may be used, for example, to treat, inhibit, reduce, alleviate, and / or prevent conditions such as complex regional pain syndrome (CRPS).

[0041] MSCs are pluripotent cells that have the ability to differentiate into a number of cell types, including muscle cells, chondrocytes, adipocytes, and osteoblasts. Typically, these cells can be found in the amniotic fluid, including the placenta, umbilical cord blood, adipose tissue, bone marrow, or perivascular tissue. As used herein, “MSC” refers to non-terminal differentiated cells, including but not limited to pluripotent stem cells, pluripotent stromal cells, stromal angiocytes, pericytes, perivascular cells, stromal cells, pluripotent cells, multipotent cells, adipocyte-derived fibroblast-like cells, adipocyte-derived stromal vascular fraction, adipocyte-derived MSCs, bone marrow-derived fibroblast-like cells, bone marrow-derived stromal vascular fraction, bone marrow-derived MSCs, tissue-derived fibroblast-like cells, adult stem cells, adult stromal cells, keratinocytes, and / or melanocytes.

[0042] In addition to their differentiation potential, MSCs possess immunomodulatory capabilities that result in the expression of many different cytokines and growth factors. As used herein, “MSC preparation” or “MSC secretome composition” refers to a composition comprising cell-free extracts of MSCs and / or MSC lysates obtained from human MSCs, fibroblast-like cells, and non-human animal MSCs, including but not limited to MSCs derived from horses, cattle, pigs, sheep, non-human primates, dogs, cats, rabbits, rats, and mice. In some embodiments, the MSCs may be derived from the patient to whom the composition is applied (autologous) or from another individual (allogeneic). MSCs may be collected in conditioned medium, cultured to increase the amount of cells relative to the lysate, or freshly used before being incorporated into the compositions of this disclosure. An MSC secretome composition (including, but not limited to, a composition comprising MSC growth factors, MSC exosomes, MSC extracts, and / or extracellular vesicles) may contain about 0.00001 to about 20% by weight, for example, about 0.01 to about 10% by weight, of mesenchymal stem cell (MSC) extracts, MSC exosomes, or MSC growth factor preparations. The MSC preparation may contain either MSC-conditioned medium or MSC lysates from cell-cultured and grown MSCs. In some embodiments, the composition may further contain about 0.01 to about 10% by weight of cell-free medium conditioned by the growth of MSCs or MSC-lineage cells, where the cells are cultured under normal hyper-oxygenated culture conditions or artificial wound healing conditions.

[0043] As disclosed herein, MSCs used to produce the MSC additives of this disclosure (including growth factor secretome compositions which are either freeze-additives or powder-additives) can be selectively stimulated to produce MSC growth factors, secretomes, cytokines, chemokines, mesenchymal stem cell proteins, peptides, glycosaminoglycans, extracellular matrix (ECM), proteoglycans, secretomes, and exosomes. The growth factors and exosomes may be derived from any cell in the human body, such as ectoderm cells, endoderm cells, or mesoderm cells. As used herein, MSC growth factors include prostaglandin E2 (PGE2), transforming growth factor 131 (TGF-(31)), hepatocyte growth factor (HGF), stromal cell-derived factor-1 (SDF-1), nitrate oxide, indoleamine 2,3-dioxygenase, interleukin-4 (IL-4), IL-6, interleukin-10 (IL-10), IL-1 receptor antagonist and soluble TMF-α receptor, insulin-like growth factor, fibroblast growth factor (FGF) 1-23 (in particular, FGF Examples of hormones include, but are not limited to, estrogen and thyroid hormones, as well as 1 and FGF2), bone morphogenetic proteins (BMPs) 1-15, epidermal growth factor (EGF), transforming growth factor-α (TGF-α), macrophage-stimulating protein (MSP), platelet-derived growth factor (PLGF), vascular endothelial growth factor (VEGF), macrophage colony-stimulating factor (M-CSF), insulin, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and other hormones.

[0044] MSC culture may be carried out under wound healing and / or hypoxic conditions. Hypoxic conditions may include approximately 1% to approximately 5% oxygen, small or zero amounts of serum, small amounts of glucose, or various combinations of these elements. The combination of small amounts of nutrients and metabolites in the environment may induce tissue healing by inducing cultured cells to produce wound healing and anti-inflammatory ECM proteins and growth factors. Direct tissue healing is likely to be in the form of growth factors and cytokines, as well as novel ECM proteins such as collagen and glycosaminoglycans (GAGs). In one embodiment, an MSC preparation (e.g., an MSC secretome composition) includes MSC growth factors, MSC exosomes, and / or cell extracts of MSCs or MSC lysates obtained from MSCs cultured under standard hyper-oxygen culture conditions (e.g., 21% oxygen) or MSCs cultured under artificial wound healing conditions (e.g., 0.1% to approximately 5% oxygen).

[0045] As disclosed herein, artificial wound healing conditions simulate growth conditions in a real wound, where there is reduced nutrient supply and reduced waste removal that normally occurs due to disruption of local blood circulation. This creates a harsh environment for cells until new blood vessels are formed and blood circulation is restored. Therefore, artificial wound healing conditions used for culturing MSCs may include one or more of the following growth conditions: reduced glucose availability, reduced oxygen tension, reduced pH, and increased temperature.

[0046] In some embodiments, glucose availability can be reduced compared to a normal control (e.g., 4.5 g / L). In modified culture media that reduce glucose without damaging cells, glucose may be reduced by 0-50%, more preferably by about 5-40%. For example, under MSC artificial wound healing culture conditions, glucose may be reduced by about 5-15%, 10-20%, 15-25%, 20-30%, or 25-35%. In some embodiments, glucose may be present in a range of about 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, or 4.0 g / L, or between any two of these values. In some embodiments, glucose is present at concentrations of less than or equal to 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, or 4.5 g / L.

[0047] In some embodiments, the oxygen tension can be reduced to oxygen levels corresponding to hypoxic conditions. Normal atmospheric oxygen is approximately 21%, and levels below this are considered hypoxic. Therefore, in one embodiment, MSCs can be cultured at oxygen levels between 0.0% and 20.9%, between approximately 1% and 0.5%, between approximately 0.1% and 2.0%, between approximately 0.1% and 5.0%, between approximately 0.5% and 5.0%, between approximately 1.0% and 1%, between approximately 5.0% and 10.0%, and between approximately 10.0% and 15.0%. Hypoxic conditions can be a form of artificial wound healing conditions.Oxygen tension is between approximately 0.5% and 20.5% of oxygen when culturing MSCs to produce therapeutic secretome compositions containing extracellular vesicles and / or MSC secreted growth factors, e.g., 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4, 10.5, 10 0.6, 10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, 15, 15.1, 15.2, 15.3, 15.4, 15.5, 15.6, 15 This could be 0.7, 15.8, 15.9, 16, 16.1, 16.2, 16.3, 16.4, 16.5, 16.6, 16.7, 16.8, 16.9, 17, 17.1, 17.2, 17.3, 17.4, 17.5, 17.6, 17.7, 17.8, 17.9, 18, 18.1, 18.2, 18.3, 18.4, 18.5, 18.6, 18.7, 18.8, 18.9, 19, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.0% oxygen, or oxygen in the range between any two of these values.

[0048] The pH can also be lowered during MSC culture. The pH can be approximately 6.0 to 7.4, for example, 6.0 to 6.4, 6.2 to 6.4, 6.2 to 6.6, 6.4 to 6.6, 6.4 to 6.8, or 6.6 to 7.0, for example, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, or 7.4.

[0049] The temperature of the culture environment may be raised above the physiological homeostasis temperature (e.g., 37°C). In one embodiment, the culture conditions for MSCs may include approximately 35°C to 39°C, approximately 35°C to 36°C, approximately 36°C to 37°C, approximately 37°C to 38°C, approximately 38°C to 39°C, and approximately 39°C to 40°C. In one embodiment, the culture temperature may be 35.0, 35.1, 35.2, 35.3, 36.4, 35.5, 35.6, 35.7, 35.8, 35.9, 36.0, 36.1, 36.2, 36.3, 36.4, 36.5, 36.6, 36.7, 36.8, 36.9, 37.0, 37.1, 37.2, 37.3, 37.4, 37 The temperature can be 0.5, 37.6, 37.7, 37.8, 37.9, 38.0, 38.1, 38.2, 38.3, 38.4, 38.5, 38.6, 38.7, 38.8, 38.9, 39.0, 39.1, 39.2, 39.3, 39.4, 39.5, 39.6, 39.7, 39.8, 39.9, or 40.0℃.

[0050] In some embodiments, the culture medium is serum-free. In some embodiments, the serum-free culture medium contains platelet lysate. In some embodiments, the platelet lysate is human platelet lysate (HPL). In some embodiments, the serum-free culture medium contains at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% HPL by volume, or HPL in a range between any two of these values. In some embodiments, the culture medium contains 8%–12%, 5%–15%, or 9%–11% HPL by volume.

[0051] In one embodiment, an MSC secretome composition (including, but not limited to, a composition comprising MSC growth factors, MSC exosomes, MSC extracts, and / or extracellular vesicles) may further include a protective coating (e.g., a cryoprotectant oligosaccharide and protein solution) that reduces the degradation of the growth factors. It is understood and intended herein that the protective coating may be designed as a polymer. "Polymer" refers to a relatively high molecular weight organic compound (natural or synthetic) whose structure can be represented by monomers, which are repeating small units. Non-limiting examples of polymers include polyethylene, rubber, and cellulose. Synthetic polymers are typically formed by addition polymerization or condensation polymerization of monomers. The term "copolymer" refers to a polymer formed from two or more different repeating units (monomer residues). By example, but not limited to, copolymers may be alternating copolymers, random copolymers, block copolymers, or graft copolymers. In certain embodiments, it is also intended that various block segments of a block copolymer themselves may constitute the copolymer. The term "polymer" encompasses all polymer forms, including but not limited to natural polymers, synthetic polymers, homopolymers, heteropolymers or copolymers, and addition polymers. In one embodiment, the gel matrix may include copolymers, block copolymers, diblock copolymers, and / or triblock copolymers. In one embodiment, the protective coating may include biocompatible polymers. In one embodiment, the biocompatible polymers may be crosslinked. Such polymers may also have the function of slowly releasing fat browning agents and / or fat modifiers into the tissue.As used herein, biocompatible polymers include polysaccharides; hydrophilic polypeptides; poly(amino acids) such as poly-L-glutamic acid (PGS), gamma-polyglutamic acid, poly-L-aspartic acid, poly-L-serine, or poly-L-lysine; polyalkylene glycols and polyalkylene oxides such as polyethylene glycol (PEG), polypropylene glycol (PPG), and poly(ethylene oxide) (PEO); poly(oxyethylated polyols); poly(olefin alcohols); polyvinylpyrrolidone); poly(hydroxyalkyl methacrylamide); poly(hydroxyalkyl methacrylate); poly(saccharides); poly(hydroxy acids); poly(vinyl alcohols), polyhydroxy acids such as poly(lactic acid), poly(glycolic acid), and poly(lactic acid-co-glycolic acid); poly-3-hydroxybutyrate or poly-4-hydroxy Examples include, but are not limited to, polyhydroxyalkanoates such as butyrates; polycaprolactones; poly(orthoesters); polyanhydrides; poly(phosphazenes); poly(lactide-co-caprolactone); polycarbonates such as tyrosine polycarbonates; polyamides (including synthetic and natural polyamides), polypeptides, and poly(amino acids); polyesteramides; polyesters; poly(dioxanone); poly(alkylene alkylates); hydrophobic polyethers; polyurethanes; polyether esters; polyacetals; polycyanoacrylates; polyacrylates; polymethyl methacrylates; polysiloxanes; poly(oxyethylene) / poly(oxyethylene) copolymers; polyketals; polyphosphates; polyhydroxyvalerates; polyalkylene oxalates; polyalkylene succinates; poly(maleic acid), and their copolymers.Biocompatible polymers also include polyamides, polycarbonates, polyalkylenes, polyalkylene glycols, polyalkylene oxides, polyalkylene terephthalates, polyvinyl alcohol (PVA), methacrylate PVA (m-PVA), polyvinyl ethers, polyvinyl esters, polyvinyl halides, polyvinylpyrrolidones, polyglycolides, polysiloxanes, polyurethanes and their copolymers, alkylcellulose, hydroxyalkylcellulose, cellulose ethers, cellulose esters, nitrocellulose, polymers of acrylic acid esters and methacrylate esters, methylcellulose, ethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxybutylmethylcellulose, cellulose acetate, cellulose propionate, cellulose acetate butyrate, cellulose acetate phthalate, carboxymethylcellulose This may also include polycellulose, cellulose triacetate, sodium cellulose sulfate, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate), polyethylene, polypropylene, poly(ethylene glycol), poly(ethylene oxide), poly(ethylene terephthalate), poly(vinyl alcohol), poly(vinyl acetate), polyvinyl chloride polystyrene and polyvinylpyrrolidone, their derivatives, their linear and branched copolymers and block copolymers, and their formulations.Examples of biodegradable polymers include polyesters, poly(orthoesters), poly(ethyleneamines), poly(caprolactones), poly(hydroxybutyrates), poly(hydroxyvalerates), polyanhydrides, poly(acrylic acids), polyglycolides, poly(urethanes), polycarbonates, polyphosphate esters, polyphospliazenes, their derivatives, their linear and branched copolymers and block copolymers, and their formulations.

[0052] In some embodiments, the protective coating comprises carbohydrate polymers such as monosaccharides, non-reducing polysaccharides, or disaccharides, and carbohydrate constructs of any combination thereof. Examples of carbohydrates that can be used for the protective coating include glucose, aldoses (D-allose, D-altrose, D-mannose, etc.), glucopyranose, pentahydroxyhexanal, aD-glucopyranosyl-D-glucose, aD-glucopyranosyl-dihydrate, polymers of PD-glycopyranosyl units, PD-fructofuranosyl aD-glucopyranosyl (anhydrous / dihydrate), f3-D-galactopyranosyl-D-glucose, aD-glucopyranosyl-aD-glucopyranosyl (anhydrous / dihydrate) In addition to starch, the product contains galactose, pentoses (ribose, xylose, lyxose), dextrose, dodecacarbon monodecahydrate, fructose, sucrose, lactose, maltose, trehalose, agarose, D-galactosyl-0-(1-4)-anhydro-L-galactosyl, cellulose, PD-glycopyranosyl unit polymers, and starch, as well as polyhydric alcohols, polyalcohols, algitol, erythritol, glycitol, glycerol, xylitol, and sorbitol.

[0053] In some embodiments, the protective coating contains biocompatible and / or biodegradable polyesters or polyanhydrides such as poly(lactic acid), poly(glycolic acid), and poly(lactic acid-co-glycolic acid). These particles may contain one or more of the following polyesters: glycolic acid units referred to herein as "PGA," lactic acid units referred to herein collectively as "PLA," such as poly-L-lactic acid, poly-D-lactic acid, poly-D,L-lactic acid, poly-L-lactide, poly-D-lactide, and poly-D,L-lactide 5, caprolactone units referred to herein collectively as "PCL," such as poly(e-caprolactone), and copolymers referred to herein collectively as "PLGA," which include lactic acid units and glycolic acid units, for example, various forms of poly(lactic acid-co-glycolic acid) and poly(lactide-co-glycolide) characterized by a lactic acid:glycolic acid ratio, and their derivatives. Exemplary polymers also include copolymers of polyethylene glycol (PEG) and the aforementioned polyesters, collectively referred to herein as “PEGylated polymers,” such as various forms of PLGA-PEG or PEA-PEG copolymers. In certain embodiments, the “PEGylated polymer” can be obtained by covalently bonding the PEG region to the polymer via a cleavable linker. In one embodiment, the polymer contains at least 60, 65, 70, 75, 80, 85, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent of acetal pendant groups.

[0054] The triblock copolymers disclosed herein include core polymers such as polyethylene glycol (PEG), polyvinyl acetate, polyvinyl alcohol, polyvinylpyrrolidone (PVP), polyethylene oxide (PEO), polyvinylpyrrolidone-co-vinyl acetate, polymethacrylate, polyoxyethylene alkyl ether, polyoxyethylene castor oil, polycaprolactam, polylactic acid, polyglycolic acid, poly(lactic-glycolic) acid, poly(lactic-co-glycolic) acid (PLGA), and cellulose derivatives such as hydroxymethylcellulose and hydroxypropylcellulose. Examples of diblock copolymers that can be used in protective coatings disclosed herein include polymers such as polyethylene glycol (PEG), polyvinyl acetate, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyethylene oxide (PEO), poly(vinylpyrrolidone-co-vinyl acetate), polymethacrylate, polyoxyethylene alkyl ether, polyoxyethylene castor oil, polycaprolactam, polylactic acid, polyglycolic acid, poly(lactic acid-glycolic acid), and poly(lactic acid-co-glycolic acid) (PLGA).

[0055] In one embodiment, the protective coating (i.e., the encapsulated composition) may further comprise lecithin or hydrolyzed lecithin as a carrier or encapsulating material. As used herein, lecithin and / or hydrolyzed lecithin coatings include coatings comprising phosphatidylcholine, phosphatidylinositol, phosphatidylethanolamine, phosphatidylserine, and phosphatidic acid. The source of lecithin may be a plant or animal source.

[0056] In one embodiment, the polymer, monosaccharide, disaccharide, or polysaccharide used to form the protective coating formed by introducing the MSC additive into the encapsulation solution can be at a concentration suitable for forming the protective coating. For example, the polymer, monosaccharide, disaccharide, or polysaccharide can be at any concentration between 0.01 mM and 10.0 M, for example, about 0.01 M to about 0.1 M, about 0.1 mM to about 1.0 M, or about 1.0 M to about 10.0 M.

[0057] In one embodiment, the MSC secretome compositions disclosed herein (including, but not limited to, compositions comprising MSC growth factors, MSC exosomes, MSC extracts, and / or extracellular vesicles) may contain any known components commonly found in the pharmaceutical field, such as free radical countermeasures, bactericides, metal ion chelating agents, preservatives, basicizing or acidifying agents, fragrances, surfactants, fillers, natural products or extracts of natural products, such as aloe or green tea extracts, vitamins, or coloring materials. Other components that may be combined with the powder may include antioxidants, which may be selected from a variety of antioxidants. Suitable antioxidants include vitamins, such as vitamin C (L-ascorbate, ascorbate-2-magnesium phosphate, ascorbyl palmitate, tetrahexyldecyl ascorbate), vitamin E (tocotrienol), vitamin A (provitamin A carotenoids such as retinol, retinal, retinoic acid, and beta-carotene), N-acetylglucosamine, or other glucosamine derivatives. Other components may include at least one essential fatty acid, such as S2-3, S2-6, and S2-9 polyunsaturated fatty acids, such as linoleic acid (LA), gamma-linoleic acid (GLA), alpha-linoleic acid (ALA), dihomo-γ-linoleic acid (DGLA), and arachidonic acid (ARA). Fatty acids may be derived from various sources, including evening primrose oil, blackcurrant oil, borage oil, or GLA-modified safflower seed oil. Other components may include a platelet-rich fibrin matrix, at least one component that supports ECM production and hyaluronic acid production, such as N-acetylglucosamine or other glucosamine derivatives, ultra-low molecular weight (ULMW) hyaluronic acid, chondroitin sulfate, or keratin sulfate.

[0058] The production of an MSC secretome composition may include culturing MSCs collected from a donor to create a culture medium under culture conditions containing reduced oxygen and nutrients (in some embodiments), stimulating cultured cells to selectively secrete desired anti-inflammatory proteins, peptides, glycosaminoglycans, proteoglycan exosomes, and secretomes by adjusting cell growth conditions, collecting aggregate mixtures and combining them with a mounting solution, freezing the aggregate mixture containing exosomes, peptides, proteins, cytokines, growth factors, extracellular matrix (ECM), proteoglycans, and glycosaminoglycans, and chemokines selected from the group consisting of human MSCs, animal MSCs, pluripotent stromal cells, fibroblasts, and fibroblasts, freezing the aggregate mixture in combination with a mounting solution such as an oligosaccharide such as a trehalose solution or a protein solution, and freeze-drying the frozen mixture to produce a dry powder. Alternatively, MSCs may be lysed in such a way that all MSCs are collected from the culture process, an extract lysate is produced, the extract lysate is concentrated, and the extract lysate is combined with a mounting solution such as an oligosaccharide such as a trehalose solution or protein solution, the mixture is frozen, and the frozen mixture is freeze-dried or lyophilized to produce a dry powder. The powder contains a highly concentrated aggregate of analgesic MSC secretomes and exosomes, as well as extracellular matrix components specific to anti-inflammatory properties.

[0059] This method may also include filtering, sterilizing, concentrating, freezing, or lyophilizing the MSC-conditioned culture medium. Furthermore, the MSC culture medium may be combined with a cryoprotectant before freezing.

[0060] Various methods exist for lysing MSCs. Lysis can be achieved by adding a hypotonic solution or by repeated freeze-thaw processes that disrupt the cell membrane. Furthermore, cells may be lysed while attached to the culture surface or in suspension. Cells may also be enzymatically released and / or lysed by mechanical homogenization.

[0061] Stimulating MSCs to selectively secrete desired anti-inflammatory proteins, peptides, glycosaminoglycans, proteoglycans, exosomes, and secretomes can be achieved by regulating cell growth conditions such as cell density, culture medium supplements, nutrient supplements, oxygen levels, length of culture under conditions, cell passage number, and combinations thereof.

[0062] III. Treatment of Complex Regional Pain Syndrome (CRPS) In some embodiments, the therapeutic compositions disclosed herein are used in a method of treating a subject's complex regional pain syndrome (CRPS). Any of the therapeutic compositions described herein may be used in such a method. In some embodiments, the subject suffers from chronic, intractable pain in one or more upper or lower limbs. In some embodiments, the subject experiences no increase or change in pain for 30 days prior to treatment. In some embodiments, the subject experiences one or more of the following in response to treatment: reduced pain, increased energy, reduced fatigue, increased social functioning, increased emotional health, increased physical functioning, decreased changes in spontaneous movement, reduced edema, reduced allodynia, reduced paresthesia, improved motor skills, and / or improved grip strength. In some embodiments, the subject experiences an increased range of motion in one or more joints in response to treatment. In some embodiments, the subject experiences a decrease in sudden pain onset, reduced swelling, and / or reduced burning sensation in response to treatment. In some embodiments, the treatment improves hair and / or nail growth in the subject's affected limb. In some embodiments, the subject experiences a reduction in pain severity and / or pain relief in response to the treatment.

[0063] In some embodiments, the investigational drug of this disclosure is administered to CRPS patients in two doses. In some embodiments, 15 mL of the investigational drug is administered per dose. In some embodiments, the investigational drug is administered by intravenous infusion. In some embodiments, the investigational drug is administered on separate days. In some embodiments, the investigational drug is administered at intervals of 1, 2, 3, 4, 5, 6, or 7 days. In some embodiments, the effect of the treatment lasts for at least 6 months or at least 1 year.

[0064] This specification also discloses a method for treating target CRPS, comprising the step of administering to the target a composition comprising secretory extracellular vesicles containing any combination of proteins and / or miRNAs selected from: ferritin, NUP85, LAMP2, GPR115, serpin Fl, OPN, PAI-1, DAPP1, cathepsin B, semaphorin 6C, PDGF R-alpha, soltirin, serpin B6, Dkk-3, thrombomodulin, PF4, MIF, periostin, furin, TIMP-1, decorin, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, lumican, TIMP-2, SLITRK5, FAP, artemin, DPPII, cIAP-1, pentraxin 3, visfatin, neprilysin, albumin, galectin-1, UNC5H3, IL-20 One or more of the following: R-beta, SREC-II, JAM-C, TNF-RI, htPAPP-A, eNOS, MSP-R, TPP1, LAMP1, B2M, NCAM-1, HIF-1-alpha, ST6GAL1, CD99-L2, Plexin-A4, EMMPRIN, p53, Semaphorin-7A, NKp80, Cystatin-B, Osteoadherin, Midoquin, Calreticulin, Osteoactivin, Regmine, TAZ, Cathepsin-L, RBP4, Serpine-A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin-3, SIRP-alpha, and Syndecane-4, as well as: Ferritin, IGFBP-4, IL-1-R6, GSTM1, NUP85, LAMP2, Meprin-A, IL-1 F10, bIG-H3, GPR115, TGFbl, Ephrin-A4, CD109, Serpine Fl, IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF R-Alpha, Sortiline, Serpine B6, Dkk-3, CNTF, TSP-1, GM-CSFRa, Thrombomodulin, Endoglycan, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periostin, Fulin, TIMP-1, PAPP-A, Decorin, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin Alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin D1, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Parcephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R Beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matryptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAKR, Nidogen-1, CD2, Kallikrein-1, TSLP R, LAMP1, TROY, VCAM-1, Siglec-11, S100A1, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R Beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF -1 Alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Nuturin, Clement-2, EMMPRIN, Activin RIB, Neuroligin 2, Epilegulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A, NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Paparin-2, Syndecane-3, Jagged1, AKR1C4, Olfactomedine-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin Alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p2'7, Siglec-6, Dectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, C A125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Regmine, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 Beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR Alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Newdecine, ILT4, uPAR, Axl, WIF-1, IL-7 R-Alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Kokurin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL-17 RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGFR, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, Serpine A4, ADAM23, NOV, Galectin-2, Neurexin 3 Beta, TLR3, Sirtuin 2, Numb, IL-28 R Alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-IRA, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpine A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFA5, Galectin-9, vWF-A2, TACE, Activin RUB, Cathepsin S, LDL R, BMPR-IA, OX40, IL-3 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDT5, ROR2, EphB4, Glypican 1, LAP(TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10Rb, XEDAR, IL-22, PILR-alpha, NRG1-bl, FABP4, RGM-A, RELT, TrkC, C5a, SREC-I, Nestin, TPO, ErbB3, Kirrel3, FLRT1, Galectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R-alpha 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurograinin, HE4, IL-23 R, Galectin-7, GALNT3, GITR L, CD14, R-Spongin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF RH, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospongin-5, IL-1 MI, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulisin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin-3, ARSB, LIF R Alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP Alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, N Kp46, MCP-3, IL-32 alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF-AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP-2, PU.1, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-Gamma-R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein-11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 At least one protein selected from the group consisting of Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, galectin-8, CA4, cystatin EM, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7, IL-13, hsa-let-7a-5p, hsa-let-7b-5p, hsa-le t-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p , hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b , hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p,hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a-5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1'7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, h sa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-30a-5p, hsa-miR-30a- 5p、hsa-miR-30b-5p、hsa-miR-30c-5p、hsa-mir-30d、hsa-miR-30d-5p、hs a-mir-30e、hsa-miR-30e-5p、hsa-miR-31-3p、hsa-miR-31-5p、hsa-miR-32 0a、hsa-miR-342-3p、hsa-miR-345-5p、hsa-miR-34a-5p、hsa-miR-361-5p、hsa-miR-376a-3p、hsa-miR-376c-3p、hsa-miR-423-3p、hsa-miR-423-5p、hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p, and hsa-miR-99b-5p. ,

[0065] In some embodiments, the therapeutic product is administered in a cellular equivalent dose range of 0.7 to 7 million cells per kg. In some embodiments, the therapeutic product is administered in a cellular equivalent dose of at least about, at most about, or about 200,000, 500,000, 700,000, 1,000,000, 1,500,000, 2,000,000, 2,500,000, 3,000,000, 3,500,000, 4,000,000, 4,500,000, 5,500,000, 6,000,000, 6,500,000, 7,000,000, 7,500,000, 8,000,000, 8,500,000, 9,000,000, 9,500,000, 10,000,000, 10,500,000, 11,000,000, 11,500,000, or 12,000,000 cells per kg. In some embodiments, the product is administered in a cellular equivalent dose of 9 × 10⁶ 11 ~1.2 × 10 12 Individual extracellular vesicles, or 5 × 10 11 ~1.5×10 12 , 6×10 11 ~1.4×10 12 , 7×10 11 ~1.3 × 10 12 , 8×10 11 ~1.2 × 10 12 , or 8 x 10 11 ~1.3 × 10 12 It is administered in a dose that yields 5 × 10⁶ extracellular vesicles. In some embodiments, the product yields at least or at most 5 × 10⁶ of the product. 11 , 6×10 11 , 7×10 11 , 8×10 11 , 9×10 11 , 1 x 10 12 , 1.1 × 10 12 , 1.2 × 10 12 , 1.3 × 1012 , 1.4×10 12 , or 1.5 × 10 12 It is administered in a dose that provides 6 × 10⁶ extracellular vesicles per ml. In some embodiments, the therapeutic product is 6 × 10⁶ per ml. 10 ~8×10 10 , 5×10 10 ~9×10 10 , 4×10 10 ~10×10 10 , 5.5×10 10 ~8.5×10 10 , or 6×10 10 ~8.5×10 10 Contains 10 cells. In some embodiments, the therapeutic product contains 6 × 10 cells per ml. 10 ~8×10 10 It contains 6 × 10⁶ extracellular vesicles or cells and is administered in doses of 10-20 ml. In some embodiments, the therapeutic product is 6 × 10⁶ per ml. 10 ~8×10 10 It contains an extracellular vesicle or cell and is administered in doses of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml, or in a range between any two of these values.

[0066] In some embodiments, the subject's visual analog pain scale (VAS) score decreases by at least 50% after administration compared to before administration. In some embodiments, the target VAS score may decrease by at least 50%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% after administration compared to before administration. In some embodiments, the minimum pre-administration VAS score in the affected limb of the target is greater than 6.

[0067] In some embodiments, the target BPI score decreases after administration compared to before administration. In some embodiments, the target BPI score decreases by at least 10%, at least 20%, at least 30%, or more than 30% after administration compared to before administration. In some embodiments, the target BPI score decreases by at least 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61% after administration compared to before administration. The reduction may be 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%. In some embodiments, the BPI pain interference score in question decreases by at least 10%, at least 20%, at least 30%, or more than 30% after administration compared to before administration. In some embodiments, the BPI pain interference score in question is at least 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 30%, 35%, 40%, 45%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 6% after administration compared to before administration. It may decrease by 1%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%.

[0068] In some embodiments, the range of motion assessment score of the subject increases by at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 95%, or at least 100% after administration compared to before administration. In some embodiments, the range of motion score of the target may increase by at least 50%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% after administration compared to before administration.

[0069] In some embodiments, the subject's Jamar dynamometer score increases after administration compared to before administration. In some embodiments, the increase in the Jamar dynamometer score is in the upper limbs. In some embodiments, the subject's grip strength is equalized after administration. In some embodiments, the subject's grip strength increases by at least 5 kg, at least 10 kg, at least 15 kg, or at least 20 kg after administration compared to before administration. In some embodiments, the subject's grip strength increases by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, or at least 210% after administration compared to before administration. In some embodiments, the grip strength of the subject is at least 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 8 The grip strength increases by 5%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 210%, 220%, 230%, 240%, 250%, 260%, 270%, 280%, 290%, or at least 300%. In some embodiments, the subject's grip strength increases in the affected limb. In some embodiments, the subject's grip strength increases in the control limb. In some embodiments, the subject experiences improvement in one or more of the following after administration compared to before administration: range of motion of the limbs, range of motion of the joints, changes in spontaneous movement, edema, allodynia, and paresthesia.

[0070] In some embodiments, the subject's Short Form 36 Health Survey Questionnaire (SF-36) score improves after administration compared to before administration. In some embodiments, the improvement in the SF-36 score includes one or more of the following: improvement in pain, improvement in energy levels, improvement in social functioning, improvement in physical functioning, positive trend in emotional health, and improvement in overall health. In some embodiments, the subject suffers from chronic, intractable pain in one or more upper or lower limbs. In some embodiments, the subject does not experience any increase or change in pain during the 30 days prior to administration. In some embodiments, the subject experiences a decrease in pain onset, a decrease in swelling, and / or a decrease in burning sensation in the affected limb after administration compared to before administration. In some embodiments, the administration improves hair and / or nail growth in the subject's affected limb.

[0071] In some embodiments, subjects that can be treated by the therapeutic products and methods described herein may include subjects that have CRPS or have been diagnosed with CRPS. In some embodiments, subjects may include mammals. In some embodiments, subjects may include humans. In some embodiments, subjects may include non-human mammals. Non-limiting examples of non-human mammals include non-human primates such as chimpanzees, other apes and monkey species, domestic animals such as cattle, horses, sheep, goats, and pigs, household animals such as rabbits, dogs, and cats, and laboratory animals such as rats, mice, and guinea pigs. [Examples]

[0072] The following examples are provided to provide a complete disclosure and description of how the compounds, compositions, articles, devices, and / or methods claimed herein are prepared and evaluated, and are intended to be purely illustrative but not intended to limit the disclosure. While efforts have been made to ensure accuracy with respect to numerical values ​​(e.g., quantities, temperatures, etc.), some error and deviation should be considered. These examples are provided for illustrative purposes only and are not intended to limit the scope of the claims provided herein.

[0073] A. Example 1 - Production of therapeutic composition The MSC secretome therapeutic composition was prepared by the following method: Human bone marrow-derived MSCs were cultured in a culture vessel containing growth medium to expand the MSC population. The growth medium was then removed and the cells were washed with PBS. Subsequently, the MSCs were cultured in reduced glucose medium with a pH of less than 7.0 under hypoxic conditions. The conditioned medium was then collected, ultrafiltered, and filler-sterilized. The production process of the therapeutic product was carried out in accordance with current Good Manufacturing Practices and current Good Tissue Practices.

[0074] The tetraspanin profile of extracellular vesicles present in the therapeutic composition was determined, and it was found that over 95% of the extracellular vesicles present in the therapeutic composition were CD63. + CD9 - CD81 - They discovered that...

[0075] The protein content of the therapeutic product was determined, and the following proteins were identified: ferritin, NUP85, LAMP2, GPR115, serpine Fl, OPN, PAI-1, DAPP1, cathepsin B, semaphorin 6C, PDGF R-alpha, sorbitin, serpine B6, Dkk-3, thrombomodulin, PF4, MIF, periostin, furin, TIMP-1, decorin, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, lumican, TIMP-2, SLITRK5, FAP, artemin, DPPII, cIAP-1, pentraxin 3, visfatin, neprilysin, albumin, galectin-1, UNC5H3, IL-20 R-beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midoquin, Calreticulin, Osteoactivin, Regmine, TAZ, Cathepsin L, RBP4, Serpine A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin-3, SIRP alpha, and Syndecane-4 (one or more), as well as: Ferritin, IGFBP-4, IL-1, R6, GSTM1, NUP85, LAMP2, Meprin A, IL-1 F10, bIG-H3, GPR115, TGFbl, Ephrin-A4, CD109, Serpine Fl, IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF R-Alpha, Sortiline, Serpine B6, Dkk-3, CNTF, TSP-1, GM-CSFRa, Thrombomodulin, Endoglycan, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periostin, Fulin, TIMP-1, PAPP-A, Decorin, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin Alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin D1, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Parcephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R Beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matryptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAKR, Nidogen-1, CD2, Kallikrein-1, TSLP R, LAMP1, TROY, VCAM-1, Siglec-11, S100A1, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R Beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF -1 Alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Nuturin, Clement-2, EMMPRIN, Activin RIB, Neuroligin 2, Epilegulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A, NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Paparin-2, Syndecane-3, Jagged1, AKR1C4, Olfactomedine-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin Alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p2'7, Siglec-6, Dectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, C A125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Regmine, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 Beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR Alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Newdecine, ILT4, uPAR, Axl, WIF-1, IL-7 R-Alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Kokurin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL-17 RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGFR, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, Serpine A4, ADAM23, NOV, Galectin-2, Neurexin 3 Beta, TLR3, Sirtuin 2, Numb, IL-28 R Alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-IRA, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpine A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP(TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirre13, FLRT1, Galectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R-alpha 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurograinin, HE4, IL-23 R, Galectin-7, GALNT3, GITR L, CD14, R-Spongin 2, CK19, Cardiotrophin-1, TREML1, HAPLIN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospongin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulisin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin-3, ARSB, LIF R Alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP Alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 Alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2 Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, insulin, syntaxin 6, GRO, Bcl-w, lipocalin-2, PDGF-AA, IL-2 Ra, angiogenin, LYVE-1, CD4, RAGE, CDNF, brevican, NAP-2, PU.1, EDAR, ADAMTS13, kynureninase, PTH1R, IFN-gamma-R1, CrkL, B7-1, PARC, draxin, VE-cadherin, procalcitonin, SOX15, kallikrein-11, BCMA, dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, resistin, HVEM, ENPP-7, syndecane-4, IL-2 We found the presence of at least one protein selected from Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, galectin-8, CA4, cystatin EM, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7, and IL-13.

[0076] Determine the nucleic acid content of the therapeutic product, and the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a-5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1’7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19’7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p,hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR -29c-3p, hsa-miR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d , hsa-miR-30d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR- 320a, hsa-miR-342-3p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-376a-3 p, hsa-miR-376c-3p, hsa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa- mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-66 We found that the following compounds exist: 3a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p, and hsa-miR-99b-5p.

[0077] Example 2 - Safety study of the treatment of complex regional pain syndrome (CRPS) using an investigational drug derived from extracellular vesicles of human bone marrow stem cells. In this example, the inventors report that an investigational drug (IP) of extracellular vesicles derived from human bone marrow stem cells (hBM-MSC EVs) is safe and effective in patients with complex regional pain syndrome. Ten subjects received two intravenous infusions of 15 mL of the investigational drug on days 1 and 4. Assessments (visual analog pain scale, simple pain assessment form, SF-36 questionnaire, range of motion analysis, jamar grip strength meter) were performed at baseline (day 0, before infusion), week 1, and at 1, 3, and 6 months after the second infusion. No serious adverse events related to the investigational drug treatment occurred. Statistically significant improvements in pain and motor assessments were observed in the subject pool. The investigational drug was found to be safe in CRPS patients. The investigational drug demonstrated effectiveness in managing CRPS symptoms. The investigational drug presents a safe and effective potential treatment for CRPS.

[0078] ExoFlo®, the investigational drug (IP) used herein, is an EV (hBM-MSC EV) preparation derived from human BM-MSCs. This IP is subject to advanced particle analysis, proteomics evaluation, and USP <71> This is a consistent extravasation product with extensive characterization, including sterility assurance. Furthermore, this IP is a biopharmacological product with consistent quality in terms of dose and biological activity, manufactured in a cGMP environment. Two peer-review studies have demonstrated the safety of intravenous infusion of this IP in patients with severe COVID-19. This IP has also demonstrated efficacy in a subpopulation of these severely ill patients.

[0079] method Ten CRPS patients who met all inclusion and exclusion criteria received 15 mL of the BM-MSC investigational drug intravenously (IV) over 30 minutes on days 1 and 4 (two doses). 15 mL of the investigational drug was mixed with 85 mL of physiological saline, and a total of 100 mL of the mixture was administered IV. This administration protocol was selected because it had been previously recognized as safe. Measurements were taken at baseline before each administration, one week after the second administration, one month after the second administration, three months after, and six months after the second administration. During the study, one patient became unfollowed, and their information was excluded from the study results and analysis.

[0080] The measurements and statistical analyses performed included the Visual Analog Scale (VAS) for pain, a simplified pain assessment form, the SF-36 questionnaire, range of motion (ROM) analysis, and the Jamar grip strength meter test. All results were subjected to one-way ANOVA with Geisser-Greenhouse correction, followed by Dunnett multiple comparison studies using GraphPad Prism 9.5.1.

[0081] Study design. Since CRPS affects women more than men (ratio range 2:3 to 4:1), the proportion of women was higher, as shown in Table 1. Diversity was not addressed in this pilot study.

[0082] [Table 1]

[0083] Inclusion Criteria: Enrolled subjects were required to have chronic, intractable pain in the upper or lower extremities secondary to complex regional pain syndrome as their primary complaint. Other possible etiologies of pain had to be excluded prior to enrollment in this study. Subjects were required to have a minimum VAS score greater than 6 in the area of ​​greatest pain and to have failed to achieve adequate pain relief with at least two previous pharmacological or interventional treatments. Subjects were over 18 years of age. Subjects' neurological function had been stable for the past 30 days. Subjects had to be psychologically appropriate in the opinion of the principal investigator. Subjects were required to be able to provide written informed consent and agree to the follow-up schedule and protocol. If enrolled in workers' compensation insurance, the status had to be permanent and steady.

[0084] Exclusion Criteria: Participants were excluded if they had shown a gradual increase or change in pain status in the past 30 days. Participants currently involved in medical malpractice litigation were excluded. Participants who had received corticosteroid therapy in the past 30 days were excluded. Participants whose pain medication dosage was unstable over 30 days were excluded. Participants with cognitive, physical, or sensory impairments that, in the opinion of the principal investigator, could limit their reporting ability were excluded. Participants with active systemic infections were excluded. Participants with significant medical complications were excluded in the opinion of the principal investigator. Participants who had participated in another clinical trial in the past 30 days were excluded. Participants diagnosed with cancer within the past two years were excluded. Participants who were pregnant, breastfeeding, or planning to become pregnant were excluded. Workers' compensation insurance coverage that did not reach the greatest medical benefit was excluded from this study.

[0085] result Patient demographics: Of the 10 subjects, 7 were female, consistent with the 3 times higher incidence rate in women compared to men (Table 1). The age range of the subjects was 22-72 years. The mean age was 55.8 years, which was consistent with the prevalence of CRPS in the elderly. One subject was lost to follow-up.

[0086] Primary endpoint: Among the 10 subjects enrolled in this study, no subjects reported serious adverse events (SAEs) related to the treatment or the investigational drug. Two subjects contracted COVID-19 during the study but continued enrollment and fully participated. One subject developed congestive heart failure, which was due to a pre-existing comorbidity and not to the treatment protocol or the investigational drug.

[0087] Secondary endpoints: At baseline, the mean VAS score for the nine subjects analyzed was 7.78 (Figure 1). The mean VAS scores were 5.56, 5.5, and 4.89 at 1 month, 3 months, and 6 months, respectively. Overall, statistically significant improvements were observed at all follow-up times. At 1 month follow-up, one subject reported a VAS reduction of ≥50%. At 3 months follow-up, three subjects reported a VAS reduction of ≥50%. Of the remaining five subjects, four patients reported a VAS reduction of ≥50% at 6 months. The remaining subjects reported a lower degree of VAS improvement.

[0088] The BPI pain score was divided into a pain interference score and a pain severity score (Figure 2). While a trend of improvement was observed during the study, statistical significance was evident only in pain interference at 3 months. The mean baseline values ​​for both were 6.99 and 7.61 for severity and interference, respectively. Compared to baseline, further decreases were observed at 1, 3, and 6 months to 5.55 and 5.66, 4.90 and 4.59, and 4.94 and 5.16, respectively. At 6 months, pain decreased by 29.23% and 32.22% in terms of severity and interference, respectively.

[0089] Calculation of the SF-36 score revealed significant improvements in pain, energy / fatigue, and social functioning metrics at 3-month and 6-month follow-up (Figure 3). Significant improvements were evident at 3 months in physical function and physical category limitations. Trends of improvement were observed in emotional limitation and emotional health during the study, but these did not reach statistical significance. Little change was observed in the overall health category.

[0090] Range of motion assessment depended on the anatomical location of the affected joint. Data points were collected at baseline, 3 months, and 6 months (Figure 4). Values ​​were recorded in degrees and compared to normal rotation of a normal joint. Of the 9 subjects in the group, 7 had impaired range of motion in the CRPS-affected limb compared to the normal range of motion. On average, subjects showed an 83.37% increase in range of motion in the affected limb, reaching 82% of the normal range of motion at the end of the study (Table 2).

[0091] [Table 2]

[0092] Of the nine subjects in this study, only three experienced an impact on upper limb grip strength (Jamar trial). Overall, grip strength improved not only on the side affected by CRPS but also on the unaffected control side (Figure 5). Grip strength was equalized for all subjects evaluated after 6 months (Table 3).

[0093] [Table 3]

[0094] Consideration This pilot study aimed to evaluate the safety of ExoFlo® investigational drug in patients with CRPS as the primary endpoint, and to explore its potential to promote improvement of CRPS-related symptoms as a secondary objective. The primary endpoint was achieved without any reported SAEs throughout the 6-month follow-up period. During the study, two subjects contracted COVID-19 and reported adverse complications related to the CRPS diagnosis. One subject contracted COVID-19 between months 3 and 6, with limb pain, color changes, and swelling returning to baseline. The other subject contracted COVID-19 between months 1 and 3, and although there was an acute onset of neurogenic pain, overall, swelling and burning decreased, and energy levels increased. At month 6, this subject reported complete resolution of burning pain. One patient who contracted COVID-19 during the study reported no pain interference or changes in pain severity on the BPI scale. Between months 1 and 3, one participant was diagnosed with congestive heart failure. However, this patient's congestive heart failure is thought to have developed due to pre-existing comorbidities and lifestyle factors, rather than as a result of the investigational drug treatment. This participant reported a 50% reduction in pain and an overall increase in range of motion.

[0095] Functional assessments showed consistently positive patient responses to treatment. Comparison of VAS scores from baseline to 6 months showed, on average, improvement in pain across all subjects. Overall, there was approximately a 50% reduction in pain by the end of the study. One subject, who rated pain as 7 / 10 at baseline, reported no pain at the end of the study. One subject reported no change in pain from baseline to 6 months, but showed a reduction in pain at the 1-month and 3-month follow-up.

[0096] When BPI scores were assessed, pain severity and pain interference decreased on average in all subjects. The greatest improvement was observed in pain interference in daily life, with pain interference decreasing by 32% overall by the end of the study. One subject reported no pain interference or severity at 3 months and at the end of the 6-month study. All but two subjects reported a decrease in pain severity throughout the study, while the remaining two subjects reported no change. All but one subject reported a decrease in pain interference at the end of the study, and their pain level remained the same but did not increase. This subject reported no change in pain severity or pain interference compared to baseline. This subject had COVID-19 between 1 and 3 months of the study. Overall, the majority of the observed benefits were reported at 3 months, with a slight recovery in pain interference at the end of the 6-month study.

[0097] In the SF-36 score assessment, all participants showed improvement at the end of the 6-month clinical trial compared to their original baseline score. The most significant improvements were observed in pain, energy levels, social functioning, and physical functioning, with a positive trend in emotional health, while improvements in overall health were the least significant. Of the 9 participants in this study, 5 reported zero emotional limitations by the end of the 6-month trial. One participant similarly reported zero physical limitations and pain limitations by 6 months.

[0098] On average, the percentage of normal range of motion in affected CRPS joints was 51% compared to the normal anatomical rotation of unaffected limbs. By 3 months, the percentage of normal range of motion was 81%, an increase of 84.41% from baseline. By 6 months, the percentage of normal range of motion had increased to 82%, an overall increase of 83.37% from baseline. Two subjects fully recovered to normal range of motion in their CRPS-affected joints at 6 months. The majority of the observed range of motion benefits were observed by 3 months. On average, all patients showed an increase in range of motion at 6 months compared to baseline. One subject did not see any benefit in range of motion or shoulder extension in a single movement from the treatment, but did show an increase in abduction range of motion with pronation of the same joint, in addition to flexion. No subjects reported a decrease in range of motion throughout this study.

[0099] At baseline, the mean grip strength measurement for the unaffected control limb was 29.33 kg, compared to 21.33 kg for the CRPS-affected limb. At 1 month, increases of 30.66 kg and 21.33 kg were observed in the control and CRPS-affected limbs, respectively. At 3 months, increases of 35.67 kg and 34.67 kg were observed, respectively. At 6 months, increases of 43.33 kg and 45 kg were observed, respectively. Overall, grip strength increased by 147.73% in the control limb and by 210.94% in the affected CRPS limb. In all subjects, grip strength increased in both the control and CRPS-affected limbs. In one subject, the CRPS-affected limb became stronger than the control limb at 6 months. These subjective and objective findings were consistent with the reported improvements in limb and joint range of motion, as well as reductions in lower limb motor changes, edema, allodynia, and paresthesia.

[0100] Two subjects reported little to no hair growth on the affected limb before the study. Both subjects experienced increased hair growth during the study. One subject reported that by the end of the study, hair growth on the affected limb had returned to normal compared to the unaffected limb. Of the two subjects who reported hair loss, one also experienced nail problems. At 6 months, both subjects reported an overall increase in nail growth rate compared to before the start of the study.

[0101] Furthermore, one participant experienced complete disappearance of CRPS symptoms and pain. Both participants reported normal range of motion, complete pain relief, no pain interference in daily life, and no pain interference to physical function, physical limitations, emotional limitations, energy deficits, emotional health, social functioning, and overall health.

[0102] conclusion The investigational drug was demonstrated to be safe and well-tolerated in patients with CRPS. On average, all subjects showed improvement after delivery of the investigational drug.

Claims

1. A method for treating complex regional pain syndrome (CRPS) in a subject requiring treatment for CRPS, comprising the step of administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles, wherein at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63 ＋ CD9 － CD81 － The method.

2. The method according to claim 1, wherein the visual analog pain scale (VAS) score of the subject decreases by at least 50% after administration compared to before administration.

3. A method for treating complex regional pain syndrome (CRPS) in a subject requiring treatment for CRPS, comprising the step of administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles, wherein the subject's visual analog pain scale (VAS) score decreases by at least 50% after administration compared to before administration.

4. At least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63 ＋ CD9 － CD81 － The method according to claim 3.

5. The method according to any one of claims 2 to 4, wherein the minimum pre-administration VAS score in the affected limb of the subject is greater than 6.

6. The method according to any one of claims 1 to 5, wherein the subject's BPI score decreases after administration compared to before administration.

7. The method according to claim 6, wherein the BPI score of the subject decreases by at least 10%, at least 20%, at least 30%, or more than 30% after administration compared to before administration.

8. The method according to claim 6 or 7, wherein the BPI pain interference score of the subject decreases by at least 10%, at least 20%, at least 30%, or more than 30% after administration compared to before administration.

9. The method according to any one of claims 1 to 8, wherein the range of motion evaluation score of the subject increases by at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 95%, or at least 100% after administration compared to before administration.

10. The method according to any one of claims 1 to 9, wherein the score of the Jamar grip strength meter test evaluation of the subject increases after administration compared to before administration.

11. The method according to claim 10, wherein the increase in the score of the Jamar grip strength meter test evaluation is in the upper limbs.

12. The method according to claim 10 or 11, wherein the grip strength of the subject is equalized after administration.

13. The method according to any one of claims 10 to 12, wherein the grip strength of the subject increases by at least 5 kg, at least 10 kg, at least 15 kg, or at least 20 kg after administration compared to before administration.

14. The method according to any one of claims 10 to 13, wherein the grip strength of the subject increases by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, or at least 210% after administration compared to before administration.

15. The method according to any one of claims 10 to 14, wherein the grip strength of the subject increases in the affected limb.

16. The method according to any one of claims 10 to 15, wherein the grip strength of the target increases in the control limb.

17. The method according to any one of claims 10 to 16, wherein, in the subject, one or more of the following improve after administration compared to before administration: range of motion of the limbs, range of motion of the joints, changes in spontaneous movement, edema, allodynia, and sensory abnormalities.

18. The method according to any one of claims 1 to 17, wherein the Short Form 36 Health Survey Questionnaire (SF-36) score of the subject improves after administration compared to before administration.

19. The method according to claim 18, wherein the improvement in the SF-36 questionnaire score includes one or more of the following: improvement in pain, improvement in energy levels, improvement in social functioning, improvement in physical functioning, positive trend in emotional health, and improvement in overall health.

20. The therapeutic MSC secretome composition contains the following proteins: ferritin, NUP85, LAMP2, GPR115, serpin Fl, OPN, PAI-1, DAPP1, cathepsin B, semaphorin 6C, PDGF-α-alpha, soltirin, serpin B6, DKK-3, thrombomodulin, PF4, MIF, periostin, furin, TIMP-1, decorin, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, lumican, TIMP-2, SLITRK5, FAP, artemin, DPPII, cIAP-1, pentraxin 3, visfatin, neprilysin, albumin, galectin-1, UNC5H3, IL-20-β-beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midoquin, Calreticulin, Osteoactivin, Regmine, TAZ, Cathepsin L, RBP4, Serpine A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin-3, SIRP alpha, and Syndecan-4 (one or more of these), as well as: Ferritin, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, Meprin A, IL-1 F10, bIG-H3, GPR115, TGFbl, Ephrin-A4, CD109, Serpine Fl, IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF R-Alpha, Sortiline, Serpine B6, DKK-3, CNTF, TSP-1, GM-CSFRa, Thrombomodulin, Endoglycan, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periostin, Fulin, TIMP-1, PAPP-A, Decorin, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin Alpha 1, EXTL3, IL-18, BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin D1, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Parcephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R-Beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matryptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAKR, Nidogen-1, CD2, Kallikrein-1, TSLP R, LAMP1, TROY, VCAM-1, Siglec-11, S100A1, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAM, OSM R Beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF -1 Alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Nuturin, Clement-2, EMMPRIN, Activin RIB, Neuroligin 2, Epilegulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A, NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Paparin-2, Syndecane-3, Jagged1, AKR1C4, Olfactomedine-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin Alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p2'7, Siglec-6, Dectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, C A125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Regmine, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 Beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR Alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1, R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Newdecine, ILT4, uPAR, Axl, WIF-1, IL-7 R-Alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Kokurin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL-17RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGFR, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, Serpine A4, ADAM23, NOV, Galectin-2, Neurexin 3 Beta, TLR3, Sirtuin 2, Numb, IL-28 R Alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2, Rb, GFR Alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-Selectin, Hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-lra, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpine A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP (TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, NES Chin, TPO, ErbB3, Kirre13, FLRT1, galectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R-Alpha 1, AMIGO 2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurogranin, HE4, IL-23R, Galectin-7, GALNT3, GITR L, CD14, R-Spongin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF-RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospongin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulisin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin-3, ARSB, LIF R Alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP Alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 Alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TRIME-2, Flt-3L, CDS, IL-6, CD229, insulin, syntaxin 6, GRO, Bcl-w, lipocalin-2, PDGF-AA, IL-2 Ra, angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP-2, PU.

1. EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-Gamma-R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein-11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 The method according to any one of claims 1 to 19, comprising at least one protein selected from Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, galectin-8, CA4, cystatin E M, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7, or IL-13.

21. The extracellular vesicles contain the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a-5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1’7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19’7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a-5p,hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR -30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-342- 3p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-376a-3p, hsa-miR-376c-3p, hsa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-4 86-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hs The method according to any one of claims 1 to 20, comprising one or more of a-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p, or hsa-miR-99b-5p.

22. The method according to any one of claims 1 to 21, wherein the subject suffers from chronic intractable pain in one or more upper or lower limbs.

23. The method according to any one of claims 1 to 22, wherein the subject does not experience any increase or change in pain during the 30 days prior to administration.

24. The method according to any one of claims 1 to 23, wherein the subject experiences a reduction in the sudden onset of pain, a reduction in swelling, and / or a reduction in burning sensation in the affected limb after administration compared to before administration.

25. The method according to any one of claims 1 to 24, wherein the administration improves hair and / or nail growth in the affected limb of the subject.

26. The composition is (a) Under the following conditions: (i) oxygen tension less than 5%, and (ii) Culture medium with a pH of less than 7 So, the process involves culturing bone marrow-derived MSCs to produce MSC-conditioned medium, (b) A step of collecting the MSC-acclimatized medium, (c) A step of formulating the MSC-conditioned medium to produce the therapeutic MSC secretome composition, wherein the therapeutic MSC secretome composition comprises proteins and extracellular vesicles produced by the bone marrow-derived MSCs in step (a) The method according to any one of claims 1 to 25, produced by...

27. The method according to claim 26, wherein the culture medium is serum-free.

28. The method according to claim 26 or 27, wherein the glucose concentration of the culture medium is less than 4.5 g / L.

29. The method according to any one of claims 1 to 28, wherein the subject is a human.

30. The method according to any one of claims 26 to 29, wherein the bone marrow-derived MSCs are derived from human bone marrow.

31. The method according to any one of claims 1 to 30, wherein the administration step includes intravenous administration.

32. The method according to any one of claims 1 to 31, wherein the dose of the therapeutic MSC secretome composition administered to the subject is a cellular equivalent dose of 0.7 to 7 million cells per kg.

33. The therapeutic MSC secretome composition contains 4 × 10 per mL. １０ ~10 x 10 １０ The method according to any one of claims 1 to 31, comprising a single cell.

34. The therapeutic MSC secretome composition is 5×10 １１ to 1.5×10 １２ The method according to any one of claims 1 to 31, comprising extracellular vesicles.

35. The method according to any one of claims 1 to 34, wherein the composition is administered monthly for two months or more, or once every one month, two months, or three months or more.

36. The method according to any one of claims 1 to 34, wherein the composition is administered twice.

37. The method according to claim 36, wherein the composition is administered at four-day intervals.

38. A method for preparing a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition for treating complex regional pain syndrome (CRPS) in subjects requiring treatment for CRPS, (a) Under the following conditions: (i) oxygen tension less than 5%, and (ii) Culture medium with a pH of less than 7 So, the process involves culturing bone marrow-derived MSCs to produce MSC-conditioned medium, (b) A step of collecting the MSC-acclimatized medium, (c) A step of formulating the MSC-conditioned medium to produce the therapeutic MSC secretome composition, wherein the therapeutic MSC secretome composition comprises proteins and extracellular vesicles produced by the bone marrow-derived MSCs in step (a) A method that includes this.

39. The method according to claim 38, wherein the culture medium is serum-free.

40. The method according to claim 38 or 39, wherein the glucose concentration of the culture medium is less than 4.5 g / L.

41. At least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63 ＋ CD9 － CD81 － The method according to any one of claims 38 to 40.

42. The method according to any one of claims 38 to 41, wherein the bone marrow-derived MSCs are derived from human bone marrow.

43. The method according to any one of claims 38 to 42, wherein the administration step includes intravenous administration.

44. The therapeutic MSC secretome composition contains the following proteins: ferritin, NUP85, LAMP2, GPR115, serpin Fl, OPN, PAI-1, DAPP1, cathepsin B, semaphorin 6C, PDGF-α-alpha, soltirin, serpin B6, DKK-3, thrombomodulin, PF4, MIF, periostin, furin, TIMP-1, decorin, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, lumican, TIMP-2, SLITRK5, FAP, artemin, DPPII, cIAP-1, pentraxin 3, visfatin, neprilysin, albumin, galectin-1, UNC5H3, IL-20-β-beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midoquin, Calreticulin, Osteoactivin, Regmine, TAZ, Cathepsin L, RBP4, Serpine A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin-3, SIRP alpha, and Syndecan-4 (one or more of these), as well as: Ferritin, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, Meprin A, IL-1 F10, bIG-H3, GPR115, TGFbl, Ephrin-A4, CD109, Serpine Fl, IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF R-Alpha, Sortiline, Serpine B6, DKK-3, CNTF, TSP-1, GM-CSFRa, Thrombomodulin, Endoglycan, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periostin, Fulin, TIMP-1, PAPP-A, Decorin, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin Alpha 1, EXTL3, IL-18, BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin D1, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Parcephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R-Beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matryptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAKR, Nidogen-1, CD2, Kallikrein-1, TSLP R, LAMP1, TROY, VCAM-1, Siglec-11, S100A1, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAM, OSM R Beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF -1 Alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Nuturin, Clement-2, EMMPRIN, Activin RIB, Neuroligin 2, Epilegulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A, NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Paparin-2, Syndecane-3, Jagged1, AKR1C4, Olfactomedine-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin Alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p2'7, Siglec-6, Dectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, C A125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Regmine, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 Beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR Alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1, R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Newdecine, ILT4, uPAR, Axl, WIF-1, IL-7 R-Alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Kokurin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL-17RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGFR, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, Serpine A4, ADAM23, NOV, Galectin-2, Neurexin 3 Beta, TLR3, Sirtuin 2, Numb, IL-28 R Alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2, Rb, GFR Alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-Selectin, Hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-lra, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpine A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP (TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, NES Chin, TPO, ErbB3, Kirre13, FLRT1, galectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R-Alpha 1, AMIGO 2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurogranin, HE4, IL-23R, Galectin-7, GALNT3, GITR L, CD14, R-Spongin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF-RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospongin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulisin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin-3, ARSB, LIF R Alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP Alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 Alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TRIME-2, Flt-3L, CDS, IL-6, CD229, insulin, syntaxin 6, GRO, Bcl-w, lipocalin-2, PDGF-AA, IL-2 Ra, angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP-2, PU.

1. EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-Gamma-R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein-11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 The method according to any one of claims 38 to 43, comprising at least one protein selected from Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, galectin-8, CA4, cystatin E M, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7, or IL-13.

45. The extracellular vesicles contain the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a-5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1’7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19’7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a-5p,hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR- 30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30d-5p, h sa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-342-3 p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-376a-3p, hsa-miR-376c-3p, h sa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-48 6-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa The method according to any one of claims 38 to 44, comprising one or more of -mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p, or hsa-miR-99b-5p.

46. Use of a composition produced by any one of claims 38 to 45 in the treatment of complex regional pain syndrome (CRPS) in subjects requiring treatment for CRPS.

47. The use according to claim 46, wherein the subject is a human.

48. The use according to claim 46 or 47, wherein the visual analog pain scale (VAS) score of the subject decreases by at least 50% after administration compared to before administration.

49. The use according to any one of claims 46 to 48, wherein the minimum pre-administration VAS score in the affected limb of the subject is greater than 6.

50. The use according to any one of claims 46 to 49, wherein the subject's BPI score decreases after administration compared to before administration.

51. The use according to claim 50, wherein the BPI score of the subject decreases by at least 10%, at least 20%, at least 30%, or more than 30% after administration compared to before administration.

52. The use according to claim 50 or 51, wherein the BPI pain interference score of the subject decreases by at least 10%, at least 20%, at least 30%, or more than 30% after administration compared to before administration.

53. The use according to any one of claims 46 to 52, wherein the range of motion evaluation score of the subject increases by at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 95%, or at least 100% after administration compared to before administration.

54. The use according to any one of claims 46 to 53, wherein the score of the Jamar grip strength meter test evaluation of the subject increases after administration compared to before administration.

55. The use according to claim 54, wherein the increase in the score of the Jamar grip strength meter test evaluation is in the upper limbs.

56. The use according to claim 54 or 55, wherein the grip strength of the subject is equalized after administration.

57. The use according to any one of claims 54 to 56, wherein the grip strength of the subject increases by at least 5 kg, at least 10 kg, at least 15 kg, or at least 20 kg after administration compared to before administration.

58. The use according to any one of claims 54 to 57, wherein the grip strength of the subject increases by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, or at least 210% after administration compared to before administration.

59. The use according to any one of claims 54 to 58, wherein the grip strength of the subject increases in the affected limb.

60. The use according to any one of claims 54 to 59, wherein the grip strength of the subject increases in the control limb.

61. The use according to any one of claims 46 to 60, wherein, in the subject, one or more of the following improve after administration compared to before administration: range of motion of the limbs, range of motion of the joints, changes in spontaneous movement, edema, allodynia, and sensory abnormalities.

62. The use according to any one of claims 46 to 61, wherein the Short Form 36 Health Survey Questionnaire (SF-36) score of the subject improves after administration compared to before administration.

63. The use according to claim 62, wherein the improvement in the SF-36 questionnaire score includes one or more of the following: improvement in pain, improvement in energy levels, improvement in social functioning, improvement in physical functioning, positive trend in emotional health, and improvement in overall health.

64. The use according to any one of claims 46 to 63, wherein the subject suffers from chronic intractable pain in one or more upper or lower limbs.

65. The use according to any one of claims 46 to 64, wherein the subject does not experience any increase or change in pain during the 30 days prior to administration.

66. The use according to any one of claims 46 to 65, wherein the subject experiences a reduction in the sudden onset of pain, a reduction in swelling, and / or a reduction in burning sensation in the affected limb after administration compared to before administration.

67. The use according to any one of claims 46 to 66, wherein the administration improves hair and / or nail growth in the affected limb of the subject.

68. The use according to any one of claims 46 to 67, wherein the dose of the therapeutic MSC secretome composition administered to the subject is a cellular equivalent dose of 0.7 to 7 million cells per kg.

69. The therapeutic MSC secretome composition contains 4 × 10 per mL. １０ ~10 x 10 １０ The method according to any one of claims 46 to 67, comprising a single cell.

70. The therapeutic MSC secretome composition is 5 × 10 １１ ~1.5 x 10 １２ The method according to any one of claims 46 to 67, comprising a number of extracellular vesicles.

71. The method according to any one of claims 38 to 70, wherein the composition is administered monthly for a period of two months or more, or once every one month, two months, or three months or more.

72. The method according to any one of claims 38 to 70, wherein the composition is administered twice.

73. The method according to claim 72, wherein the composition is administered at four-day intervals.

74. The use of a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles in the treatment of complex regional pain syndrome (CRPS) in subjects requiring treatment for CRPS, wherein at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63 ＋ CD9 － CD81 － It is used.

75. The use according to claim 74, wherein the composition is administered intravenously to the subject.

76. The use according to claim 74 or 75, wherein the visual analog pain scale (VAS) score of the subject decreases by at least 50% after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

77. The use according to any one of claims 74 to 76, wherein the minimum VAS score in the affected limb of the subject before administration of the therapeutic MSC secretome composition is greater than 6.

78. The use according to any one of claims 74 to 77, wherein the subject's BPI score decreases after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

79. Use of a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles in the treatment of complex regional pain syndrome (CRPS) in a subject requiring treatment for CRPS, wherein the subject's visual analog pain scale (VAS) score decreases by at least 50% after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

80. The use according to claim 79, wherein the minimum VAS score in the affected limb of the subject before administration of the therapeutic MSC secretome composition is greater than 6.

81. The use according to claim 79 or 80, wherein the subject's BPI score decreases after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

82. At least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63 ＋ CD9 － CD81 － The use according to any one of claims 79 to 81.

83. The therapeutic MSC secretome composition contains the following proteins: ferritin, NUP85, LAMP2, GPR115, serpin Fl, OPN, PAI-1, DAPP1, cathepsin B, semaphorin 6C, PDGF-α-alpha, soltirin, serpin B6, DKK-3, thrombomodulin, PF4, MIF, periostin, furin, TIMP-1, decorin, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, lumican, TIMP-2, SLITRK5, FAP, artemin, DPPII, cIAP-1, pentraxin 3, visfatin, neprilysin, albumin, galectin-1, UNC5H3, IL-20-β-beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midoquin, Calreticulin, Osteoactivin, Regmine, TAZ, Cathepsin L, RBP4, Serpine A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin-3, SIRP alpha, and Syndecan-4 (one or more of these), as well as: Ferritin, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, Meprin A, IL-1 F10, bIG-H3, GPR115, TGFbl, Ephrin-A4, CD109, Serpine Fl, IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF R-Alpha, Sortiline, Serpine B6, DKK-3, CNTF, TSP-1, GM-CSFRa, Thrombomodulin, Endoglycan, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periostin, Fulin, TIMP-1, PAPP-A, Decorin, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin Alpha 1, EXTL3, IL-18, BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin D1, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Parcephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R-Beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matryptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAKR, Nidogen-1, CD2, Kallikrein-1, TSLP R, LAMP1, TROY, VCAM-1, Siglec-11, S100A1, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAM, OSM R Beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF -1 Alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Nuturin, Clement-2, EMMPRIN, Activin RIB, Neuroligin 2, Epilegulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A, NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Paparin-2, Syndecane-3, Jagged1, AKR1C4, Olfactomedine-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin Alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p2'7, Siglec-6, Dectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, C A125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Regmine, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 Beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR Alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1, R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Newdecine, ILT4, uPAR, Axl, WIF-1, IL-7 R-Alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Kokurin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL-17RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGFR, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, Serpine A4, ADAM23, NOV, Galectin-2, Neurexin 3 Beta, TLR3, Sirtuin 2, Numb, IL-28 R Alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2, Rb, GFR Alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-Selectin, Hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-lra, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpine A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP (TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, NES Chin, TPO, ErbB3, Kirre13, FLRT1, galectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R-Alpha 1, AMIGO 2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurogranin, HE4, IL-23R, Galectin-7, GALNT3, GITR L, CD14, R-Spongin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF-RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospongin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulisin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin-3, ARSB, LIF R Alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP Alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 Alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TRIME-2, Flt-3L, CDS, IL-6, CD229, insulin, syntaxin 6, GRO, Bcl-w, lipocalin-2, PDGF-AA, IL-2 Ra, angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP-2, PU.

1. EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-Gamma-R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein-11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 The use according to any one of claims 79 to 82, comprising at least one protein selected from Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, galectin-8, CA4, cystatin E M, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7, or IL-13.

84. The extracellular vesicles contain the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a-5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1’7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19’7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a-5p,hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR- 30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30d-5p, h sa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-342-3 p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-376a-3p, hsa-miR-376c-3p, h sa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-48 6-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa The use according to any one of claims 79 to 83, comprising one or more of -mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p, or hsa-miR-99b-5p.

85. The composition is (a) Under the following conditions: (i) oxygen tension less than 5%, and (ii) Culture medium with a pH of less than 7 So, the process involves culturing bone marrow-derived MSCs to produce MSC-conditioned medium, (b) A step of collecting the MSC-acclimatized medium, (c) A step of formulating the MSC-conditioned medium to produce the therapeutic MSC secretome composition, wherein the therapeutic MSC secretome composition comprises proteins and extracellular vesicles produced by the bone marrow-derived MSCs in step (a) The use according to any one of claims 79 to 84, produced by...

86. The use according to claim 85, wherein the culture medium is serum-free.

87. The use according to claim 85 or 86, wherein the glucose concentration of the culture medium is less than 4.5 g / L.

88. The use according to any one of claims 85 to 87, wherein the BPI score of the subject decreases by at least 10%, at least 20%, at least 30%, or more than 30% after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

89. The use according to claim 88, wherein the BPI pain interference score of the subject decreases by at least 10%, at least 20%, at least 30%, or more than 30% after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

90. The use according to any one of claims 85 to 89, wherein the range of motion evaluation score of the subject increases by at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 95%, or at least 100% after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

91. The use according to any one of claims 85 to 90, wherein the score of the Jamar grip strength meter test evaluation of the subject increases after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

92. The use according to claim 91, wherein the increase in the score of the Jamar grip strength meter test evaluation is in the upper limbs.

93. The use according to claim 91 or 92, wherein the grip strength of the subject is equalized after administration of the therapeutic MSC secretome composition.

94. The use according to any one of claims 91 to 93, wherein the grip strength of the subject increases by at least 5 kg, at least 10 kg, at least 15 kg, or at least 20 kg after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

95. The use according to any one of claims 91 to 94, wherein the grip strength of the subject increases by at least 50%, at least 75%, at least 100%, at least 125%, at least 150%, at least 175%, at least 200%, or at least 210% after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

96. The use according to any one of claims 91 to 95, wherein the grip strength of the subject increases in the affected limb.

97. The use according to any one of claims 91 to 96, wherein the grip strength of the subject increases in the control limb.

98. The use according to any one of claims 79 to 97, wherein, in the subject, one or more of the range of motion of the limbs, range of motion of the joints, changes in spontaneous movement, edema, allodynia, and sensory abnormalities are improved after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

99. The use according to any one of claims 79 to 98, wherein the Short Form 36 Health Survey Questionnaire (SF-36) score of the subject improves after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

100. The use according to claim 99, wherein the improvement in the SF-36 questionnaire score includes one or more of the following: improvement in pain, improvement in energy levels, improvement in social functioning, improvement in physical functioning, positive trend in emotional health, and improvement in overall health.

101. The use according to any one of claims 79 to 100, wherein the subject suffers from chronic intractable pain in one or more upper or lower limbs.

102. The use according to any one of claims 79 to 101, wherein the subject does not experience any increase or change in pain during the 30 days prior to administration of the therapeutic MSC secretome composition.

103. The use according to any one of claims 79 to 102, wherein the subject experiences a reduction in the sudden onset of pain, a reduction in swelling, and / or a reduction in burning sensation in the affected limb after administration of the therapeutic MSC secretome composition compared to before administration of the therapeutic MSC secretome composition.

104. The use according to any one of claims 79 to 103, wherein administration of the therapeutic MSC secretome composition improves hair and / or nail growth in the affected limb of the subject.

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