Treatment of idiopathic facial paralysis with extracellular vesicle composition

Abstract

Disclosed are methods of treating idiopathic facial paralysis in a subject by administering a therapeutic MSC secretome product made by a method comprising culturing bone marrow-derived MSCs under conditions that include oxygen tension below 5% and a culture media with a pH below 7.

Description

WSGR Ref. No. 66309-733.601TREATMENT OF IDIOPATHIC FACIAL PARALYSIS WITH EXTRACELLULAR VESICLE COMPOSITIONCROSS REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 698,959 filed on September 25, 2024, the entirety of which is hereby incorporated by reference herein.BACKGROUND

[0002] Paralysis of the facial nerve (CN VII) is an often debilitating condition. Bell’s Palsy is a commonly seen mononeuropathy affecting the facial nerves. Although usually self-limited, symptomatology can persist for decades in chronic cases. Currently, therapies for idiopathic facial paralysis tend to rely on surgical interventions, and any non-invasive treatments are often ineffective. There exists a need for safe, non-surgical alternative therapies for idiopathic facial paralysis and Bell’s Palsy.SUMMARY

[0003] An aspect of the present disclosure is a method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein at least 80% of the EVs are CD63+CD9 CD81 . An aspect of the present disclosure is a method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the composition comprises one or more factors, the one or more factors comprising 2B4, 4-1BB, 4-1BB Ligand, 6Ckine, ACE, Activin A, Activin RIB, Activin RIIA, Activin RIM, ADA, ADAM23, ADAMS, ADAMTS13, ADAMTSL-1, Adiponectin, Adipsin, Aggrecan, AgRP, AIF, AKR1C4, Albumin, ALCAM, AMIGO, AMIG02, Aminopeptidase LRAP, Aminopeptidase P2, AMSH, ANG-1, ANG-2, ANG-4, Angiogenin, Angiotensinogen, ANGPTL4, ANGPTL7, APRIL, Arginase 1, ARSB, Artemin, Arylsulfatase A, ASAH2, ASAHL, Axl, B2M, B7-1, B7-H3, B7-H4, BAFF R, Bcl-10, BCL-2, Bcl-w, BCMA, BID, Biglycan, BLAME, BMP-2, BMP-8, BMP-9, BMPR-IA, Brevican, C-myc, CA125, CA13, CA4, CA8, Cadherin-13, Cadherin-4, Calreticulin, Calreticulin-2, Cardiotrophin-1, CASA, Caspr2, Cathepsin E, Cathepsin L, Cathepsin S, Cathepsin V, CD14, CD155, CD157, CD2, CD200, CD229, CD27, CD27 Ligand, CD28, CD30, CD300f, CD314, CD34, CD36, CD39L2, CD4, CD40, CD40L, CD42b, CD48, CD51, CD58, CD6, CD73, CD83, CD97, CD99-L2, CDC25B, CDNF, CDS, CEACAM-1, CEACAM-3, CEACAM-5, Cerberus 1, CES1, CES2, CF VII, CF XIV, Chemerin, CHI3L1, CHST2, CHST3, CHST4, cIAP-1, CILP-1, CK19, CL-P1, CLEC-1, CLEC-2, CLEC10A, Clq, ClqTNF9,WSGR Ref. No. 66309-733.601 cMASP3, CMG-2, Cochlin, COMT, Contactin-1, Contactin-2, CPB1, CPE, CREG, CRELD2, CRIM1, CrkL, CSa, CTACK, CTRC, CXCL14, CXCL16, Cyr61, Cystatin B, Cystatin C, Cy statin D, Cystatin E M, Cystatin S, Cystatin SA, Cystatin SN, Cytokeratin-8, DAPP1, DCTN1, DDR1, Dectin- 1, Dectin-2, Desmin, Desmoglein 2, Desm oglein-3, DKK-1, DLL4, DNAM-1, DNMT3 A, Dopa Decarboxylase, DPPII, DPPIV, DR3, DR6, Draxin, DSCAM, DSPG3, Dtk, E-Selectin, ED AR, EDIL3, EG-VEGF, EGF R, EMMPRIN, ENA-78, Endocan, Endoglin, eNOS, ENPP-2, ENPP-7, Enteropeptidase, EpCAM, EphA2, EphAl, EphB3, EphB4, EphB6, Ephrin-A4, Ephrin-B3, Epimorphin, Epiregulin, ErbB3, ErbB4, Erythropoietin, ESAM, EXTL3, FABP1, FABP2, FABP4, FABP6, FAP, Fas, FCAR, FcERI, FCRL1, FCRL3, FCRLB, FCRLS, FGF R5, FGF-12, FGF-16, FGF-17, FGF-19, FGF-20, FGF-21, FGF-23, FGF-3, FGF- 5, Ficolin-1, FKBP51, FLRG, FLRT1, FLRT2, Flt-3, Flt-3L, Follistatin, Follistatin-like 1, FUT8, Galanin, Galectin-1, Galectin-2, Galectin-3, Galectin-4, Galectin-7, Galectin-8, Galectin- 9, GALNT10, GALNT2, GALNT3, Gash, GATA-4, GBA3, GCP-2, GDF-11, GDF-15, GDF-9, GFAP, GFR alpha-1, GFR alpha-2, GITR, GITR L, GKN1, GLP-1, Glycoprotein V, Glyoxalase II, Glypican 1, Glypican 2, Glypican 5, gpl30, GPR56, GPVI, Granulysin, Granzyme A, Granzyme B, Granzyme H, GRAP2, GRKS, GRO, GSTM1, GUSB, HAO-1, HAPLN1, HCC-1, HCC-4, hCGb, HE4, Hepsin, HGF, HGF R, HIF-1 alpha, HIF-1 beta, HO-1, HS3ST1, HS3ST3B1, HSD17B1, htPAPP-A, HTRA2, HVEM, I-TAC, ICAM-1, ICAM-2, ICAM-3, ICOS, IDO, IFN-gamma Rl, IFNab R2, IFNb, IGF-2R, IGFBP-5, IGSF3, IGSF4B, IL-1 F10, IL-1 R3, IL-1 R4, IL-1 RI, IL-1 RII, IL-10 Rb, IL-12p40, IL-13 R2, IL-15, IL-15 R, IL-17 RC, IL- 17 RD, IL-17B, IL-17F, IL- 18, IL- 18 BPa, IL- 19, IL-2 Ra, IL-2 Rb, IL-2 Rg, IL-20 R beta, IL-20 Ra, IL-21R, IL-22, IL-22 R alpha 1, IL-22BP, IL-23, IL-23 R, IL-24, IL-27, IL-27 Ra, IL- 28 R alpha, IL-28A, IL-31, IL-31 RA, IL-32 alpha, IL-33, IL-4 Ra, IL-6, IL-6R, IL-7 R alpha, IL-8, IL-9, IL-lra, ILT2, ILT4, Inhibin A, Insulin, Insulin R, Integrin alpha 1, Integrin alpha 5, IP-10, Jagged 1, JAM-A, JAM-B, JAM-C, Kallikrein 1, Kallikrein 11, Kallikrein 12, Kallikrein 14, Kallikrein 5, Kallikrein 7, Kell, KIR2DL3, Kirrel3, KLF4, Kremen-2, Kynureninase, L- Selectin, L1CAM-2, LAIR2, LAMA4, LAMP, LAMP1, Langerin, LAP(TGFbl), LDL R, LEDGF, Legumain, LH, LIF, LIF R alpha, LIGHT, LILRB4, LIMPII, Lin28, Lipocalin-2, LOX-1, LRP-6„ST3GAL1, LRRC4, LRRTM4, LSECtin, LTbR, Lymphotactin, LYVE-1, MAP1D, Matrilin-2, Matrilin-3, Matriptase, MBL, Mcl-1, MCEMP1, MCP-2, MCP-3, MCP-4, MCSF, MDC, MDL-1, MEF2C, MEP1B, MEPE, MeprinA, Mer, Mesothelin, METAP2, MFRP, MIA, MICA, MICB, Midkine, MIP-3a, MIP-3b, MIP-lb, MIS RII, MMP-1, MMP-10, MMP-12, MMP-13, MMP-2, MOG, MPIF-1, MSP R, N-Cadherin, NAP-2, NCAM-1, NCAM-1 (CD56), NCK1, Nectin-1, Nectin-2, Nectin-3, Nectin-4, Neprilysin, Nestin, Netrin-4, Neudesin, Neurexin 3 beta, Neurogranin, Neuroligin 2, Neuropilin-2, Neurturin, NG2, NGF R, Nidogen-1,WSGR Ref. No. 66309-733.601Nidogen-2, NKp30, NKp44, NKp46, NKp80, Nogo Receptor, Nogo-A, Norrin, Notch- 1, Notch - 3, NOV, NPDC-1, NPTXR, NQO-1, Nrf2, NRG1-131, NSE, NT AL, NUDTS, Numb, NUP85, Olfactomedin-2, Olig2, OPG, or IL-13, OSCAR, OSM R beta, Osteoactivin, Osteoadherin, 0X40, 0X40 Ligand, P1GF-2, p27, p53, p63, PAPP-A, Pappalysin-2, PARI, PARC, Pax3, PCSK2, PD-1, PD-ECGF, PD-L2, PDGF R alpha, PDGF Rb, PDGF-AA, PDGF-AB, PDGF- CC, PDX-1, PEAR1, PECAM-1, Pentraxin 3, Pepsinogen II, Persephin, PGRP-S, PIGF, PILR- alpha, Plexin A4, Plexin B3, Plexin DI, Podocalyxin, Podoplanin, POGLUT1, PON1, Pref-1, PRELP, Presenilin 1, Procalcitonin, Progranulin, proGRP, Prolactin, Prolactin R, PRX2, PSA- total, PSMA, PSMA1, PTH, PTH1R, PTP1B, PU.l, PYY, R-Spondin 2, RAGE, RalA, RANK, RANTES, RBP4, RCOR1, Reg3A, Reg4, RELT, Renin, Resistin, Ret Midkine, RGM-A, RGM- B, ROBO2, ROBO3, ROBO4, R0R1, R0R2, S100A1, S100A13, SALM4, SCCA2, SCF, SCF R, SDF-la, SDF-lb, Semaphorin 4C, Semaphorin 4D, Semaphorin 4G, Semaphorin 6 A, Semaphorin 6B, Semaphorin 6D, Semaphorin 7A, Serpin A4, Serpin A5, sFRP-3, SH2D1A, Shh-N, SHP-1, Siglec-10, Siglec-11, Siglec-2, Siglec-6, Siglec-7, Siglec-9, SIRP alpha, Sirtuin1, Sirtuin 2, Sirtuin 5, SLAM, SLITRK5, Smad4, SMPD1, SorCSl, SorCS2, SorCS3, SOST, SOX15, SOX2, SOX7, SOX9, SP-D, SPHK1, Spinesin, SPINK1, SR-AI, SREC-I, SREC-II, ST6GAL1, ST8SIA1, Stabilin-2, SULT2A1, Syndecan-3, Syndecan-4, Syntaxin 4, Syntaxin 6, TACE, TAFA1, TAFA2, TAFAS, TARC, TAZ, TC-PTP, TCN2, TECK, Tenascin R, Testican2, TFF3, TFPI, TFPI-2, TGFa, TGFb2, TGFb3 FOLR2, TGFbl, TGM3, THAP11, Thrombospondin-2, Thrombospondin-5, THSD1, Thyroglobulin, Thyroid Peroxidase, Tie-1, Tie-2, TIM-1, TIM-3, TLR1, TLR2, TLR3, TLR4, TNF RI, TNF RII, TNF4, TNFb, TPO, TPP1, TPST2, TRACP, TRAIL, TRAIL Rl, TRAIL R3, TRAIL R4, TRANCE, TREM-1, TREM-2, TREML1, TrkC, TROP-2, Troponin C, TROY, Trypsin 1, Trypsin 3, TSH, TSK, TSLP, TSLP R, TWEAK R, Ubiquitin+1, UCH-L3, ULBP-1, ULBP-2, ULBP-3, ULBP-4, UNC5H3, UNC5H4, uPA, uPAR, Uromodulin, VAMP-1, VAMP-2, VAP-A, VCAM-1, VE-Cadherin, VEGF, VEGF Rl, VEGF R2, VEGF R3, VEGF-C, Visfatin, vWF-A2, WIF-1, WISP-1, Wnt-4, XEDAR, or XIAP, or a combination of two or more thereof; optionally wherein the EVs comprise the one or more factors. An aspect of the present disclosure is a method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the composition comprises one or more factors, the one or more factors comprising TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), LAMB1 (Laminin Subunit Beta 1), FBN1 (Fibrillin 1), HSPG2 (Heparin Sulphate Proteoglycan 2), BGN (Biglycan), or FN1 (Fibronectin 1), or a combinationWSGR Ref. No. 66309-733.601 of two or more thereof; optionally wherein the EVs comprise the one or more factors. In some embodiments, the composition further comprises beta-IG-H3 (Transforming growth factor-betainduced protein ig-H3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD81 antigen, CD9 antigen, CD44 antigen, CD99 antigen, CD 109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Furin, GM-CSF Ra (Granulocytemacrophage colony-stimulating factor receptor subunit alpha), GPR115 (Adhesion G protein- coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF -2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein-3), IGFBP-6 (Insulin-like growth factor binding protein-6), IL18BP (Interleukin- 18 Binding Protein), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome- associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor- 1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin Fl (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein-glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), TSP-2 (Thrombospondin 2), CAD11 (Cadherin 11), JAM3 (Junctional Adhesion Molecule 3), VIM (Vimentin), FBLN5 (Fibulin 5), FBN2 (Fibrillin 2), Hyaluronic Acid, COL1A1 (Collagen Type 1 Alpha 1), COL1 A2 (Collagen Type 1 Alpha 2), or Transferrin, or a combination of two or more thereof. An aspect of the present disclosure is a method of treating idiopathic facial paralysis in a subject, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the composition comprises hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, or let-7, or a combination of two or more thereof; optionally wherein the EVs comprise the hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, or let-7, or a combination of two or more thereof. In some embodiments, the composition further comprises hsa-miR-125b-5p, hsa-miR-132-3p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-221-3p, hsa-miR-22-3p, hsa- miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31- 5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, or hsa-miR-940, or a combination of two or more thereof. In some embodiments, the composition is produced by: culturing bone marrow-derived MSCs (BM-MSCs) under the following conditions to produce an MSC conditioned media: oxygen tension below 5%; and culture media having a pH below 7; harvesting the MSC conditioned media; and formulating the MSC conditioned media to produce a therapeutic MSC secretome, wherein the therapeutic MSC secretome comprises proteins andWSGR Ref. No. 66309-733.601EVs produced by the BM-MSCs in (a). In some embodiments, the subject exhibits one or more of the following symptoms prior to the administering: extracranial paralysis, intracranial paralysis, epiphora, inability to close one or two eyes, drooping of the oral commissure, salivary incompetence, facial tingling, fasciculations of one or more facial muscles, hyperacusis, taste disturbance, inability to elevate one or two eyebrows, difficulty communicating, inability to communicate, inability to smile, difficulty with social interaction, psychological disturbance, depression, headache, earache, tinnitus, dizziness, loss of balance, and / or secondary facial paralysis. In some embodiments, the extracranial paralysis, intracranial paralysis, or secondary facial paralysis affects one side of the face. In some embodiments, the onset of the one or more symptoms is acute. In some embodiments, the one or more symptoms do not improve or do not improve fully in response to one or more of surgery, facial reconstruction, oral steroids, antiviral medications, facial exercises, laser treatment, herbal remedy, acupuncture, botulinum toxin, massage, physical therapy, prednisone, gabapentin, potassium, valaciclovir, vitamin B-12, and / or diuretics. In some embodiments, the subject has experienced the one or more symptoms for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months,10 months, 11 months, 12 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or longer. In some embodiments, the results of the subject’s radiological studies are unremarkable before and / or after the administering. In some embodiments, the administering comprises one or more injections. In some embodiments, at least one of the one or more injections is injected into tissue around the facial nerve. In some embodiments, at least one of the one or more injections comprises 1 cc, 2 cc, 3 cc, 4 cc, or 5 cc of the composition. In some embodiments, at least of the one or more injections comprises 5 cc, 6 cc, 7 cc, 8 cc, 9 cc, 10 cc,11 cc, 12 cc, 13 cc, 14 cc, 15 cc, 16 cc, 17 cc, 18 cc, 19 cc, or 20 cc of the composition. In some embodiments, at least one of the one or more injections is injected intravenously. In some embodiments, the administering restores facial nerve function in the subject as compared to before administration. In some embodiments, the administering reduces House-Brackmann score in the subject as compared to before administration. In some embodiments, the decrease in House-Brackmann score is at least 1, at least 2, at least 3, at least 4, at least 5, or at least 6. In some embodiments, the administering improves a Facial Disability Index score in a subject as compared to before administration. In some embodiments, the improvement in the Facial Disability Index score is an improvement in a physical index. In some embodiments, the improvement in the Facial Disability Index score is an improvement in a social index. In some embodiments, the improvement in the Facial Disability Index score is an increase in at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, atWSGR Ref. No. 66309-733.601 least 80%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%. In some embodiments, the administering restores one or more of brow motion, eyelid motion, buccal control, and / or facial expressiveness in the subject as compared to before administration. In some embodiments, the administering results in improvement in wellbeing, reduction in tingling, reduction in fasciculations, relaxation of muscles, reduction in pain, restoration of ability to wink, and / or improvement in symmetrical facial movements.

[0004] An aspect of the present disclosure is a method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome comprising extracellular vesicles, wherein the subject exhibits an improvement in House-Brackmann score after administration compared to before administration. An aspect of the present disclosure is a method of making a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome for treating idiopathic facial paralysis in a subject in need thereof, the method comprising: culturing bone marrow-derived MSCs (BM-MSCs) under the following conditions to produce an MSC conditioned media: oxygen tension below 5%; and culture media having a pH below 7; harvesting the MSC conditioned media; and formulating the MSC conditioned media to produce the therapeutic MSC secretome, wherein the therapeutic MSC secretome comprises proteins and extracellular vesicles produced by the BM-MSCs in (a). In some embodiments, the idiopathic facial paralysis comprises Bell’s Palsy, facial palsy, and / or secondary facial paralysis. In some embodiments, the culture media is serum-free. In some embodiments, the culture media has a glucose concentration below 4.5 g / L. In some embodiments, formulating the MSC conditioned media comprises exchanging the conditioned media for a pharmaceutically acceptable formulation. In some embodiments, the pharmaceutically acceptable formulation comprises saline. In some embodiments, the composition comprises at least 6xlO10to 8xl010extracellular vesicles per mL and is administered at a dose of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml. In some embodiments, the dose is administered in combination with normal saline at a final volume of 100 mL. In some embodiments, the composition comprises saline (0.9% sodium chloride). In some embodiments, the saline is present in the composition at about 80% to about 95% saline. In some embodiments, the composition comprises sodium chloride, sodium lactate, potassium chloride, and / or calcium chloride. In some embodiments, the molecular weight of any non-excipient component of the composition is greater than about 2 kDa (kilodaltons), 8 kDa, 10 kDa, 50 kDa, or 100 kDa. In some embodiments, the composition comprises an oligosaccharide. In some embodiments, the oligosaccharide is present in the composition at from about 0.2 M to about 0.6 M. In some embodiments, any non-excipient component of the composition has a sizeWSGR Ref. No. 66309-733.601 of less than about 0.2 microns. In some embodiments, the composition is sterile per USP <71>. In some embodiments, the composition is endotoxin free per USP <85>. In some embodiments, the composition is negative for mycoplasma DNA. In some embodiments, the composition is cell-free. In some embodiments, the composition is stored between -80°C and -60°C. In some embodiments, the composition is administered within 6 hours of thaw when maintained at ambient temperature. In some embodiments, the composition is freeze-dried (lyophilized) to a powder cake and stored at or below room temperature. In some embodiments, the composition is reconstituted with water prior to administration. In some embodiments, the composition is present in a glass vial. In some embodiments, the composition is formulated for intravenous administration. In some embodiments, the composition has a pH of about 6 to about 7.5. In some embodiments, the BM-MSCs are negative for CD14, CD31, CD34, and CD45. In some embodiments, the BM-MSCs are positive for CD73, CD105, CD166, and CD90. In some embodiments, the BM-MSCs are capable of undergoing trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes. In some embodiments, the BM-MSCs are obtained from an iliac crest aspiration of a single donor. In some embodiments, a concentration of the one or more EVs in the composition is measured by nanoparticle tracking analysis (NTA). In some embodiments, the NTA comprises light scatter and fluorescence evaluation. In some embodiments, the concentration of the one or more EVs is about 10 billion to about 250 billion EVs per mL of the composition. In some embodiments, the concentration of the one or more EVs is at about 1 billion to about 40 billion EVs per ml of the composition. In some embodiments, the one or more EVs have an average diameter of about 30 nm to about 170 nm. In some embodiments, a total protein concentration of the composition is about 10 to about 40 pg per ml of the composition. In some embodiments, a total protein concentration of the composition is about 1.5 to about 6 pg per ml of the composition. In some embodiments, the total protein concentration is measured by ELISA.BRIEF DESCRIPTION OF THE DRAWINGS

[0005] The features of the present disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings (also “Figure” and “FIG.” herein), of which:

[0006] FIG. 1 shows images of Patient #1. The left image shows Patient #1 two years after the onset of right facial paralysis, post acoustic neuroma surgery. The right image shows Patient #1WSGR Ref. No. 66309-733.601 one month after completion of treatment with an MSC secretome comprising extracellular vesicles.

[0007] FIG. 2 shows images of Patient #4. The left image shows Patient #4 three years after developing left-sided facial paralysis post-temporomandibular joint (TMJ) surgery. The right image shows Patient #2 1 month after completion of treatment with an MSC secretome comprising extracellular vesicles.

[0008] FIG. 3 shows a graph of average House-Brackmann scores for all 7 patients before (top trace) and after (bottom trace) treatment with the MSC secretome comprising extracellular vesicles.

[0009] FIG. 4 shows a graph depicting a summary of Facial Disability Index before and after treatment with the MSC secretome comprising extracellular vesicles, including both physical and social indices.

[0010] FIG. 5 shows a graph depicting a comparison of time since onset of symptoms in each patient and their grade manifestation of disease versus responsivity of treatment. For each patient #l-#7, the first bar represents the House Brackmann grade before, and the second bar represents the House Brackmann grade after treatment with the MSC secretome comprising extracellular vesicles.DETAILED DESCRIPTIONI. Compositions

[0011] Example compositions herein comprise an extracellular vesicle (EV) and / or a protein. The EV may originate from a mesenchymal stem cell (MSC). The protein may originate from a MSC. In an exemplary embodiment, the MSC is a bone marrow MSC (BM-MSC). The EV and / or protein may be purified or otherwise separated from the MSC growth and / or culturing condition from which the EV and / or protein was secreted into. Purified may include partially purified, such that some of the MSC growth and / or culturing condition is present in the composition. The composition may be formulated into an aqueous solution for intravenous administration.

[0012] Extracellular Vesicles

[0013] Extracellular vesicles (EV) are small membrane bound spheres containing proteins and RNA (of which exosomes are a subset). Exosomes are small lipid bilayer vesicles secreted by cells that lack a nucleus and cannot replicate. Other EV populations are derived directly from the plasma membrane or are formed during apoptosis (apoptotic bodies). Disclosed herein are compositions comprising an EV. In example embodiments, the EV is an exosome. EmbodimentsWSGR Ref. No. 66309-733.601 of an EV herein have a diameter of about 20 nm to about 200 nm. In some embodiments, the diameter is measured by nanoparticle tracking analysis (NTA).

[0014] The number of EVs within a composition may be about 10 billion to about 250 billion EVs per mL when suspended. The suspension may be diluted for intravenous administration, wherein the EVs within the composition may be about 1 billion to about 40 billion EVs per mL.

[0015] In some embodiments, the EV has a phenotype of CD63+CD9 and CD81 . In some embodiments, at least 70, 75, 80, 85, 90, 91, 92, 93, 94, or 95% of the EVs are CD63+CD9 and CD81 . In some embodiments, at least 50, 60, 70, 80, 85, 90, 91, 92, 93, 94, or 95% of the EVs are CD9 . In some embodiments, at least 50, 60, 70, 80, 85, 90, 91, 92, 93, 94, or 95% of the EVs are CD81 .

[0016] In some embodiments, the EV is produced from a MSC. The MSC may be a bone marrow MSC. The MSC may be a human MSC. In some embodiments, the EV is produced from a MSC that has the capacity to undergo trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes. In some embodiments, the MSC is positive for CD73, CD105, CD166, and CD90 and is negative for CD14, CD31, CD34, and CD45.

[0017] In some embodiments, EVs are analyzed via light scatter and fluorescence evaluation (e.g., NanoSight, Malvern Panalytical Ltd., United Kingdom). In some embodiments, the EVs are characterized by single particle interferometric reflectance imaging sensor technology to visualize and quantify fluorescent antibody-labeled particles (e.g., Nano View Biosciences, Boston, MA).

[0018] In some embodiments, the EV comprises a peptide or protein. In some embodiments, the EV comprises a nucleic acid. Nucleic acids include ribonucleic acids (RNA), such as siRNA, shRNA, and microRNA (miRNA).

[0019] In some embodiments, the EV comprises one or more factors (e.g., 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). Non-limiting examples of the one or more factors include: Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), IL-1 R6 (Interleukin 1 Receptor 6), LAMP2 (Lysosome-associated membrane glycoprotein 2), bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), GPR115 (Adhesion G protein-coupled receptor F4), CD63 antigen, CD 109 antigen, Serpin Fl (Pigment epithelium-derived factor), IGFBP-6 (Insulin-like growth factor binding protein-6), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor- 1 or SERPINE 1), Cathepsin B, IGFBP-2 (Insulin-like growth factor binding protein-2), Semaphorin 6C, IGF-2 (Insulin-like growth factor-2), Sortilin, Serpin B6, Dkk-3 (Dickkopf-related protein 3), CNTF (Ciliary neurotrophic factor), TSP-1 (Thrombospondin 1), GM-CSF Ra (Granulocyte-WSGR Ref. No. 66309-733.601 macrophage colony-stimulating factor receptor subunit alpha), Thrombomodulin, Endoglycan, (podocalyxin-like protein 2) IGFBP-3 (Insulin-like binding protein-3), RGM-C (Hemojuvelin), PF4 (Platelet Factor 4), MIF (Macrophage migration inhibitory factor), TGM4 (Protein- glutamine gamma-glutamyltransf erase 4), Periostin, Furin, TIMP-1 (Tissue inhibitor of MMPs 1), Decorin, PCK1 (Phosphoenol pyruvate carboxykinase, cytosolic) CD9 antigen, CD99 antigen, CA2 (Carbonic anhydrase 2), PRDX4 (Peroxidredoxin-4), Transferrin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), GP73 (Golgi membrane protein 1), CD81 antigen, Lumican, or TIMP-2 (Tissue Inhibitor of MMPs 2), and the factors of Table 1. An EV that comprises the one or more of the factors may include the one or more factors within the EV. An EV that comprises one or more of the factors may include the one or more factors anchored within the EV. An EV that comprises the one or more of the factors may be associated with the outside of the EV. In some embodiments, a factor may comprise a protein. In some embodiments, a factor may comprise a hormone. In some embodiments, a factor may comprise a growth factor. In some embodiments, a factor may comprise a nucleic acid.

[0020] Non-limiting example factors that may be present in the a composition of the present disclosure include: 2B4, 4-1BB, 4-1BB Ligand, 6Ckine, ACE, Activin A, Activin RIB, Activin RIIA, Activin RIM, ADA, ADAM23, ADAMS, ADAMTS13, ADAMTSL-1, Adiponectin, Adipsin, Aggrecan, AgRP, AIF, AKR1C4, Albumin, ALCAM, AMIGO, AMIG02, Aminopeptidase LRAP, Aminopeptidase P2, AMSH, ANG-1, ANG-2, ANG-4, Angiogenin, Angiotensinogen, ANGPTL4, ANGPTL7, APRIL, Arginase 1, ARSB, Artemin, Arylsulfatase A, ASAH2, ASAHL, Axl, B2M, B7-1, B7-H3, B7-H4, BAFF R, Bcl-10, BCL-2, Bcl-w, BCMA, BID, Biglycan, BLAME, BMP-2, BMP-8, BMP-9, BMPR-IA, Brevican, C-myc, CA125, CA13, CA4, CA8, Cadherin-13, Cadherin-4, Calreticulin, Calreticulin-2, Cardiotrophin-1, CASA, Caspr2, Cathepsin E, Cathepsin L, Cathepsin S, Cathepsin V, CD14, CD155, CD157, CD2, CD200, CD229, CD27, CD27 Ligand, CD28, CD30, CD300f, CD314, CD34, CD36, CD39L2, CD4, CD40, CD40L, CD42b, CD48, CD51, CD58, CD6, CD73, CD83, CD97, CD99- L2, CDC25B, CDNF, CDS, CEACAM-1, CEACAM-3, CEACAM-5, Cerberus 1, CES1, CES2, CF VII, CF XIV, Chemerin, CHI3L1, CHST2, CHST3, CHST4, cIAP-1, CILP-1, CK19, CL-P1, CLEC-1, CLEC-2, CLEC10A, Clq, ClqTNF9, cMASP3, CMG-2, Cochlin, COMT, Contactin- 1, Contactin-2, CPB1, CPE, CREG, CRELD2, CRIM1, CrkL, CSa, CTACK, CTRC, CXCL14, CXCL16, Cyr61, Cystatin B, Cystatin C, Cystatin D, Cystatin E M, Cystatin S, Cystatin SA,Cy statin SN, Cytokeratin-8, DAPP1, DCTN1, DDR1, Dectin- 1, Dectin-2, Desmin, Desmoglein2, Desm oglein-3, DKK-1, DLL4, DNAM-1, DNMT3A, Dopa Decarboxylase, DPPII, DPPIV, DR3, DR6, Draxin, DSCAM, DSPG3, Dtk, E-Selectin, ED AR, EDIL3, EG-VEGF, EGF R, EMMPRIN, ENA-78, Endocan, Endoglin, eNOS, ENPP-2, ENPP-7, Enteropeptidase, EpCAM,WSGR Ref. No. 66309-733.601EphA2, EphAl, EphB3, EphB4, EphB6, Ephrin-A4, Ephrin-B3, Epimorphin, Epiregulin, ErbB3, ErbB4, Erythropoietin, ESAM, EXTL3, FABP1, FABP2, FABP4, FABP6, FAP, Fas, FC AR, FcERI, FCRL1, FCRL3, FCRLB, FCRLS, FGF R5, FGF-12, FGF-16, FGF-17, FGF-19, FGF- 20, FGF-21, FGF-23, FGF-3, FGF-5, Ficolin-1, FKBP51, FLRG, FLRT1, FLRT2, Flt-3, Flt-3L, Follistatin, Follistatin-like 1, FUT8, Galanin, Galectin-1, Galectin-2, Galectin-3, Galectin-4, Galectin-7, Galectin-8, Galectin-9, GALNT10, GALNT2, GALNT3, Gash, GATA-4, GBA3, GCP-2, GDF-11, GDF-15, GDF-9, GFAP, GFR alpha-1, GFR alpha-2, GITR, GITR L, GKN1, GLP-1, Glycoprotein V, Glyoxalase II, Glypican 1, Glypican 2, Glypican 5, gpl30, GPR56, GPVI, Granulysin, Granzyme A, Granzyme B, Granzyme H, GRAP2, GRKS, GRO, GSTM1, GUSB, HAO-1, HAPLN1, HCC-1, HCC-4, hCGb, HE4, Hepsin, HGF, HGF R, HIF-1 alpha, HIF-1 beta, HO-1, HS3ST1, HS3ST3B1, HSD17B1, htPAPP-A, HTRA2, HVEM, I-TAC, ICAM-1, ICAM-2, ICAM-3, ICOS, IDO, IFN-gamma Rl, IFNab R2, IFNb, IGF-2R, IGFBP-5, IGSF3, IGSF4B, IL-1 F10, IL-1 R3, IL-1 R4, IL-1 RI, IL-1 RII, IL-10 Rb, IL-12p40, IL-13 R2, IL-15, IL-15 R, IL-17 RC, IL-17 RD, IL-17B, IL-17F, IL-18, IL-18 BPa, IL-19, IL-2 Ra, IL-2 Rb, IL-2 Rg, IL-20 R beta, IL-20 Ra, IL-21R, IL-22, IL-22 R alpha 1, IL-22BP, IL-23, IL-23 R, IL-24, IL-27, IL-27 Ra, IL-28 R alpha, IL-28A, IL-31, IL-31 RA, IL-32 alpha, IL-33, IL-4 Ra, IL-6, IL-6R, IL-7 R alpha, IL-8, IL-9, IL-lra, ILT2, ILT4, Inhibin A, Insulin, Insulin R, Integrin alpha 1, Integrin alpha 5, IP-10, Jagged 1, JAM-A, JAM-B, JAM-C, Kallikrein 1, Kallikrein 11, Kallikrein 12, Kallikrein 14, Kallikrein 5, Kallikrein 7, Kell, KIR2DL3, Kirrel3, KLF4, Kremen-2, Kynureninase, L-Selectin, LI CAM-2, LAIR2, LAMA4, LAMP, LAMP1, Langerin, LAP(TGFbl), LDL R, LEDGF, Legumain, LH, LIF, LIF R alpha, LIGHT, LILRB4, LIMPII, Lin28, Lipocalin-2, LOX-1, LRP-6„ST3GAL1, LRRC4, LRRTM4, LSECtin, LTbR, Lymphotactin, LYVE-1, MAP ID, Matrilin-2, Matrilin-3, Matriptase, MBL, Mcl-1, MCEMP1, MCP-2, MCP-3, MCP-4, MCSF, MDC, MDL-1, MEF2C, MEP1B, MEPE, MeprinA, Mer, Mesothelin, METAP2, MFRP, MIA, MICA, MICB, Midkine, MIP-3a, MIP-3b, MIP-lb, MIS RII, MMP-1, MMP-10, MMP-12, MMP-13, MMP-2, MOG, MPIF-1, MSP R, N-Cadherin, NAP-2, NCAM-1, NCAM-1 (CD56), NCK1, Nectin-1, Nectin-2, Nectin-3, Nectin-4, Neprilysin, Nestin, Netrin-4, Neudesin, Neurexin 3 beta, Neurogranin, Neuroligin 2, Neuropilin- 2, Neurturin, NG2, NGF R, Nidogen-1, Nidogen-2, NKp30, NKp44, NKp46, NKp80, Nogo Receptor, Nogo- A, Norrin, Notch- 1, Notch-3, NOV, NPDC-1, NPTXR, NQO-1, Nrf2, NRG1- 131, NSE, NTAL, NUDTS, Numb, NUP85, Olfactomedin-2, Olig2, OPG, or IL-13, OSCAR, OSM R beta, Osteoactivin, Osteoadherin, 0X40, 0X40 Ligand, P1GF-2, p27, p53, p63, PAPP- A, Pappaly sin-2, PARI, PARC, Pax3, PCSK2, PD-1, PD-ECGF, PD-L2, PDGF R alpha, PDGF Rb, PDGF-AA, PDGF-AB, PDGF-CC, PDX-1, PEAR1, PECAM-1, Pentraxin 3, Pepsinogen II, Persephin, PGRP-S, PIGF, PILR-alpha, Plexin A4, Plexin B3, Plexin DI, Podocalyxin,WSGR Ref. No. 66309-733.601Podoplanin, POGLUT1, PON1, Pref-1, PRELP, Presenilin 1, Procalcitonin, Progranulin, proGRP, Prolactin, Prolactin R, PRX2, PSA-total, PSMA, PSMA1, PTH, PTH1R, PTP1B, PU.l, PYY, R-Spondin 2, RAGE, Rai A, RANK, RANTES, RBP4, RCOR1, Reg3A, Reg4, RELT, Renin, Resistin, Ret Midkine, RGM-A, RGM-B, ROBO2, ROBO3, ROBO4, ROR1, ROR2, S100A1, S100A13, SALM4, SCCA2, SCF, SCF R, SDF-la, SDF-lb, Semaphorin 4C, Semaphorin 4D, Semaphorin 4G, Semaphorin 6A, Semaphorin 6B, Semaphorin 6D, Semaphorin 7A, Serpin A4, Serpin A5, sFRP-3, SH2D1A, Shh-N, SHP-1, Siglec-10, Siglec-11, Siglec-2, Siglec-6, Siglec-7, Siglec-9, SIRP alpha, Sirtuin 1, Sirtuin 2, Sirtuin 5, SLAM, SLITRK5, Smad4, SMPD1, SorCSl, SorCS2, SorCS3, SOST, SOX15, SOX2, SOX7, SOX9, SP-D, SPHK1, Spinesin, SPINK1, SR-AI, SREC-I, SREC-II, ST6GAL1, ST8SIA1, Stabilin-2, SULT2A1, Syndecan-3, Syndecan-4, Syntaxin 4, Syntaxin 6, TACE, TAFA1, TAFA2, TAFAS, TARC, TAZ, TC-PTP, TCN2, TECK, Tenascin R, Testican 2, TFF3, TFPI, TFPI-2, TGFa, TGFb2, TGFb3 FOLR2, TGFbl, TGM3, THAP11, Thrombospondin-2, Thrombospondin-5, THSD1, Thyroglobulin, Thyroid Peroxidase, Tie-1, Tie-2, TIM-1, TIM-3, TLR1, TLR2, TLR3, TLR4, TNF RI, TNF RII, TNF4, TNFb, TPO, TPP1, TPST2, TRACP, TRAIL, TRAIL Rl, TRAIL R3, TRAIL R4, TRANCE, TREM-1, TREM-2, TREML1, TrkC, TROP-2, Troponin C, TROY, Trypsin 1, Trypsin 3, TSH, TSK, TSLP, TSLP R, TWEAK R, Ubiquitin+1, UCH-L3, ULBP-1, ULBP-2, ULBP-3, ULBP-4, UNC5H3, UNC5H4, uPA, uPAR, Uromodulin, VAMP- 1, VAMP-2, VAP-A, VCAM-1, VE-Cadherin, VEGF, VEGF Rl, VEGF R2, VEGF R3, VEGF- C, Visfatin, vWF-A2, WIF-1, WISP-1, Wnt-4, XEDAR, and XIAP.

[0021] Table 1 below provides additional information regarding non-limiting examples of factors that may be comprised within an EV or EV composition of the present disclosure. In some embodiments, the factors may be present internally in an EV. In some embodiments, the factors may be present external to an EV within the composition.Table 1WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGRRef. No.66309-733.601WSGR Ref. No. 66309-733.601

[0022] In some embodiments, the EV comprises one or more nucleic acids (e.g., 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). Non-limiting examples of the one or more nucleic acids include: hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR- 199a-3p, hsa-miR-21-5p,hsa-miR-221-3p, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa- miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, and hsa-miR- 940. miRNA sequences may be obtained from mirbase.org. An EV that comprises the one or more of the nucleic acids may include the one or more nucleic acids within the EV. In some embodiments, the EV comprises the one or more nucleic acids internally. In some embodiments, the one or more nucleic acids are bound, attached, or embedded in the EV. In some embodiments, the one or more nucleic acids are present in the composition separate from the EV. In some embodiments, the one or more nucleic acids are both present internally in and externally to the EV within the composition.

[0023] Non-limiting example miRNA that may be present in the a composition of the present disclosure include: 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-10b-5p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-miR- 16-5p, hsa-miR-17-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-miR-21-3p, hsa- miR-21-5p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-24-3p, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-WSGR Ref. No. 66309-733.60130a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-34a-5p, hsa-miR-93-5p, hsa-miR-99a-5p, hsa-miR-99b-5p, hsa-miR- 100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa- miR-138-5p, hsa-miR-139-5p, hsa-miR-140-3p, hsa-miR-145-5p, hsa-miR-146a-5p, hsa-miR- 148a-3p, hsa-miR-152-3p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b- 3p, hsa-miR-197-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-miR-221-3p, hsa-miR-222-3p, hsa-miR-320a, hsa-miR-342-3p, hsa-miR- 345-5p, 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-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR- 663a, hsa-miR-874-3p, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-miR-940.

[0024] Non-limiting example miRNA that may be present in a composition of the present disclosure include: 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, and combinations of two or more thereof. In some embodiments, the miRNA may bind a sequence encoding TMPRSS2.

[0025] Proteins

[0026] Disclosed herein are compositions comprising a protein. The protein may be independent of an EV. For instance, the protein may not be present within the EV or within the membrane of the EV. The composition may comprises one or more proteins (e.g., 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). The protein may be present as a monomer or multimer. The protein may have a molecular weight (as a monomer or multimer as applicable) of at least about 10 kDa (kilodalton). In some embodiments, the protein may have a molecular weight greater than about 2 kDa (kilodaltons), 8 kDa, 10 kDa, 50 kDa, or 100 kDa. In some embodiments, a composition disclosed herein may comprise non-excipient components. In some embodiments, a non-excipient component may comprise a protein. In some embodiments, the molecular weight of any non-excipient component of the composition is greater than about 2 kDa (kilodaltons), 8 kDa, 10 kDa, 50 kDa, or 100 kDa. In some embodiments, any non-excipient component of the composition has a size of less than about 0.2 microns.

[0027] Non-limiting examples of the one or more proteins include: Ferritin, IGFBP-4 (Insulinlike growth factor binding protein-4), IL-1 R6 (Interleukin 1 Receptor 6), LAMP2 (Lysosome- associated membrane glycoprotein 2), bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), GPR115 (Adhesion G protein-coupled receptor F4), CD63 antigen, CD109 antigen, Serpin Fl (Pigment epithelium-derived factor), IGFBP-6 (Insulin-like growth factor binding protein-6), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), OPNWSGR Ref. No. 66309-733.601(Osteopontin), PAI-1 (Plasminogen activator inhibitor- 1 or SERPINE 1), Cathepsin B, IGFBP-2 (Insulin-like growth factor binding protein-2), Semaphorin 6C, IGF-2 (Insulin-like growth factor-2), Sortilin, Serpin B6, Dkk-3 (Dickkopf-related protein 3), CNTF (Ciliary neurotrophic factor), TSP-1 (Thrombospondin 1), GM-CSF Ra (Granulocyte-macrophage colony-stimulating factor receptor subunit alpha), Thrombomodulin, Endoglycan, (podocalyxin-like protein 2) IGFBP-3 (Insulin-like binding protein-3), RGM-C (Hemojuvelin), PF4 (Platelet Factor 4), MIF (Macrophage migration inhibitory factor), TGM4 (Protein-glutamine gammaglutamyltransferase 4), Periostin, Furin, TIMP-1 (Tissue inhibitor of MMPs 1), Decorin, PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic) CD9 antigen, CD99 antigen, CA2 (Carbonic anhydrase 2), PRDX4 (Peroxidredoxin-4), Transferrin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), GP73 (Golgi membrane protein 1), CD81 antigen, Lumican, or TEMP- 2 (Tissue Inhibitor of MMPs 2).

[0028] In some embodiments, the one or more proteins comprises TEMPI. In some embodiments, the TEMPI is present at about 200 pg / mL to about 80 ng / mL of the composition or about 30 pg / mL to about 12 ng / mL. In some embodiments, the one or more proteins comprises OPN. In some embodiments, the OPN is present at about 200 pg / mL to about 80 ng / mL of the composition or about 30 pg / mL to about 12 ng / mL. In some embodiments, the one or more proteins comprises IGFBP4. In some embodiments, the IGFBP4 is present at about 200 pg / mL to about 80 ng / mL of the composition or about 30 pg / mL to about 12 ng / mL. In some embodiments, the one or more proteins comprises osteonectin. In some embodiments, the osteonectin is present at about 200 pg / mL to about 80 ng / mL of the composition or about 30 pg / mL to about 12 ng / mL. In some embodiments, the concentration of the one or more proteins is measured by ELISA.

[0029] In some embodiments, the total protein concentration of the one or more proteins is about 10 to about 40 pg per ml of the composition. For example, the concentration in frozen product. In some embodiments, the total protein concentration of the one or more proteins is about 1.5 to about 6 pg per ml of the composition. For example, the concentration in an intravenous solution for administration.

[0030] In some embodiments, compositions described herein can safely target one or more (e.g., more than hundreds) different biomolecular interactions or signaling pathways. In some embodiments, compositions described herein can treat one or more injuries or diseases caused by multiple etiologies. In some embodiments, compositions described herein can be used for treating a disease or condition without identifying the pathogen underlying the disease or condition, thus offering an advantage for treating a disease or condition caused by an emerging or previously unknown pathogen.WSGR Ref. No. 66309-733.601A. Pharmaceutical Compositions

[0031] Compositions described herein may be administered in vivo in a pharmaceutically acceptable carrier. A pharmaceutically acceptable carrier may be not biologically or otherwise undesirable, z.e., the material may be administered to a subject, along with the EV and / or protein, without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained. As a non-limiting example, pharmaceutically acceptable carrier may comprise sodium chloride. Additional examples of pharmaceutically acceptable carriers include but are not limited to: microcrystalline cellulose, povidone, lactose, mannitol, dicalcium phosphate, croscarmellose sodium, sodium starch glycolate, crospovidone, magnesium stearate, stearic acid, colloidal silicon dioxide, talc, hydroxypropyl methylcellulose, ethylcellulose, polyethylene glycol, triethyl citrate, methylparaben, propylparaben, benzalkonium chloride, butylated hydroxytoluene, ascorbic acid, sucralose, saccharin, menthol, acacia, alginic acid, carboxymethylcellulose sodium, hypromellose, polyvinyl alcohol, sodium lauryl sulfate, titanium dioxide, starch, pregelatinized starch, xanthan gum, gelatin, glycerin, sorbitol, mannitol, maltitol, maltodextrin, polysorbate 20, polysorbate 80, propylene glycol, citric acid, sodium citrate, potassium sorbate, sodium benzoate, disodium edetate (EDTA), sodium chloride, calcium carbonate, silicified microcrystalline cellulose, carnauba wax, shellac, lecithin, dimethyl sulfoxide (DMSO), cyclodextrins (e.g., P-cyclodextrin, hydroxypropyl-P-cyclodextrin), mannitol, trehalose, sucrose, dextrose, polyethylene oxide, polyvinylpyrrolidone, and stearyl alcohol. As defined herein, “carrier” and “excipient” may be used interchangeably.

[0032] Parenteral administration of the composition, if used, is generally characterized by injection. The injection may be an intravenous injection. The injection may involve administration of the composition over a period of about 60 minutes. The composition for injection may comprise saline, e.g., 0.9% saline (NaCl). The 0.9% saline may be about 85% to about 99% or more of the composition by volume. For instance, the composition comprises about 85% of 0.9% saline and about 15% of an EV and / or protein component. The EV and / or protein component may be formulated in an isotonic infusion solution. The EV and / or protein component may comprise sodium chloride. The EV and / or protein component may comprise sodium chloride, sodium lactate, potassium chloride, and calcium chloride. The EV and / or protein component may comprise sodium chloride (NaCl) at 100-150 mEq / L, sodium lactate (CsHsNaOs) at 25-35 mEq / L, potassium chloride (KC1) at 2-6 mEq / L, calcium chloride (CaCL) at 1-5 mEq / L, and chloride ions (CL) at 80-130 mEq / L. The EV and / or protein component may comprise 0.1 M to 0.8 M polysaccharide. Non-limiting example polysaccharides include, but are not limited to, glucose, aldose (D-allose, D-altrose, D-mannose, etc.), glucopyranose,WSGR Ref. No. 66309-733.601 pentahydroxyhexanal, alpha-D-glucopyranoside dihydrate, a-D-glucopyranosyl-D-glucose, a-D- glucopyranosyl-dihydrate, polymer of P-D-glycopyranosyl units, P-D-fructofuranosyl a-D- glucopyranoside (anhydrous / dihydrate), P-D-galactopyranosyl-D-glucose, a-D-glucopyranosyl- a-D-glucopyranoside (anhydrous / dihydrate), galactose, pentoses (ribose, xylose, lyxose), dextrose, dodecacarbon monodecahydrate, fructose, sucrose, lactose, maltose, trehalose, agarose, D-galactosyl-0-(l-4)-anhydro-L-galactosyl, cellulose, starch, polyhydric alcohol, polyalcohol, alditol, erythritol, glycitol, glycerol, xylitol, and sorbitol. In some embodiments, the composition has a pH of about 6 to about 7.5.

[0033] In some embodiments, the composition is sterile by USP <71>. In some embodiments, the composition is endotoxin USP <85> free. In some embodiments, the composition is negative for mycoplasma DNA. In some embodiments, the composition is cell-free. In some embodiments, the composition is stored between -80 °C and -60 °C. In some embodiments, the composition is administered within 6 hours of thaw when maintained at ambient temperature. In some embodiments, the composition is present in a glass vial.B. Methods of Composition Production

[0034] Compositions herein comprising an EV and a factor may include components produced from a MSC. The EV of the composition may be produced from a MSC. The factor of the composition may be produced from a MSC. The EV and / or factor may be produced from a MSC cultured at one or more of the following conditions: about 0.1% to about 5% oxygen, reduced or no serum, pH of about 5-7.5, reduced glucose, increased temperature, or these elements in various combinations. The factor may comprise a protein. The MSC may be cultured under the aforementioned one or more conditions after the cell achieves confluency. The MSC may be cultured under the aforementioned one or more conditions after the cell is cultured at 37°C and 5% CO2. The EV and / or factor of the composition may be obtained from a method comprising: (1) growing the MSCs to 50-80% confluency, then (2) culturing the MSCs at one or more of the following conditions: about 1% to about 5% oxygen, reduced or no serum, reduced glucose, or increased temperature. The growing at step (1) may be performed at about 37°C and about 5% CO2. Culturing the MSCs may comprise introducing the cells to a culture media. The culture media may comprise basal media. For instance, basal media comprising amino acids, vitamins, and inorganic salts, and optionally a carbon source such as glucose. The culture media may comprise an isotonic infusion solution. In a non-limiting example embodiment, the culture media comprises a basal media and an isotonic infusion solution. The culture media may comprise basal media and sodium chloride. The culture media may comprise basal media, sodium chloride, sodium lactate, potassium chloride, and calcium chloride. The culture mediaWSGR Ref. No. 66309-733.601 may comprise a basal media and an isotonic infusion solution. The culture media may comprise basal media and sodium chloride, without serum. The culture media may comprise basal media, sodium chloride, sodium lactate, potassium chloride, and calcium chloride, without serum. The culturing at step (2) may occur over a period of 1, 2, 3, 4, 5, 6, or 7 days, wherein the oxygen and pH may change over time. At any time during the culturing step, the pH may be about 6.5 to about 7. At any time during the culturing step, the oxygen may be at about 0.5%, 1%, 1.5%, or 2%.

[0035] In some embodiments, the EV and / or factor may be buffer exchanged into a concentrated solution. In some embodiments, the concentrated solution may comprise electrolytes. In some embodiments, the EV and / or factor solution comprising the concentrated solution may be diluted using, for example, saline prior to administration. In some embodiments, the EV and / or factor may be separated from the MSC. In some embodiments, the separation may comprise but is not limited to centrifugation, elutriation, filtering, sedimentation, decantation, sieving, ultrafiltration, microfiltration, cross-flow filtration, dialysis, precipitation, ion-exchange chromatography, sizeexclusion chromatography, affinity chromatography, electrophoresis, density gradient separation, differential centrifugation, flotation, magnetic separation, flocculation, membrane filtration, vacuum filtration, tangential -flow filtration, and / or field-flow fractionation.

[0036] The EV and / or factor produced in the second culturing step (2) may be purified (e.g., partially or entirely), from the culture media of the second culturing step (2). The purification may comprise formulating the EV and / or the factor into an EV and / or factor component. The EV and / or factor component may comprise an isotonic infusion solution. The EV and / or factor component may comprise sodium chloride. The EV and / or factor component may comprise sodium chloride, sodium lactate, potassium chloride, and calcium chloride. The EV and / or factor component may comprise sodium chloride (NaCl) at 100-150 mEq / L, sodium lactate (CsHsNaCh) at 25-35 mEq / L, potassium chloride (KC1) at 2-6 mEq / L, calcium chloride (CaCL) at 1-5 mEq / L, and chloride ions (CL) at 80-130 mEq / L. The EV and / or factor component may comprise a polysaccharide, e.g., 0.1 M to 0.8 M polysaccharide. Non-limiting example polysaccharides include, but are not limited to, glucose, aldose (D-allose, D-altrose, D-mannose, etc.), glucopyranose, pentahydroxyhexanal, alpha-D-glucopyranoside dihydrate, a-D- glucopyranosyl-D-glucose, a-D-glucopyranosyl-dihydrate, polymer of P-D-glycopyranosyl units, P-D-fructofuranosyl a-D-glucopyranoside (anhydrous / dihydrate), P-D-galactopyranosyl-D- glucose, a-D-glucopyranosyl-a-D-glucopyranoside (anhydrous / dihydrate), galactose, pentoses (ribose, xylose, lyxose), dextrose, dodecacarbon monodecahydrate, fructose, sucrose, lactose, maltose, trehalose, agarose, D-galactosyl-0-(l-4)-anhydro-L-galactosyl, cellulose, starch, polyhydric alcohol, polyalcohol, alditol, erythritol, glycitol, glycerol, xylitol, and sorbitol. TheWSGR Ref. No. 66309-733.601 amount of polysaccharide in the EV and / or factor component may be about 0.2 M to about 0.6 M, or about 0.4 M. In some embodiments, the EV and / or factor component is frozen. The frozen material may be thawed and combined with saline (e.g., 0.9% saline or sodium chloride) to generate an IV formulation for IV administration. The amount of polysaccharide in the IV formulation may be about 40 mM to about 80 mM, or about 60 mM.

[0037] The EV and / or factor component may be filter-sterilized. The EV and / or factor component may be filter sterilized after and / or during formulation from the culture media into the EV and / or factor component. The EV and / or factor may be concentrated. The concentration may occur during and / or after purification and / or exchange from the culture media into the EV and / or factor component. The EV and / or factor may be frozen, e.g., after and / or during purification. If frozen, the EV and / or factor may be formulated with a cryoprotectant prior to freezing. The cryoprotectant may comprises a polysaccharide, e.g., as described above. In some embodiments, the EV and / or factor component may be buffer exchanged into a concentrated solution. In some embodiments, the concentrated solution may be diluted prior to administration. In some embodiments, the concentrated solution may be diluted with saline.

[0038] In some embodiments, reduced glucose is less than a normal control (e.g., 4.5 g / L). For example, the glucose reduction may be about 5% to about 15%, from about 10% to about 20%, from about 15% to about 25%, from about 20% to about 30%, or from about 25% to about 35% of the glucose in a normal control. In some embodiments, the reduced glucose is present in the MSC culture media (e.g., at step (2) as noted above) at a concentration of about 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, or 4.0 g / L, or a range between any two of these values. In some embodiments, glucose is present at a concentration of less than or no more than 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.

[0039] In some embodiments, the MSC is cultured, e.g., at step (2) at about 0.1% to about 5% oxygen, for example, about 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, or 5% oxygen. In an example embodiment, the oxygen is about 0.5% to about 1.5%, or about 1%.

[0040] The pH at which the MSC is cultured, e.g., at step (2), can be from about 6.0 to about 7.4, for example, from 6.5 to about 7, or about 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.

[0041] The temperature of the culture environment, e.g., at step (2), may be raised relative to physiologic homeostasis temperature (e.g., 37°C). In one aspect, the temperature of the culture can 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,WSGR Ref. No. 66309-733.60136.6, 36.7, 36.8, 36.9, 37.0, .37.1, 37.2, 37.3, 37.4, 37.5, 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°C.

[0042] In some embodiments, compositions described herein can be lyophilized for packaging and storing. In some embodiments, compositions described herein can be stored at ambient or room temperature (e.g., between 60°F and 75°F or between 15°C and 24°C). In some embodiments, the stability of compositions described herein at ambient or room temperature can provide advantages for logistics and / or delivery.

[0043] Example therapeutic compositions herein comprise MSC secretome compositions and / or components derived from mesenchymal stem cells (MSCs) (e.g., biomolecules such as nucleic acids like miRNA and peptides like growth factors). In one aspect, disclosed herein are MSC secretome compositions (including, but not limited to MSC factor, MSC protein, MSC protein factor, MSC growth factor, MSC exosome, MSC extracts and / or extracellular vesicle comprising compositions) for the treatment, inhibition, decrease, reduction, amelioration, and / or prevention of conditions such as, for example, idiopathic facial paralysis.

[0044] MSCs are multipotent cells that have the ability to differentiate into a multitude of cell types including myocytes, chondrocytes, adipocytes, and osteoblasts. Typically, these cells can be found in the placenta, umbilical cord blood, adipose tissue, bone marrow, or amniotic fluid, including perivascular tissue. As used herein, "MSC" refers to non-terminally differentiated cells including but not limited to multipotential stem cell, multipotential stromal cell, stromal vascular cells, pericytes, perivascular cells, stromal cells, pluripotent cells, multipotent cells, adipose- derived fibroblast-like cells, adipose-derived stromal vascular fraction, adipose-derived MSC, bone marrow-derived fibroblast-like cells, bone marrow-derived stromal vascular fraction, bone marrow-derived MSC, tissue-derived fibroblast-like cells, adult stem cells, adult stromal cells, keratinocytes, and / or melanocytes.

[0045] In some embodiments, an MSC secretome composition comprises MSC factors, MSC proteins, MSC protein factors, MSC growth factors, MSC exosomes, extracellular vesicles, extracellular vesicle isolate product (EVIP), acellular extract of MSC or a MSC lysate, or a combination of two or more thereof, obtained from an MSC. The MSC may be a human MSC, fibroblast-like cell, or a non-human animal MSC including, but not limited to an MSC from horses, cows, pigs, sheep, non-human primates, dogs, cats, rabbits, rats, or mice. In embodiments, the MSC may be derived from the patient to which the composition will be applied (autologous) or derived from another individual (allogeneic). The MSC may be culture expanded to collect the conditioned media and / or to increase the quantity of cells for the lysate or used freshly prior to incorporation into a therapeutic composition of the present disclosure.WSGR Ref. No. 66309-733.601

[0046] The MSC secretome compositions (including, but not limited to MSC factors, protein factors, MSC exosomes, MSC extracts and / or extracellular vesicle comprising compositions) may comprise about 0.00001 to about 20 wt.%, such as from about 0.01 to about 10 wt.%, of a mesenchymal stem cell (MSC) extract, MSC exosome, or MSC factor, or MSC protein factor preparation.

[0047] MSCs can be selectively stimulated to produce the MSC secretome compositions or components of the MSC secretome compositions disclosed herein. The stimulated MSCs may product MSC factors, MSC proteins, MSC protein factors, MSC growth factors, secretomes, mesenchymal stem cell secretome, cytokines, chemokines, mesenchymal stem cell proteins, peptides, glycosaminoglycans, extracellular matrix (ECM), proteoglycans, or extracellular vesicles (e.g., exosomes), or a combination of two or more thereof. MSC factors include, but are not limited to, prostaglandin E2 (PGE2), transforming growth factor 131 (TGF-131), hepatocyte growth factor (HGF), stromal cell derived factor-1 (SDF-1), nitric oxide, indoleamine 2,3- dioxygenase, interleukin-4 (IL-4), IL-6, interleukin- 10 (IL-10), IL-1 receptor antagonist and soluble TNF-a receptor, insulin-like growth factors, fibroblast growth factors (FGF) 1-23 (especially, FGF1 and FGF2), bone morphogenetic proteins (BMPs) 1-15, epidermal growth factor (EGF), transforming growth factor-a (TGF-a) 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), as well as hormones including estrogen, and thyroid hormones.

[0048] Culturing the MSCs may occur under wound healing and / or hypoxic conditions. Wound healing conditions may comprise about 1% to about 5% oxygen, reduced or no serum, reduced glucose, or these elements in various combinations. The combined reduced nutrient and metabolite environment may trigger the cultured cells to produce wound healing and antiinflammatory ECM proteins and growth factors to direct tissue healing. In one aspect, the MSC secretome composition comprises MSC growth factors, MSC exosomes, and / or cellular extracts of MSCs or MSC lysates obtained from MSCs cultured under standard hyperoxic culturing conditions (for example, 21% oxygen) or MSCs cultured under artificial wound healing conditions (such as, for example, 0.1% to about 5% oxygen).

[0049] Artificial wound healing conditions used to culture MSCs may include one or more of the following growth conditions reduction in glucose availability, reduction in oxygen tension, reduction in pH, and increased temperature.

[0050] In some embodiments, the glucose availability can be reduced relative to normal control (e.g., 4.5 g / L). Modified culture media to reduce glucose, but not damage the cells can beWSGR Ref. No. 66309-733.601 between 0 and 50% reduction in glucose, more preferably between about 5% and 40% reduction in glucose. For example, MSC artificial wound healing culture conditions can comprise glucose reduction of about 5% to about 15%, from about 10% to about 20%, from about 15% to about 25%, from about 20% to about 30%, or from about 25% to about 35%. In some embodiments, glucose is present at a concentration of about 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, or 4.0 g / L, or a range between any two of these values. In some embodiments, glucose is present at a concentration of less than or no more than 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2,I.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.

[0051] In some embodiments, oxygen tension can be reduced to oxygen levels to hypoxic conditions. Normal atmospheric oxygen is approximately 21% and any reduction is considered hypoxic. Thus, in one aspect, MSCs can be cultured at between 0.0% and 20.9% oxygen, from about 0.1% to about 0.5% oxygen, from about 0.1% to about 2.0%, from about 0.1% to about 5.0% oxygen, from about 0.5% to 5.0%, from about 1.0% to about 10% oxygen, about 5.0% to about 10.0% oxygen; and from about 10.0% to about 15.0%. The hypoxic oxygen conditions may be an aspect of artificial wound healing conditions. Oxygen tension may be between about 0.5% and 20.5% oxygen when culturing MSCs to produce a therapeutic secretome composition comprising extracellular vesicles and / or MSC-secreted factors, such as, for example, 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, 10.6, 10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3,I I.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, 15.7, 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, or 20.0% oxygen, or a range between any two of these values.

[0052] The pH can also be reduced during MSC culturing. The pH can be from about 6.0 to about 7.4, for example, from 6.0 to about 6.4, from about 6.2 to about 6.4, from about 6.2 to about 6.6, from about 6.4 to about 6.6, from about 6.4 to about 6.8, or from about 6.6 to about 7.0, such as 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.

[0053] The temperature of the culture environment may be raised relative to physiologic homeostasis temperature (e.g., 37°C). In one aspect, the culture conditions for the MSCs canWSGR Ref. No. 66309-733.601 comprise from about 35°C to about 39°C, from about 35°C to about 36°C, from about 36°C to about 37°C, from about 37°C to about 38°C, from about 38°C to about 39°C, from about 39°C to about 40°C. In one aspect, the temperature of the culture can 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, 37.5, 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°C.

[0054] In some embodiments, the culture media is serum free. In some embodiments, the serum free culture media comprises platelet lysate. In some embodiments, the platelet lysate is human platelet lysate (HPL). In some embodiments, the serum free culture media comprises at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% of HPL by volume, or a range between any two of these values. In some embodiments, the culture media comprises from 8% to 12%, 5% to 15%, or 9% to 11% of HPL by volume.

[0055] In one aspect, the therapeutic composition, e.g., MSC secretome compositions (including, but not limited to MSC growth factor, MSC exosome, MSC extracts and / or extracellular vesicle comprising compositions) comprise an additive. The additive can serve as a protective coating. The additive can reduce degradation of components within the composition, such as a growth factor. The additive may be a cryoprotectant. The cryoprotectant may be an oligosaccharide. The additive may be a polymer. Polymers include a relatively high molecular weight organic compound, natural or synthetic, whose structure can be represented by a repeated small unit, the monomer. Non-limiting examples of polymers include polyethylene, rubber, cellulose. Synthetic polymers are typically formed by addition or condensation polymerization of monomers. The term copolymer may refer to a polymer formed from two or more different repeating units (monomer residues). By way of example and without limitation, a copolymer can be an alternating copolymer, a random copolymer, a block copolymer, or a graft copolymer. The polymer may be a natural polymer, synthetic polymer, homopolymer, heteropolymer or copolymer. In one aspect, the polymer can comprise a copolymer, block copolymer, diblock copolymer, and / or triblock copolymer. In one aspect, the additive may comprise a biocompatible polymer. In one aspect, the biocompatible polymer is crosslinked. Biocompatible polymers include, but are not limited to, 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 polyethylene oxide) (PEO); poly(oxyethylated polyol); poly(olefinic alcohol); polyvinylpyrrolidone); poly(hydroxyalkylmethacrylamide); poly(hydroxyalkylmethacrylate); poly(saccharides); poly(hydroxy acids); poly(vinyl alcohol), polyhydroxyacids such as poly(lactic acid), poly (glyWSGR Ref. No. 66309-733.601 colic acid), and poly (lactic acid-co-glycolic acids); polyhydroxyalkanoates such as poly3- hydroxybutyrate or poly4-hydroxybutyrate; polycaprolactones; poly(orthoesters); polyanhydrides; poly(phosphazenes); poly(lactide-co-caprolactones); polycarbonates such as tyrosine polycarbonates; polyamides (including synthetic and natural polyamides), polypeptides, and poly(amino acids); polyesteramides; polyesters; poly(dioxanones); poly(alkylene alkylates); hydrophobic polyethers; polyurethanes; polyetheresters; polyacetals; polycyanoacrylates; polyacrylates; polymethylmethacrylates; polysiloxanes; poly(oxyethylene) / poly(oxypropylene) copolymers; polyketals; polyphosphates; polyhydroxyvalerates; polyalkylene oxalates; polyalkylene succinates; poly(maleic acids), as well as copolymers thereof. Biocompatible polymers can also include polyamides, polycarbonates, polyalkylenes, polyalkylene glycols, polyalkylene oxides, polyalkylene terepthalates, polyvinyl alcohols (PVA), methacrylate PVA(m- PVA), polyvinyl ethers, polyvinyl esters, polyvinyl halides, polyvinylpyrrolidone, polyglycolides, polysiloxanes, polyurethanes and copolymers thereof, alkyl cellulose, hydroxyalkyl celluloses, cellulose ethers, cellulose esters, nitro celluloses, polymers of acrylic and methacrylic esters, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxybutyl methyl cellulose, cellulose acetate, cellulose propionate, cellulose acetate butyrate, cellulose acetate phthalate, carboxylethyl cellulose, cellulose triacetate, cellulose sulphate sodium salt, poly (methyl methacrylate), poly(ethylmethacrylate), poly(butylmethacrylate), poly(isobutylmethacrylate), poly(hexlmethacrylate), poly(isodecylmethacrylate), poly(lauryl methacrylate), poly (phenyl methacrylate), poly(methyl acrylate), poly (isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate), polyethylene, polypropylene, poly(ethylene glycol), polyethylene oxide), poly(ethylene terephthalate), poly(vinyl alcohols), poly(vinyl acetate, poly vinyl chloride polystyrene and polyvinylpryrrolidone, derivatives thereof, linear and branched copolymers and block copolymers thereof, and blends thereof. Exemplary biodegradable polymers include polyesters, poly(ortho esters), poly(ethylene amines), poly(caprolactones), poly(hydroxybutyrates), poly(hydroxyvalerates), polyanhydrides, poly(acrylic acids), polyglycolides, poly(urethanes), polycarbonates, polyphosphate esters, polyphospliazenes, derivatives thereof, linear and branched copolymers and block copolymers thereof, and blends thereof.

[0056] In some embodiments the protective coating comprises carbohydrate construction of monosaccharides as well as carbohydrate polymers such as di saccharides or polysaccharides including but not limited to non-reducing poly or disaccharides as well as any combination thereof. Examples of carbohydrates that can be used in the protective coating comprise Glucose, Aldoses (D-Allose, D-Altrose, D-Mannose, etc.), Glucopyranose, Pentahydroxyhexanal, a-D- Glucopyranosyl-D-glucose, a-D-Glucopyranosyl-dihydrate, Polymer of P-D-GlycopyranosylWSGR Ref. No. 66309-733.601 units, P-D-Fructofuranosyl a-D-glucopyranoside (anhydrous / dihydrate), f3-D-Galactopyranosyl- D-glucose, a-D-Glucopyranosyl-a-D-glucopyranoside (anhydrous / dihydrate), Galactose, Pentoses (Ribose, xylose, lyxose), Dextrose, Dodecacarbon monodecahydrate, Fructose, Sucrose, Lactose, Maltose, Trehalose, Agarose, D-galactosyl-0-(l-4)-anhydro-L-galactosyl, Cellulose, Polymer of P-D-Glycopyranosyl units, and Starch, as well as, Polyhydric alcohols, Polyalcohols, Alditols, Erythritol, Glycitols, Glycerol, Xylitol, and Sorbitol.

[0057] 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-co-gly colic acid). The particles can contain one more of the following polyesters: homopolymers including glycolic acid units, referred to herein as "PGA", and lactic acid units, 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-lactide5 collectively referred to herein as "PLA", and caprolactone units, such as poly(e-caprolactone), collectively referred to herein as "PCL"; and copolymers including lactic acid and glycolic acid units, such as various forms of poly(lactic acid-co-glycolic acid) and poly(lactide-co-glycolide) characterized by the ratio of lactic acid:gly colic acid, collectively referred to herein as "PLGA"; and polyacrylates, and derivatives thereof. Exemplary polymers also include copolymers of polyethylene glycol (PEG) and the aforementioned polyesters, such as various forms of PLGA-PEG or PLA-PEG copolymers, collectively referred to herein as "PEGylated polymers". In certain embodiments, the PEG region can be covalently associated with polymer to yield "PEGylated polymers" by a cleavable linker. In one aspect, the polymer comprises at least 60, 65, 70, 75, 80, 85, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent acetal pendant groups.

[0058] The triblock copolymers disclosed herein comprise a core polymer such as, example, polyethylene glycol (PEG), polyvinyl acetate, polyvinyl alcohol, polyvinyl pyrrolidone (PVP), polyethyleneoxide (PEO), poly(vinyl pyrrolidone-co-vinyl acetate), polymethacrylates, polyoxyethylene alkyl ethers, polyoxyethylene castor oils, polycaprolactam, polylactic acid, polyglycolic acid, poly(lactic-glycolic) acid, poly(lactic co-glycolic) acid (PLGA), cellulose derivatives, such as hydroxymethylcellulose, hydroxypropylcellulose and the like. Examples of diblock copolymers that can be used in the protective coatings disclosed herein comprise a polymer such as, example, polyethylene glycol (PEG), polyvinyl acetate, polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP), polyethyleneoxide (PEO), poly(vinyl pyrrolidone-co-vinyl acetate), polymethacrylates, polyoxyethylene alkyl ethers, polyoxyethylene castor oils, polycaprolactam, polylactic acid, polyglycolic acid, poly(lactic-glycolic) acid, poly(lactic co- glycolic) acid (PLGA).WSGR Ref. No. 66309-733.601

[0059] In one aspect, the protective coating contains (i.e., the encapsulated, the encapsulated compositions can further comprise lecithin or hydrolyzed lecithin as a carrier or as encapsulation material. As used herein, lecithin and / or hydrolyzed lecithin coatings include coatings comprising phosphatidyl choline, phosphatidyl inositol, phosphatidyl ethanolamine, phosphatidyl serine, and phosphatidic acid. Sources of the lecithin can be plant or animal sources.

[0060] In one aspect, any of the polymers, monosaccharides, disaccharides, or polysaccharides used to form the protective coating formed by placing the MSC additive in an encapsulating solution can be at an appropriate concentration for form the protective coating. For example, polymers, monosaccharides, disaccharides, or polysaccharides can be at any concentration between 0.01 mM and 10.0 M concentration, for example, from about 0.01 M to about 0.1 M, from about 0.1 mM to about 1.0 M, from about 1.0 M to about 10.0 M.

[0061] In one aspect, the MSC secretome compositions (including, but not limited to MSC growth factor, MSC factor, MSC protein, MSC exosome, MSC extracts and / or extracellular vesicle comprising compositions) disclosed herein may comprise any known ingredients typically found pharmaceutical fields such as agents for combating free radicals; bactericides; sequestering agents; preservatives; basifying or acidifying agents; fragrances; surfactants; fillers; natural products or extracts of natural product, such as aloe or green tea extract; vitamins; or coloring materials. Other ingredients that may be combined with the powder may include an antioxidant, which can be selected from a variety of antioxidants. Suitable antioxidants include vitamins, such as Vitamin C (L- Ascorbate, Ascorbate-2 Phosphate magnesium salt, Ascorbyl Palmitate, Tetrahexyl decyl Ascorbate), Vitamin E (Tocotrienol), Vitamin A (retinol, retinal, retinoic acid, provitamin A carotenoids, such as beta-carotene), N-acetyl glucosamine, or other derivatives of glucosamine. Other ingredients 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-y-linolenic acid (DGLA), arachidonic acid (ARA), and others. The fatty acids may be derived from various sources including evening primrose oil, black currant oil, borage oil, or GLA modified safflower seeds. Other ingredients may include a platelet rich fibrin matrix, at least one ingredient to support ECM production and production of hyaluronic acid, such as N-acetyl glucosamine or other derivatives of glucosamine, ultra-low molecular weight (ULMW) hyaluronic acid, chondroitin sulfate, or keratin sulfate.

[0062] The MSC secretome compositions may be produced by a method comprising culturing MSCs collected from a donor under stimulation conditions; collecting and combining the secretome with an additive, and freezing the combination, wherein the combination is formulated into a therapeutic composition. The frozen combination may be a dry powder. TheWSGR Ref. No. 66309-733.601 therapeutic composition may comprise the dry powder reconstituted into a delivery vehicle suitable for human administration, e.g., saline. The therapeutic composition may comprise factors, exosomes, peptides, proteins, cytokines, growth factors, extracellular matrix (ECM), proteoglycans, glycosaminoglycans, chemokines, or a combination of two or more thereof. The MSCs may be bone marrow MSCs. The MSCs may be human MSCs, animal MSCs, multipotential stromal cells, fibroblasts, or fibroblast cells. The MSC secretome compositions may be produced by a method comprising culturing MSCs collected from a donor under stimulation conditions, lysing the MSCs to generate an extracted lysate, concentrating the extracted lysate, and combining the extracted lysate with an additive, and freezing the mixture to generate a dry powder. In example embodiments, the additive is a saccharide. In some embodiments, freezing includes lyophilization. The powder may comprise a concentrated collection of analgesic MSC secretomes components (e.g., exosomes, extracellular matrix) that are specific to anti-inflammation.

[0063] The methods may also include filter-sterilizing, concentrating, freezing, or freeze drying the MSC culture medium, e.g., after culturing under wound healing conditions. Additionally, the MSC culture medium may be combined with a cryoprotectant prior to freezing. The cryoprotectant may be the additive, such as an oligosaccharide.

[0064] Lysing may be achieved by the addition of a hypotonic solution or repeated freeze-thaw processes to disrupt the cell membranes. The cells may be lysed while attached to the culture surface or in suspension. The cells may be enzymatically released and / or lysed by mechanical homogenization.

[0065] Stimulation conditions include stimulating the MSC to selectively secrete desired antiinflammatory proteins, peptides, glycosaminoglycans, proteoglycans, exosomes and / or secretomes. Stimulation may be achieved by adjusting the cell growth conditions, such as cell confluency, culture media supplements, nutritional supplements, oxygen levels, length of culture in those conditions, cell passage number or combinations of those, and the like. Stimulation may comprise growth under wound healing conditions. Wound healing conditions may comprise about 1% to about 5% oxygen, reduced or no serum, reduced glucose, or these elements in various combinations.C. Pharmaceutical carriers / Delivery of pharmaceutical products

[0066] Therapeutic compositions, such as MSC secretome compositions, may be administered in vivo in a pharmaceutically acceptable carrier. By "pharmaceutically acceptable" is meant a material that is not biologically or otherwise undesirable, i.e., the material may be administered to a subject, along with the nucleic acid or vector, without causing any undesirable biologicalWSGR Ref. No. 66309-733.601 effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained. The carrier may be selected to minimize any degradation of the active ingredient and to minimize any adverse side effects in the subject. The compositions may be administered parenterally (e.g., intravenously).

[0067] Parenteral administration of the composition, if used, is generally characterized by injection. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution of suspension in liquid prior to injection, or as emulsions. Parenteral administration may involve use of a slow release or sustained release system such that a constant dosage is maintained. Preparations for parenteral administration may include a sodium chloride solution.D. Therapeutic Uses

[0068] In some embodiments, therapeutic compositions herein, such as MSC secretome compositions comprise a factor and / or exosome. The factor and / or exosome may induce cell proliferation and / or angiogenesis. The factor may comprise but is not limited to a protein, a peptide, a growth factor, a nucleic acid, and / or hormone. The therapeutic compositions herein may comprise mitogenic proteins such as transforming growth factor-alpha (TGF-a), TGFP, hepatocyte growth factor (HGF), epithelial growth factor (EGF), basic fibroblast growth factor (FGF-2) and / or insulin-like growth factor-1 (IGF-1). These increase fibroblast, epithelial and endothelial cell division. The therapeutic compositions herein may comprise vascular endothelial growth factor (VEGF), IGF-1, EGF and / or angiopoietin-1, which recruit endothelial lineage cells and initiate vascularization. The therapeutic compositions herein may comprise components that are anti-inflammatory. The therapeutic compositions herein may comprise components that are immunomodulatory. Non-limiting example anti-inflammatory proteins include prostaglandin 2, TGF-131, HGF, SDF-1, nitrous oxide, indoleamine 2, 3 -di oxygenase, IL-4, IL- 10, IL-1 receptor antagonist and soluble tumor necrosis factor-a receptor. The therapeutic compositions herein may comprise components that prevent proliferation and function of inflammatory immune cells, including T-cells, natural killer cells, B-cells, monocytes, macrophages, and dendritic cells.

[0069] A characteristic of chronically inflamed environments is a persistent imbalance in the types of helper T-cells and macrophages. In some embodiments, therapeutic compositions, e.g., MSC secretome compositions, indirectly promote the transition of TH1 to TH2 cells by reducing INF-y and increasing IL-4 and IL- 10. The restored TH1 / TH2 balance may improve tissue regeneration in cartilage, muscle, and other soft tissue. The reduction in INF-y and secretion of IL-4 may promote a shift in macrophages from Ml (proinflammatory, anti-angiogenic and tissueWSGR Ref. No. 66309-733.601 growth inhibition) to M2 (anti-inflammatory, pro-remodeling and tissue healing) type, for skeletal, muscular, and neural healing and regeneration.II. Methods of Treating Idiopathic Facial Paralysis

[0070] In some embodiments, the therapeutic compositions, e.g., MSC secretome compositions, disclosed herein are used in methods of treating idiopathic facial paralysis in a subject. Any of the therapeutic compositions described herein may be used in such a method. In some embodiments, the subject exhibits idiopathic facial paralysis. In some embodiments, the subject exhibits Bell’s Palsy. In some embodiments, the subject exhibits facial palsy. In some embodiments, the subject exhibits secondary facial paralysis. In some embodiments, paralysis affects one or more extracranial and / or intracranial components. In some embodiments, paralysis affects the facial nerve (VII). In some embodiments, the subject exhibits extracranial paralysis, intracranial paralysis, epiphora or a dry eye, inability to close one or both eyes, drooping of the oral commissure, salivary incompetence, facial tingling, fasciculations of one or more facial muscles, inability to smile, taste disturbance, hyperacusis, earache, dizziness, loss of balance, tinnitus, hearing loss, taste disturbances, and / or inability to raise one or both eyebrows. In some embodiments, onset of any of the symptoms disclosed herein is acute. In some embodiments, persistence of any of the symptoms disclosed herein is chronic. In some embodiments, one or more of the symptoms disclosed herein persist for 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or longer.

[0071] In some embodiments, the subject is treated with one or more of surgery, facial reconstruction, oral steroids, anti-viral medications, acupuncture, massage, vitamin B-12, diuretics, gabapentin, potassium, valaciclovir, prednisone, physical therapy, botulinum toxin, facial exercises, laser treatment, and / or herbal medicine prior to administration of the therapeutic composition. In some embodiments, the subject experiences depression, psychological disturbance, difficulty with social interaction, difficulty communicating, inability to communicate, and / or inability to express emotion. In some embodiments, the subject is evaluated for cerebrovascular accident prior to treatment. In some embodiments, the subject’s radiological studies are unremarkable before and / or after the administration of the therapeutic composition.

[0072] In some embodiments, idiopathic facial paralysis comprises Bell’s Palsy. In some embodiments, idiopathic facial paralysis comprises secondary facial paralysis. In some embodiments, vascular secondary and tertiary ischemia cause the facial nerve sheath to thickenWSGR Ref. No. 66309-733.601 and inhibit nerve function in cases of facial paralysis. In some embodiments, facial paralysis present with viral involvement. Viruses implicated in facial paralysis but not limited to include Herpes simplex, Herpes zoster, Epstein-Barr virus, Coxsackie virus, rubella and cytomegalovirus. In some embodiments, Lyme disease contributes to facial paralysis. In some embodiments, facial paralysis is associated with or follows the onset of pregnancy. In some embodiments, demyelination contributes to facial paralysis. In some embodiments, autoimmune involvement is associated with facial paralysis. In some embodiments, high levels of interleukins such as interleukin-6 (IL-6) and IL-8 and / or elevated levels of tumor necrosis factor gamma (TNF-gamma) and TNF-alpha are associated with facial paralysis. In some embodiments, round inflammatory cells are present in the facial nerves in cases of facial paralysis. Demyelination may or may not be present. Inflammatory cells have been reported between nerve bundles with normal blood vessels between the internal acoustic meatus and the stylomastoid foramen. A segmental breakdown of the myelin sheath may be observed. In some embodiments, treatment with the therapeutic product disclosed herein reduces inflammation. Nerve recovery following treatment with the therapeutic product disclosed herein may be partial or complete. In some embodiments, patient experience a lower quality of life before treatment as compared to after treatment. In some embodiments, an improvement in conditions not associated with facial paralysis is reported following treatment. In some embodiments, improvement in conditions including but not limited to psoriatic arthritis and systemic sclerosis is reported following treatment.

[0073] In some embodiments, the therapeutic product is administered via intravenous infusion. In some embodiments, the therapeutic product is administered on separate days. In some embodiments, the therapeutic product is administered 1, 2, 3, 4, 5, 6, or 7 days apart. In some embodiments, the effects of treatment persist for at least 6 months or at least one year. In some embodiments, the therapeutic product is administered in 100 cc of normal saline. In some embodiments, 13 cc of therapeutic product in 100 cc of normal saline is administered to a subject. In some embodiments, 13 cc of therapeutic product in a total volume of 100 cc when combined with normal saline is administered to a subject. In some embodiments, the therapeutic product is administered intravenously. In some embodiments, 13 cc of therapeutic product in 100 cc of normal saline is administered intravenously. In some embodiments, 13 cc of therapeutic product in 100 cc of normal saline is administered intravenously once a week. In some embodiments, 1 cc, 2 cc, 3 cc, 4 cc, 5 cc, 6 cc, 7 cc, 8 cc, 9 cc, 10 cc, 11 cc, 12 cc, 13 cc, 14 cc, 15 cc, 16 cc, 17 cc, 18 cc, 19 cc, or 20 cc of therapeutic product is administered to a subject. In some embodiments, the administering is intravenous. In some embodiments, the therapeutic product is diluted in normal saline to a final volume of 100 cc for intravenousWSGR Ref. No. 66309-733.601 administration. In some embodiments, the therapeutic product is administered intravenously and directly into facial tissue. In some embodiments, the therapeutic product is administered into facial tissue. In some embodiments, the facial tissue surrounds the facial nerve. In some embodiments, 2 cc of therapeutic product is administered into the facial tissue around the facial nerve. In some embodiments, the therapeutic product is administered more than once into the facial tissue around the facial nerve. In some embodiments, 1 cc, 2 cc, 3 cc, 4 cc, or 5 cc of the therapeutic product is administered to the facial tissue around the facial nerve.

[0074] Disclosed herein is a method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising 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-. In some embodiments, the subject exhibits one or more of the following symptoms prior to the administering: extracranial paralysis, intracranial paralysis, epiphora, inability to close one or two eyes, drooping of the oral commissure, salivary incompetence, facial tingling, fasciculations of one or more facial muscles, hyperacusis, taste disturbance, inability to elevate one or two eyebrows, difficulty communicating, inability to communicate, inability to smile, difficulty with social interaction, psychological disturbance, depression, headache, earache, tinnitus, dizziness, loss of balance, and / or secondary facial paralysis. In some embodiments, the extracranial paralysis, intracranial paralysis, or secondary facial paralysis affects one side of the face. In some embodiments, the onset of the one or more symptoms is acute. In some embodiments, the one or more symptoms do not improve or do not improve fully in response to one or more of surgery, facial reconstruction, oral steroids, antiviral medications, facial exercises, laser treatment, herbal remedy, acupuncture, botulinum toxin, massage, physical therapy, prednisone, gabapentin, potassium, valaciclovir, vitamin B-12, and / or diuretics. In some embodiments, the subject has experienced the one or more symptoms for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months,10 months, 11 months, 12 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or longer. In some embodiments, results of the subject’s radiological studies are unremarkable before and / or after the administering. In some embodiments, the administering comprises one or more injections. In some embodiments, at least one of the one or more injections is injected into tissue around the facial nerve. In some embodiments, at least one of the one or more injections comprises 1 cc, 2 cc, 3 cc, 4 cc, or 5 cc of the composition. In some embodiments, at least one of the one or more injections is injected intravenously. In some embodiments, at least of the one or more injections comprises 5 cc, 6 cc, 7 cc, 8 cc, 9 cc, 10 cc,11 cc, 12 cc, 13 cc, 14 cc, 15 cc, 16 cc, 17 cc, 18 cc, 19 cc, or 20 cc of the composition. In someWSGR Ref. No. 66309-733.601 embodiments, the administering restores facial nerve function in the subject as compared to before administration. In some embodiments, the administering reduces House-Brackmann score in the subject as compared to before administration. In some embodiments, the decrease in House-Brackmann score is at least 1, at least 2, at least 3, at least 4, at least 5, or at least 6. In some embodiments, wherein the administering improves a Facial Disability Index score in a subject as compared to before administration. In some embodiments, the improvement in the Facial Disability Index score is an improvement in a physical index. In some embodiments, the improvement in the Facial Disability Index score is an improvement in a social index. In some embodiments, the improvement in the Facial Disability Index score is an increase in at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%. In some embodiments, the administering restores one or more of brow motion, eyelid motion, buccal control, and / or facial expressiveness in the subject as compared to before administration. In some embodiments, the administering results in improvement in wellbeing, reduction in tingling, reduction in fasciculations, relaxation of muscles, reduction in pain, restoration of ability to wink, and / or improvement in symmetrical facial movements.

[0075] Disclosed herein are methods and compositions for treating idiopathic facial paralysis in a subject in need thereof, wherein the methods comprise administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition comprising extracellular vesicles, wherein the subject exhibits an improvement in House- Brackmann score after administration as compared to before administration. In some embodiments, the subject exhibits one or more of the following symptoms prior to the administering: extracranial paralysis, intracranial paralysis, epiphora, inability to close one or two eyes, drooping of the oral commissure, salivary incompetence, facial tingling, fasciculations of one or more facial muscles, hyperacusis, taste disturbance, inability to elevate one or two eyebrows, difficulty communicating, inability to communicate, inability to smile, difficulty with social interaction, psychological disturbance, depression, headache, earache, tinnitus, dizziness, loss of balance, and / or secondary facial paralysis. In some embodiments, the extracranial paralysis, intracranial paralysis, or secondary facial paralysis affects one side of the face. In some embodiments, the onset of the one or more symptoms of the idiopathic facial paralysis is acute. In some embodiments, the one or more symptoms do not improve or do not improve fully in response to one or more of surgery, facial reconstruction, oral steroids, anti-viral medications, facial exercises, laser treatment, herbal remedy, acupuncture, botulinum toxin, massage, physical therapy, prednisone, gabapentin, potassium, valaciclovir, vitamin B-12, and / orWSGR Ref. No. 66309-733.601 diuretics. In some embodiments, the subject has experienced the one or more symptoms for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or longer. In some embodiments, results of the subject’s radiological studies are unremarkable before and / or after the administering. In some embodiments, the administering comprises one or more injections. In some embodiments, at least one of the one or more injections is injected into tissue around the facial nerve. In some embodiments, at least one of the one or more injections comprises 1 cc, 2 cc, 3 cc, 4 cc, or 5 cc of the composition. In some embodiments, at least one of the one or more injections is injected intravenously. In some embodiments, at least of the one or more injections comprises 5 cc, 6 cc, 7 cc, 8 cc, 9 cc, 10 cc, 11 cc, 12 cc, 13 cc, 14 cc, 15 cc, 16 cc, 17 cc, 18 cc, 19 cc, or 20 cc of the composition. In some embodiments, the administering restores facial nerve function in the subject as compared to before administration. In some embodiments, the administering reduces House-Brackmann score in the subject as compared to before administration. In some embodiments, the decrease in House-Brackmann score is at least 1, at least 2, at least 3, at least 4, at least 5, or at least 6. In some embodiments, wherein the administering improves a Facial Disability Index score in a subject as compared to before administration. In some embodiments, the improvement in the Facial Disability Index score is an improvement in a physical index. In some embodiments, the improvement in the Facial Disability Index score is an improvement in a social index. In some embodiments, the improvement in the Facial Disability Index score is an increase in at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%. In some embodiments, the administering restores one or more of brow motion, eyelid motion, buccal control, and / or facial expressiveness in the subject as compared to before administration. In some embodiments, the administering results in improvement in wellbeing, reduction in tingling, reduction in fasciculations, relaxation of muscles, reduction in pain, restoration of ability to wink, and / or improvement in symmetrical facial movements.

[0076] Also disclosed herein are methods and compositions for treating idiopathic facial paralysis in a subject, wherein the methods comprise administering to the subject a therapeutic composition, e.g., MSC secretome composition comprising any combination of factors and / or miRNAs, selected from the following: 2B4, 4-1BB, 4-1BB Ligand, 6Ckine, ACE, Activin A, Activin RIB, Activin RIIA, Activin RIM, ADA, ADAM23, ADAMS, ADAMTS13, ADAMTSL-1, Adiponectin, Adipsin, Aggrecan, AgRP, AIF, AKR1C4, Albumin, ALCAM, AMIGO, AMIG02, Aminopeptidase LRAP, Aminopeptidase P2, AMSH, ANG-1, ANG-2,WSGR Ref. No. 66309-733.601ANG-4, Angiogenin, Angiotensinogen, ANGPTL4, ANGPTL7, APRIL, Arginase 1, ARSB, Artemin, Arylsulfatase A, ASAH2, ASAHL, Axl, B2M, B7-1, B7-H3, B7-H4, BAFF R, Bcl-10, BCL-2, Bcl-w, BCMA, BID, Biglycan, BLAME, BMP-2, BMP-8, BMP-9, BMPR-IA, Brevican, C-myc, CA125, CA13, CA4, CA8, Cadherin-13, Cadherin-4, Calreticulin, Calreticulin-2, Cardiotrophin-1, CASA, Caspr2, Cathepsin E, Cathepsin L, Cathepsin S, Cathepsin V, CD14, CD155, CD157, CD2, CD200, CD229, CD27, CD27 Ligand, CD28, CD30, CD300f, CD314, CD34, CD36, CD39L2, CD4, CD40, CD40L, CD42b, CD48, CD51, CD58, CD6, CD73, CD83, CD97, CD99-L2, CDC25B, CDNF, CDS, CEACAM-1, CEACAM-3, CEACAM-5, Cerberus 1, CES1, CES2, CF VII, CF XIV, Chemerin, CHI3L1, CHST2, CHST3, CHST4, cIAP-1, CILP-1, CK19, CL-P1, CLEC-1, CLEC-2, CLEC10A, Clq, ClqTNF9, CMASP3, CMG-2, Cochlin, COMT, Contactin-1, Contactin-2, CPB1, CPE, CREG, CRELD2, CRIM1, CrkL, CSa, CTACK, CTRC, CXCL14, CXCL16, Cyr61, Cystatin B, Cystatin C, Cy statin D, Cystatin E M, Cystatin S, Cystatin SA, Cystatin SN, Cytokeratin-8, DAPP1, DCTN1, DDR1, Dectin- 1, Dectin-2, Desmin, Desmoglein 2, Desm oglein-3, DKK-1, DLL4, DNAM-1, DNMT3 A, Dopa Decarboxylase, DPPII, DPPIV, DR3, DR6, Draxin, DSCAM, DSPG3, Dtk, E-Selectin, ED AR, EDIL3, EG-VEGF, EGF R, EMMPRIN, ENA-78, Endocan, Endoglin, eNOS, ENPP-2, ENPP-7, Enteropeptidase, EpCAM, EphA2, EphAl, EphB3, EphB4, EphB6, Ephrin-A4, Ephrin-B3, Epimorphin, Epiregulin, ErbB3, ErbB4, Erythropoietin, ESAM, EXTL3, FABP1, FABP2, FABP4, FABP6, FAP, Fas, FCAR, FcERI, FCRL1, FCRL3, FCRLB, FCRLS, FGF R5, FGF-12, FGF-16, FGF-17, FGF-19, FGF-20, FGF-21, FGF-23, FGF-3, FGF- 5, Ficolin-1, FKBP51, FLRG, FLRT1, FLRT2, Flt-3, Flt-3L, Follistatin, Follistatin-like 1, FUT8, Galanin, Galectin-1, Galectin-2, Galectin-3, Galectin-4, Galectin-7, Galectin-8, Galectin- 9, GALNT10, GALNT2, GALNT3, Gash, GATA-4, GBA3, GCP-2, GDF-11, GDF-15, GDF-9, GFAP, GFR alpha-1, GFR alpha-2, GITR, GITR L, GKN1, GLP-1, Glycoprotein V, Glyoxalase II, Glypican 1, Glypican 2, Glypican 5, gpl30, GPR56, GPVI, Granulysin, Granzyme A, Granzyme B, Granzyme H, GRAP2, GRKS, GRO, GSTM1, GUSB, HAO-1, HAPLN1, HCC-1, HCC-4, hCGb, HE4, Hepsin, HGF, HGF R, HIF-1 alpha, HIF-1 beta, HO-1, HS3ST1, HS3ST3B1, HSD17B1, htPAPP-A, HTRA2, HVEM, LT AC, ICAM-1, ICAM-2, ICAM-3, ICOS, IDO, IFN-gamma Rl, IFNab R2, IFNb, IGF-2R, IGFBP-5, IGSF3, IGSF4B, IL-1 F10, IL-1 R3, IL-1 R4, IL-1 RI, IL-1 RII, IL-10 Rb, IL-12p40, IL-13 R2, IL-15, IL-15 R, IL-17 RC, IL- 17 RD, IL-17B, IL-17F, IL- 18, IL- 18 BPa, IL- 19, IL-2 Ra, IL-2 Rb, IL-2 Rg, IL-20 R beta, IL-20 Ra, IL-21R, IL-22, IL-22 R alpha 1, IL-22BP, IL-23, IL-23 R, IL-24, IL-27, IL-27 Ra, IL- 28 R alpha, IL-28A, IL-31, IL-31 RA, IL-32 alpha, IL-33, IL-4 Ra, IL-6, IL-6R, IL-7 R alpha, IL-8, IL-9, IL-lra, ILT2, ILT4, Inhibin A, Insulin, Insulin R, Integrin alpha 1, Integrin alpha 5, IP-10, Jagged 1, JAM-A, JAM-B, JAM-C, Kallikrein 1, Kallikrein 11, Kallikrein 12, KallikreinWSGR Ref. No. 66309-733.60114, Kallikrein 5, Kallikrein 7, Kell, KIR2DL3, Kirrel3, KLF4, Kremen-2, Kynureninase, L- Selectin, L1CAM-2, LAIR2, LAMA4, LAMP, LAMP1, Langerin, LAP(TGFbl), LDL R, LEDGF, Legumain, LH, LIF, LIF R alpha, LIGHT, LILRB4, LIMPII, Lin28, Lipocalin-2, LOX-1, LRP-6„ST3GAL1, LRRC4, LRRTM4, LSECtin, LTbR, Lymphotactin, LYVE-1, MAP1D, Matrilin-2, Matrilin-3, Matriptase, MBL, Mcl-1, MCEMP1, MCP-2, MCP-3, MCP-4, MCSF, MDC, MDL-1, MEF2C, MEP1B, MEPE, MeprinA, Mer, Mesothelin, METAP2, MFRP, MIA, MICA, MICB, Midkine, MIP-3a, MIP-3b, MIP-lb, MIS RII, MMP-1, MMP-10, MMP-12, MMP-13, MMP-2, MOG, MPIF-1, MSP R, N-Cadherin, NAP-2, NCAM-1, NCAM-1 (CD56), NCK1, Nectin-1, Nectin-2, Nectin-3, Nectin-4, Neprilysin, Nestin, Netrin-4, Neudesin, Neurexin 3 beta, Neurogranin, Neuroligin 2, Neuropilin-2, Neurturin, NG2, NGF R, Nidogen-1, Nidogen-2, NKp30, NKp44, NKp46, NKp80, Nogo Receptor, Nogo-A, Norrin, Notch- 1, Notch - 3, NOV, NPDC-1, NPTXR, NQO-1, Nrf2, NRG1-131, NSE, NT AL, NUDTS, Numb, NUP85, Olfactomedin-2, Olig2, OPG, or IL-13, OSCAR, OSM R beta, Osteoactivin, Osteoadherin, 0X40, 0X40 Ligand, P1GF-2, p27, p53, p63, PAPP-A, Pappalysin-2, PARI, PARC, Pax3, PCSK2, PD-1, PD-ECGF, PD-L2, PDGF R alpha, PDGF Rb, PDGF-AA, PDGF-AB, PDGF- CC, PDX-1, PEAR1, PECAM-1, Pentraxin 3, Pepsinogen II, Persephin, PGRP-S, PIGF, PILR- alpha, Plexin A4, Plexin B3, Plexin DI, Podocalyxin, Podoplanin, POGLUT1, PON1, Pref-1, PRELP, Presenilin 1, Procalcitonin, Progranulin, proGRP, Prolactin, Prolactin R, PRX2, PSA- total, PSMA, PSMA1, PTH, PTH1R, PTP1B, PU.l, PYY, R-Spondin 2, RAGE, RalA, RANK, RANTES, RBP4, RCOR1, Reg3A, Reg4, RELT, Renin, Resistin, Ret Midkine, RGM-A, RGM- B, ROBO2, ROBO3, ROBO4, R0R1, R0R2, S100A1, S100A13, SALM4, SCCA2, SCF, SCF R, SDF-la, SDF-lb, Semaphorin 4C, Semaphorin 4D, Semaphorin 4G, Semaphorin 6 A, Semaphorin 6B, Semaphorin 6D, Semaphorin 7A, Serpin A4, Serpin A5, sFRP-3, SH2D1A, Shh-N, SHP-1, Siglec-10, Siglec-11, Siglec-2, Siglec-6, Siglec-7, Siglec-9, SIRP alpha, Sirtuin1, Sirtuin 2, Sirtuin 5, SLAM, SLITRK5, Smad4, SMPD1, SorCSl, SorCS2, SorCS3, SOST, SOX15, SOX2, SOX7, SOX9, SP-D, SPHK1, Spinesin, SPINK1, SR-AI, SREC-I, SREC-II, ST6GAL1, ST8SIA1, Stabilin-2, SULT2A1, Syndecan-3, Syndecan-4, Syntaxin 4, Syntaxin 6, TACE, TAFA1, TAFA2, TAFAS, TARC, TAZ, TC-PTP, TCN2, TECK, Tenascin R, Testican2, TFF3, TFPI, TFPI-2, TGFa, TGFb2, TGFb3 FOLR2, TGFbl, TGM3, THAP11, Thrombospondin-2, Thrombospondin-5, THSD1, Thyroglobulin, Thyroid Peroxidase, Tie-1, Tie-2, TIM-1, TIM-3, TLR1, TLR2, TLR3, TLR4, TNF RI, TNF RII, TNF4, TNFb, TPO, TPP1, TPST2, TRACP, TRAIL, TRAIL Rl, TRAIL R3, TRAIL R4, TRANCE, TREM-1, TREM-2, TREML1, TrkC, TROP-2, Troponin C, TROY, Trypsin 1, Trypsin 3, TSH, TSK, TSLP, TSLP R, TWEAK R, Ubiquitin+1, UCH-L3, ULBP-1, ULBP-2, ULBP-3, ULBP-4, UNC5H3, UNC5H4, uPA, uPAR, Uromodulin, VAMP-1, VAMP-2, VAP-A, VCAM-1, VE-Cadherin,WSGR Ref. No. 66309-733.601VEGF, VEGF Rl, VEGF R2, VEGF R3, VEGF-C, Visfatin, vWF-A2, WIF-1, WISP-1, Wnt-4, XEDAR, XIAP, 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-lOb, 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-l'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-486-2, hsa-miR-486-5p, hsa-miR- 570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-l, 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] In some embodiments, the subject exhibits a decrease in House-Brackmann score after administration as compared to before administration. In some embodiments, the subject exhibits a decrease in House-Brackmann score of 1, 2, 3, 4, or 5 after administration as compared to before administration.

[0078] In some embodiments, the subject exhibits an increase in physical Facial Disability Index score after administration as compared to before administration. For example, the subject can have an increase of at least 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%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%,46%, 47%, 48%, 49%, 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%, 99%, or at least 100% in physical Facial Disability Index score after administration compared to before administration.WSGR Ref. No. 66309-733.601

[0079] In some embodiments, the subject exhibits an increase in social Facial Disability Index score after administration as compared to before administration. For example, the subject can have an increase of at least 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%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%,46%, 47%, 48%, 49%, 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%, 99%, or at least 100% in social Facial Disability Index score after administration compared to before administration.

[0080] In some embodiments, a subject that can be treated with the therapeutic product and methods described herein can comprise any subject suffering from or diagnosed with idiopathic facial paralysis. In some embodiments, a subject can comprise a mammal. In some embodiments, a subject can comprise a human. In some embodiments, a subject can comprise a non-human mammal. Non-limiting examples of a non-human mammal can include a non-human primate such as chimpanzee, and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice and guinea pigs, and the like.

[0081] An aspect of the present disclosure is a method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising 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 . In some embodiments, the subject exhibits one or more of the following symptoms prior to the administering: extracranial paralysis, intracranial paralysis, epiphora, inability to close one or two eyes, drooping of the oral commissure, salivary incompetence, facial tingling, fasciculations of one or more facial muscles, hyperacusis, taste disturbance, inability to elevate one or two eyebrows, difficulty communicating, inability to communicate, inability to smile, difficulty with social interaction, psychological disturbance, depression, headache, earache, tinnitus, dizziness, loss of balance, and / or secondary facial paralysis. In some embodiments, the extracranial paralysis, intracranial paralysis, or secondary facial paralysis affects one side of the face. In some embodiments, the onset of the one or more symptoms is acute. In some embodiments, the one or more symptoms do not improve or do not improve fully in response to one or more of surgery, facial reconstruction, oral steroids, antiviral medications, facial exercises, laser treatment, herbal remedy, acupuncture, botulinum toxin, massage, physical therapy, prednisone, gabapentin, potassium, valaciclovir, vitamin B-12,WSGR Ref. No. 66309-733.601 and / or diuretics. In some embodiments, the subject has experienced the one or more symptoms for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months,10 months, 11 months, 12 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or longer. In some embodiments, results of the subject’s radiological studies are unremarkable before and / or after the administering. In some embodiments, the administering comprises one or more injections. In some embodiments, at least one of the one or more injections is injected into tissue around the facial nerve. In some embodiments, at least one of the one or more injections comprises 1 cc, 2 cc, 3 cc, 4 cc, or 5 cc of the composition. In some embodiments, at least one of the one or more injections is injected intravenously. In some embodiments, at least of the one or more injections comprises 5 cc, 6 cc, 7 cc, 8 cc, 9 cc, 10 cc,11 cc, 12 cc, 13 cc, 14 cc, 15 cc, 16 cc, 17 cc, 18 cc, 19 cc, or 20 cc of the composition. In some embodiments, the administering restores facial nerve function in the subject as compared to before administration. In some embodiments, the administering reduces House-Brackmann score in the subject as compared to before administration. In some embodiments, the decrease in House-Brackmann score is at least 1, at least 2, at least 3, at least 4, at least 5, or at least 6. In some embodiments, the administering improves a Facial Disability Index score in a subject as compared to before administration. In some embodiments, the improvement in the Facial Disability Index score is an improvement in a physical index. In some embodiments, the improvement in the Facial Disability Index score is an improvement in a social index. In some embodiments, the improvement in the Facial Disability Index score is an increase in at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%. In some embodiments, the administering restores one or more of brow motion, eyelid motion, buccal control, and / or facial expressiveness in the subject as compared to before administration. In some embodiments, the administering results in improvement in wellbeing, reduction in tingling, reduction in fasciculations, relaxation of muscles, reduction in pain, restoration of ability to wink, and / or improvement in symmetrical facial movements. In some embodiments, the idiopathic facial paralysis comprises Bell’s Palsy, facial palsy, and / or secondary facial paralysis.

[0082] An aspect of the present disclosure is a method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition comprising extracellular vesicles, wherein the subject exhibits an improvement in House-Brackmann score after administration compared to before administration. In some embodiments, the compositionWSGR Ref. No. 66309-733.601 is produced by: (a) culturing bone marrow-derived MSCs under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media 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 (a). In some embodiments, the subject exhibits one or more of the following symptoms prior to the administering: extracranial paralysis, intracranial paralysis, epiphora, inability to close one or two eyes, drooping of the oral commissure, salivary incompetence, facial tingling, fasciculations of one or more facial muscles, hyperacusis, taste disturbance, inability to elevate one or two eyebrows, difficulty communicating, inability to communicate, inability to smile, difficulty with social interaction, psychological disturbance, depression, headache, earache, tinnitus, dizziness, loss of balance, and / or secondary facial paralysis. In some embodiments, the extracranial paralysis, intracranial paralysis, or secondary facial paralysis affects one side of the face. In some embodiments, the onset of the one or more symptoms is acute. In some embodiments, the one or more symptoms do not improve or do not improve fully in response to one or more of surgery, facial reconstruction, oral steroids, antiviral medications, facial exercises, laser treatment, herbal remedy, acupuncture, botulinum toxin, massage, physical therapy, prednisone, gabapentin, potassium, valaciclovir, vitamin B-12, and / or diuretics. In some embodiments, the subject has experienced the one or more symptoms for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months,10 months, 11 months, 12 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or longer. In some embodiments, results of the subject’s radiological studies are unremarkable before and / or after the administering. In some embodiments, the administering comprises one or more injections. In some embodiments, at least one of the one or more injections is injected into tissue around the facial nerve. In some embodiments, at least one of the one or more injections comprises 1 cc, 2 cc, 3 cc, 4 cc, or 5 cc of the composition. In some embodiments, at least one of the one or more injections is injected intravenously. In some embodiments, at least of the one or more injections comprises 5 cc, 6 cc, 7 cc, 8 cc, 9 cc, 10 cc,11 cc, 12 cc, 13 cc, 14 cc, 15 cc, 16 cc, 17 cc, 18 cc, 19 cc, or 20 cc of the composition. In some embodiments, the administering restores facial nerve function in the subject as compared to before administration. In some embodiments, the administering reduces House-Brackmann score in the subject as compared to before administration. In some embodiments, the decrease in House-Brackmann score is at least 1, at least 2, at least 3, at least 4, at least 5, or at least 6. In some embodiments, the administering improves a Facial Disability Index score in a subject as compared to before administration. In some embodiments, the improvement in the FacialWSGR Ref. No. 66309-733.601Disability Index score is an improvement in a physical index. In some embodiments, the improvement in the Facial Disability Index score is an improvement in a social index. In some embodiments, the improvement in the Facial Disability Index score is an increase in at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%. In some embodiments, the administering restores one or more of brow motion, eyelid motion, buccal control, and / or facial expressiveness in the subject as compared to before administration. In some embodiments, the administering results in improvement in wellbeing, reduction in tingling, reduction in fasciculations, relaxation of muscles, reduction in pain, restoration of ability to wink, and / or improvement in symmetrical facial movements. In some embodiments, the idiopathic facial paralysis comprises Bell’s Palsy, facial palsy, and / or secondary facial paralysis. In some embodiments, the idiopathic facial paralysis comprises Bell’s Palsy, facial palsy, and / or secondary facial paralysis.

[0083] An aspect of the present disclosure is a method of making a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition for treating idiopathic facial paralysis in a subject in need thereof, the method comprising: (a) culturing bone marrow- derived MSCs under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media 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 (a). In some embodiments, the subject exhibits one or more of the following symptoms prior to the administering: extracranial paralysis, intracranial paralysis, epiphora, inability to close one or two eyes, drooping of the oral commissure, salivary incompetence, facial tingling, fasciculations of one or more facial muscles, hyperacusis, taste disturbance, inability to elevate one or two eyebrows, difficulty communicating, inability to communicate, inability to smile, difficulty with social interaction, psychological disturbance, depression, headache, earache, tinnitus, dizziness, loss of balance, and / or secondary facial paralysis. In some embodiments, the extracranial paralysis, intracranial paralysis, or secondary facial paralysis affects one side of the face. In some embodiments, the onset of the one or more symptoms is acute. In some embodiments, the one or more symptoms do not improve or do not improve fully in response to one or more of surgery, facial reconstruction, oral steroids, anti-viral medications, facial exercises, laser treatment, herbal remedy, acupuncture, botulinum toxin, massage, physical therapy, prednisone, gabapentin, potassium, valaciclovir, vitamin B-12, and / or diuretics. In some embodiments, theWSGR Ref. No. 66309-733.601 subject has experienced the one or more symptoms for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or longer. In some embodiments, results of the subject’s radiological studies are unremarkable before and / or after the administering. In some embodiments, the administering comprises one or more injections. In some embodiments, at least one of the one or more injections is injected into tissue around the facial nerve. In some embodiments, at least one of the one or more injections comprises 1 cc, 2 cc, 3 cc, 4 cc, or 5 cc of the composition. In some embodiments, at least one of the one or more injections is injected intravenously. In some embodiments, at least of the one or more injections comprises 5 cc, 6 cc, 7 cc, 8 cc, 9 cc, 10 cc, 11 cc, 12 cc, 13 cc, 14 cc, 15 cc, 16 cc, 17 cc, 18 cc, 19 cc, or 20 cc of the composition. In some embodiments, the administering restores facial nerve function in the subject as compared to before administration. In some embodiments, the administering reduces House-Brackmann score in the subject as compared to before administration. In some embodiments, the decrease in House-Brackmann score is at least 1, at least 2, at least 3, at least 4, at least 5, or at least 6. In some embodiments, the administering improves a Facial Disability Index score in a subject as compared to before administration. In some embodiments, the improvement in the Facial Disability Index score is an improvement in a physical index. In some embodiments, the improvement in the Facial Disability Index score is an improvement in a social index. In some embodiments, the improvement in the Facial Disability Index score is an increase in at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%. In some embodiments, the administering restores one or more of brow motion, eyelid motion, buccal control, and / or facial expressiveness in the subject as compared to before administration. In some embodiments, the administering results in improvement in wellbeing, reduction in tingling, reduction in fasciculations, relaxation of muscles, reduction in pain, restoration of ability to wink, and / or improvement in symmetrical facial movements. In some embodiments, the idiopathic facial paralysis comprises Bell’s Palsy, facial palsy, and / or secondary facial paralysis. In some embodiments, the idiopathic facial paralysis comprises Bell’s Palsy, facial palsy, and / or secondary facial paralysis.III. Definitions

[0084] As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the content clearly dictates otherwise. It should also be noted that the term “or” is generally employed in its sense including “and / or” unless the contentWSGR Ref. No. 66309-733.601 clearly dictates otherwise. The terms “and / or” and “any combination thereof’ and their grammatical equivalents as used herein, can be used interchangeably.

[0085] The term “about” or “approximately” can mean within an acceptable error range for the particular value, which may depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, “about” can mean within 1 or more than 1 standard deviation. Alternatively, “about” can 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 can mean within an order of magnitude, within 5-fold, or within 2-fold, of a value. Where particular values are described in the application and claims, unless otherwise stated the term “about” meaning within an acceptable error range for the particular value should be assumed.

[0086] As used in this specification and claim(s), the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.IV. Numbered Embodiments

[0087] Embodiment 1. A method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising 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 .

[0088] Embodiment 2. The method of embodiment 1, wherein the subject exhibits one or more of the following symptoms prior to the administering: extracranial paralysis, intracranial paralysis, epiphora, inability to close one or two eyes, drooping of the oral commissure, salivary incompetence, facial tingling, fasciculations of one or more facial muscles, hyperacusis, taste disturbance, inability to elevate one or two eyebrows, difficulty communicating, inability to communicate, inability to smile, difficulty with social interaction, psychological disturbance, depression, headache, earache, tinnitus, dizziness, loss of balance, and / or secondary facial paralysis.

[0089] Embodiment 3. The method of embodiment 2, wherein the extracranial paralysis, intracranial paralysis, or secondary facial paralysis affects one side of the face.

[0090] Embodiment 4. The method of embodiment 2, wherein the onset of the one or more symptoms is acute.WSGR Ref. No. 66309-733.601

[0091] Embodiment 5. The method of embodiment 2, wherein the one or more symptoms do not improve or do not improve fully in response to one or more of surgery, facial reconstruction, oral steroids, anti-viral medications, facial exercises, laser treatment, herbal remedy, acupuncture, botulinum toxin, massage, physical therapy, prednisone, gabapentin, potassium, valaciclovir, vitamin B-12, and / or diuretics.

[0092] Embodiment 6. The method of embodiment 2, wherein the subject has experienced the one or more symptoms for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or longer.

[0093] Embodiment 7. The method of embodiment 1, wherein results of the subject’s radiological studies are unremarkable before and / or after the administering.

[0094] Embodiment 8. The method of embodiment 1, wherein the administering comprises one or more injections.

[0095] Embodiment 9. The method of embodiment 8, wherein at least one of the one or more injections is injected into tissue around the facial nerve.

[0096] Embodiment 10. The method of embodiment 9, wherein at least one of the one or more injections comprises 1 cc, 2 cc, 3 cc, 4 cc, or 5 cc of the composition.

[0097] Embodiment 11. The method of embodiment 8, wherein at least one of the one or more injections is injected intravenously.

[0098] Embodiment 12. The method of embodiment 11, wherein at least of the one or more injections comprises 5 cc, 6 cc, 7 cc, 8 cc, 9 cc, 10 cc, 11 cc, 12 cc, 13 cc, 14 cc, 15 cc, 16 cc, 17 cc, 18 cc, 19 cc, or 20 cc of the composition.

[0099] Embodiment 13. The method of embodiment 1, wherein the administering restores facial nerve function in the subject as compared to before administration.

[0100] Embodiment 14. The method of embodiment 1, wherein the administering reduces House-Brackmann score in the subject as compared to before administration.

[0101] Embodiment 15. The method of embodiment 14, wherein the decrease in House- Brackmann score is at least 1, at least 2, at least 3, at least 4, at least 5, or at least 6.

[0102] Embodiment 16. The method of embodiment 1, wherein the administering improves a Facial Disability Index score in a subject as compared to before administration.

[0103] Embodiment 17. The method of embodiment 16, wherein the improvement in the Facial Disability Index score is an improvement in a physical index.

[0104] Embodiment 18. The method of embodiment 16, wherein the improvement in the Facial Disability Index score is an improvement in a social index.WSGR Ref. No. 66309-733.601

[0105] Embodiment 19. The method of embodiment 16, wherein the improvement in the Facial Disability Index score is an increase in at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%.

[0106] Embodiment 20. The method of embodiment 1, wherein the administering restores one or more of brow motion, eyelid motion, buccal control, and / or facial expressiveness in the subject as compared to before administration.

[0107] Embodiment 21. The method of embodiment 1, wherein the administering results in improvement in wellbeing, reduction in tingling, reduction in fasciculations, relaxation of muscles, reduction in pain, restoration of ability to wink, and / or improvement in symmetrical facial movements.

[0108] Embodiment 22. A method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition comprising extracellular vesicles, wherein the subject exhibits an improvement in House-Brackmann score after administration compared to before administration.

[0109] Embodiment 23. The method of any one of embodiments 1 to 21, wherein the composition is produced by:(a) culturing bone marrow-derived MSCs under the following conditions to produce an MSC conditioned media:(i) oxygen tension below 5%; and(ii) culture media having a pH below 7;(b) harvesting the MSC conditioned media; and(c) formulating the MSC conditioned media 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 (a).

[0110] Embodiment 24. A method of making a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition for treating idiopathic facial paralysis in a subject in need thereof, the method comprising:(a) culturing bone marrow-derived MSCs under the following conditions to produce an MSC conditioned media:(i) oxygen tension below 5%; and(ii) culture media having a pH below 7;(b) harvesting the MSC conditioned media; andWSGR Ref. No. 66309-733.601(c) formulating the MSC conditioned media 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 (a).[OHl] Embodiment 25. The method of any one of the preceding embodiments, wherein the idiopathic facial paralysis comprises Bell’s Palsy, facial palsy, and / or secondary facial paralysis.V. Examples

[0112] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how the compounds, compositions, articles, devices and / or methods claimed herein are made and evaluated and are intended to be purely exemplary and are not intended to limit the disclosure. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. These examples are provided for illustrative purposes only and not to limit the scope of the claims provided herein.Example 1 - Production of Therapeutic Composition

[0113] An MSC secretome therapeutic composition (investigational medicinal product or “IMP”) was made by the following method: human bone marrow-derived MSCs (hBM-MSCs) were cultured in culture vessels with growth media to expand the MSC population. Growth media was then removed, and the cells were washed with PBS. The hBM-MSCs were then cultured at a pH below 7.0, less than 5% oxygen (e.g., to a final concentration of about 1% oxygen), and in a culture media comprising basal media and sodium chloride. The culture media comprising the MSC secretome was formulated into an isotonic infusion solution, such as sodium chloride, and filter sterilized. The production process for the IMP was done under current Good Manufacturing Practices and Current Good Tissue Practices.

[0114] The IMP was manufactured from the banked hBM-MSCs of a single donor under CGMP conditions and according to FDA Master File protocols. Each lot of the IMP was characterized by proteomic and miRNA characterization. Additionally, the size and quantity of EVs and the presence of a specific surface marker expression profile were confirmed.

[0115] The IMP comprises extracellular vesicles (EVs) that are acellular and nonimmunogenic, containing no nucleus or deoxyribonucleic acid (DNA).

[0116] The tetraspanin profile of extracellular vesicles present in the IMP was determined, and it was found that greater than 95% of the extracellular vesicles present in the composition were CD63+CD9 CD8 I . The EVs were measured via Nanoparticle tracking analysis (NT A), having a median diameter of about 100 nm. At least 10 billion EVs per mL were calculated using NTA with fluorescent staining of EV membranes.WSGR Ref. No. 66309-733.601

[0117] Protein content of the IMP was determined, and the following proteins were found to be present. Proteins were detected using an antibody -based sandwich ELISAs or by Luminex® or Nanoview / Unchained Labs-based methods. The total concentration of certain proteins in the IMP was measured via ELISA, with each protein having a concentration of about 200 pg / mL to about 80 ng / mL. For instance, TEMPI, OPN, IGFBP4, and osteonectin were characterized. Nonlimiting examples of proteins present in the IMP are shown in Table 2 below.Table 2WSGR Ref. No. 66309-733.601

[0118] The nucleic acid content of the IMP was determined, and the following nucleic acids were found to be present: hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a- 3p, hsa-miR-21-5p,hsa-miR-221-3p, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa- miR-23b-3p, hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31- 5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, and hsa-miR-940. RNA were measured using semi-quantitative PCR. Total RNA was analyzed by UV detection using a ClarioStar Plus microplate reader and BMG Labtech LVis plate adapter for nucleic acid quantification. Using this method, the amount of each RNA may be less than the limit of detection, or about 50 ng / mL.

[0119] Protein content of the therapeutic product was determined, and the following proteins were found to be present: 2B4, 4-1BB, 4-1BB Ligand, 6Ckine, ACE, Activin A, Activin RIB, Activin RIIA, Activin RIM, ADA, ADAM23, ADAMS, ADAMTS13, ADAMTSL-1,WSGR Ref. No. 66309-733.601Adiponectin, Adipsin, Aggrecan, AgRP, AIF, AKR1C4, Albumin, ALCAM, AMIGO, AMIGO2, Aminopeptidase LRAP, Aminopeptidase P2, AMSH, ANG-1, ANG-2, ANG-4, Angiogenin, Angiotensinogen, ANGPTL4, ANGPTL7, APRIL, Arginase 1, ARSB, Artemin, Arylsulfatase A, ASAH2, ASAHL, Axl, B2M, B7-1, B7-H3, B7-H4, BAFF R, Bcl-10, BCL-2, Bcl-w, BCMA, BID, Biglycan, BLAME, BMP -2, BMP-8, BMP-9, BMPR-IA, Brevican, C-myc, CA125, CA13, CA4, CA8, Cadherin-13, Cadherin-4, Calreticulin, Calreticulin-2, Cardiotrophin-1, CASA, Caspr2, Cathepsin E, Cathepsin L, Cathepsin S, Cathepsin V, CD14, CD155, CD157, CD2, CD200, CD229, CD27, CD27 Ligand, CD28, CD30, CD300f, CD314, CD34, CD36, CD39L2, CD4, CD40, CD40L, CD42b, CD48, CD51, CD58, CD6, CD73, CD83, CD97, CD99- L2, CDC25B, CDNF, CDS, CEACAM-1, CEACAM-3, CEACAM-5, Cerberus 1, CES1, CES2, CF VII, CF XIV, Chemerin, CHI3L1, CHST2, CHST3, CHST4, cIAP-1, CILP-1, CK19, CL-P1, CLEC-1, CLEC-2, CLEC10A, Clq, ClqTNF9, cMASP3, CMG-2, Cochlin, COMT, Contactin- 1, Contactin-2, CPB1, CPE, CREG, CRELD2, CRIM1, CrkL, CSa, CTACK, CTRC, CXCL14, CXCL16, Cyr61, Cystatin B, Cystatin C, Cystatin D, Cystatin E M, Cystatin S, Cystatin SA,Cy statin SN, Cytokeratin-8, DAPP1, DCTN1, DDR1, Dectin- 1, Dectin-2, Desmin, Desmoglein2, Desm oglein-3, DKK-1, DLL4, DNAM-1, DNMT3A, Dopa Decarboxylase, DPPII, DPPIV, DR3, DR6, Draxin, DSCAM, DSPG3, Dtk, E-Selectin, ED AR, EDIL3, EG-VEGF, EGF R, EMMPRIN, ENA-78, Endocan, Endoglin, eNOS, ENPP-2, ENPP-7, Enteropeptidase, EpCAM, EphA2, EphAl, EphB3, EphB4, EphB6, Ephrin-A4, Ephrin-B3, Epimorphin, Epiregulin, ErbB3, ErbB4, Erythropoietin, ESAM, EXTL3, FABP1, FABP2, FABP4, FABP6, FAP, Fas, FC AR, FcERI, FCRL1, FCRL3, FCRLB, FCRLS, FGF R5, FGF-12, FGF-16, FGF-17, FGF-19, FGF- 20, FGF-21, FGF-23, FGF-3, FGF-5, Ficolin-1, FKBP51, FLRG, FLRT1, FLRT2, Flt-3, Flt-3L, Follistatin, Follistatin-like 1, FUT8, Galanin, Galectin-1, Galectin-2, Galectin-3, Galectin-4, Galectin-7, Galectin-8, Galectin-9, GALNT10, GALNT2, GALNT3, Gash, GATA-4, GBA3, GCP-2, GDF-11, GDF-15, GDF-9, GFAP, GFR alpha-1, GFR alpha-2, GITR, GITR L, GKN1, GLP-1, Glycoprotein V, Glyoxalase II, Glypican 1, Glypican 2, Glypican 5, gpl30, GPR56, GPVI, Granulysin, Granzyme A, Granzyme B, Granzyme H, GRAP2, GRKS, GRO, GSTM1, GUSB, HAO-1, HAPLN1, HCC-1, HCC-4, hCGb, HE4, Hepsin, HGF, HGF R, HIF-1 alpha, HIF-1 beta, HO-1, HS3ST1, HS3ST3B1, HSD17B1, htPAPP-A, HTRA2, HVEM, I-TAC, ICAM-1, ICAM-2, ICAM-3, ICOS, IDO, IFN-gamma Rl, IFNab R2, IFNb, IGF-2R, IGFBP-5, IGSF3, IGSF4B, IL-1 F10, IL-1 R3, IL-1 R4, IL-1 RI, IL-1 RII, IL-10 Rb, IL-12p40, IL-13 R2, IL-15, IL-15 R, IL-17 RC, IL-17 RD, IL-17B, IL-17F, IL-18, IL-18 BPa, IL-19, IL-2 Ra, IL-2 Rb, IL-2 Rg, IL-20 R beta, IL-20 Ra, IL-21R, IL-22, IL-22 R alpha 1, IL-22BP, IL-23, IL-23 R, IL-24, IL-27, IL-27 Ra, IL-28 R alpha, IL-28A, IL-31, IL-31 RA, IL-32 alpha, IL-33, IL-4 Ra, IL-6, IL-6R, IL-7 R alpha, IL-8, IL-9, IL-lra, ILT2, ILT4, Inhibin A, Insulin, Insulin R, IntegrinWSGR Ref. No. 66309-733.601 alpha 1, Integrin alpha 5, IP-10, Jagged 1, JAM-A, JAM-B, JAM-C, Kallikrein 1, Kallikrein 11, Kallikrein 12, Kallikrein 14, Kallikrein 5, Kallikrein 7, Kell, KIR2DL3, Kirrel3, KLF4, Kremen-2, Kynureninase, L-Selectin, LI CAM-2, LAIR2, LAMA4, LAMP, LAMP1, Langerin, LAP(TGFbl), LDL R, LEDGF, Legumain, LH, LIF, LIF R alpha, LIGHT, LILRB4, LIMPII, Lin28, Lipocalin-2, LOX-1, LRP-6„ST3GAL1, LRRC4, LRRTM4, LSECtin, LTbR, Lymphotactin, LYVE-1, MAP ID, Matrilin-2, Matrilin-3, Matriptase, MBL, Mcl-1, MCEMP1, MCP-2, MCP-3, MCP-4, MCSF, MDC, MDL-1, MEF2C, MEP1B, MEPE, MeprinA, Mer, Mesothelin, METAP2, MFRP, MIA, MICA, MICB, Midkine, MIP-3a, MIP-3b, MIP-lb, MIS RII, MMP-1, MMP-10, MMP-12, MMP-13, MMP-2, MOG, MPIF-1, MSP R, N-Cadherin, NAP-2, NCAM-1, NCAM-1 (CD56), NCK1, Nectin-1, Nectin-2, Nectin-3, Nectin-4, Neprilysin, Nestin, Netrin-4, Neudesin, Neurexin 3 beta, Neurogranin, Neuroligin 2, Neuropilin- 2, Neurturin, NG2, NGF R, Nidogen-1, Nidogen-2, NKp30, NKp44, NKp46, NKp80, Nogo Receptor, Nogo- A, Norrin, Notch- 1, Notch-3, NOV, NPDC-1, NPTXR, NQO-1, Nrf2, NRG1- 131, NSE, NTAL, NUDTS, Numb, NUP85, Olfactomedin-2, Olig2, OPG, or IL-13, OSCAR, OSM R beta, Osteoactivin, Osteoadherin, 0X40, 0X40 Ligand, P1GF-2, p27, p53, p63, PAPP- A, Pappaly sin-2, PARI, PARC, Pax3, PCSK2, PD-1, PD-ECGF, PD-L2, PDGF R alpha, PDGF Rb, PDGF-AA, PDGF-AB, PDGF-CC, PDX-1, PEAR1, PECAM-1, Pentraxin 3, Pepsinogen II, Persephin, PGRP-S, PIGF, PILR-alpha, Plexin A4, Plexin B3, Plexin DI, Podocalyxin, Podoplanin, POGLUT1, PON1, Pref-1, PRELP, Presenilin 1, Procalcitonin, Progranulin, proGRP, Prolactin, Prolactin R, PRX2, PSA-total, PSMA, PSMA1, PTH, PTH1R, PTP1B, PU.l, PYY, R-Spondin 2, RAGE, Rai A, RANK, RANTES, RBP4, RCOR1, Reg3A, Reg4, RELT, Renin, Resistin, Ret Midkine, RGM-A, RGM-B, ROBO2, ROBO3, ROBO4, R0R1, R0R2, S100A1, S100A13, SALM4, SCCA2, SCF, SCF R, SDF-la, SDF-lb, Semaphorin 4C, Semaphorin 4D, Semaphorin 4G, Semaphorin 6A, Semaphorin 6B, Semaphorin 6D, Semaphorin 7A, Serpin A4, Serpin A5, sFRP-3, SH2D1A, Shh-N, SHP-1, Siglec-10, Siglec-11, Siglec-2, Siglec-6, Siglec-7, Siglec-9, SIRP alpha, Sirtuin 1, Sirtuin 2, Sirtuin 5, SLAM, SLITRK5, Smad4, SMPD1, SorCSl, SorCS2, SorCS3, SOST, SOX15, SOX2, SOX7, SOX9, SP-D, SPHK1, Spinesin, SPINK1, SR-AI, SREC-I, SREC-II, ST6GAL1, ST8SIA1, Stabilin-2, SULT2A1, Syndecan-3, Syndecan-4, Syntaxin 4, Syntaxin 6, TACE, TAFA1, TAFA2, TAFAS, TARC, TAZ, TC-PTP, TCN2, TECK, Tenascin R, Testican 2, TFF3, TFPI, TFPI-2, TGFa, TGFb2, TGFb3 FOLR2, TGFbl, TGM3, THAP11, Thrombospondin-2, Thrombospondin-5, THSD1, Thyroglobulin, Thyroid Peroxidase, Tie-1, Tie-2, TIM-1, TIM-3, TLR1, TLR2, TLR3, TLR4, TNF RI, TNF RII, TNF4, TNFb, TPO, TPP1, TPST2, TRACP, TRAIL, TRAIL Rl, TRAIL R3, TRAIL R4, TRANCE, TREM-1, TREM-2, TREML1, TrkC, TROP-2, Troponin C, TROY, Trypsin 1, Trypsin 3, TSH, TSK, TSLP, TSLP R, TWEAK R, Ubiquitin+1, UCH-L3,WSGR Ref. No. 66309-733.601ULBP-1, ULBP-2, ULBP-3, ULBP-4, UNC5H3, UNC5H4, uPA, uPAR, Uromodulin, VAMP-1, VAMP-2, VAP-A, VCAM-1, VE-Cadherin, VEGF, VEGF Rl, VEGF R2, VEGF R3, VEGF- C, Visfatin, vWF-A2, WIF-1, WISP-1, Wnt-4, XEDAR, XIAP.

[0120] The nucleic acid content of the therapeutic product was determined, and the following nucleic acids were found to be present: 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-lOb, 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-l'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-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir- 92a-l, 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.Example 2 - Safety Study for Treatment of Idiopathic Facial Paralysis (Bell’s Palsy) and Secondary Facial Paralysis using Human Bone Marrow Stem Cell Derived Extracellular Vesicle Therapeutic Product

[0121] In this Example, a new and safe treatment was developed to restore facial function using extracellular vesicles (EVs) in patients who have been unable to recover from idiopathic facial paralysis. A pilot safety study was performed on 7 patients with idiopathic and secondary facial paralysis to determine if any functional restoration was possible. Each patient had symptomology for varying periods of time, with diverse House-Brackmann scores. All patients were treated with the same protocol of EVs over a 4-week period of time and were evaluated before, during, and after treatment. All patients received treatment by a private physician prior toWSGR Ref. No. 66309-733.601 entering the study. A record review was completed, with independent physical examinations. House-Brackmann scores and Facial Disability Indices were obtained prior to and after completing the study. EVs were injected into the area of the main trunk of the facial nerve on the affected side, and an intravenous drip of EVs on visits during weeks 1, 2, and 4 was administered. All 7 patients showed improvement in response to the treatment protocol. After the second week of treatment, a progression of independent motion was observed in the affected eyelid, brow, and commissure. All patients completed 4-week the study with a similar House- Brackmann endpoint score. No adverse effects were reported.

[0122] The therapeutic product (IP) used herein is a human BM-MSC derived EV (hBM-MSC EV) preparation. The IP was prepared by culturing BM-MSC at an oxygen tension below 5% in a media having a pH below 7. The secretome was separated from the BM-MSCs and formulated for administration in humans. For example, the formulation involves dilution and / or buffer exchange into a saline solution for IV administration. The IP is a consistent EV product with extensive characterization including advanced particle analysis, proteomic evaluation and USP <71> sterility assurance. The IP comprises various factors and nucleic acids including but not limited to SCF1, M-CSF, BMP-1, IGF-2, TGF- 1, SCF1, GALC, TUBB3, PDGFRA, PDGFRB, COL6A3, TNC, HSP90AB1, HSP90B1, RAC1, TGM2, VEGF, miR-199a-3p, let-7b-5p, miR- 26a-5p, miR-125b-5p, let-7a-5p, miR-21, miR-125b, and miR-26a. The IP comprises EVs which are at least 80% CD63+CD9 CD8 I . Additionally, through manufacturing in a cGMP environment, the IP is a quality bio-pharmacological product that is consistent regarding dose and biological activity. Two peer reviewed studies have demonstrated the safety of administering IV infusions of the IP to severe CO VID-19 patients. The IP has also demonstrated efficacy in a subset of these severely ill patients.Methods

[0123] 7 subjects with idiopathic facial paralysis were examined. None of the subjects were pregnant. None of the subjects had reconstructive surgery to correct the paralysis prior to treatment. All study procedures and protocols were approved in advance by an independent review board at the Institute of Regenerative and Cellular Medicine. The IRB approval number is IRCM-2021-304. Imaging was performed on each subject along with routine laboratory analyses and independent medical screening examinations. Laboratory analyses included but were not limited to complete blood count (CBC), comprehensive metabolic panel (CMP), and urinalysis (U / A). Baseline assessments were obtained at the start of the study. Facial disability index and House-Brackmann score were obtained at the start of the study. Radiographical studies were reviewed and / or CT scans were ordered. Any supportive therapy such as treatmentWSGR Ref. No. 66309-733.601 with botulinum toxin was ceased prior to entering the study. Informed consent was obtained from all subjects prior to entering the study.

[0124] During Week 1 of the study, ultrasound of the facial nerve and / or parotid was performed. 13 cc of the therapeutic product was administered in 100 cc of normal saline intravenously. 2 cc of therapeutic product were injected directly into tissue around the facial nerve. Week 1 protocol was repeated during Week 2. No intervention was administered during Week 3. Week 1 protocol was repeated during Week 4. Final assessments, facial disability index, and House-Brackmann score were assessed during Week 4. Patients were re-evaluated during Week 4, and follow up calls were conducted during Week 5.

[0125] All patients were examined by an independent physician prior to entering the study. All medical records were reviewed and additional studies were ordered as needed to ensure a safe and reasonable standard of compliance and a baseline to proceed with the study. All protocols were discussed for more detailed patient understanding as well as informed consent details. All cases had been seen and treated by their personal physicians and had exhausted treatment options. The time since the onset of symptoms was different in each case, but the treatment protocol was the same in all cases. There were no excluded cases with the manifestation of facial palsy in the study as patient selection was thorough. At the time of entering the study, all patients were off Botox except for one (#7) who used it to control epiphora and was free of any other contributary modalities that could interfere with the study.

[0126] Each patient was scored before treatment and upon completion of the study with the House-Brackmann scale, as well as a Facial Disability Index score. The therapeutic product was administered in 100 cc normal saline intravenously with a synchronous injection of 2 cc therapeutic product directly into tissue around the facial nerve with ultrasound guidance during Weeks 1, 2, and 4. Before treatment, subjects presented with severe and moderately severe state of paralysis, with a maximum House-Brackmann score of 5 and a minimum score of 3.Patients

[0127] Patient #1 : A 61-year-old male had an acoustic neuroma removed from his right side 2 years prior. His postop recovery was uneventful for twenty-four hours. Then he spontaneously developed an acute onset of right facial paralysis. He was treated with gabapentin, potassium, and diuretics without any improvement. Radiological studies were unremarkable. Mild to moderate symptoms of depression were experienced. (FIG. 1).

[0128] Patient #2: A 38-year-old female developed sudden onset of left-sided facial paralysis in July 2018 when she was 32 weeks pregnant. She was treated with 10 days of prednisone and valaciclovir unsuccessfully. After treatment with massage, physical therapy, and acupuncture, she had a slight improvement nine months later in that she could blink her affected left eye.WSGR Ref. No. 66309-733.601Radiological studies were unremarkable. Her symptoms persisted for 4 years prior to entering the study. Severe symptoms of depression were experienced.

[0129] Patient #3: A 54-year-old male basketball player developed a spontaneous Bell’s Palsy while touring in Asia. He developed spontaneous acute left-sided facial nerve paralysis. He was treated with acupuncture with unremarkable results. He has had Bell’s Palsy for 10 years with no improvement. MRI of the facial nerve was normal. Mild to moderate symptoms of depression were experienced.

[0130] Patient #4: A 32-year-old female with a history of a narrow palate and an open bite with a receded chin had a LeFort I osteotomy 13 years prior. She developed bilateral condylar reabsorption and had TMJ replacement 3 years prior. She awoke in the recovery room with bilateral facial paralysis. The right side recovered quickly, but the left-sided facial paralysis persisted. She displayed limited progress with facial exercises and steroids. She had facial paralysis with synkinesis for 3 years. She had a history of psoriatic arthritis. Radiological studies were unremarkable for facial nerve pathology. Moderate symptoms of depression were experienced. (FIG. 2).

[0131] Patient #5: A 51 -year-old female who awoke from her sleep 2 years prior with a drooping right lip commissure. She was treated with prednisone and valaciclovir for 10 days and a repeat dose of prednisone for eight weeks. She received acupuncture treatment because of her frequent headaches with her palsy. Bell’s Palsy symptoms were present for 10 months with no improvement. Similarly, radiological studies were unremarkable She had a history of systemic sclerosis affecting her ankles that made ambulation painful. Mild to moderate symptoms of depression were experienced.

[0132] Patient #6: A 58-year-old female with a history of surgery for a 3 cm right-sided acoustic neuroma 8 years prior and developed Bell’s Palsy 24 h postoperatively. Radiological studies were normal. Treatment consisted of physical therapy and botulinum toxin. No improvement was seen on this regimen. Her symptoms have been stable and unchanged for 8 years prior to entering the study. Mild to moderate symptoms of depression were experienced.

[0133] Patient #7: A 55-year-old female developed a spontaneous left earache, headache, and epiphora 1 year ago. Immediately a left-sided Bell’s Palsy developed. After treatment with prednisone and valaciclovir for one week, she received vitamin B-12 shots, facial massage, and botulinum toxin with an improvement in her left-sided epiphora only. She had left-sided Bell’s Palsy for 11 months. Mild to moderate symptoms of depression were experienced.Results

[0134] Each patient, before treatment and upon completion of the study was scored with the House-Brackmann scale, as well as a Facial Disability Index score (Table 3). Before treatment,WSGR Ref. No. 66309-733.601 cases appeared with severe and moderately severe states of paralysis, with a maximum House- Brackmann score of 5 and minimum score of 3. After treatment completion, there was a decrease of House-Brackmann scores in all patients (FIG. 3). They all decreased to a minimum House-Brackmann score of 2 except for one (Patient #7), displaying mild impairment of the disease, as FIG. 3 shows. House-Brackmann scores before and after treatment were analyzed statistically for the total number of patients using a paired t-test to determine whether, on average, there is a change between these two variables for the same subject. The statistic t-test = 4.804, with 6 degrees of freedom for a 95% confidence interval, and significance level a = 0.003, less than a = 0.05 shows there is significant evidence that the House-Brackmann index decreased after the treatment by an average of 1.429 (Table 5 and Table 6). The Facial Disability Index has two components, the Physical Index and Social Index, which were measured before and after the treatment for each patient as is shown in FIG. 4. Every patient scored a significant increase in Physical Index and Social Index values at the end of the treatment, except for one (Patient #6). None of them showed a decrease in functionality. The analyzed data for a total of 7 variables showed that the Physical Index before the treatment was registered in a minimum value of 44 and after treatment reached a maximum value of 137 (Table 4). Also, a statistical analysis using a paired t-test was conducted to determine whether, on average, there was a change in Physical Index before and after the treatment. The statistic test t = -5.542, with 6 degrees of freedom for 95% confidence interval, and significance level a = 0.001, less than a = 0.05. This shows evidence of significant changes of Physical Index after the treatment, by an average of 28.21 (Table 5 and Table 6). The Social Index before treatment was scored at its minimum of 44, and after treatment, it had an increase to a maximum value of 84 (Table 4). A statistic-paired t-test was made to determine whether there was a change of this parameter before and after the treatment. The statistic test t = -2.497, with 6 degrees of freedom for a 95% confidence interval, and significance level a = 0.047, less than a = 0.05 shows significant changes in Social Index after the treatment, by an average of 14.11 (Table 5 and Table 6). After treatment, each of the patients noticed an increase in face functionality, where there was significant impairment prior to treatment. Their social exposure had a significant improvement, and they claimed to feel more confident and socially interactive. Data analysis after study completion showed a trend, and it is displayed in FIG. 5. Some patients manifested the disease symptomatology for eight to ten years. Others had them for several months.Although the subjects had different lengths of time since the onset of the symptoms and different House-Brackmann grade points, they all responded almost to the same point after treatment, respectively over a one-month period of time. The symptoms decreased from severe and moderately severe to mild. The post-study Zoom calls revealed that all patients felt that theyWSGR Ref. No. 66309-733.601 made significant improvements. Patient #3 gained independent brow and eyelid motion, which he totally lacked prior to treatment. He also developed buccal control and gained overall facial expression. All patients developed a sequence of functional restoration beginning with the brow, then the eye, and finally the commissure.

[0135] Table 3 below shows a table of clinical assessments in each patient before and after treatment with the MSC secretome comprising extracellular vesicles.Table 3

[0136] Table 4 below shows a table of the analysis of descriptive statistics from the 7 treated patients (N=7), such as mean values, minimum values, and maximum values for each parameter assessed in the study disclosed herein.Table 4WSGR Ref. No. 66309-733.601

[0137] Table 5 and Table 6 below shows a table of statistical t-test values and significance level values (2 -tailed) for a 95% confidence interval conducted for each pair of variables.Table 5Table 6Discussion

[0138] No adverse events were observed over the course of the study disclosed herein. No bruising or pain was reported. Each patient was closely monitored for medical progress, change, and adverse events. Independent medical evaluations were performed on all patients prior to entering the study. All seven patients enrolled in the study improved with the treatment protocol disclosed herein. Four reported a sensation of nerve “tingling “, mild fasciculations, or “coming alive “of the nerve in the temple area, moving progressively to the buccal musculature, and to the angle of the mandible. Then a general relaxation was reported. Two patients reported theWSGR Ref. No. 66309-733.601 relaxation directly without fasciculations, as they did not feel the need to massage their faces every morning, as many patients do. After the second week of treatment, some patients developed independent motion of the affected eyelid - the restoration of the ability to wink. By the end of the second week into the third week, most developed early brow motion. Patients were allowed to rest on Week 3, to allow the EVs an opportunity to work. In some embodiments, EVs are effective in a treatment protocol for 4 weeks or more. After the third treatment, (Week 4), commissure motion improved. Although all patients began at different House-Brackman starting points, almost all ended at the same endpoint on the scale over the same period of time - four weeks. The duration of symptomology did not appear to affect the final outcome in terms of the House-Brackmann scoring.

[0139] In some embodiments, idiopathic facial paralysis comprises Bell’s Palsy. In some embodiments, idiopathic facial paralysis comprises secondary facial paralysis. In some embodiments, vascular secondary and tertiary ischemia cause the facial nerve sheath to thicken and inhibit nerve function in cases of facial paralysis. In some embodiments, facial paralysis present with viral involvement. Viruses implicated in facial paralysis but not limited to include Herpes simplex, Herpes zoster, Epstein-Barr virus, Coxsackie virus, rubella and cytomegalovirus. In some embodiments, Lyme disease contributes to facial paralysis. In some embodiments, facial paralysis is associated with or follows the onset of pregnancy. In some embodiments, demyelination contributes to facial paralysis. In some embodiments, autoimmune involvement is associated with facial paralysis. In some embodiments, high levels of interleukins such as interleukin-6 (IL-6) and IL-8 and / or elevated levels of tumor necrosis factor gamma (TNF-gamma) and TNF-alpha are associated with facial paralysis. In some embodiments, round inflammatory cells are present in the facial nerves in cases of facial paralysis. Demyelination may or may not be present. Inflammatory cells have been reported between nerve bundles with normal blood vessels between the internal acoustic meatus and the stylomastoid foramen. A segmental breakdown of the myelin sheath may be observed. In some embodiments, treatment with the therapeutic product disclosed herein reduces inflammation. Nerve recovery following treatment with the therapeutic product disclosed herein may be partial or complete. In some embodiments, patient experience a lower quality of life before treatment as compared to after treatment. In some embodiments, an improvement in conditions not associated with facial paralysis is reported following treatment. In some embodiments, improvement in conditions including but not limited to psoriatic arthritis and systemic sclerosis is reported following treatment. In some embodiments, no return of severe symptomology is reported after the cessation of treatment. In some embodiments, continued improvement is noted over the course of 2 months following treatment.WSGR Ref. No. 66309-733.601

[0140] The examples and embodiments described herein are for illustrative purposes only and various modifications or changes suggested to persons skilled in the art are to be included within the spirit and purview of this application.

Claims

1. WSGR Ref. No. 66309-733.601CLAIMSWHAT IS CLAIMED IS:

1. A method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein at least 80% of the EVs are CD63+CD9" CD81 .

2. A method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the composition comprises one or more factors, the one or more factors comprising 2B4, 4-1BB, 4-1BB Ligand, 6Ckine, ACE, Activin A, Activin RIB, Activin RIIA, Activin RIM, ADA, ADAM23, ADAMS, ADAMTS13, ADAMTSL-1, Adiponectin, Adipsin, Aggrecan, AgRP, AIF, AKR1C4, Albumin, ALCAM, AMIGO, AMIG02, Aminopeptidase LRAP, Aminopeptidase P2, AMSH, ANG-1, ANG-2, ANG-4, Angiogenin, Angiotensinogen, ANGPTL4, ANGPTL7, APRIL, Arginase 1, ARSB, Artemin, Arylsulfatase A, ASAH2, ASAHL, Axl, B2M, B7-1, B7-H3, B7-H4, BAFF R, Bcl-10, BCL-2, Bcl-w, BCMA, BID, Biglycan, BLAME, BMP-2, BMP-8, BMP-9, BMPR-IA, Brevican, C-myc, CA125, CA I 3, CA4, CA8, Cadherin-13, Cadherin-4, Calreticulin, Calreticulin-2, Cardiotrophin-1, CASA, Caspr2, Cathepsin E, Cathepsin L, Cathepsin S, Cathepsin V, CD14, CD155, CD157, CD2, CD200, CD229, CD27, CD27 Ligand, CD28, CD30, CD300f, CD314, CD34, CD36, CD39L2, CD4, CD40, CD40L, CD42b, CD48, CD51, CD58, CD6, CD73, CD83, CD97, CD99-L2, CDC25B, CDNF, CDS, CEACAM-1, CEACAM-3, CEACAM-5, Cerberus 1, CES1, CES2, CF VII, CF XIV, Chemerin, CHI3L1, CHST2, CHST3, CHST4, cIAP-1, CILP-1, CK19, CL-P1, CLEC-1, CLEC-2, CLEC10A, Clq, ClqTNF9, cMASP3, CMG-2, Cochlin, COMT, Contactin- 1, Contactin-2, CPB1, CPE, CREG, CRELD2, CRIM1, CrkL, CSa, CT ACK, CTRC, CXCL14, CXCL16, Cyr61, Cy statin B, Cy statin C,Cy statin D, Cy statin E M, Cy statin S, Cy statin SA, Cy statin SN, Cytokeratin-8, DAPP1, DCTN1, DDR1, Dectin- 1, Dectin-2, Desmin, Desmoglein 2, Desm oglein-3, DKK-1, DLL4, DNAM-1, DNMT3A, Dopa Decarboxylase, DPPII, DPPIV, DR3, DR6, Draxin, DSCAM, DSPG3, Dtk, E-Selectin, ED AR, EDIL3, EG-VEGF, EGF R, EMMPRIN, ENA-78, Endocan, Endoglin, eNOS, ENPP-2, ENPP-7, Enteropeptidase, EpCAM, EphA2, EphAl, EphB3, EphB4, EphB6, Ephrin-A4, Ephrin-B3, Epimorphin, Epiregulin, ErbB3, ErbB4, Erythropoietin, ESAM, EXTL3, FABP1, FABP2, FABP4, FABP6, FAP, Fas, FCAR, FcERI, FCRL1, FCRL3, FCRLB, FCRLS, FGF R5, FGF-12, FGF-16, FGF- 17, FGF-19, FGF-20, FGF-21, FGF-23, FGF-3, FGF-5, Ficolin-1, FKBP51, FLRG,WSGR Ref. No. 66309-733.601FLRT1, FLRT2, Flt-3, Flt-3L, Follistatin, Folli statin-like 1, FUT8, Galanin, Galectin-1, Galectin-2, Galectin-3, Galectin-4, Galectin-7, Galectin-8, Galectin-9, GALNT10, GALNT2, GALNT3, Gash, GATA-4, GBA3, GCP-2, GDF-11, GDF-15, GDF-9, GFAP, GFR alpha- 1, GFR alpha-2, GITR, GITR L, GKN1, GLP-1, Glycoprotein V, Glyoxalase II, Glypican 1, Glypican 2, Glypican 5, gpl30, GPR56, GPVI, Granulysin, Granzyme A, Granzyme B, Granzyme H, GRAP2, GRKS, GRO, GSTM1, GUSB, HAO-1, HAPLN1, HCC-1, HCC-4, hCGb, HE4, Hepsin, HGF, HGF R, HIF-1 alpha, HIF-1 beta, HO-1, HS3ST1, HS3ST3B1, HSD17B1, htPAPP-A, HTRA2, HVEM, I-TAC, ICAM-1, ICAM- 2, ICAM-3, ICOS, IDO, IFN-gamma Rl, IFNab R2, IFNb, IGF-2R, IGFBP-5, IGSF3, IGSF4B, IL-1 F10, IL-1 R3, IL-1 R4, IL-1 RI, IL-1 RII, IL- 10 Rb, IL-12p40, IL- 13 R2, IL-15, IL-15 R, IL-17 RC, IL-17 RD, IL-17B, IL-17F, IL-18, IL-18 BPa, IL-19, IL-2 Ra, IL-2 Rb, IL-2 Rg, IL-20 R beta, IL-20 Ra, IL-21R, IL-22, IL-22 R alpha 1, IL-22BP, IL- 23, IL-23 R, IL-24, IL-27, IL-27 Ra, IL-28 R alpha, IL-28A, IL-31, IL-31 RA, IL-32 alpha, IL-33, IL-4 Ra, IL-6, IL-6R, IL-7 R alpha, IL-8, IL-9, IL-lra, ILT2, ILT4, InhibinA, Insulin, Insulin R, Integrin alpha 1, Integrin alpha 5, IP-10, Jagged 1, JAM-A, JAM-B, JAM-C, Kallikrein 1, Kallikrein 11, Kallikrein 12, Kallikrein 14, Kallikrein 5, Kallikrein 7, Kell, KIR2DL3, Kirrel3, KLF4, Kremen-2, Kynureninase, L-Selectin, L1CAM-2, LAIR2, LAMA4, LAMP, LAMP1, Langerin, LAP(TGFbl), LDL R, LEDGF, Legumain, LH, LIF, LIF R alpha, LIGHT, LILRB4, LIMP II, Lin28, Lipocalin- 2, LOX-1, LRP-6„ST3GAL1, LRRC4, LRRTM4, LSECtin, LTbR, Lymphotactin, LYVE-1, MAP1D, Matrilin-2, Matrilin-3, Matriptase, MBL, Mcl-1, MCEMP1, MCP-2, MCP-3, MCP-4, MCSF, MDC, MDL-1, MEF2C, MEP1B, MEPE, MeprinA, Mer, Mesothelin, METAP2, MFRP, MIA, MICA, MICB, Midkine, MIP-3a, MIP-3b, MIP-lb, MIS RII, MMP-1, MMP-10, MMP-12, MMP-13, MMP-2, MOG, MPIF-1, MSP R, N- Cadherin, NAP-2, NCAM-1, NCAM-1 (CD56), NCK1, Nectin-1, Nectin-2, Nectin-3, Nectin-4, Neprilysin, Nestin, Netrin-4, Neudesin, Neurexin 3 beta, Neurogranin, Neuroligin 2, Neuropilin-2, Neurturin, NG2, NGF R, Nidogen-1, Nidogen-2, NKp30, NKp44, NKp46, NKp80, Nogo Receptor, Nogo-A, Norrin, Notch- 1, Notch-3, NOV, NPDC-1, NPTXR, NQO-1, Nrf2, NRG1-131, NSE, NTAL, NUDTS, Numb, NUP85, Olfactomedin-2, Olig2, OPG, or IL-13, OSCAR, OSM R beta, Osteoactivin, Osteoadherin, 0X40, 0X40 Ligand, P1GF-2, p27, p53, p63, PAPP -A, Pappalysin-2, PARI, PARC, Pax3, PCSK2, PD-1, PD-ECGF, PD-L2, PDGF R alpha, PDGF Rb, PDGF-AA, PDGF-AB, PDGF-CC, PDX-1, PEAR1, PECAM-1, Pentraxin 3, Pepsinogen II, Persephin, PGRP-S, PIGF, PILR-alpha, Plexin A4, Plexin B3, Plexin DI, Podocalyxin, Podoplanin, POGLUT1, PON1, Pref-1, PRELP, Presenilin 1,WSGR Ref. No. 66309-733.601Procalcitonin, Progranulin, proGRP, Prolactin, Prolactin R, PRX2, PSA-total, PSMA, PSMA1, PTH, PTH1R, PTP1B, PU. l, PYY, R-Spondin 2, RAGE, RalA, RANK, RANTES, RBP4, RCOR1, Reg3A, Reg4, RELT, Renin, Resistin, Ret Midkine, RGM-A, RGM-B, ROBO2, ROBO3, ROBO4, ROR1, ROR2, S100A1, S100A13, SALM4, SCCA2, SCF, SCF R, SDF-la, SDF-lb, Semaphorin 4C, Semaphorin 4D, Semaphorin 4G, Semaphorin 6A, Semaphorin 6B, Semaphorin 6D, Semaphorin 7A, Serpin A4, Serpin A5, sFRP-3, SH2D1A, Shh-N, SHP-1, Siglec-10, Siglec-11, Siglec-2, Siglec-6, Siglec-7, Siglec-9, SIRP alpha, Sirtuin 1, Sirtuin 2, Sirtuin 5, SLAM, SLITRK5, Smad4, SMPD1, SorCSl, SorCS2, SorCS3, SOST, SOX15, SOX2, SOX7, SOX9, SP-D, SPHK1, Spinesin, SPINK1, SR-AI, SREC-I, SREC-II, ST6GAL1, ST8SIA1, Stabilin-2, SULT2A1, Syndecan-3, Syndecan-4, Syntaxin 4, Syntaxin 6, TACE, TAFA1, TAFA2, TAFAS, TARC, TAZ, TC-PTP, TCN2, TECK, Tenascin R, Testican 2, TFF3, TFPI, TFPI-2, TGFa, TGFb2, TGFb3 FOLR2, TGFbl, TGM3, THAP11, Thrombospondin-2, Thrombospondin-5, THSD1, Thyroglobulin, Thyroid Peroxidase, Tie-1, Tie-2, TIM-1, TIM-3, TLR1, TLR2, TLR3, TLR4, TNF RI, TNF RII, TNF4, TNFb, TPO, TPP1, TPST2, TRACP, TRAIL, TRAIL Rl, TRAIL R3, TRAIL R4, TRANCE, TREM-1, TREM-2, TREML1, TrkC, TROP-2, Troponin C, TROY, Trypsin 1, Trypsin 3, TSH, TSK, TSLP, TSLP R, TWEAK R, Ubiquitin+1, UCH-L3, ULBP-1, ULBP-2, ULBP-3, ULBP-4, UNC5H3, UNC5H4, uPA, uPAR, Uromodulin, VAMP-1, VAMP-2, VAP-A, VCAM-1, VE-Cadherin, VEGF, VEGF Rl, VEGF R2, VEGF R3, VEGF-C, Visfatin, vWF-A2, WIF-1, WISP-1, Wnt-4, XEDAR, or XIAP, or a combination of two or more thereof; optionally wherein the EVs comprise the one or more factors.

3. A method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the composition comprises one or more factors, the one or more factors comprising TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP- 2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulinlike growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), LAMB1 (Laminin Subunit Beta 1), FBN1 (Fibrillin 1), HSPG2 (Heparin Sulphate Proteoglycan 2), BGN (Biglycan), or FN1 (Fibronectin 1), or a combination of two or more thereof; optionally wherein the EVs comprise the one or more factors.

4. The method of claim 3, wherein the composition further comprises beta-IG-H3 (Transforming growth factor-beta-induced protein ig-H3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD81 antigen, CD9 antigen, CD44 antigen, CD99 antigen, CD 109 antigen,WSGR Ref. No. 66309-733.601CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Furin, GM-CSF Ra (Granulocyte-macrophage colonystimulating factor receptor subunit alpha), GPR115 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF -2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein-3), IGFBP-6 (Insulin-like growth factor binding protein-6), IL18BP (Interleukin- 18 Binding Protein), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein- 4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor- 1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin Fl (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein-glutamine gammaglutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), TSP-2 (Thrombospondin 2), CAD11 (Cadherin 11), JAM3 (Junctional Adhesion Molecule 3), VIM (Vimentin), FBLN5 (Fibulin 5), FBN2 (Fibrillin 2), Hyaluronic Acid, COL 1 Al (Collagen Type 1 Alpha 1), COL1 A2 (Collagen Type 1 Alpha 2), or Transferrin, or a combination of two or more thereof.

5. A method of treating idiopathic facial paralysis in a subject, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the composition comprises hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, or let-7, or a combination of two or more thereof; optionally wherein the EVs comprise the hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, or let-7, or a combination of two or more thereof.

6. The method of claim 5, wherein the composition further comprises hsa-miR-125b-5p, hsa-miR-132-3p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-221-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa- miR-424-5p, or hsa-miR-940, or a combination of two or more thereof.

7. The method of any one of claims 1 to 6, wherein the composition is produced by:(a) culturing bone marrow-derived MSCs (BM-MSCs) under the following conditions to produce an MSC conditioned media:WSGR Ref. No. 66309-733.601(i) oxygen tension below 5%; and(ii) culture media having a pH below 7;(b) harvesting the MSC conditioned media; and(c) formulating the MSC conditioned media to produce a therapeutic MSC secretome, wherein the therapeutic MSC secretome comprises proteins and EVs produced by the BM-MSCs in (a).

8. The method of any one of claims 1 to 7, wherein the subject exhibits one or more of the following symptoms prior to the administering: extracranial paralysis, intracranial paralysis, epiphora, inability to close one or two eyes, drooping of the oral commissure, salivary incompetence, facial tingling, fasciculations of one or more facial muscles, hyperacusis, taste disturbance, inability to elevate one or two eyebrows, difficulty communicating, inability to communicate, inability to smile, difficulty with social interaction, psychological disturbance, depression, headache, earache, tinnitus, dizziness, loss of balance, and / or secondary facial paralysis.

9. The method of claim 8, wherein the extracranial paralysis, intracranial paralysis, or secondary facial paralysis affects one side of the face.

10. The method of claim 8 or 9, wherein the onset of the one or more symptoms is acute.

11. The method of any one of claims 8 to 10, wherein the one or more symptoms do not improve or do not improve fully in response to one or more of surgery, facial reconstruction, oral steroids, anti-viral medications, facial exercises, laser treatment, herbal remedy, acupuncture, botulinum toxin, massage, physical therapy, prednisone, gabapentin, potassium, valaciclovir, vitamin B-12, and / or diuretics.

12. The method of any one of claims 8 to 11, wherein the subject has experienced the one or more symptoms for 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, or longer.

13. The method of any one of claims 1 to 12, wherein results of the subject’s radiological studies are unremarkable before and / or after the administering.

14. The method of any one of claims 1 to 13, wherein the administering comprises one or more injections.WSGR Ref. No. 66309-733.60115. The method of claim 14, wherein at least one of the one or more injections is injected into tissue around the facial nerve.

16. The method of claim 14 or 15, wherein at least one of the one or more injections comprises 1 cc, 2 cc, 3 cc, 4 cc, or 5 cc of the composition.

17. The method of claim 14 or 15, wherein at least of the one or more injections comprises 5 cc, 6 cc, 7 cc, 8 cc, 9 cc, 10 cc, 11 cc, 12 cc, 13 cc, 14 cc, 15 cc, 16 cc, 17 cc, 18 cc, 19 cc, or 20 cc of the composition.

18. The method of any one of claims 14 to 17, wherein at least one of the one or more injections is injected intravenously.

19. The method of any one of claims 1 to 18, wherein the administering restores facial nerve function in the subject as compared to before administration.

20. The method of any one of claims 1 to 19, wherein the administering reduces House- Brackmann score in the subject as compared to before administration.

21. The method of claim 20, wherein the decrease in House-Brackmann score is at least 1, at least 2, at least 3, at least 4, at least 5, or at least 6.

22. The method of any one of claims 1 to 21, wherein the administering improves a Facial Disability Index score in a subject as compared to before administration.

23. The method of claim 22, wherein the improvement in the Facial Disability Index score is an improvement in a physical index.

24. The method of claim 22, wherein the improvement in the Facial Disability Index score is an improvement in a social index.

25. The method of any one of claims 22 to 24, wherein the improvement in the Facial Disability Index score is an increase in at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%.

26. The method of any one of claims 1 to 25, wherein the administering restores one or more of brow motion, eyelid motion, buccal control, and / or facial expressiveness in the subject as compared to before administration.WSGR Ref. No. 66309-733.60127. The method of any one of claims 1 to 26, wherein the administering results in improvement in wellbeing, reduction in tingling, reduction in fasciculations, relaxation of muscles, reduction in pain, restoration of ability to wink, and / or improvement in symmetrical facial movements.

28. A method of treating idiopathic facial paralysis in a subject in need thereof, the method comprising administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome comprising extracellular vesicles, wherein the subject exhibits an improvement in House-Brackmann score after administration compared to before administration.

29. A method of making a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome for treating idiopathic facial paralysis in a subject in need thereof, the method comprising:(a) culturing bone marrow-derived MSCs (BM-MSCs) under the following conditions to produce an MSC conditioned media:(i) oxygen tension below 5%; and(ii) culture media having a pH below 7;(b) harvesting the MSC conditioned media; and(c) formulating the MSC conditioned media to produce the therapeutic MSC secretome, wherein the therapeutic MSC secretome comprises proteins and extracellular vesicles produced by the BM-MSCs in (a).

30. The method of any one of claims 1 to 29, wherein the idiopathic facial paralysis comprises Bell’s Palsy, facial palsy, and / or secondary facial paralysis.

31. The method of claim 29 or 30, wherein the culture media is serum-free.

32. The method of any one of claims 29 to 31, wherein the culture media has a glucose concentration below 4.5 g / L.

33. The method of any one of claims 29 to 32, wherein formulating the MSC conditioned media comprises exchanging the conditioned media for a pharmaceutically acceptable formulation.WSGR Ref. No. 66309-733.60134. The method of claim 33, wherein the pharmaceutically acceptable formulation comprises saline.

35. The method of any one of claims 1 to 34, wherein the composition comprises at least 6xlO10to 8xlO10extracellular vesicles per mL and is administered at a dose of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml.

36. The method of claim 35, wherein the dose is administered in combination with normal saline at a final volume of 100 mL.

37. The method of any one of claims 1 to 36, wherein the composition comprises saline (0.9% sodium chloride).

38. The method of claim 37, wherein the saline is present in the composition at about 80% to about 95% saline.

39. The method of any one of claims 1 to 38, wherein the composition comprises sodium chloride, sodium lactate, potassium chloride, and / or calcium chloride.

40. The method of any one of claims 1 to 39, wherein the molecular weight of any nonexcipient component of the composition is greater than about 2 kDa (kilodaltons), 8 kDa, 10 kDa, 50 kDa, or 100 kDa.

41. The method of any one of claims 1 to 40, wherein the composition comprises an oligosaccharide.

42. The method of claim 41, wherein the oligosaccharide is present in the composition at from about 0.2 M to about 0.6 M.

43. The method of any one of claims 1 to 42, wherein any non-excipient component of the composition has a size of less than about 0.2 microns.

44. The method of any one of claims 1 to 43, wherein the composition is sterile per USP <71>.

45. The method of any one of claims 1 to 44, wherein the composition is endotoxin free per USP <85>.

46. The method of any one of claims 1 to 45, wherein the composition is negative for mycoplasma DNA.WSGR Ref. No. 66309-733.60147. The method of any one of claims 1 to 46, wherein the composition is cell-free.

48. The method of any one of claims 1 to 47, wherein the composition is stored between - 80°C and -60°C.

49. The method of claim 48, wherein the composition is administered within 6 hours of thaw when maintained at ambient temperature.

50. The method of any one of claims 1 to 49, wherein the composition is freeze-dried (lyophilized) to a powder cake and stored at or below room temperature.

51. The method of any one of claims 1 to 50, wherein the composition is reconstituted with water prior to administration.

52. The method of any one of claims 1 to 51, wherein the composition is present in a glass vial.

53. The method of any one of claims 1 to 52, wherein the composition is formulated for intravenous administration.

54. The method of any one of claims 1 to 53, wherein the composition has a pH of about 6 to about 7.5.

55. The method of any one of claims 7 to 54, wherein the BM-MSCs are negative for CD 14, CD31, CD34, and CD45.

56. The method of any one of claims 7 to 55, wherein the BM-MSCs are positive for CD73, CD 105, CD 166, and CD90.

57. The method of any one of claims 7 to 56, wherein the BM-MSCs are capable of undergoing trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes.

58. The method of any one of claims 7 to 57, wherein the BM-MSCs are obtained from an iliac crest aspiration of a single donor.

59. The method of any one of claims 1 to 58, wherein a concentration of the one or more EVs in the composition is measured by nanoparticle tracking analysis (NTA).WSGR Ref. No. 66309-733.60160. The method of claim 59, wherein the NTA comprises light scatter and fluorescence evaluation.

61. The method of claim 59 or 60, wherein the concentration of the one or more EVs is about 10 billion to about 250 billion EVs per mL of the composition.

62. The method of any one of claims 59 to 61, wherein the concentration of the one or more EVs is at about 1 billion to about 40 billion EVs per ml of the composition.

63. The method of any one of claims 1 to 62, wherein the one or more EVs have an average diameter of about 30 nm to about 170 nm.

64. The method of any one of claims 1 to 63, wherein a total protein concentration of the composition is about 10 to about 40 pg per ml of the composition.

65. The method of any one of claims 1 to 64, wherein a total protein concentration of the composition is about 1.5 to about 6 pg per ml of the composition.

66. The method of claim 64 or 65, wherein the total protein concentration is measured by ELISA.

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