Bone marrow mesenchymal stem cell derived extracellular vesicles to improve perioral area and face fat graft retention

Abstract

Disclosed are methods of improving perioral and face fat graft retention 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

BONE MARROW MESENCHYMAL STEM CELL DERIVED EXTRACELLULAR VESICLES TO IMPROVE PERIORAL AREA AND FACE FAT GRAFT RETENTIONCROSS REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 638,207 filed on April 24, 2024, the entirety of which is hereby incorporated by reference herein.BACKGROUND

[0002] Patients pursue facial rejuvenation procedures to improve skin general appearance and restore lost tissue volume. Due to age-associated skeletal changes and reduction in facial fat compartments, structural support is decreased, resulting in skin wrinkling and folding. Among the procedures available is the grafting of adipose tissue to restore structural support to critical areas of the face. Microfat graft retention in the perioral area is highly variable, leading to failure to improve facial volume over the long term. This leads to repeated procedures that in turn causes a poor aesthetic result. There exists a need for safe and effective methods for improving graft retention.SUMMARY

[0003] This Summary introduces a selection of concepts that are described further below in the Detailed Description. This Summary is not intended to limit the scope of the claimed subject matter.

[0004] Disclosed herein are methods and compositions for improving perioral face fat graft retention in a subject. Disclosed herein is the use of a composition comprising a mesenchymal stem cell (MSC) secretome in improving perioral face fat graft retention. Disclosed herein are methods and compositions for treating wrinkles in a subject. Disclosed herein are methods and compositions for treating age-related facial volume loss in a subject. Disclosed herein are methods and compositions for improving facial appearance in a subject. Disclosed herein are methods and compositions for improving fat graft retention in a subject. In some embodiments, a method disclosed herein comprises administering to the subject a composition comprising a mesenchymal stem cell (MSC) secretome. In some embodiments, a method disclosed herein comprises administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the composition comprises NUP85, DAPP1, PDGF R alpha, SLITRK5, FAP, Artemin, DPPII, cIAP-1, Pentraxin 3, Visfatin, Neprilysin, Albumin, Galectin-1, UNC5H3, IL-20 R beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin7A, NKp80, Cystatin B, Osteoadherin, Midkine, Calreticulin, Osteoactivin, Legumain, TAZ, Cathepsin L, RBP4, Serpin A4, JAM- A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin 3, SIRP alpha, and Syndecan-4, GSTM1, NUP85, MeprinA, IL-1 F10, TGFbl, Ephrin- A4, Aminopeptidase LRAP, GDF-9, PDGF R alpha, PAPP -A, Arylsulfatase A, LAIR2, ULBP- 4, TFPI, SOX2, SLITRK5, Spinesin, ENPP-2, CD97, CTACK, Integrin alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin DI, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Persephin, TRAIL Rl, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, P0N1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, Norrin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gpl30, C-myc, LILRB4, BMP-2, MIA, CD34, IFNab R2, Glypican 2, MSP R, DSCAM, Matriptase, KIR2DL3, CD30, Sigi ec- 10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nidogen-1, CD2, Kallikrein 1, TSLP R, TROY, VCAM-1, Siglec-11, S100A1, PARI, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD- ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 alpha, 0X40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, Semaphorin 4D, ROBO2, PDX-1, APRIL, Neurturin, Kremen-2, EMMPRIN, Activin RIB, Neuroligin 2, Epiregulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF Rl, DSPG3, SorCSl, Matrilin-2, sFRP-3, EphB3, NCK1, Prolactin, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Pappalysin-2, Syndecan-3, Jagged 1, AKR1C4, Olfactomedin-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p27, Siglec-6, Dectin- 1, CD51, Notch- 1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, CA125, HAO-1, PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, R0R1, CMG-2, 4-1BB Ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Legumain, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 beta, TAFA1, Podocalyxin, Rai A, CRELD2, GRAP2, SP-D, BID, GFR alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR,Contactin-2, CD83, IL-1 R3, SALM4, GBA3, R0B04, OSCAR, VEGF, IGSF3, Biglycan, Neudesin, ILT4, uPAR, Axl, WIF-1, IL-7 R alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Cochlin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL- 17 RC, MIP-3b, Cy statin C, Cy statin D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, ADAM23, NOV, Galectin-2, Neurexin 3 beta, TLR3, Sirtuin 2, Numb, IL-28 R alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM- A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA I3, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-Selectin, Hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, LTAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-lra, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpin A5, PTH, FGF-19, IL-28 A, FGF-12, METAP2, AS AHL, EDIL3, NT AL, EGF R, TAFAS, Galectin- 9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, 0X40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, R0R2, EphB4, Glypican 1, LAP(TGFbl), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL- 10 Rb, XEDAR, IL-22, PILR-alpha, NRG1- 131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirrel3, FLRT1, Galectin-3, CXCL16, JAM-B, DR6, Nogo Receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R alpha 1, AMIG02, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurogranin, HE4, IL-23 R, Galectin-7, GALNT3, GITR L, CD14, R- Spondin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospondin-5, IL- 1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulysin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin 3, ARSB, LIF R alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF-AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP- 2, PU.l, ED AR, ADAMTS13, Kynureninase, PTH1R, IFN-gamma Rl, CrkL, B7-1, PARC,Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 Rg, MICA, Dopa Decarboxylase, NPDC-1, MCP-4, EG-VEGF, Glycoprotein V, Semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, Endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, Galectin-8, CA4, Cystatin E M, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, Kallikrein 7, or IL-13, or a combination of two or more thereof. In some embodiments, a method disclosed herein comprises administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the composition comprises TIMP-1 (tissue inhibitor of metalloproteinases 1), TEMP -2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), LAMB 1 (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. In some embodiments, 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, 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 (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), PALI (Plasminogen activator inhibitor- 1 or SERPINE 1), PCK1 (Phosphoenol pyruvate 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, COL1 Al (Collagen Type 1 Alpha 1), COL1 A2 (Collagen Type 1 Alpha 2), or Transferrin, or a combination of two or more thereof. In some embodiments, the composition further comprises one or more of COL2A1, COL3A1, COL5A1, COL5A2, COL6A1, COL11 Al, or COL14A1, or a combination of two or more thereof. In some embodiments, a method disclosed herein comprises administering to the subjecta 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. 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, a method disclosed herein comprises administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein at least 80% of the one or more EVs are CD63+, CD9-, CD81-. In some embodiments, the perioral face fat graft is administered to the cheek, temple, under-eye area, nasolabial folds, lips, or any combination thereof. In some embodiments, the perioral face fat graft is obtained from the abdomen, flank, outer thigh, inner thigh, lower back, and / or buttocks. In some embodiments, the subject exhibits improved skin texture, improved skin quality, improve facial aesthetics, and / or any combination thereof after the administering. In some embodiments, the subject experiences a volumetric increase of a perioral or face area of 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60% after the administering. In some embodiments, the subject exhibits an increase of 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, 100, 101, 102, 103, 104, 105, 106, 107, 108,109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, or 130 mm3after administration of the composition. In some embodiments, the subject has skin rhytids. In some embodiments, the subject has periorbital or facial volume loss. In some embodiments, the administering comprises administering 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, or 3.0 mL of the composition. In some embodiments, the composition is administered after administration of a fat graft. In some embodiments, 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, or 3.0 mL of the fat graft is administered. In some embodiments, the fat graft is administered to more than one area of the face. In some embodiments, the fat graft is administered to a left side and a right side of the face. In some embodiments, the fat graft is administered to the dermis. In some embodiments, the fat graft is administered to the perioral dermis. In some embodiments, the composition is prepared by a process comprising: (a) culturing bone marrow mesenchymal stem cells (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 composition. In some embodiments, a method disclosed herein comprises preparing the composition prior to the administering, wherein the preparing the composition occurs by a process comprising: (a) culturing bone marrow mesenchymal stem cells (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 composition. 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 8xlO10extracellular vesicles per mL and is administered at a dose of 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 chlori de. In some embodiments, the molecular weight of any nonexcipient component of the composition is greater than about 2 kDa (kilodaltons), 8 kDa, 10 kDa, 50kDa, or lOOkDa. 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 size 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 towardadipocyte, 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. In some embodiments, the composition comprises 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 combination of two or more thereof. In some embodiments, 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, 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 (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 (Phosphoenol pyruvate 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, COL1 Al (Collagen Type 1 Alpha 1),C0L1 A2 (Collagen Type 1 Alpha 2), or Transferrin, or a combination of two or more thereof. In some embodiments, the composition further comprises one or more of COL2A1, COL3A1, COL5A1, COL5A2, COL6A1, COL11 Al, or COL14A1, or a combination of two or more thereof. In some embodiments, 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. 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, at least 80% of the one or more EVs are CD63+, CD9-, CD81-.

[0005] Provided herein are methods and compositions for grafting microfat into a subject’s face or perioral area. In some embodiments, a method disclosed herein comprises: (a) injecting a fat graft into the dermis of the subject’s face or perioral area; and (b) injecting a therapeutic bone marrow-derived MSC secretome composition into the dermis at or near the location of the microfat graft. In some embodiments, (b) is carried out before, after, or concurrently with (a). In some embodiments, the fat graft is injected into the upper or lower lip area. In some embodiments, the fat graft is injected at a volume of 0.5 mL, 1.0 mL, 1.5 mL, 2.0 mL, 2.5 mL, or 3.0 mL. In some embodiments, the subject receives multiple fat graft injections. In some embodiments, the MSC secretome composition is injected at a volume of 0.5 mL, 1.0 mL, 1.5 mL, 2.0 mL, 2.5 mL, or 3.0 mL. In some embodiments, the MSC secretome composition is collected from MSC conditioned media of bone marrow-derived MSCs cultured under conditions comprising oxygen tension below 5% and culture media having a pH below 7.INCORPORATION BY REFERENCE

[0006] Each patent, publication, and non-patent literature cited in the application is hereby incorporated by reference in its entirety as if each was incorporated by reference individually. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and / or take precedence over any such contradictory material.BRIEF DESCRIPTION OF THE DRAWINGS

[0007] 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 illustrativeembodiments, in which the principles of the disclosure are utilized, and the accompanying drawings (also “Figure” and “FIG.” herein), of which:

[0008] FIG. 1 shows images of subjects at baseline and nine months after treatment with the therapeutic product of the present disclosure.DETAILED DESCRIPTION

[0009] Provided herein are compositions and methods for improving perioral area and face fat graft retention.

[0010] 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. The composition may be referred to as a therapeutic composition.I. Therapeutic CompositionsA. Extracellular Vesicles

[0011] 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. Embodiments 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).

[0012] 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.

[0013] 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 .

[0014] 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 froma 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.

[0015] 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).

[0016] In some embodiments, the EV comprises a peptide or protein. In some embodiments, the peptide or protein is within the EV. In some embodiments, the peptide or protein is outside the EV. In some embodiments, the peptide or protein is embedded in the EV. In some embodiments, the peptide or protein is attached to the EV. In some embodiments, the EV comprises a nucleic acid. Nucleic acids include ribonucleic acids (RNA), such as siRNA, shRNA, and microRNA (miRNA). In some embodiments, the nucleic acid is within the EV. In some embodiments, the nucleic acid is outside the EV. In some embodiments, the nucleic acid is embedded in the EV. In some embodiments, the nucleic acid is attached to the EV.

[0017] In some embodiments, the EV 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). Non-limiting examples of the one or more proteins include: beta-IG-H3 (Transforming growth factor-beta-induced protein ig-H3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD63 antigen, 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), Ferritin, Furin, GM-CSF Ra (Granulocyte-macrophage 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-4 (Insulin-like growth factor binding protein-4), IGFBP-6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), IL18BP (Interleukin- 18 Binding Protein), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, MIF (Macrophage migration inhibitory factor), OPN (Osteopontin), PALI (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), TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), TSP-2 (Thrombospondin 2), CAD11 (Cadherin 11), JAM3 (Junctional Adhesion Molecule 3), VIM (Vimentin), FBLN5 (Fibulin 5), FBN2 (Fibrillin 2), Hyaluronic Acid, C0L1A1 (Collagen Type 1 Alpha 1), COL1A2 (Collagen Type 1 Alpha 2), Transferrin, LAMB1 (Laminin Subunit Beta 1), FBN1 (Fibrillin 1), HSPG2 (Heparin Sulphate Proteoglycan 2), BGN (Biglycan), and FN1 (Fibronectin 1). An EV that comprises the one or more of the proteins may include the one or more proteins within the EV. An EV that comprises one or more of the proteins may include the one or more proteins anchored within the EV. An EV that comprises the one or more of the proteins may be associated with the outside of the EV.

[0018] 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-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, let-7, 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, and hsa-miR-940. miRNA sequences may be obtained from www.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.B. Proteins

[0019] 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 comprise 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). Non-limiting examples of the one or more proteins include: LAMB1 (Laminin Subunit Beta 1), FBN1 (Fibrillin 1), HSPG2 (Heparin Sulphate Proteoglycan 2), BGN (Biglycan), FN1 (Fibronectin 1), IL18BP (Interleukin- 18 Binding Protein), 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), COL1A2 (Collagen Type 1 Alpha 2), beta-IG-H3 (Transforming growth factor-beta-induced protein ig-H3), CA2 (Carbonic anhydrase 2),Cathepsin B, CD63 antigen, 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), Ferritin, Furin, GM-CSF Ra (Granulocyte-macrophage 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-4 (Insulin-like growth factor binding protein-4), IGFBP- 6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, MIF (Macrophage migration inhibitory factor), 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), TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), and Transferrin. Additional non-limiting examples of the one or more proteins include: NUP85, DAPP1, PDGF R alpha, SLITRK5, FAP, Artemin, DPPII, cIAP-1, Pentraxin 3, Visfatin, Neprilysin, Albumin, Galec-tin-1, UNC5H3, IL-20 R beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7 A, NKp80, Cy statin B, Osteoadherin, Midkine, Calreticulin, Osteoactivin, Legumain, TAZ, Cathepsin L, RBP4, Serpin A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Ga-lectin-3, MOG, Trypsin 3, SIRP alpha, and Syndecan-4, GSTM1, NUP85, MeprinA, IL-1 F10, TGFbl, Ephrin-A4, Aminopeptidase LRAP, GDF-9, PDGF R alpha, PAPP-A, Aryl-sulfatase A, LAIR2, ULBP-4, TFPI, SOX2, SLITRK5, Spinesin, ENPP-2, CD97, CTACK, Integrin alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin DI, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angio-tensinogen, Follistatin, CF VII, Persephin, TRAIL Rl, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Nepri-lysin, Stabilin-2, IL-17 RD, Albumin, Folli statin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, Nor-rin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9,MMP-1, TC-PTP, IL-24, gpl30, C-myc, LILRB4, BMP-2,, MIA, CD34, IFNab R2, Glypican 2, MSP R, DSCAM, Matriptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nidogen-1, CD2, Kallikrein 1, TSLP R, TROY, VCAM-1, Siglec-11, S100A1, PARI, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R beta, Throm-bospondin-2, SMPD1, B2M, MFRP, LRP-6„ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF- 23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 alpha, 0X40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, Semaphorin 4D, ROBO2, PDX-1, APRIL, Neurturin, Kremen-2, EMMPRIN, Activin RIB, Neuroligin 2, Epiregulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF Rl, DSPG3, SorCSl, Matrilin-2, sFRP-3, EphB3, NCK1, Prolactin, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Pappalysin-2, Syndecan-3, Jag-ged 1, AKR1C4, Olfactomedin-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p27, Siglec-6, Dectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, CA125, HAO-1, PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, R0R1, CMG-2, 4-1BB Ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Leg-umain, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Con-tactin-2, CD83, IL-1 R3, SALM4, GBA3, R0B04, OSCAR, VEGF, IGSF3, Biglycan, Neudesin, ILT4, uPAR, Axl, WIF-1, IL-7 R alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Cochlin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR- Al, TAFA2, N-Cadherin, IL-17B, IL- 17 RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, Prolactin R, FGF-20, CD28, Nogo- A, HSD17B1, IL- 19, Enteropeptidase, Cathep-sin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, ADAM23, NOV, Galectin-2, Neu-rexin 3 beta, TLR3, Sirtuin 2, Numb, IL-28 R alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL- 27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR alpha- 1, Smad4, ICAM-1, MEF2C, TREM-1, L- Selectin, Hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, LTAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-lra, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4,Pepsin-ogen II, Desmoglein-3, Nectin-4, SCF, Serpin A5, PTH, FGF-19, IL-28A, FGF-12, METAP2, AS AHL, EDIL3, NT AL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, 0X40, IL- 13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, R0R2, EphB4, Glypican 1, LAP(TGFbl), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RC0R1, IL- 10 Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirrel3, FLRT1, Galec-tin-3, CXCL16, JAM- B, DR6,Nogo Receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R alpha 1, AMIG02, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurogranin, HE4, IL-23 R, Galectin-7, GALNT3, GITR L, CD14, R-Spondin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombos-pondin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulysin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin 3, ARSB, LIF R alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM- 2, Flt-3L, CDS, IL-6, CD229, Insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF-AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP-2, PU.l, ED AR, ADAMTS13, Kynureninase, PTH1R, IFN-gamma Rl, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP- 1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 Rg, MICA, Dopa Decarboxylase, NPDC-1, MCP-4, EG- VEGF, Glycoprotein V, Semaphorin 4G, IL-12p40, PSA- total, IL-15, MAP1D, Clq, TNF4, Dtk, Endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL- 8, Galectin-8, CA4, Cystatin E M, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, Kallikrein 7, and IL-13. The one or more proteins may comprise a collagen protein.

[0020] In some embodiments, the composition may comprise one or more collagen proteins. The one or more collagen proteins may comprise but are not limited to: COL1A1, COL1 A2, COL2A1, COL3A1, COL5A1, COL5A2, COL6A1, COL11A1, or COL14A1, or a combination of two or more thereof.

[0021] 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 80ng / 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.

[0022] 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.C. Pharmaceutical Applications

[0023] Disclosed herein are compositions for use in pharmaceutical applications. 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.

[0024] Compositions described herein may be administered in vivo in a pharmaceutically acceptable carrier. A pharmaceutically acceptable carrier may biologically suitable, 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 therapeutic composition in which it is contained. As a non-limiting example, the pharmaceutically acceptable carrier may comprise sodium chloride.

[0025] Compositions described herein may be administered to a patient. Administration may comprise parenteral, oral, intravenous, intramuscular, subcutaneous, inhalation, topical, sublingual, rectal, and / or transdermal administration. Compositions described herein may be administered via a variety of routes and formulations including but not limited to: oral, intravenous, intramuscular, subcutaneous, inhalation, topical, sublingual, rectal, transdermal, sublingual, buccal, intranasal, intrathecal, epidural, ocular, otic, vaginal, transbuccal, intraocular, intraperitoneal, intravesical, intradermal, intraventricular, intracardiac, intrapulmonary,sublingual tablet, oral solution, oral suspension, effervescent tablets, oral film, oral powder, chewable tablet, extended-release tablet, enteric-coated tablet, intramuscular injection, intravenous infusion, inhaler, nebulizer, transdermal patch, buccal tablet, film strip, lozenge, suppository, cream, ointment, gel, aerosol, nasal spray, enema, eye drop, ear drop, and implant. Parenteral administration of a 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 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60 minutes.

[0026] In some embodiments, the composition for administration may comprise an excipient. In some embodiments, the excipient may comprise saline. 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 (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 protein component may comprise 0.1 M to 0.8 M oligosaccharide. Non-limiting example oligosaccharides 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. In some embodiments, the composition has a pH of from about 6.0 to about 7.5. In some embodiments, the composition has a pH of 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, 7.4, or 7.5. In some embodiments, the molecular weight of any non-excipient component of the composition may be greater than about 2 kDa (kilodaltons), 3 kDa, 4 kDa, 5 kDa, 6 kDa, 7 kDa, 8 kDa, 9 kDa, 10 kDa, 20 kDa, 30 kDa, 40 kDa, 50 kDa, 60 kDa, 70 kDa, 80 kDa, 90 kDa, or 100 kDa. 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, 50kDa, or lOOkDa.

[0027] 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 freeze-dried into a powder cake and stored at room temperature. In some embodiments, the composition will have the powdered cake reconstituted with water prior to administration. In some embodiments, the composition is administered within 6 hours of thaw when maintained at ambient temperature. In some embodiments, the composition is stored in a glass vial. In some embodiments, the composition comprises a certain % of water as determined by Karl Fisher measurement. In some embodiments, the composition comprises less than 2.0%, less than 2.5%, less than 3.0%, less than 3.5%, less than 4.0%, less than 4.5%, or less than 5.0% water as determined by Karl Fisher measurement.D. Methods of Composition Production

[0028] Compositions herein comprising an EV and a protein may include components produced from a MSC. The EV of the composition may be produced from a MSC. The protein of the composition may be produced from a MSC. The EV and / or protein 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 any of these elements in various combinations. 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 protein 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 media 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%.

[0029] The EV and / or protein 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 protein into an EV and / or protein component. The EV and / or protein component may comprise 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 (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 protein component may comprise an oligosaccharide, e.g., 0.1 M to 0.8 M oligosaccharide. Non-limiting example oligosaccharides 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. The amount of oligosaccharide in the EV and / or protein component may be about 0.2 M to about 0.6 M, or about 0.4 M. In some embodiments, the amount of oligosaccharide in the compositions disclosed herein may be about 0.10 M, 0.15 M, 0.20 M, 0.25 M, 0.30 M, 0.35 M, 0.40 M, 0.45 M, 0.50 M, 0.55 M, 0.60 M, 0.65 M, or 0.70 M. In some embodiments, the EV and / or protein 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 oligosaccharide in the IV formulation may be about 40 mM to about 80 mM, or about 60 mM.

[0030] The EV and / or protein component may be filter-sterilized. The EV and / or protein component may be filter sterilized after and / or during formulation from the culture media into the EV and / or protein component. The EV and / or protein may be concentrated. The concentration may occur during and / or after purification and / or exchange from the culture media into the EV and / or protein component. The EV and / or protein may be frozen, e.g., after and / orduring purification. If frozen, the EV and / or protein may be formulated with a cryoprotectant prior to freezing. The cryoprotectant may comprises an oligosaccharide, e.g., as described above.

[0031] 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.

[0032] 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%.

[0033] 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.

[0034] 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, 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.

[0035] In some embodiments, compositions described herein can be lyophilized (freeze-dried) 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.II. Methods of Improving Perioral Area and Face Fat Graft Retention

[0036] In some embodiments, the therapeutic compositions disclosed herein are used in methods of improving perioral area and face fat graft retention in a subject who has received a face fatgraft. In some embodiments, the therapeutic compositions disclosed herein are used in methods of treating wrinkles in a subject. In some embodiments, the therapeutic compositions disclosed herein are used in methods of treating age-related facial volume loss in a subject. In some embodiments, the therapeutic compositions disclosed herein are used in methods of improving facial appearance in a subject. In some embodiments, the therapeutic compositions disclosed herein are used in methods of improving fat graft retention in a subject. Any of the therapeutic compositions described herein may be used in such a method.

[0037] In some embodiments, a subject treated with a therapeutic composition disclosed herein experiences improved graft retention and / or acceptance after administration of the therapeutic composition described herein. In some embodiments, the subject experiences an increase in facial volume. In some embodiments, the subject experiences improved skin texture, improved skin quality, improved facial aesthetics, or any combination thereof. In some embodiments, a therapeutic composition is administered to a subject. The administering may comprise one or more injections. The one or more injections may be to the dermis. In some embodiments, the subject may have periorbital or facial volume loss. In some embodiments, the subject may have skin rhytids.

[0038] In some embodiments, a therapeutic composition of the present disclosure may be administered to a subject who has received a face or perioral area fat graft. In some embodiments, the fat graft is obtained from the subject who is receiving the fat graft. In some embodiments, the fat graft is obtained from a donor who is not receiving the fat graft. In some embodiments, the fat graft is obtained using liposuction. In some embodiments, the fat graft is obtained from the abdomen, flank, outer thigh, inner thigh, lower back, and / or buttocks. In some embodiments, the fat graft is administered to the cheek, temple, under-eye area, nasolabial folds, and / or lips. In some embodiments, more than one administration of the fat graft is performed. In some embodiments, more than one administration of the therapeutic composition is performed. In some embodiments, the fat graft and therapeutic composition are administered concurrently. In some embodiments, the therapeutic composition is administered after the fat graft. In some embodiments, 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, or 3.0 mL of the fat graft is administered per area of interest. In some embodiments, 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, or 3.0 mL of the therapeutic product is administered per area of interest. In some embodiments, the fat graft is administered to the dermis. In some embodiments, the therapeutic composition is administered to the dermis. In some embodiments, the fat graft is administered to the perioral dermis. In some embodiments, the therapeutic composition is administered to the perioral dermis.

[0039] In some embodiments, administration of a therapeutic composition of the present disclosure to a subject who has received a perioral or face fat graft results in improved retention of the perioral or face fat graft. In some embodiments, the administration results in a volumetric increase of the affected perioral or face area as measured by facial imaging and volumetric quantification. In some embodiments, the volumetric increase of the perioral or face area is 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60% as compared to before treatment. In some embodiments, the volumetric increase of the perioral or face area is 0, 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, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, or 150 mm3after treatment. In some embodiments, the facial imaging comprises 3 -dimensional photographic imaging. In some embodiments, the subject exhibits improve aesthetic appearance after treatment. In some embodiments, the subject exhibits positive textural changes after treatment. In some embodiments, the subject exhibits fewer wrinkles after treatment. In some embodiments, the subject exhibits smoother skin after treatment.

[0040] Also disclosed herein are methods of improving perioral area and face fat graft retention in a subject who has received a face fat graft comprising administering to the subject a composition comprising secreted extracellular vesicles that contain a composition that includes any combination of composition proteins and / or miRNAs, selected from the following: Ferritin, NUP85, LAMP2, GPR115, Serpin Fl, OPN, PAI-1, DAPP1, Cathepsin B, Semaphorin 6C, PDGF R alpha, Sortilin, Serpin B6, Dkk-3, Thrombomodulin, PF4, MIF, Periostin, Furin, TIMP- 1, Decorin, PCK1, CD99, CD63, CD9, CD81, Transferrin, DcR3, Lumican, TIMP-2, SLITRK5, FAP, Artemin, DPPII, cIAP-1, Pentraxin 3, Visfatin, Neprilysin, Albumin, Galectin-1, UNC5H3, IL-20 R beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midkine, Calreticulin, Osteoactivin, Legumain, TAZ, Cathepsin L, RBP4, Serpin A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Tiypsin 3, SIRP alpha, and Syndecan-4, and at least one protein selected from the group consisting of: Ferritin, IGFBP-4 IL-1 R6 GSTM1, NUP85, LAMP2, MeprinA, IL-1 F10, bIG-H3, GPR115, TGFbl, Ephrin-A4, CD 109, Serpin Fl, IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF R alpha, Sortilin, Serpin B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, Thrombomodulin, Endoglycan, IGFBP-3, RGM-C, PF4,MIF, TGM4, Periostin, Furin, TIMP-1, PAPP -A, Decorin, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin DI, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP- 1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Persephin, TRAIL Rl, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, P0N1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Folli statin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, Norrin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP- 1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gpl3O, C-myc, LILRB4, BMP-2,, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matriptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nidogen-1, CD2, Kallikrein 1, TSLP R, LAMP1, TROY, VCAM-1, Siglec-11, S100A1, PARI, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM Rbeta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 alpha, 0X40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Neurturin, Kremen-2, EMMPRIN, Activin RIB, Neuroligin 2, Epiregulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF Rl, DSPG3, SorCSl, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A,NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Pappalysin-2, Syndecan-3, Jagged 1, AKR1C4, Olfactomedin-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p2'7, Siglec-6, Dectin-1, CD51, Notch- 1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, CA125, HAO-1, PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, R0R1, CMG-2, 4-1BB Ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Legumain, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 beta, TAFA1, Podocalyxin, Rai A, CRELD2, GRAP2, SP-D, BID, GFR alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Neudesin, ILT4, uPAR, Axl, WIF-1, IL-7 R alpha,GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Cochlin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR- AI, TAFA2, N-Cadherin, IL-17B, IL- 17 RC, MIP-3b, Cy statin C, Cy statin D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, Prolactin R, FGF-20, CD28, Nogo- A, HSD17B1, IL- 19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, Serpin A4, ADAM23, NOV, Galectin-2, Neurexin 3 beta, TLR3, Sirtuin 2, Numb, IL-28 R alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM- A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-Selectin, Hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-lra, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpin A5, PTH, FGF-19, IL-28 A, FGF- 12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFA5, Galectin-9, vWF-A2, TACE, Activin RUB, Cathepsin S, LDL R, BMPR-IA, 0X40, IL- 3 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDT5, R0R2, EphB4, Glypican 1, LAP(TGFbl), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL- 10 Rb, XEDAR, IL-22, PILR-alpha, NRGl-bl, FABP4, RGM-A, RELT, TrkC, C5a, SREC-I, Nestin, TPO, ErbB3, Kirrel3, FLRT1, Galectin-3, CXCL16, JAM-B, DR6,Nogo Receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R alpha 1, AMIG02, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurogranin, HE4, IL-23 R, Galectin-7, GALNT3, GITR L, CD14, R-Spondin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF RH, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospondin-5, IL-1 MI, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulysin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin 3, ARSB, LIF R alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF-AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP-2, PU.l, ED AR, ADAMTS13, Kynureninase, PTH1R, IFN-gamma Rl, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF,LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 Rg, MICA, Dopa Decarboxylase, NPDC-1, MCP-4, EG-VEGF, Glycoprotein V, Semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, Endoglin, ENA-78, Reg3A, MIP- lb, FGF-17, IL-6R, IL-8, Galectin-8, CA4, Cystatin E M, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, Kallikrein 7, IL-13, 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.

[0041] Also disclosed herein are methods of grafting microfat into a subject’s face or perioral area. In some embodiments, the method comprises injecting a microfat graft into the dermis of the subject’s face or perioral area. In some embodiments, the method further comprises injecting a therapeutic bone marrow-derived MSC secretome composition of the present disclosure into the dermis. In some cases, the therapeutic bone marrow-derived MSC secretome composition is injected at or near the location of the microfat graft. In some embodiments, the microfat graft and MSC secretome composition are injected concurrently. In some embodiments, the MSC secretome composition is injected after the microfat graft. In some embodiments, the MSC secretome is injected before the microfat graft. In some embodiments, the MSC secretome composition is injected concurrently with the microfat graft. In some embodiments, the microfat graft is injected into the upper or lower lip area. In some embodiments, the MSC secretomecomposition is injected at a volume of 0.5 mL, 1.0 mL, 1.5 mL, 2.0 mL, 2.5 mL, 3.0 mL or any amount in between. In some embodiments, the microfat graft is injected at a volume of 0.5 mL, 1.0 mL, 1.5 mL, 2.0 mL, 2.5 mL, 3.0 mL or any amount in between. In some embodiments, the subject receives multiple microfat graft injections on the same day. In some embodiments, the subject receives microfat graft injections on one or more days. In some embodiments, the MSC secretome composition is injected 1 or more times. In some embodiments, the MSC secretome composition is injected several times over the course of 1, 2, 3, 4, 5, or 6 months. In some embodiments, the MSC secretome composition is collected from MSC conditioned media of bone marrow-derived MSCs cultured under conditions comprising oxygen tension below 5% and culture media having a pH below 7. The MSC secretome composition may comprise proteins and extracellular vesicles produced by the bone-marrow derived MSCs. The dosage of each injection of the MSC secretome composition administered to the subject may be a cellequivalent dosage of 0.7 to 7 million cells / kg. The culture media may be serum-free. In some embodiments, at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63+CD9 CD81". In some embodiments, the MSC secretome composition comprises one or more of the following proteins: Ferritin, NUP85, LAMP2, GPR115, Serpin Fl, OPN, PAI-1, DAPP1, Cathepsin B, Semaphorin 6C, PDGF R alpha, Sortilin, Serpin B6, Dkk-3, Thrombomodulin, PF4, MIF, Periostin, Furin, TIMP-1, Decorin, PCK1, CD99, CD63, CD9, CD81, Transferrin, DcR3, Lumican, TIMP-2, SLITRK5, FAP, Artemin, DPPII, cIAP-1, Pentraxin 3, Visfatin, Neprilysin, Albumin, Galectin-1, UNC5H3, IL-20 R beta, SREC-II, JAM- C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7 A, NKp80, Cy statin B, Osteoadherin, Midkine, Calreticulin, Osteoactivin, Legumain, TAZ, Cathepsin L, RBP4, Serpin A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin 3, SIRP alpha, and Syndecan-4, and at least one protein selected from the following: Ferritin, IGFBP-4 IL-1 R6 GSTM1, NUP85, LAMP2, MeprinA, IL-1 F10, bIG-H3, GPR115, TGFbl, Ephrin-A4, CD109, Serpin Fl, IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF R alpha, Sortilin, Serpin B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, Thrombomodulin, Endoglycan, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periostin, Furin, TIMP-1, PAPP- A, Decorin, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin alpha 1, EXTL3, IL- 18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin DI, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Persephin, TRAIL Rl, THAP11, CD200,CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Folli statin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, Norrin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gpl3O, C-myc, LILRB4, BMP-2,, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matriptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nidogen-1, CD2, Kallikrein 1, TSLP R, LAMP1, TROY, VCAM-1, Siglec-11, S100A1, PARI, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 alpha, 0X40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, S0X9, ST6GAL1, MEP1B, CD99- L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Neurturin, Kremen-2, EMMPRIN, Activin RIB, Neuroligin 2, Epiregulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF RI, DSPG3, SorCSl, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A,NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Pappalysin-2, Syndecan-3, Jagged 1, AKR1C4, Olfactomedin-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p2'7, Siglec-6, Dectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, CA125, HAO-1, PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, R0R1, CMG-2, 4-1BB Ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Legumain, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Neudesin, ILT4, uPAR, Axl, WIF-1, IL-7 R alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Cochlin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL- 17 RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, Serpin A4, ADAM23, NOV, Galectin-2, Neurexin 3 beta, TLR3, Sirtuin 2, Numb, IL-28 R alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin,Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, 01ig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR alpha- 1, Smad4, ICAM-1, MEF2C, TREM-1, L-Selectin, Hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL- 31 RA, Arginase 1, POGLUT1, IL-lra, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP- 2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmogl ein-3, Nectin-4, SCF, Serpin A5, PTH, FGF-19, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NT AL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, 0X40, IL- 13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, R0R2, EphB4, Glypican 1, LAP(TGFbl), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirrel3, FLRT1, Galectin-3, CXCL16, JAM-B, DR6, Nogo Receptor, TLR4, VEGF R2, Tie-2, IL- 15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R alpha 1, AMIG02, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurogranin, HE4, IL-23 R, Galectin-7, GALNT3, GITRL, CD14, R-Spondin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospondin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulysin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin 3, ARSB, LIF R alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF-AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP -2, PU.l, ED AR, ADAMTS13, Kynureninase, PTH1R, IFN-gamma Rl, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP- 10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 Rg, MICA, Dopa Decarboxylase, NPDC-1, MCP-4, EG- VEGF, Glycoprotein V, Semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, Endoglin, ENA-78, Reg3 A, MIP-lb, FGF-17, IL-6R, IL-8, Galectin-8, CA4, Cystatin E M, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, Kallikrein 7, and IL-1, or any combination thereof. In some embodiments, the MSC secretome composition comprises one or more of the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-lOb, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR- 1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-13Oa-3p, hsa-mir-13Ob, hsa-miR-13Ob-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-3Oa-5p, hsa-miR-3Oa-5p, hsa-miR-3Ob-5p, hsa-miR-3Oc-5p, hsa-mir- 3Od, hsa-miR-3Od-5p, hsa-mir-3Oe, hsa-miR-3Oe-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, or any combination thereof.

[0042] In some embodiments, the therapeutic product of the present disclosure is administered at a cell-equivalent dose range of 0.7 to 7 million cells / kg. In some embodiments, the therapeutic product is administered at a cell-equivalent dose of at least about, at most about, or about 0.2, 0.5, 0.7, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, or 12.0 million cells / kg, or a range between any two of these values. In some embodiments, the product is administered at a dose that provides 9xlOnto 1.2xl012extracellular vesicles or 5xl0nto 1.5xl012, 6xlOnto 1.4xl012, 7xlOnto 1.3xl012, 8xl0nto 1.2xl012, or 8xl0nto 1.3xl012extracellular vesicles. In some embodiments, the product is administered at a dose that provides at least or at most 5xl0n, 6xlOn, 7xlOn, 8xl0n, 9xlOn, IxlO12, l.lxlO12, 1.2xl012, 1.3xl012, 1.4xl012, or 1.5xl012extracellular vesicles. In some embodiments, the therapeutic product comprises 6xlO10to 8xl010, 5xl010to 9xlO10, 4xlO10to 10xl010, 5.5xl010to 8.5xl010, 6xlO10to 8.5xl010cells / ml. In some embodiments, the therapeutic product comprises 6xlO10to 8xl010extracellular vesicles per ml and is administered at a dose of 10 to 20 ml. In some embodiments, the therapeutic product comprises 6xlO10to 8xl010extracellular vesicles per ml and is administered at a dose of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml, or a range between any two of these values. In some embodiments, the therapeutic product is safe toadminister at a dosage of 5 million cells / kg. In some embodiments, the therapeutic product dosage ceiling is 10 million cells / kg.III. Examples

[0043] 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.A. Example 1 - Production of Therapeutic Composition

[0044] An MSC secretome therapeutic composition (referred to herein as investigational product or “IP” as well as investigational medicinal product or “IMP”) was made by the following method: human bone marrow-derived 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 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.

[0045] 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.

[0046] The IMP comprises extracellular vesicles (EVs) that are acellular and nonimmunogenic, containing no nucleus or deoxyribonucleic acid (DNA).

[0047] 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.

[0048] 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 orNanoview / 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 1 below.Table 1

[0049] The nucleic acid content of the IMP was determined, and the following nucleic acids were found to be present: hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, let-7, 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, 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.

[0050] Additional proteins that may be present in the IMP include: NUP85, DAPP1, PDGF R alpha, SLITRK5, FAP, Artemin, DPPII, cIAP-1, Pentraxin 3, Visfatin, Neprilysin, Albumin, Galectin-1, UNC5H3, IL-20 R beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midkine, Calreticulin, Osteoactivin,Legumain, TAZ, Cathepsin L, RBP4, Serpin A4, JAM- A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin 3, SIRP alpha, and Syndecan-4, GSTM1, NUP85, MeprinA, IL-1 F10, TGFbl, Ephrin-A4, Aminopeptidase LRAP, GDF-9, PDGF R alpha, PAPP -A, Arylsulfatase A, LAIR2, ULBP-4, TFPI, SOX2, SLITRK5, Spinesin, ENPP-2, CD97, CTACK, Integrin alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF- 2R, ADAMTSL-1, Erythropoietin, Plexin DI, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Persephin, TRAIL Rl, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, Norrin, JAM-C, Bcl- 10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gpl30, C- myc, LILRB4, BMP-2,, MIA, CD34, IFNab R2, Glypican 2, MSP R, DSCAM, Matriptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nidogen-1, CD2, Kallikrein 1, TSLP R, TROY, VCAM-1, Siglec-11, S100A1, PARI, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6„ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD- ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 alpha, 0X40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, Semaphorin 4D, ROBO2, PDX-1, APRIL, Neurturin, Kremen-2, EMMPRIN, Activin RIB, Neuroligin 2, Epiregulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF Rl, DSPG3, SorCSl, Matrilin-2, sFRP-3, EphB3, NCK1, Prolactin, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Pappalysin-2, Syndecan-3, Jagged 1, AKR1C4, Olfactomedin-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p27, Siglec-6, Dectin- 1, CD51, Notch- 1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, CA125, HAO-1, PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, R0R1, CMG-2, 4-1BB Ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Legumain, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 beta, TAFA1, Podocalyxin, Rai A, CRELD2, GRAP2, SP-D, BID, GFR alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan,Neudesin, ILT4, uPAR, Axl, WIF-1, IL-7 R alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Cochlin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL- 17 RC, MIP-3b, Cy statin C, Cy statin D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, ADAM23, NOV, Galectin-2, Neurexin 3 beta, TLR3, Sirtuin 2, Numb, IL-28 R alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM- A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA I3, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-Selectin, Hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, LTAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-lra, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpin A5, PTH, FGF-19, IL-28 A, FGF-12, METAP2, AS AHL, EDIL3, NT AL, EGF R, TAFAS, Galectin- 9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, 0X40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, R0R2, EphB4, Glypican 1, LAP(TGFbl), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL- 10 Rb, XEDAR, IL-22, PILR-alpha, NRG1- 131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirrel3, FLRT1, Galectin-3, CXCL16, JAM-B, DR6,Nogo Receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R alpha 1, AMIG02, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, Neurogranin, HE4, IL-23 R, Galectin-7, GALNT3, GITR L, CD14, R- Spondin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospondin-5, IL- 1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulysin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin 3, ARSB, LIF R alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF-AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP- 2, PU.l, ED AR, ADAMTS13, Kynureninase, PTH1R, IFN-gamma Rl, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dectin-2, EpCAM, HCC-4,TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 Rg, MICA, Dopa Decarboxylase, NPDC-1, MCP-4, EG-VEGF, Glycoprotein V, Semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, Endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, Galectin-8, CA4, Cystatin E M, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, Kallikrein 7, and IL-13, and combinations of two or more thereof.B. Example 2 - Prospective Study of the Use of Bone Marrow Mesenchymal Stem Cell Derived Extracellular Vesicles to Improve Perioral Area and Face Fat Graft Retention

[0051] In this Example, the human bone marrow-derived extracellular vesicle (hBM-MSC EV) therapeutic product of Example 1 above was evaluated for safety and efficacy in 10 subjects undergoing perioral microfat grafting using injection of the therapeutic product of the present disclosure into the graft. Graft retention was improved as evidenced by an average 43% volume increase over nine months in subjects evaluated by facial imaging and volumetric quantification. Subjective assessment revealed improvement in the skin texture and quality.

[0052] The therapeutic product used in the study is a consistent biologic drug candidate preparation manufactured under CGMP processes per FDA guidance and a Quality Management System, with USP<71> sterility assurance, lot-specific master batch records and specified lot release criteria including advanced particle analysis, surface protein expression and multiple quantitative bioactivity assays. The purpose of the study was to evaluate the safety of combining the therapeutic product with mini fat grafting into the dermis of the perioral area and face. The study also evaluated the improvement of aesthetics with improved graft retention. The study format was a prospective non-randomized study and was conducted at a single site.

[0053] Materials and Methods: The primary aim of this study was to evaluate the safety of utilizing a single 2 mL injection of the therapeutic product after a mini fat graft injection into the dermis under local anesthesia of 10 female subjects, 30 to 70 years of age, who had clinically significant peri-orbital or facial volumetric tissue loss with skin rhytids. The secondary aim was to probe the potential for efficacy of the composition of the present disclosure in the same subjects. Written informed consent was obtained from all subjects. The protocol was approved by the American Academy of Stem Cell Physicians Institutional Review Board (IRB Approval Number: IRCM-2021-284).

[0054] The proposed study was a single site and single investigator study. The Inclusion and Exclusion criteria are listed below and the schedule of events is described as follows. Informed consent was acquired at baseline. Baseline was established prior to treatment. A medical history and list of current medications was acquired at baseline and at 9 months after initial treatment (±1 month). A physical exam was conducted at baseline. Subjects who met the Inclusion Criteria were enrolled before the start of initial treatment. Objective data was collected at baseline and at 3, 6, and 9 months after initial treatment (± 1 month). Assessment of any events observed over the course of the study was conducted at the time of initial treatment as well as at 2, 3, and 6 weeks after the start of initial treatment (± 1 week) and at 3, 6, and 9 months after the start of initial treatment (± 1 month).INCLUSION CRITERIA

[0055] All candidates for this study met ALL of the following Inclusion Criteria to be eligible for enrollment:1. Voluntary signature of the approved Informed Consent2. Females 30 to 50 years old3. Females with clinical volumetric peri-oral volume loss with peri-oral skin rhytids4. Agree to NOT become pregnant during the StudyEXCLUSION CRITERIA

[0056] Candidates who met ANY of the following Exclusion Criteria at the time of the study procedure were not eligible for enrollment in the study:1. The subj ect is unable to conform to the study protocol follow-up procedures and visits.2. The subject has major risk factors such as a history of narcotic abuse, paucity of family support, unemployed, history of previous physical or mental abuse or severe medical comorbidities.3. Subjects with any auto-immune disorder.

[0057] The 10 subjects received a mini fat graft placed into the abnormal peri-oral dermis under local anesthesia. The fat preparation was obtained using kits from Tulip Medical (San Diego, CA) as per manufacturer’s instructions. The initial volume of micro-fat injected was patient specific and carefully documented. After the fat grafting procedure 2 mL of the therapeutic product was injected into the graft area. Each patient was monitored for a total of nine months.

[0058] Safety assessment was based upon evaluation of all events reported by the patient. Primary outcome measurements included a daily monitoring by the subject and documentation of any and all events observed during the 2 mL injection of therapeutic product and at all follow up visits specified in Table 1, whether directly or indirectly associated with the therapeutic product treatment. The clinical and objective changes were captured and evaluated with the Canfield Vectra H2 imaging system and volume analysis software to provide objective volume changes. All data obtained at three-, six- and nine-month follow-up were compared to the pretreatment values.

[0059] Results: To quantify retention of the microfat graft, initial 3 -dimensional photographic data was acquired by the Canfield, Vectra system prior to treatment and then again nine months later. Subjects showed visible improvement in skin texture and overall skin quality. Subjects reported they experienced a positive outcome and that they would repeat the procedure at a later date, if needed. Subjects received 1.5 mL of microfat in each area of interest (AO I). Example images of subjects obtained at baseline and again at nine months after treatment are shown in Figure 1. The region treated, quantified volume increase, and the percent volume increase as compared to baseline Vectra measurements are summarized in Table 2 below. All three subjects exhibited volume increases in both sides of the perioral areas treated, and the average volume increase across the six AOIs was 43.2%. The careful volume changes improved overall aesthetic appearance with the added benefit of positive textural changes.

[0060] Table 2: Three-dimensional analysis of microfat graft retention over nine months.

[0061] Conclusion: The results of this study indicate that the therapeutic treatment is safe for use in combination with microfat when administered by injection into the perioral area. The therapeutic treatment is efficacious for the use of facial rejuvenation. In subjects evaluated using three-dimensional capture of facial images, there was a perceptible change in both the retained volume (43.2% increase vs baseline) and a subjectively perceived improvement in skin quality.

[0062] The high mobility of the perioral and face area causes grafts to be absorbed over time. In practice, microfat volume retention in the perioral area is very low and the fat graft procedure is primarily expected to improve skin texture and appearance. Thus, this aesthetically important area has been over-treated with grafting in the past in an effort to increase graft acceptance and retention. In turn, this can cause an extremely poor cosmetic appearance. The results of this study indicate that an average 43.2% improvement in micro graft acceptance can be expected, clearly more than what has been previously observed. The success of the application of the therapeutic product of Example 1 in graft retention is surprising and unexpected. This study demonstrates that the CGMP BM-MSC EV product of the present disclosure safely improves microfat graft retention. This study also represents a biopharmaceutical quality product that is consistent, standardized and quality tested regarding dose and biological activity and offers ameans to reduce one source of variability (i.e., inconsistent biological material preparation) in the success rate of perioral graft success.

[0063] While some embodiments have been shown and described herein, such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the disclosure provided herein. It should be understood that various alternatives to the embodiments described herein can be employed.

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A method of improving perioral face fat graft retention in a subject, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the composition comprises NUP85, DAPP1, PDGF R alpha, SLITRK5, FAP, Artemin, DPPII, cIAP-1, Pentraxin 3, Visfatin, Neprilysin, Albumin, Galectin-1, UNC5H3, IL-20 R beta, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 alpha, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midkine, Calreticulin, Osteoactivin, Legumain, TAZ, Cathepsin L, RBP4, Serpin A4, JAM- A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin 3, SIRP alpha, and Syndecan-4, GSTM1, NUP85, MeprinA, IL-1 F10, TGFbl, Ephrin-A4, Aminopeptidase LRAP, GDF-9, PDGF R alpha, PAPP -A, Arylsulfatase A, LAIR2, ULBP-4, TFPI, SOX2, SLITRK5, Spinesin, ENPP-2, CD97, CTACK, Integrin alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin DI, DNMT3A, BCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Persephin, TRAIL Rl, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, P0N1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Folli statin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC- II, Norrin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gpl30, C-myc, LILRB4, BMP-2, MIA, CD34, IFNab R2, Glypican 2, MSP R, DSCAM, Matriptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nidogen-1, CD2, Kallikrein 1, TSLP R, TROY, VCAM-1, Siglec- 11, S100A1, PARI, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6„ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 alpha, 0X40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, Semaphorin 4D, ROBO2, PDX-1, APRIL, Neurturin, Kremen-2, EMMPRIN, Activin RIB, Neuroligin 2, Epiregulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF Rl, DSPG3, SorCSl, Matrilin-2, sFRP-3, EphB3, NCK1, Prolactin, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, Pappalysin-2, Syndecan-3,Jagged 1, AKR1C4, Olfactomedin-2, Osteoadherin, NKp44, Thyroglobulin, IL-21R, Chemerin, EphAl, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H, PRX2, p27, Siglec- 6, Dectin- 1, CD51, Notch- 1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, CA125, HAO-1, PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, R0R1, CMG-2, 4-1BB Ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5, CD6, Siglec-2, Legumain, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 beta, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR alpha-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Cathepsin L, 6Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1 R3, SALM4, GBA3, R0B04, OSCAR, VEGF, IGSF3, Biglycan, Neudesin, ILT4, uPAR, Axl, WIF-1, IL-7 R alpha, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Cochlin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen- 2, IL-17F, SR-AI, TAFA2, N-Cadherin, IL-17B, IL-17 RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, Prolactin R, FGF-20, CD28, Nogo- A, HSD17B1, IL-19, Enteropeptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, ADAM23, NOV, Galectin-2, Neurexin 3 beta, TLR3, Sirtuin 2, Numb, IL-28 R alpha, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA I3, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR alpha-1, Smad4, ICAM-1, MEF2C, TREM-1, L-Selectin, Hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-lra, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LRRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin- 4, SCF, Serpin A5, PTH, FGF-19, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NT AL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR- IA, 0X40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, R0R2, EphB4, Glypican 1, LAP(TGFbl), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirrel3, FLRT1, Galectin-3, CXCL16, JAM-B, DR6, Nogo Receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R alpha 1, AMIG02, HCC-1, TFPI-2, ULBP-2, Desm oglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2,Neurogranin, HE4, IL-23 R, Galectin-7, GALNT3, GITR L, CD14, R-Spondin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-la, PDGF-AB, GPVI, CD40, SCF R, Thrombospondin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulysin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin 3, ARSB, LIF R alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF-AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP -2, PU.1, ED AR, ADAMTS13, Kynureninase, PTH1R, IFN-gamma Rl, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP- 10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 Rg, MICA, Dopa Decarboxylase, NPDC-1, MCP-4, EG- VEGF, Glycoprotein V, Semaphorin 4G, IL-12p40, PSA-total, IL-15, MAP1D, Clq, TNF4, Dtk, Endoglin, ENA-78, Reg3A, MIP-lb, FGF-17, IL-6R, IL-8, Galectin-8, CA4, Cystatin E M, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, Kallikrein 7, or IL-13, or a combination of two or more thereof.

2. A method of improving perioral face fat graft retention in a subject, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the composition comprises TIMP-1 (tissue inhibitor of metalloproteinases 1), TEMP -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.

3. The method of claim 2, 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, 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 (Granulocyte-macrophage 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 (Phosphoenol pyruvate 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, C0L1A1 (Collagen Type 1 Alpha 1), C0L1A2 (Collagen Type 1 Alpha 2), or Transferrin, or a combination of two or more thereof.

4. The method of claim 2 or 3, wherein the composition further comprises one or more of C0L2A1, C0L3A1, C0L5A1, COL5A2, C0L6A1, C0L11A1, or C0L14A1, or a combination of two or more thereof.

5. A method of improving perioral face fat graft retention 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.

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. A method of improving perioral face fat graft retention in a subject, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein at least 80% of the one or more EVs are CD63+, CD9-, CD81-.

8. The method of any one of claims 1-7, wherein the perioral face fat graft is administered to the cheek, temple, under-eye area, nasolabial folds, lips, or any combination thereof.

9. The method of any one of claims 1-8, wherein the perioral face fat graft is obtained from the abdomen, flank, outer thigh, inner thigh, lower back, and / or buttocks.

10. The method of any one of claims 1-9, wherein the subject exhibits improved skin texture, improved skin quality, improve facial aesthetics, and / or any combination thereof after the administering.

11. The method of any one of claims 1-10, wherein the subject experiences a volumetric increase of a perioral or face area of 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, or 60% after the administering.

12. A method of treating wrinkles in a subject, the method comprising administering to the subject a composition comprising a mesenchymal stem cell (MSC) secretome.

13. A method of treating age-related facial volume loss in a subject, the method comprising administering to the subject a composition comprising a mesenchymal stem cell (MSC) secretome.

14. A method of improving facial appearance in a subject, the method comprising administering to the subject a composition comprising a mesenchymal stem cell (MSC) secretome.

15. A method of improving fat graft retention in a subject, the method comprising administering to the subject a composition comprising a mesenchymal stem cell (MSC) secretome.

16. The method of any one of claims 1-15, wherein the subject exhibits an increase of 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, 100, 101, 102, 103, 104, 105,106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, or 130 mm3after administration of the composition.

17. The method of any one of claims 1-16, wherein the subject has skin rhytids.

18. The method of any one of claims 1-17, wherein the subject has periorbital or facial volume loss.

19. The method of any one of claims 1-18, wherein the administering comprises administering 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, or 3.0 mL of the composition.

20. The method of claim 19, wherein the composition is administered after administration of a fat graft.

21. The method of claim 20, wherein 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, or 3.0 mL of the fat graft is administered.

22. The method of claims 21, wherein the fat graft is administered to more than one area of the face.

23. The method of claim 22, wherein the fat graft is administered to a left side and a right side of the face.

24. The method of any one of claims 20-23, wherein the fat graft is administered to the dermis.

25. The method of claim 24, wherein the fat graft is administered to the perioral dermis.

26. The method of any one of claims 1-25, wherein the composition is prepared by a process comprising: (a) culturing bone marrow mesenchymal stem cells (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 composition.

27. The method of any one of claims 1-25, the method comprising preparing the composition prior to the administering, wherein the preparing the composition occurs by a process comprising: (a) culturing bone marrow mesenchymal stem cells (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 composition.

28. The method of claim 26 or 27, wherein the culture media is serum-free.

29. The method of any one of claims 26-28, wherein the culture media has a glucose concentration below 4.5 g / L.

30. The method of any one of claims 26-29, wherein formulating the MSC conditioned media comprises exchanging the conditioned media for a pharmaceutically acceptable formulation.

31. The method of claim 30, wherein the pharmaceutically acceptable formulation comprises saline.

32. The method of any one of claims 1-31, wherein the composition comprises at least 6xlO10to 8xlO10extracellular vesicles per mL and is administered at a dose of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml.

33. The method of claim 32, wherein the dose is administered in combination with normal saline at a final volume of 100 mL.

34. The method of any one of claims 1-33, wherein the composition comprises saline (0.9% sodium chloride).

35. The method of claim 34, wherein the saline is present in the composition at about 80% to about 95% saline.

36. The method of any one of claims 1-35, wherein the composition comprises sodium chloride, sodium lactate, potassium chloride, and / or calcium chloride.

37. The method of any one of claims 1-36, wherein the molecular weight of any non-excipient component of the composition is greater than about 2 kDa (kilodaltons), 8 kDa, 10 kDa, 50kDa, or lOOkDa.

38. The method of any one of claims 1-37, wherein the composition comprises an oligosaccharide.

39. The method of claim 38, wherein the oligosaccharide is present in the composition at from about 0.2 M to about 0.6 M.

40. The method of any one of claims 1-39, wherein any non-excipient component of the composition has a size of less than about 0.2 microns.

41. The method of any one of claims 1-40, wherein the composition is sterile per USP <71>.

42. The method of any one of claims 1-41, wherein the composition is endotoxin free per USP <85>.

43. The method of any one of claims 1-42, wherein the composition is negative for mycoplasma DNA.

44. The method of any one of claims 1-43, wherein the composition is cell-free.

45. The method of any one of claims 1-44, wherein the composition is stored between -80 °C and -60 °C.

46. The method of claim 45, wherein the composition is administered within 6 hours of thaw when maintained at ambient temperature.

47. The method of any one of claims 1-46, wherein the composition is freeze-dried (lyophilized) to a powder cake and stored at or below room temperature.

48. The method of any one of claims 1-47, wherein the composition is reconstituted with water prior to administration.

49. The method of any one of claims 1-48, wherein the composition is present in a glass vial.

50. The method of any one of claims 1-49, wherein the composition is formulated for intravenous administration.

51. The method of any one of claims 1-50, wherein the composition has a pH of about 6 to about 7.5.

52. The method of any one of claims 26-51, wherein the BM-MSCs are negative for CD14, CD31, CD34, and CD45.

53. The method of any one of claims 26-52, wherein the BM-MSCs are positive for CD73, CD 105, CD 166, and CD90.

54. The method of any one of claims 26-53, wherein the BM-MSCs are capable of undergoing trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes.

55. The method of any one of claims 26-54, wherein the BM-MSCs are obtained from an iliac crest aspiration of a single donor.

56. The method of any one of claims 1-55, wherein a concentration of the one or more EVs in the composition is measured by nanoparticle tracking analysis (NTA).

57. The method of claim 56, wherein the NTA comprises light scatter and fluorescence evaluation.

58. The method of claim 56 or 57, wherein the concentration of the one or more EVs is about 10 billion to about 250 billion EVs per mL of the composition.

59. The method of any one of claims 56-58, wherein the concentration of the one or more EVs is at about 1 billion to about 40 billion EVs per ml of the composition.

60. The method of any one of claims 1-59, wherein the one or more EVs have an average diameter of about 30 nm to about 170 nm.

61. The method of any one of claims 1-60, wherein a total protein concentration of the composition is about 10 to about 40 pg per ml of the composition.

62. The method of any one of claims 1-61, wherein a total protein concentration of the composition is about 1.5 to about 6 pg per ml of the composition.

63. The method of claim 61 or 62, wherein the total protein concentration is measured by ELISA.

64. The method of any one of claims 4-63, wherein the composition comprises 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 combination of two or more thereof.

65. The method of any one of claims 4-64, 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, 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 (Granulocyte-macrophage 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 (Phosphoenol pyruvate 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), COL1A2 (Collagen Type 1 Alpha 2), or Transferrin, or a combination of two or more thereof.

66. The method of any one of claims 1 or 5-65, wherein the composition further comprises one or more of COL2A1, COL3A1, COL5A1, COL5A2, COL6A1, COL11A1, or COL14A1, or a combination of two or more thereof.

67. The method of any one of claims 1-4 or 7-66, 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.

68. The method of any one of claims 1-4 or 7-67, 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.

69. The method of any one of claims 1-6 or 8-68, wherein at least 80% of the one or more EVs are CD63+, CD9-, CD81-.

70. A method of grafting microfat into a subject’s face or perioral area, the method comprising:(a) injecting a fat graft into the dermis of the subject’s face or perioral area; and(b) injecting a therapeutic bone marrow-derived MSC secretome composition into the dermis at or near the location of the microfat graft.

71. The method of claim 70, wherein (b) is carried out before, after, or concurrently with (a).

72. The method of claim 70 or 71, wherein the fat graft is injected into the upper or lower lip area.

73. The method of any one of claims 70-72, wherein the fat graft is injected at a volume of 0.5 mL, 1.0 mL, 1.5 mL, 2.0 mL, 2.5 mL, or 3.0 mL.

74. The method of any one of claims 70-73, wherein the subject receives multiple fat graft injections.

75. The method of any one of claims 70-74, wherein the MSC secretome composition is injected at a volume of 0.5 mL, 1.0 mL, 1.5 mL, 2.0 mL, 2.5 mL, or 3.0 mL.

76. The method of any one of claims 70-75, wherein the MSC secretome composition is collected from MSC conditioned media of bone marrow-derived MSCs cultured under conditions comprising oxygen tension below 5% and culture media having a pH below 7.

77. Use of a composition comprising a mesenchymal stem cell (MSC) secretome in improving retention of a perioral face fat graft.

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