Treatment of traumatic brain injury with extracellular vesicle composition

Abstract

Disclosed are methods of treating traumatic brain injury 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

Attorney Docket No.66309-729.601 TREATMENT OF TRAUMATIC BRAIN INJURY WITH EXTRACELLULAR VESICLE COMPOSITION CROSS REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 638,077 filedon April 24, 2024, the entirety of which is hereby incorporated by reference herein. BACKGROUND

[0002] Traumatic brain injury (TBI) outcomes may vary depending on injury severity andtreatment provided. Following traumatic tissue damage, a chronic response characterized by hypoxia, inflammation, free radical generation, apoptosis, and diffuse axonal injury contributes to sustained TBI deficits. These deficits include impaired memory and attention, changes in executive function, emotional instability, and sensorimotor deficits. Routine therapies have yielded variable results. There exists a need for a safe and effective treatment for TBI. SUMMARY

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

[0004] Disclosed herein are methods and compositions for treating traumatic brain injury or asymptom thereof in a subject. Disclosed herein is the use of a composition comprising a mesenchymal stem cell (MSC) secretome in treating traumatic brain injury. Disclosed herein are methods and compositions for treating a head injury or a symptom thereof in a subject. Disclosed herein are methods and compositions for treating a cognitive impairment or a symptom thereof in a subject. Disclosed herein are methods and compositions for improving independence in a subject. Disclosed herein are methods and compositions for improving self- care, sphincter control, mobility, locomotion, communication, psychosocial adjustment, and / or cognitive function 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, Semaphorin 7A, NKp80, Cystatin B, Osteoadherin, Midkine, Calreticulin, Osteoactivin, Legumain, TAZ, Cathepsin L, RBP4, SerpinAttorney Docket No.66309-729.601 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 D1, 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 R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP- 1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, 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, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6„ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF- 23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2,Insulin R, HIF-1 alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, Semaphorin 4D, ROBO2, PDX-1, APRIL, Neurturin, Kremen-2, EMMPRIN, Activin RIB, Neuroligin 2, Epiregulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, 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, ROR1, 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,Attorney Docket No.66309-729.601 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, 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-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP(TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirre13, FLRT1, Galectin-3, CXCL16, JAM- B, DR6,Nogo Receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R alpha 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, 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, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-gamma R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10,Attorney Docket No.66309-729.601 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), 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, CD109 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 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (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. In some embodiments, the composition further comprises one or more of COL2A1, COL3A1, COL5A1, COL5A2, COL6A1, COL11A1, or COL14A1, 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 compositionAttorney Docket No.66309-729.601 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 traumatic brain injury is acute. In some embodiments, the traumatic brain injury is chronic. In some embodiments, the subject exhibits impaired self- care, sphincter control, mobility, locomotion, communication, psychosocial adjustment, and / or cognitive function. In some embodiments, the subject exhibits one or more of headache, dizziness, nausea, confusion, memory problems, difficulty concentrating, blurred vision, sensitivity to light or noise, sleep disturbances, mood swings, irritability, fatigue, loss of balance, ringing in the ears, speech difficulties, anxiety, depression, seizures, loss of coordination, changes in taste or smell, blurred vision, slurred speech, weakness, numbness, trouble finding words, trouble swallowing, difficulty with problem-solving, changes in personality, impulsivity, sensitivity to touch, difficulty multitasking, inability to focus, coordination issues, reduced concentration, poor judgment, hyperactivity, decreased attention span, apathy, headaches that worsen over time, and / or an inability to perform routine tasks. In some embodiments, the subject exhibits an improvement of at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% in Functional Independent Measure (FIM) score, Functional Assessment Measure (FAM) score, and / or FIM+FAM score after administration of the composition. In some embodiments, the subject has a traumatic head injury. In some embodiments, the subject has difficulty ambulating. In some embodiments, the administering comprises administering 15 mL of the composition. In some embodiments, the composition is administered with 85 mL of saline. In some embodiments, three doses of the composition are administered per month for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In some embodiments, the composition is administered during non-consecutive months. In some embodiments, the three doses are administered over the course of one week. In some embodiments, each dose of the three doses is administered within 48 hours of another dose. In some embodiments, the second dose is administered 48 hours after administration of the first dose. In some embodiments, the third dose is administered 48 hours after administration of the second dose. In some embodiments, the composition is administered at least once monthly over the course of 36 weeks. In some embodiments, the administering comprises intravenousAttorney Docket No.66309-729.601 administration. In some embodiments, the intravenous administration is carried out over the course of 30, 35, 40, 45, 50, 55, or 60 minutes. 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 6x1010to 8x1010extracellular 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 chloride. In some embodiments, the molecular weight of any non- excipient component of the composition is greater than about 2 kDa (kilodaltons), 8 kDa, 10 kDa, 50kDa, or 100kDa. 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 isAttorney Docket No.66309-729.601 present in a glass vial. In some embodiments, the composition is formulated for intravenous administration. In some embodiments, the composition has a pH of about 6 to about 7.5. In some embodiments, the BM-MSCs are negative for CD14, CD31, CD34, and CD45. In some embodiments, the BM-MSCs are positive for CD73, CD105, CD166, and CD90. In some embodiments, the BM-MSCs are capable of undergoing trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes. In some embodiments, the BM-MSCs are obtained from an iliac crest aspiration of a single donor. In some embodiments, a concentration of the one or more EVs in the composition is measured by nanoparticle tracking analysis (NTA). In some embodiments, the NTA comprises light scatter and fluorescence evaluation. In some embodiments, the concentration of the one or more EVs is about 10 billion to about 250 billion EVs per mL of the composition. In some embodiments, the concentration of the one or more EVs is at about 1 billion to about 40 billion EVs per ml of the composition. In some embodiments, the one or more EVs have an average diameter of about 30 nm to about 170 nm. In some embodiments, a total protein concentration of the composition is about 10 to about 40 µg per ml of the composition. In some embodiments, a total protein concentration of the composition is about 1.5 to about 6 µg 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, CD109 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 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4),Attorney Docket No.66309-729.601 Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (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. In some embodiments, the composition further comprises one or more of COL2A1, COL3A1, COL5A1, COL5A2, COL6A1, COL11A1, 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-. INCORPORATION BY REFERENCE

[0005] Each patent, publication, and non-patent literature cited in the application is herebyincorporated 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

[0006] The features of the present disclosure are set forth with particularity in the appendedclaims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings (also “Figure” and “FIG.” herein), of which:

[0007] FIG. 1 is a graph showing Functional Independence Measure (FIM), FunctionalAssessment Measure (FAM) and FIM+FAM summed scores over 36 weeks. Treatments were administered over a 36-week period. FIM and FAM instruments were administered weekly and the individual weekly FIM, FAM, and FIM+FAM summed scores were plotted vs protocol week (WK).Attorney Docket No.66309-729.601

[0008] FIG. 2 is a graph showing FIM and FAM scores summed by outcome category. FIM andFAM instruments were administered weekly. The scores for each of seven functional categories were summed for each week and summed category scores were plotted vs protocol week (WK). The maximum possible score for each category is shown in parentheses. DETAILED DESCRIPTION

[0009] Provided herein are compositions and methods for treating and managing symptoms oftraumatic brain injury (TBI).

[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 andRNA (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 billionEVs 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 someembodiments, 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 bonemarrow MSC. The MSC may be a human MSC. In some embodiments, the EV is produced fromAttorney Docket No.66309-729.601 a MSC that has the capacity to undergo trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes. In some embodiments, the MSC is positive for CD73, CD105, CD166, and CD90 and is negative for CD14, CD31, CD34, and CD45.

[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., NanoView Biosciences, Boston, MA).

[0016] In some embodiments, the EV comprises a peptide or protein. In some embodiments, thepeptide 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, CD109 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), 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 F1 (Pigment epithelium-derived factor),Attorney Docket No.66309-729.601 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, COL1A1 (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 independentof 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),Attorney Docket No.66309-729.601 Cathepsin B, CD63 antigen, CD81 antigen, CD9 antigen, CD44 antigen, CD99 antigen, CD109 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 F1 (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 7A, NKp80, Cystatin 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 D1, 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 R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Nepri-lysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Nor-rin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9,Attorney Docket No.66309-729.601 MMP-1, TC-PTP, IL-24, gp130, 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, PAR1, 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, OX40 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 R1, DSPG3, SorCS1, 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, ROR1, 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, 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, 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, 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,Attorney Docket No.66309-729.601 Pepsin-ogen II, Desmoglein-3, Nectin-4, SCF, Serpin A5, PTH, FGF-19, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP(TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirre13, FLRT1, 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, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, 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.1, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-gamma R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalci- tonin, 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, COL1A2, 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 TIMP1. In someembodiments, the TIMP1 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 80Attorney Docket No.66309-729.601 ng / mL of the composition or about 30 pg / mL to about 12 ng / mL. In some embodiments, the one or more proteins comprises IGFBP4. In some embodiments, the IGFBP4 is present at about 200 pg / mL to about 80 ng / mL of the composition or about 30 pg / mL to about 12 ng / mL. In some embodiments, the one or more proteins comprises osteonectin. In some embodiments, the osteonectin is present at about 200 pg / mL to about 80 ng / mL of the composition or about 30 pg / mL to about 12 ng / mL. In some embodiments, the concentration of the one or more proteins is measured by ELISA.

[0022] In some embodiments, the total protein concentration of the one or more proteins isabout 10 to about 40 µg 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 µg 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 someembodiments, 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 pharmaceuticallyacceptable carrier. A pharmaceutically acceptable carrier may biologically suitable, i.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 maycomprise 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,Attorney Docket No.66309-729.601 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. Insome 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 (C₃H₅NaO₃) at 25-35 mEq / L, potassium chloride (KCl) at 2-6 mEq / L, calcium chloride (CaCl₂) 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), β-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-galactosy1-0-(1-4)-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 100kDa.Attorney Docket No.66309-729.601

[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 producedfrom 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,Attorney Docket No.66309-729.601 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 (C₃H₅NaO₃) at 25-35 mEq / L, potassium chloride (KCl) at 2-6 mEq / L, calcium chloride (CaCl₂) 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), β-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-galactosy1-0-(1-4)-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 proteincomponent 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 / orAttorney Docket No.66309-729.601 during 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). Forexample, 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 about7.4, 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 tophysiologic 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 Treating Traumatic Brain Injury

[0036] In some embodiments, the therapeutic compositions disclosed herein are used in methodsof treating traumatic brain injury in a subject. In some embodiments, the therapeuticAttorney Docket No.66309-729.601 compositions disclosed herein are used in a method of treating a head injury or symptom thereof in a subject. In some embodiments, the therapeutic compositions disclosed herein are used in a method of treating cognitive impairment or a symptom thereof in a subject. In some embodiments, the therapeutic compositions disclosed herein are used in a method of improving independence in a subject. In some embodiments, the therapeutic compositions disclosed herein are used in a method of improving self-care, sphincter control, mobility, locomotion, communication, psychosocial adjustment, and / or cognitive function in a subject. Any of the therapeutic compositions described herein may be used in the methods described herein.

[0037] Examples of traumatic brain injury include, but are not limited to, closed brain injury,penetrating brain injury, diffuse axonal injury (DAI), extra-axial hematoma, traumatic subarachnoid hemorrhage (SAH), concussion, contusion, acute traumatic brain injury, or non- inflammatory traumatic brain injury. In some embodiments, closed brain injuries can happen when there is a nonpenetrating injury to the brain with no break in the skull. In one example, a closed brain injury can be caused by a rapid forward or backward movement and shaking of the brain inside the bony skull that results in bruising and tearing of brain tissue and blood vessels. In some embodiments, closed brain injuries can be caused by motor vehicle-related collisions, falls, sports injuries, violence, shaking a baby (e.g., shaken baby syndrome), explosive blasts and other combat injuries, or any combinations thereof. In another example, penetrating or open head injuries can happen when there is a break in the skull. Another example, diffuse axonal injury (DAI), shearing or tearing of axons, can happen when the brain is injured as it shifts and rotates inside the bony skull. DAI usually can cause coma and injury to many different parts of the brain. In some embodiments, the changes in the brain can be microscopic and may not be evident on computed tomography (CT scan) or magnetic resonance imaging (MRI) scans. In some embodiments, extra-axial hematoma can include epidural hematomas (EDH), subdural hematomas (SDH), or both. In some embodiments, EDH can result from bleeding from the middle meningeal artery and its branches or a fracture. In some embodiments, EDH can be acute. In some embodiments, SDH can result from the bleeding of a bridging vein. In some embodiments, SDH can be acute or chronic. In some embodiments, subarachnoid hemorrhage (SAH) can be caused by trauma. In some embodiments, SAH can result from the tearing of small capillaries with blood subsequently entering into the subarachnoid space. In some embodiments, SAH can occur over the convexity. In some embodiments, SAH secondary to aneurysmal rupture can occur in the basal cisterns. In some embodiments, contusion, bruising of the brain, can comprise a coup type contusion or contrecoup type contusion. In some embodiments, coup contusions can occur at the site of impact. In some embodiments, contrecoup contusions can occur on the contralateral side of impact (e.g., the basi-frontal lobeAttorney Docket No.66309-729.601 and anterior temporal lobe). In some embodiments, concussion can be a mild TBI without any gross structural damage and can occur secondary to a nonpenetrating TBI. In some embodiments, concussion can result from acceleration / deceleration forces occurring secondary to a direct blow to the head. In some embodiments, concussion can cause a transient altered mental status, which can range from confusion to loss of consciousness. In some embodiments, concussion cannot be diagnosed with a routine computed tomogram (CT) scan or magnetic resonance imaging (MRI) scan. In some embodiments, special sequence MRI like diffusion tensor imaging and functional MRI may result in earlier diagnosis of concussion. In some embodiments, concussion can comprise second impact syndrome or chronic traumatic encephalopathy (CTE). In some embodiments, second impact syndrome can occur when a subject or a patient (e.g., an athlete) starts to play without fully recovering from a previous concussion and sustains another injury. In some embodiments, second impact syndromed can comprise a rapid evolution of malignant cerebral edema, ensuing over a short-time course of time. In some embodiments, CTE can occur when there is a delayed manifestation of repetitive mild TBI. In some embodiments, CTE can be common in athletes and can lead to psychiatric disturbances, suicidal behavior, attention deficits, and / or derangements in memory and executive functions.

[0038] In some embodiments, traumatic brain injury can have wide-ranging physical andpsychological effects. In some embodiments, signs or symptoms may appear immediately after the traumatic event. In some embodiments, signs or symptoms may appear days or weeks later. Non-limiting examples of physical symptoms of mild traumatic brain injury can include headache, nausea, vomiting, fatigue, drowsiness, problems with speech, dizziness, or loss of balance. Non-limiting examples of sensory symptoms of mild traumatic brain injury can include blurred vision, ringing in the ears, a bad taste in the mouth, changes in the ability to smell, sensitivity to light, or sensitivity to sound. Non-limiting examples of cognitive, behavioral, or mental symptoms of mild traumatic brain injury can include loss of consciousness for a few seconds to a few minutes, a state of being dazed, confused, or disoriented without loss of consciousness, memory problems, concentration problems, mood changes, mood swings, feeling depressed, feeling anxious, difficulty sleeping, or sleeping more than usual.

[0039] In some embodiments, moderate to severe traumatic brain injury can include any of thesigns and symptoms of mild injury described herein, and symptoms that may appear within the first hours to days after a head injury. Non-limiting examples of physical symptoms of moderate to severe traumatic brain injury can include loss of consciousness (e.g., loss of consciousness from several minutes to hours), persistent headache or headache that worsens over time, repeated vomiting, repeated nausea, convulsions, seizures, dilation of one or both pupils of theAttorney Docket No.66309-729.601 eyes, clear fluids draining from the nose or ears, inability to awaken from sleep, weakness or numbness in fingers and toes, or loss of coordination. Non-limiting examples of cognitive, behavioral, or mental symptoms of moderate or severe traumatic brain injury can include profound confusion, agitation, combativeness, any unusual behavior, slurred speech, coma, or any disorders of consciousness.

[0040] In some embodiments, infants and / or young children may suffer traumatic brain injuries.In some embodiments, infants and / or young children with brain injuries might not be able to communicate headaches, sensory problems, confusion, and / or similar symptoms. In some embodiments, an infant and / or a child with traumatic brain injury may present change in eating or nursing habits, unusual or easy irritability, persistent crying and / or inability to be consoled, change in ability to pay attention, change in sleep habits, seizures, sad or depressed mood, drowsiness, or loss of interest in favorite toys or activities. Symptoms of traumatic brain injury in children or adults include but are not limited to headache, dizziness, nausea, confusion, memory problems, difficulty concentrating, blurred vision, sensitivity to light or noise, sleep disturbances, mood swings, irritability, fatigue, loss of balance, ringing in the ears, speech difficulties, anxiety, depression, seizures, loss of coordination, changes in taste or smell, blurred vision, slurred speech, weakness, numbness, trouble finding words, trouble swallowing, difficulty with problem-solving, changes in personality, impulsivity, sensitivity to touch, difficulty multitasking, inability to focus, coordination issues, reduced concentration, poor judgment, hyperactivity, decreased attention span, apathy, headaches that worsen over time, and / or an inability to perform routine tasks. In some embodiments, the traumatic brain injury is acute. In some embodiments, the traumatic brain injury is chronic.

[0041] In some embodiments, the traumatic brain injury comprises acute traumatic brain injury.In some embodiments, the traumatic brain injury comprises non-inflammatory traumatic brain injury. In some embodiments, treating traumatic brain injury comprises treating one or more symptoms of the traumatic brain injury. In some embodiments, the one or more symptoms of traumatic injury comprises non-inflammatory symptoms of the traumatic brain injury. In some embodiments, the subject suffered or is suffering from intracranial hemorrhage. In some embodiments, the subject has difficulty ambulating. In some embodiments, the subject has severe impairment in cognition, speech, ambulation, activities of daily living, mobility, coordination, memory, or combinations thereof. In some embodiments, the subject does not respond to or has a delayed response to a conventional treatment or has no change in symptoms after a conventional treatment. In some embodiments, methods described herein can be used to treat a subject that is experiencing a non-inflammatory traumatic brain injury. In someAttorney Docket No.66309-729.601 embodiments, methods described herein can be used to treat a subject that is not experiencing an inflammatory traumatic brain injury.

[0042] In some embodiments, the subject experiences improvement in Functional IndependenceMeasure (FIM) scores, Functional Assessment Measure (FAM) scores, or combinations thereof, after treatment with a composition disclosed herein. FIM is an instrument developed as a measure of disability for a variety of populations and may not be specific to any diagnosis. It is designed to assess areas of dysfunction in activities that commonly occur in subjects with any progressive, reversible or stable neurologic, musculoskeletal, or other disorder (e.g., patients with functional mobility impairments). In some embodiments, FIM can be used by healthcare practitioners or subjects in need thereof to assess and grade the functional status of the subject based on the level of assistance the subject requires. In some embodiments, the FIM instrument can include measures of independence for self-care, including sphincter control, transfers, locomotion, communication, and / or social cognition. In some embodiments, the FIM instrument can comprise an 18-item, seven-level, ordinal scale intended to be sensitive to changes over the course of a comprehensive inpatient medical rehabilitation program. In some embodiments, the FIM instrument can use the level of assistance a subject needs to grade functional status from total independence to total assistance. In some embodiments, the FIM instrument can be used to assess a subject's level of disability as well as a change in subject status in response to rehabilitation or medical intervention. In some embodiments, the FIM instrument can comprise 18 items comprising 13 motor tasks and 5 cognitive tasks. In some embodiments, the 18 items of the FIM instrument can comprise eating, grooming, bathing, upper body dressing, lower body dressing, toileting, bladder management, bowel management, bed to chair transfer, toilet transfer, shower transfer, locomotion (ambulatory or wheelchair level), stairs, cognitive comprehension, expression, social interaction, problem solving, and memory. The FAM instrument comprises 12 additional items added on to the 18 items of the FIM to enhance the utility for the brain injury. In some embodiments, the 12 items of the FAM instrument can comprise swallowing, car transfer, community access, reading, writing, speech intelligibility, emotional status, adjustability to limitations, employability, orientation, attention, and safety judgement. In some embodiments, the total 30 item scale combination can be referred to as the FIM+FAM.

[0043] In some embodiments, the FIM+FAM scores may take into consideration one or more ofthe following items: self-care, sphincter control, mobility, locomotion, communication, psychosocial adjustment, and / or cognitive function. In some embodiments, self-care may be scored out of a maximal score of 49. In some embodiments, a self-care score may be 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,Attorney Docket No.66309-729.601 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49. In some embodiments, a self-care score may improve by 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%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 186%, 187%, 188%, 189%, 190%, 191%, 192%, 193%, 194%, 195%, 196%, 197%, 198%, 199%, 200%, 201%, 202%, 203%, 204%, 205%, 206%, 207%, 208%, 209%, 210%, 211%, 212%, 213%, 214%, 215%, 216%, 217%, 218%, 219%, 220%, 221%, 222%, 223%, 224%, 225%, 226%, 227%, 228%, 229%, 230%, 231%, 232%, 233%, 234%, 235%, 236%, 237%, 238%, 239%, 240%, 241%, 242%, 243%, 244%, 245%, 246%, 247%, 248%, 249%, 250%, 251%, 252%, 253%, 254%, 255%, 256%, 257%, 258%, 259%, 260%, 261%, 262%, 263%, 264%, 265%, 266%, 267%, 268%, 269%, 270%, 271%, 272%, 273%, 274%, 275%, 276%, 277%, 278%, 279%, 280%, 281%, 282%, 283%, 284%, 285%, 286%, 287%, 288%, 289%, 290%, 291%, 292%, 293%, 294%, 295%, 296%, 297%, 298%, 299%, or 300% from baseline in response to treatment with a composition of the present disclosure. In some embodiments, a sphincter control score may be scored out of a maximal score of 14. In some embodiments, a sphincter control score may be 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14. In some embodiments, a sphincter control score may improve by 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%,Attorney Docket No.66309-729.601 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 186%, 187%, 188%, 189%, 190%, 191%, 192%, 193%, 194%, 195%, 196%, 197%, 198%, 199%, 200%, 201%, 202%, 203%, 204%, 205%, 206%, 207%, 208%, 209%, 210%, 211%, 212%, 213%, 214%, 215%, 216%, 217%, 218%, 219%, 220%, 221%, 222%, 223%, 224%, 225%, 226%, 227%, 228%, 229%, 230%, 231%, 232%, 233%, 234%, 235%, 236%, 237%, 238%, 239%, 240%, 241%, 242%, 243%, 244%, 245%, 246%, 247%, 248%, 249%, 250%, 251%, 252%, 253%, 254%, 255%, 256%, 257%, 258%, 259%, 260%, 261%, 262%, 263%, 264%, 265%, 266%, 267%, 268%, 269%, 270%, 271%, 272%, 273%, 274%, 275%, 276%, 277%, 278%, 279%, 280%, 281%, 282%, 283%, 284%, 285%, 286%, 287%, 288%, 289%, 290%, 291%, 292%, 293%, 294%, 295%, 296%, 297%, 298%, 299%, or 300% from baseline in response to treatment with a composition of the present disclosure. In some embodiments, a mobility score may be scored out of a maximal score of 28. In some embodiments, a mobility score may be 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, or 28. In some embodiments, a mobility score may improve by 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%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 186%, 187%, 188%, 189%, 190%, 191%, 192%, 193%, 194%, 195%, 196%, 197%, 198%, 199%, 200%, 201%, 202%, 203%, 204%, 205%, 206%, 207%, 208%, 209%, 210%, 211%, 212%, 213%, 214%, 215%, 216%, 217%, 218%, 219%, 220%, 221%, 222%, 223%, 224%, 225%, 226%, 227%, 228%, 229%, 230%, 231%, 232%, 233%, 234%, 235%, 236%, 237%, 238%, 239%, 240%, 241%, 242%, 243%, 244%, 245%, 246%, 247%, 248%, 249%, 250%, 251%, 252%,Attorney Docket No.66309-729.601 253%, 254%, 255%, 256%, 257%, 258%, 259%, 260%, 261%, 262%, 263%, 264%, 265%, 266%, 267%, 268%, 269%, 270%, 271%, 272%, 273%, 274%, 275%, 276%, 277%, 278%, 279%, 280%, 281%, 282%, 283%, 284%, 285%, 286%, 287%, 288%, 289%, 290%, 291%, 292%, 293%, 294%, 295%, 296%, 297%, 298%, 299%, or 300% from baseline in response to treatment with a composition of the present disclosure. In some embodiments, a locomotion score may be scored out of a maximal score of 21. In some embodiments, a locomotion score may be 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21. In some embodiments, a locomotion score may improve by 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%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 186%, 187%, 188%, 189%, 190%, 191%, 192%, 193%, 194%, 195%, 196%, 197%, 198%, 199%, 200%, 201%, 202%, 203%, 204%, 205%, 206%, 207%, 208%, 209%, 210%, 211%, 212%, 213%, 214%, 215%, 216%, 217%, 218%, 219%, 220%, 221%, 222%, 223%, 224%, 225%, 226%, 227%, 228%, 229%, 230%, 231%, 232%, 233%, 234%, 235%, 236%, 237%, 238%, 239%, 240%, 241%, 242%, 243%, 244%, 245%, 246%, 247%, 248%, 249%, 250%, 251%, 252%, 253%, 254%, 255%, 256%, 257%, 258%, 259%, 260%, 261%, 262%, 263%, 264%, 265%, 266%, 267%, 268%, 269%, 270%, 271%, 272%, 273%, 274%, 275%, 276%, 277%, 278%, 279%, 280%, 281%, 282%, 283%, 284%, 285%, 286%, 287%, 288%, 289%, 290%, 291%, 292%, 293%, 294%, 295%, 296%, 297%, 298%, 299%, or 300% from baseline in response to treatment with a composition of the present disclosure. In some embodiments, a communication score may be scored out of a maximal score of 35. In some embodiments, a communication score may be 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, or 35. In some embodiments, a communication score may improve by 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%,Attorney Docket No.66309-729.601 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%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 186%, 187%, 188%, 189%, 190%, 191%, 192%, 193%, 194%, 195%, 196%, 197%, 198%, 199%, 200%, 201%, 202%, 203%, 204%, 205%, 206%, 207%, 208%, 209%, 210%, 211%, 212%, 213%, 214%, 215%, 216%, 217%, 218%, 219%, 220%, 221%, 222%, 223%, 224%, 225%, 226%, 227%, 228%, 229%, 230%, 231%, 232%, 233%, 234%, 235%, 236%, 237%, 238%, 239%, 240%, 241%, 242%, 243%, 244%, 245%, 246%, 247%, 248%, 249%, 250%, 251%, 252%, 253%, 254%, 255%, 256%, 257%, 258%, 259%, 260%, 261%, 262%, 263%, 264%, 265%, 266%, 267%, 268%, 269%, 270%, 271%, 272%, 273%, 274%, 275%, 276%, 277%, 278%, 279%, 280%, 281%, 282%, 283%, 284%, 285%, 286%, 287%, 288%, 289%, 290%, 291%, 292%, 293%, 294%, 295%, 296%, 297%, 298%, 299%, or 300% from baseline in response to treatment with a composition of the present disclosure. In some embodiments, a psychosocial adjustment score may be scored out of a maximal score of 28. In some embodiments, a psychosocial adjustment score may be 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, or 28. In some embodiments, a psychosocial adjustment score may improve by 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%, 150%, 151%, 152%, 153%, 154%,Attorney Docket No.66309-729.601 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 186%, 187%, 188%, 189%, 190%, 191%, 192%, 193%, 194%, 195%, 196%, 197%, 198%, 199%, 200%, 201%, 202%, 203%, 204%, 205%, 206%, 207%, 208%, 209%, 210%, 211%, 212%, 213%, 214%, 215%, 216%, 217%, 218%, 219%, 220%, 221%, 222%, 223%, 224%, 225%, 226%, 227%, 228%, 229%, 230%, 231%, 232%, 233%, 234%, 235%, 236%, 237%, 238%, 239%, 240%, 241%, 242%, 243%, 244%, 245%, 246%, 247%, 248%, 249%, 250%, 251%, 252%, 253%, 254%, 255%, 256%, 257%, 258%, 259%, 260%, 261%, 262%, 263%, 264%, 265%, 266%, 267%, 268%, 269%, 270%, 271%, 272%, 273%, 274%, 275%, 276%, 277%, 278%, 279%, 280%, 281%, 282%, 283%, 284%, 285%, 286%, 287%, 288%, 289%, 290%, 291%, 292%, 293%, 294%, 295%, 296%, 297%, 298%, 299%, or 300% from baseline after treatment with a composition of the present disclosure. In some embodiments, a cognitive function score may be scored out of a maximal score of 35. In some embodiments, a cognitive function score may be 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, or 35. In some embodiments, a cognitive function score may improve by 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%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 186%, 187%, 188%, 189%, 190%, 191%, 192%, 193%, 194%, 195%, 196%, 197%, 198%, 199%, 200%, 201%, 202%, 203%, 204%, 205%, 206%, 207%, 208%, 209%, 210%, 211%, 212%, 213%, 214%, 215%, 216%, 217%, 218%, 219%, 220%, 221%, 222%, 223%, 224%, 225%, 226%, 227%, 228%, 229%, 230%, 231%, 232%, 233%, 234%, 235%, 236%, 237%, 238%, 239%, 240%, 241%, 242%, 243%, 244%, 245%, 246%, 247%, 248%, 249%, 250%, 251%, 252%, 253%, 254%, 255%, 256%, 257%, 258%, 259%, 260%, 261%, 262%, 263%, 264%, 265%, 266%, 267%, 268%, 269%, 270%,Attorney Docket No.66309-729.601 271%, 272%, 273%, 274%, 275%, 276%, 277%, 278%, 279%, 280%, 281%, 282%, 283%, 284%, 285%, 286%, 287%, 288%, 289%, 290%, 291%, 292%, 293%, 294%, 295%, 296%, 297%, 298%, 299%, or 300% from baseline in response to treatment with a composition of the present disclosure. In some embodiments, an improvement may be observed after 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, 49 weeks, 50 weeks, 51 weeks, or 52 weeks after an initial administration of a composition of the present disclosure. In some embodiments, an improvement may be steady and continue over time. In some embodiments, an improvement may be sudden. In some embodiments, a FIM, FAM, or FIM+FAM score of the present disclosure may plateau in response to treatment with a composition of the present disclosure. In some embodiments, an improvement may be delayed by 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 weeks, 49 weeks, 50 weeks, 51 weeks, or 52 weeks. In some embodiments, a FIM score may improve by 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%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 186%, 187%, 188%, 189%, 190%, 191%, 192%, 193%, 194%, 195%, 196%, 197%, 198%, 199%, 200%, 201%, 202%, 203%, 204%, 205%, 206%,Attorney Docket No.66309-729.601 207%, 208%, 209%, 210%, 211%, 212%, 213%, 214%, 215%, 216%, 217%, 218%, 219%, 220%, 221%, 222%, 223%, 224%, 225%, 226%, 227%, 228%, 229%, 230%, 231%, 232%, 233%, 234%, 235%, 236%, 237%, 238%, 239%, 240%, 241%, 242%, 243%, 244%, 245%, 246%, 247%, 248%, 249%, 250%, 251%, 252%, 253%, 254%, 255%, 256%, 257%, 258%, 259%, 260%, 261%, 262%, 263%, 264%, 265%, 266%, 267%, 268%, 269%, 270%, 271%, 272%, 273%, 274%, 275%, 276%, 277%, 278%, 279%, 280%, 281%, 282%, 283%, 284%, 285%, 286%, 287%, 288%, 289%, 290%, 291%, 292%, 293%, 294%, 295%, 296%, 297%, 298%, 299%, or 300% from baseline in response to treatment with a composition of the present disclosure. In some embodiments, a FAM score may improve by 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%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 186%, 187%, 188%, 189%, 190%, 191%, 192%, 193%, 194%, 195%, 196%, 197%, 198%, 199%, 200%, 201%, 202%, 203%, 204%, 205%, 206%, 207%, 208%, 209%, 210%, 211%, 212%, 213%, 214%, 215%, 216%, 217%, 218%, 219%, 220%, 221%, 222%, 223%, 224%, 225%, 226%, 227%, 228%, 229%, 230%, 231%, 232%, 233%, 234%, 235%, 236%, 237%, 238%, 239%, 240%, 241%, 242%, 243%, 244%, 245%, 246%, 247%, 248%, 249%, 250%, 251%, 252%, 253%, 254%, 255%, 256%, 257%, 258%, 259%, 260%, 261%, 262%, 263%, 264%, 265%, 266%, 267%, 268%, 269%, 270%, 271%, 272%, 273%, 274%, 275%, 276%, 277%, 278%, 279%, 280%, 281%, 282%, 283%, 284%, 285%, 286%, 287%, 288%, 289%, 290%, 291%, 292%, 293%, 294%, 295%, 296%, 297%, 298%, 299%, or 300% from baseline after treatment with a composition of the present disclosure. In some embodiments, a FIM+FAM score may improve by 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%,Attorney Docket No.66309-729.601 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%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 186%, 187%, 188%, 189%, 190%, 191%, 192%, 193%, 194%, 195%, 196%, 197%, 198%, 199%, 200%, 201%, 202%, 203%, 204%, 205%, 206%, 207%, 208%, 209%, 210%, 211%, 212%, 213%, 214%, 215%, 216%, 217%, 218%, 219%, 220%, 221%, 222%, 223%, 224%, 225%, 226%, 227%, 228%, 229%, 230%, 231%, 232%, 233%, 234%, 235%, 236%, 237%, 238%, 239%, 240%, 241%, 242%, 243%, 244%, 245%, 246%, 247%, 248%, 249%, 250%, 251%, 252%, 253%, 254%, 255%, 256%, 257%, 258%, 259%, 260%, 261%, 262%, 263%, 264%, 265%, 266%, 267%, 268%, 269%, 270%, 271%, 272%, 273%, 274%, 275%, 276%, 277%, 278%, 279%, 280%, 281%, 282%, 283%, 284%, 285%, 286%, 287%, 288%, 289%, 290%, 291%, 292%, 293%, 294%, 295%, 296%, 297%, 298%, 299%, or 300% from baseline after treatment with a composition of the present disclosure. In some embodiments, the subject experiences improvement after treatment in one or more of the following aspects selected from the group consisting of: communication, sphincter control, locomotion, mobility, cognitive function, psychosocial adjustment, self-care, or combinations thereof. In some embodiments, the subject exhibits an improvement of at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% in FIM score, FAM score, and / or FIM+FAM score after administration of the composition.

[0044] Also disclosed herein are methods of treating traumatic brain injury in a subjectcomprising 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,Attorney Docket No.66309-729.601 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, 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, 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 D1, 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 R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref- 1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2 MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, 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, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6„ ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF- 23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2,Insulin R, HIF-1 alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Neurturin, Kremen-2, EMMPRIN, Activin RIB, Neuroligin 2, Epiregulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A,NKp80, Prolactin, Cystatin B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, 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,Attorney Docket No.66309-729.601 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, ROR1, 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, 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-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFA5, Galectin-9, vWF-A2, TACE, Activin RUB, Cathepsin S, LDL R, BMPR-IA, OX40, IL- 3 R2, B7- H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF- 11, Nrf2, TROP-2, NUDT5, ROR2, EphB4, Glypican 1, LAP(TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-alpha, NRG1- bl, FABP4, RGM-A, RELT, TrkC, C5a, SREC-I, Nestin, TPO, ErbB3, Kirrel3, FLRT1, Galectin- 3, CXCL16, JAM-B, DR6,Nogo Receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R alpha 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, 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,Attorney Docket No.66309-729.601 TRAIL R3, Trypsin 3, ARSB, LIF R alpha, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP alpha, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-lb, NKp46, MCP-3, IL-32 alpha, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF- AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP-2, PU.1, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-gamma R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 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-10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR- 125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145- 5p, hsa-mir-146a, hsa-miR-146a- 5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa- miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1'7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa- miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir- 203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir- 221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR- 23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a- 5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR- 30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30d-5p, hsa- mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-342-3p, hsa- miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-376a-3p, hsa-miR-376c-3p, hsa-miR- 423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa- miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p, and hsa-miR-99b-5p. In some embodiments, methods of treating traumatic brain injury described herein can comprise administering to a subject in need thereof a composition comprising secreted extracellular vesicles that contain a composition that includes any combination of composition proteins but does not include any RNAs described herein.Attorney Docket No.66309-729.601

[0045] In some embodiments, the therapeutic product is administered intravenously. In someembodiments, the therapeutic product 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 9x1011to 1.2x1012extracellular vesicles or 5x1011to 1.5x1012, 6x1011to 1.4x1012, 7x1011to 1.3x1012, 8x1011to 1.2x1012, or 8x1011to 1.3x1012extracellular vesicles. In some embodiments, the product is administered at a dose that provides at least or at most 5x1011, 6x1011, 7x1011, 8x1011, 9x1011, 1x1012, 1.1x1012, 1.2x1012, 1.3x1012, 1.4x1012, or 1.5x1012extracellular vesicles. In some embodiments, the therapeutic product comprises 6x1010to 8x1010, 5x1010to 9x1010, 4x1010to 10x1010, 5.5x1010to 8.5x1010, or 6x1010to 8.5x1010cells / ml. In some embodiments, the therapeutic product comprises 6x1010to 8x1010extracellular vesicles per ml and is administered at a dose of 10 to 20 ml. In some embodiments, the therapeutic product comprises 6x1010to 8x1010extracellular 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 to administer at a dosage of 5 million cells / kg. In some embodiments, the therapeutic product dosage ceiling is 10 million cells / kg.

[0046] In some embodiments, the administration may comprise an intravenous infusion. In someembodiments, the administration may be carried out over the course of several minutes. In some embodiments, the administration may be carried out over the course of 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60 minutes. In some embodiments, the administration may be carried out over the course of greater than 60 minutes. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 milliliters (mL) of the compositions disclosed herein may be administered at a time. In some embodiments, a dose of a composition disclosed herein may be administered. In some embodiments, a dose of a composition disclosed herein may comprise 1, 2, 3, 4, or 5 milliliters. In some embodiments, a dose of a composition disclosed herein may be combined with normal saline. In some embodiments, a dose of a composition disclosed herein may be combined with normal saline for a final volume of 45 milliliters. In some embodiments, a dose of 15 milliliters of a composition disclosed herein may be combined with 30 milliliters of normal saline. In some embodiments, a dose of a composition disclosed herein may be combined with normal saline for a final volume of 100 milliliters. In some embodiments, a dose of 5 milliliters of a composition disclosed herein may be combined with 95 milliliters of normal saline. In some embodiments, a dose of a composition disclosed herein may be combined withAttorney Docket No.66309-729.601 normal saline for a final volume of 100 milliliters. In some embodiments, a dose of 15 milliliters of a composition disclosed herein may be combined with 85 milliliters of normal saline. In some embodiments, the compositions disclosed herein may be administered once. In some embodiments, the compositions disclosed herein may be administered more than once.

[0047] In some embodiments, a composition of the present disclosure may be administered viaintravenous infusion in combination with saline. In some embodiments, 15 mL of the composition may be administered in combination with 85 mL of saline. In some embodiments, one dose of the composition may be administered at one time. In some embodiments, more than one dose of the composition may be administered at one time. In some embodiments, one, two, or three doses of the composition may be administered at one time. In some embodiments, one, two, or three doses of the composition may be administered over the course of one week. In some embodiments, one dose may be administered every 48 hours. In some embodiments, three doses may be administered per week. In some embodiments, three doses may be administered per month.III. Definitions

[0048] As used in this specification and the appended claims, the singular forms “a,” “an,” and“the” include plural referents unless the content clearly dictates otherwise. It should also be noted that the term “or” is generally employed in its sense including “and / or” unless the content clearly dictates otherwise. The terms “and / or”, “a combination of two or more thereof”, and “any combination thereof” and their grammatical equivalents as used herein, can be used interchangeably. These terms can convey that any combination is specifically contemplated. Solely for illustrative purposes, the following phrases “A, B, and / or C” or “A, B, C, or any combination thereof” can mean “A individually; B individually; C individually; A and B; B and C; A and C; and A, B, and C.” The term “or” can be used conjunctively or disjunctively, unless the context specifically refers to a disjunctive use.

[0049] The term “about” or “approximately” can mean within an acceptable error range for theparticular value, which may depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, “about” can mean within 1 or more than 1 standard deviation. Alternatively, “about” can mean a range of up to 20%, up to 10%, up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, within 5-fold, or within 2-fold, of a value. Where particular values are described in the application and claims, unless otherwise stated the term “about” meaning within an acceptable error range for the particular value should be assumed.Attorney Docket No.66309-729.601

[0050] As used in this specification and claim(s), the words “comprising” (and any form ofcomprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method or composition of the present disclosure, and vice versa. Furthermore, compositions of the present disclosure can be used to achieve methods of the present disclosure.

[0051] Reference in the specification to “some embodiments,” “an embodiment,” “oneembodiment” or “other embodiments” means that a particular feature, structure, or characteristic described in connection with the embodiments is included in at least some embodiments, but not necessarily all embodiments, of the present disclosures. To facilitate an understanding of the present disclosure, a number of terms and phrases are defined below.

[0052] Certain specific details of this description are set forth in order to provide a thoroughunderstanding of various embodiments. However, one skilled in the art will understand that the present disclosure may be practiced without these details. In other instances, well-known techniques or methods have not been shown or described in detail to avoid unnecessarily obscuring descriptions of the embodiments. Unless the context requires otherwise, throughout the specification and claims which follow, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense, that is, as “including, but not limited to.” Further, headings provided herein are for convenience only and do not interpret the scope or meaning of the claimed disclosure.

[0053] Although methods and materials similar or equivalent to those described herein can beused in the practice or testing of the present disclosure, suitable methods, and materials are described below.IV. Examples

[0054] The following examples are put forth so as to provide those of ordinary skill in the artwith 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.Attorney Docket No.66309-729.601A. Example 1 – Production of Therapeutic Composition

[0055] An MSC secretome therapeutic composition (referred to herein as investigationalproduct 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.

[0056] The IMP was manufactured from the banked hBM-MSCs of a single donor under cGMPconditions 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.

[0057] The IMP comprises extracellular vesicles (EVs) that are acellular and nonimmunogenic,containing no nucleus or deoxyribonucleic acid (DNA).

[0058] The tetraspanin profile of extracellular vesicles present in the IMP was determined, andit was found that greater than 95% of the extracellular vesicles present in the composition were CD63+CD9–CD81–. The EVs were measured via Nanoparticle tracking analysis (NTA), 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.

[0059] Protein content of the IMP was determined, and the following proteins were found to bepresent. Proteins were detected using an antibody-based sandwich ELISAs or by Luminex or Nanoview / Unchained Labs-based methods. The total concentration of certain proteins in the IMP was measured via ELISA, with each protein having a concentration of about 200 pg / mL to about 80 ng / mL. For instance, TIMP1, OPN, IGFBP4, and osteonectin were characterized. Non- limiting examples of proteins present in the IMP are shown in Table 1 below. Table 1Attorney Docket No.66309-729.601Attorney Docket No.66309-729.601

[0060] The nucleic acid content of the IMP was determined, and the following nucleic acidswere 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.

[0061] Additional proteins that may be present in the IMP include: NUP85, DAPP1, PDGF Ralpha, 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 D1, 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 R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1,Attorney Docket No.66309-729.601 Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC-II, Norrin, JAM-C, Bc1- 10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C- myc, LILRB4, BMP-2„ MIA, CD34, 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, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6„ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD- ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2,Insulin R, HIF-1 alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, Semaphorin 4D, ROBO2, PDX-1, APRIL, Neurturin, Kremen-2, EMMPRIN, Activin RIB, Neuroligin 2, Epiregulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, 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, ROR1, 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, 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,Attorney Docket No.66309-729.601 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, NTAL, EGF R, TAFAS, Galectin- 9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP(TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-alpha, NRG1- 131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirre13, FLRT1, Galectin-3, CXCL16, JAM-B, DR6,Nogo Receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R alpha 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2, 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, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-gamma R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 Rg, MICA, 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.Attorney Docket No.66309-729.601B. Example 2 – Clinical Study of Bone Marrow Derived Mesenchymal Stem CellExtracellular Vesicle Isolate as a Successful Treatment for Severe, Chronic Traumatic Brain Injury

[0062] In this Example, treatment of a severe TBI patient with a human bone marrow derivedmesenchymal stem cell (MSC) extracellular vesicle (EV) preparation is described. Also described herein is an evaluation of the safety and potential efficacy of a human bone marrow derived MSC EV in a patient with severe acquired traumatic brain injury (ATBI). Given the severity of injury, slow progress of conventional rehabilitation, and compelling improvement in the patient’s Functional Independence Measure (FIM) and Functional Assessment Measure (FAM) scores following treatment, the results described herein demonstrate that human bone marrow derived MSC EVs offer a novel and safe means to improve ATBI patient outcomes.

[0063] Patient

[0064] A 33-year-old male was struck by a motor vehicle (approximately 45 mph). Hesustained severe intracranial hemorrhage and sheer force damage, requiring surgical decompression. He was in a medical coma for two months, and had over three months of intense hospital rehabilitation afterward. He was suffering from severe impairment in cognition, speech, ambulation, and activities of daily living at the first visit, which was two years after the motor vehicle incident. Moderate gains were achieved with rehabilitation cycling of three months on and three months off. While unable to live independently or cook, he was able to dress, bathe, and feed himself. He used a wheelchair for mobility but was able to stand. He exhibited greater weakness on the left versus right, coordination difficulty and poor memory for history details. He and the family were counseled regarding conventional versus regenerative medicine options.

[0065] After 3 months, his symptoms were stable with no changes from his previous visit. Hehad consented to move forward with a therapeutic exemption application to receive treatment of mesenchymal stem cell (MSC) derived extracellular vesicles (EVs). The United States Food and Drug Administration (FDA) approved a single patient Investigational New Drug (IND) intravenous (IV) treatment of MSC derived EVs for him given he had experienced no change in his baseline symptoms with conventional therapy. The Institutional Review Board (IRB) approved treatment of the patient (Protocol ID 28,349), and the patient consented to treatment with MSC derived EVs.

[0066] Materials and methods

[0067] The investigational product (IP) comprising MSC EVs was generated according toCurrent Good Manufacturing Practices (cGMP), with all manufacturing processes under a Quality Management System and with implementation.Attorney Docket No.66309-729.601

[0068] Procedure

[0069] Prior to the infusion, the patient was educated about alternative treatment options, andthe potential infusion related and investigational product related risks and benefits. The patient provided written consent to move forward with treatment. An IV was started, and 15 mL of IP mixed with 85 ML of Normal Saline was infused over one hour. Blood pressure (BP), heart rate (HR), respiratory rate (RR) and pulse oximetry were monitored throughout the procedure and 15 minutes afterwards; all vitals remained stable at all time points. The patient tolerated the procedure without difficulty and showed no treatment-related adverse events (AEs) or serious adverse events (SAEs). The patient received three infusions (Q48 hours or every other day) during the first week of six of nine months for a total of 18 infusions with no AEs or SAEs. FIM and FAM instruments were administered immediately prior to treatment and weekly for 36 weeks. Safety monitoring was also performed every week for nine months.

[0070] Outcome

[0071] The patient’s progress indicated by FIM, FAM and FIM+FAM summed scores areshown in FIG.1. Three IP treatments were administered every month (indicated by arrows) in May, June, July, September and November of 2022 and again in January of 2023 covering 36 weeks. FIM and FAM instruments were administered weekly and the individual weekly FIM, FAM and FIM+FAM summed scores are plotted vs protocol week (WK).

[0072] Within 2 weeks, improvement was apparent and continued through week 9, followed bya near plateau of all scores. From week 0 to 36, scores improved by 48% (FIM), 55% (FAM) and 50% (FIM+FAM).

[0073] Patient scores were also summarized according to instrument outcome categories (FIG.2). FIM and FAM instruments were administered weekly, the scores for each of seven functional items were summed for each week and summed item scores plotted vs protocol week (WK). The maximum possible score for each item is shown in parentheses. IP treatment events are indicated by arrows. All specific outcome categories assessed by FIM and FAM that were initially low showed sustained improvements ranging from 41% to 233%, with the greatest improvements seen in locomotion, mobility and cognitive function. Some categories had already approachedtheir maximum score (Table 2), and improvements were less apparent (e.g., communication:10.3% improvement; sphincter control: 0% improvement). Other categories showed clear improvements from baseline including locomotion (233%), mobility (117%), cognitive function (90%), psychosocial adjustment (71%), and self-care (41%). Regarding attainment of the maximal possible category score, percentages of the maximum at week 36 included sphincter control (100%), communication (91%), self-care (98%), mobility (93%), cognitive function (81%), psychosocial adjustment (73%), and locomotion (48%). Certain categories showedAttorney Docket No.66309-729.601 improvements within 1 week (e.g., cognitive function, self-care) while others were delayed by up to 3 weeks (e.g., psychosocial adjustment, communication).

[0074] Table 2 below shows a partial list of FIM and FAM scores.Table 2

[0075] Discussion

[0076] These results highlight safety and efficacy of human bone marrow-derived MSC EVs forthe treatment of TBI. Notable is that, after several months of plateaued response prior to rehabilitative therapy, this patient exhibited substantial and sustained improvements in several response categories within months of MSC-derived EV treatment.

[0077] The examples and embodiments described herein are for illustrative purposes only andvarious modifications or changes suggested to persons skilled in the art are to be included within the spirit and purview of this application.

Claims

Attorney Docket No.66309-729.601 CLAIMS WHAT IS CLAIMED IS:

1. A method of treating traumatic brain injury or a symptom thereof in a subject, the methodcomprising 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 D1, 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 R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like 1, MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 Ligand, IL-20 R beta, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mc1-1, DPPIV, SREC- II, Norrin, JAM-C, Bc1-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, ClqTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, 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, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R beta, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6„ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2,Insulin R, HIF-1 alpha, OX40 Ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, Semaphorin 4D, ROBO2, PDX-1, APRIL, Neurturin, Kremen-2, EMMPRIN, Activin RIB, Neuroligin 2, Epiregulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, 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,Attorney Docket No.66309-729.601 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, ROR1, 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, 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-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Cathepsin S, LDL R, BMPR- IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP(TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-alpha, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirre13, FLRT1, Galectin-3, CXCL16, JAM-B, DR6,Nogo Receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R alpha 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4, VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, P1GF-2,Attorney Docket No.66309-729.601 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, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-gamma R1, CrkL, B7-1, PARC, Draxin, VE-Cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 Rg, MICA, 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 treating traumatic brain injury or a symptom thereof in a subject, the methodcomprising administering to the subject a composition comprising one or more extracellular vesicles (EVs), 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.

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, CD109 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),Attorney Docket No.66309-729.601 IGFBP-3 (Insulin-like binding protein-3), IGFBP-6 (Insulin-like growth factor binding protein-6), IL18BP (Interleukin-18 Binding Protein), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (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.

4. The method of claim 2 or claim 3, wherein the composition further comprises one or more ofCOL2A1, COL3A1, COL5A1, COL5A2, COL6A1, COL11A1, or COL14A1, or a combination of two or more thereof.

5. A method of treating traumatic brain injury or a symptom thereof in a subject, the methodcomprising 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 treating traumatic brain injury or a symptom thereof in a subject, the methodcomprising 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 traumatic brain injury is acute.

9. The method of any one of claims 1-7, wherein the traumatic brain injury is chronic.

10. The method of any one of claims 1-9, wherein the subject exhibits impaired self-care,sphincter control, mobility, locomotion, communication, psychosocial adjustment, and / or cognitive function.

11. The method of any one of claims 1-10, wherein the subject exhibits one or more ofheadache, dizziness, nausea, confusion, memory problems, difficulty concentrating, blurred vision, sensitivity to light or noise, sleep disturbances, mood swings, irritability, fatigue, lossAttorney Docket No.66309-729.601 of balance, ringing in the ears, speech difficulties, anxiety, depression, seizures, loss of coordination, changes in taste or smell, blurred vision, slurred speech, weakness, numbness, trouble finding words, trouble swallowing, difficulty with problem-solving, changes in personality, impulsivity, sensitivity to touch, difficulty multitasking, inability to focus, coordination issues, reduced concentration, poor judgment, hyperactivity, decreased attention span, apathy, headaches that worsen over time, and / or an inability to perform routine tasks.

12. A method of treating a head injury or a symptom thereof in a subject, the method comprisingadministering to the subject a composition comprising a mesenchymal stem cell (MSC) secretome.

13. A method of treating a cognitive impairment or a symptom thereof in a subject, the methodcomprising administering to the subject a composition comprising a mesenchymal stem cell (MSC) secretome.

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

15. A method of improving self-care, sphincter control, mobility, locomotion, communication,psychosocial adjustment, and / or cognitive function 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 improvement of atleast 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% in Functional Independent Measure (FIM) score, Functional Assessment Measure (FAM) score, and / or FIM+FAM score after administration of the composition.

17. The method of any one of claims 1-16, wherein the subject has a traumatic head injury.

18. The method of any one of claims 1-17, wherein the subject has difficulty ambulating.

19. The method of any one of claims 1-18, wherein the administering comprises administering15 mL of the composition.

20. The method of claim 19, wherein the composition is administered with 85 mL of saline.

21. The method of any one of claims 1-20, wherein three doses of the composition areadministered per month for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

22. The method of claim 21, wherein the composition is administered during non-consecutivemonths.

23. The method of claim 21 or 22, wherein the three doses are administered over the course ofone week.Attorney Docket No.66309-729.60124. The method of claim 23, wherein each dose of the three doses is administered within 48hours of another dose.

25. The method of any one of claims 21-24, wherein the second dose is administered 48 hoursafter administration of the first dose.

26. The method of any one of claims 21-25, wherein the third dose is administered 48 hoursafter administration of the second dose.

27. The method of any one of claims 21-26, wherein the composition is administered at leastonce monthly over the course of 36 weeks.

28. The method of any one of claims 1-27, wherein the administering comprises intravenousadministration.

29. The method of claim 28, wherein the intravenous administration is carried out over thecourse of 30, 35, 40, 45, 50, 55, or 60 minutes.

30. The method of any one of claims 1-29, wherein the composition is prepared by a processcomprising: (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.

31. The method of any one of claims 1-29, the method comprising preparing the compositionprior 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.

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

33. The method of any one of claims 30-32, wherein the culture media has a glucoseconcentration below 4.5 g / L.

34. The method of any one of claims 30-33, wherein formulating the MSC conditioned mediacomprises exchanging the conditioned media for a pharmaceutically acceptable formulation.

35. The method of claim 34, wherein the pharmaceutically acceptable formulation comprisessaline.

36. The method of any one of claims 1-35, wherein the composition comprises at least 6x1010 to8x1010extracellular 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.

37. The method of claim 36, wherein the dose is administered in combination with normal salineat a final volume of 100 mL.Attorney Docket No.66309-729.60138. The method of any one of claims 1-37, wherein the composition comprises saline (0.9%sodium chloride).

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

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

41. The method of any one of claims 1-40, wherein the molecular weight of any non-excipientcomponent of the composition is greater than about 2 kDa (kilodaltons), 8 kDa, 10 kDa, 50kDa, or 100kDa.

42. The method of any one of claims 1-41, wherein the composition comprises anoligosaccharide.

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

44. The method of any one of claims 1-43, wherein any non-excipient component of thecomposition has a size of less than about 0.2 microns.

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

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

47. The method of any one of claims 1-46, wherein the composition is negative for mycoplasmaDNA.

48. The method of any one of claims 1-47, wherein the composition is cell-free.

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

50. The method of claim 49, wherein the composition is administered within 6 hours of thawwhen maintained at ambient temperature.

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

52. The method of any one of claims 1-51, wherein the composition is reconstituted with waterprior to administration.

53. The method of any one of claims 1-52, wherein the composition is present in a glass vial.Attorney Docket No.66309-729.60154. The method of any one of claims 1-53, wherein the composition is formulated forintravenous administration.

55. The method of any one of claims 1-54, wherein the composition has a pH of about 6 to about7.5.

56. The method of any one of claims 30-55, wherein the BM-MSCs are negative for CD14,CD31, CD34, and CD45.

57. The method of any one of claims 30-56, wherein the BM-MSCs are positive for CD73,CD105, CD166, and CD90.

58. The method of any one of claims 30-57, wherein the BM-MSCs are capable of undergoingtrilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes.

59. The method of any one of claims 30-58, wherein the BM-MSCs are obtained from an iliaccrest aspiration of a single donor.

60. The method of any one of claims 1-59, wherein a concentration of the one or more EVs inthe composition is measured by nanoparticle tracking analysis (NTA).

61. The method of claim 60, wherein the NTA comprises light scatter and fluorescenceevaluation.

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

63. The method of any one of claims 60-62, wherein the concentration of the one or more EVs isat about 1 billion to about 40 billion EVs per ml of the composition.

64. The method of any one of claims 1-63, wherein the one or more EVs have an averagediameter of about 30 nm to about 170 nm.

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

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

67. The method of claim 65 or 66, wherein the total protein concentration is measured byELISA.

68. The method of any one of claims 4-67, wherein the composition comprises TIMP-1 (tissueinhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (MacrophageAttorney Docket No.66309-729.601 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.

69. The method of any one of claims 4-68, 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, CD109 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 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (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.

70. The method of any one of claims 1 or 5-69, wherein the composition further comprises oneor more of COL2A1, COL3A1, COL5A1, COL5A2, COL6A1, COL11A1, or COL14A1, or a combination of two or more thereof.

71. The method of any one of claims 1-4 or 7-70, 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.

72. The method of any one of claims 1-4 or 7-71, wherein the composition further compriseshsa-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-Attorney Docket No.66309-729.601 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.

73. The method of any one of claims 1-6 or 8-72, wherein at least 80% of the one or more EVsare CD63+, CD9-, CD81-.

74. Use of a composition comprising a mesenchymal stem cell (MSC) secretome in treatingtraumatic brain injury.

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