Chronic wounds including diabetic foot ulcers are clinical emergencies that need careful management. Exosomes from human adipose-derived mesenchymal stem cells (hADSCs-Ex) are a new promising cell-free therapy for the regeneration of dermal wounds. We established a delayed wound healing model using diabetic female mice. A 1.5 cm2 full-thickness cutaneous wound was made ventrally in 6-week-old db/db mice. After treatment with phosphate-buffered saline, recombinant human epidermal growth factor, hADSCs-CM, or hADSCs-Ex three times a day for 2 weeks, we measured wound healing closure rates and performed histological analysis. Human dermal fibroblasts (WS1) were evaluated by PKH26-Exo co-localization test, CCK-8 test, cell scratch test, and the transwell test, while the expression of matrix metalloproteinase-1 (MMP1), MMP3, Collagen I, and Collagen III were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Wound closure and re-epithelialization were accelerated by hADSCs-Ex. Besides, hADSCs-Ex enhanced skin collagen production, angiogenesis, cell proliferation, inhibited apoptosis, promoted skin barrier function repair, and reduced inflammation in skin lesions. Furthermore, negative regulation of MMP1 and MMP3 enhanced collagen synthesis wound healing-promoting effects of hADSCs-Ex. hADSCs-Ex treatment for diabetic wounds provided a novel cell-free therapeutic strategy.
Get full access to this article
View all access options for this article.
References
1.
ZhaoR, LiangH, ClarkeE, JacksonC and XueM. (2016). Inflammation in chronic wounds. Int J Mol Sci, 17:2085.
2.
BasiriR, SpicerMT, LevensonCW, OrmsbeeMJ, LedermannT and ArjmandiBH. (2020). Nutritional supplementation concurrent with nutrition education accelerates the wound healing process in patients with diabetic foot ulcers. Biomedicines, 8:263.
3.
NelsonEA and AdderleyU. (2016). Venous leg ulcers. BMJ Clin Evid, 2016:1902.
4.
MervisJS and PhillipsTJ. (2019). Pressure ulcers: prevention and management. J Am Acad Dermatol, 81:893–902.
5.
YaoB, HuangS, GaoD, XieJ, LiuN and FuX. (2016). Age-associated changes in regenerative capabilities of mesenchymal stem cell: impact on chronic wounds repair. Int Wound J, 13:1252–1259.
6.
TsourdiE, BarthelA, RietzschH, ReichelA and BornsteinSR. (2013). Current aspects in the pathophysiology and treatment of chronic wounds in diabetes mellitus. Biomed Res Int, 2013:385641.
7.
OlssonM, JarbrinkK, DivakarU, BajpaiR, UptonZ, SchmidtchenA and CarJ. (2019). The humanistic and economic burden of chronic wounds: a systematic review. Wound Repair Regen, 27:114–125.
8.
EverettE and MathioudakisN. (2018). Update on management of diabetic foot ulcers. Ann N Y Acad Sci, 1411:153–165.
9.
HingoraniA, LaMuragliaGM, HenkeP, MeissnerMH, LoretzL, ZinszerKM, DriverVR, FrykbergR, CarmanTL, et al. (2016). The management of diabetic foot: a clinical practice guideline by the Society for Vascular Surgery in collaboration with the American Podiatric Medical Association and the Society for Vascular Medicine. J Vasc Surg, 63:3S–21S.
10.
Nik HisamuddinNAR, Wan Mohd ZahiruddinWN, Mohd YazidB and RahmahS. (2019). Use of hyperbaric oxygen therapy (HBOT) in chronic diabetic wound—a randomised trial. Med J Malaysia, 74:418–424.
11.
BorysS, HohendorffJ, FrankfurterC, Kiec-WilkB and MaleckiMT. (2019). Negative pressure wound therapy use in diabetic foot syndrome-from mechanisms of action to clinical practice. Eur J Clin Invest, 49:e13067.
12.
CaoY, GangX, SunC and WangG. (2017). Mesenchymal stem cells improve healing of diabetic foot ulcer. J Diabetes Res, 2017:9328347.
13.
MoonKC, SuhHS, KimKB, HanSK, YoungKW, LeeJW and KimMH. (2019). Potential of allogeneic adipose-derived stem cell-hydrogel complex for treating diabetic foot ulcers. Diabetes, 68:837–846.
14.
YangJ, ChenZ, PanD, LiH and ShenJ. (2020). Umbilical cord-derived mesenchymal stem cell-derived exosomes combined Pluronic F127 hydrogel promote chronic diabetic wound healing and complete skin regeneration. Int J Nanomedicine, 15:5911–5926.
15.
MathewSA, NaikC, CahillPA and BhondeRR. (2020). Placental mesenchymal stromal cells as an alternative tool for therapeutic angiogenesis. Cell Mol Life Sci, 77:253–265.
16.
TsuchiyaA, KojimaY, IkarashiS, SeinoS, WatanabeY, KawataY and TeraiS. (2017). Clinical trials using mesenchymal stem cells in liver diseases and inflammatory bowel diseases. Inflamm Regen, 37:16.
17.
OhtaH, LiuX and MaedaM. (2020). Autologous adipose mesenchymal stem cell administration in arteriosclerosis and potential for anti-aging application: a retrospective cohort study. Stem Cell Res Ther, 11:538.
18.
ShenT, ZhengQQ, ShenJ, LiQS, SongXH, LuoHB, HongCY and YaoK. (2018). Effects of adipose-derived mesenchymal stem cell exosomes on corneal stromal fibroblast viability and extracellular matrix synthesis. Chin Med J (Engl), 131:704–712.
19.
PhinneyDG and PittengerMF. (2017). Concise review: MSC-derived exosomes for cell-free therapy. Stem Cells, 35:851–858.
20.
HuL, WangJ, ZhouX, XiongZ, ZhaoJ, YuR, HuangF, ZhangH and ChenL. (2016). Exosomes derived from human adipose mensenchymal stem cells accelerates cutaneous wound healing via optimizing the characteristics of fibroblasts. Sci Rep, 6:32993.
21.
GolchinA, HosseinzadehS and ArdeshirylajimiA. (2018). The exosomes released from different cell types and their effects in wound healing. J Cell Biochem, 119:5043–5052.
22.
SzatanekR, Baj-KrzyworzekaM, ZimochJ, LekkaM, SiedlarM and BaranJ. (2017). The methods of choice for extracellular vesicles (EVs) characterization. Int J Mol Sci, 18:1153.
23.
Marshall CB. (2016). Rethinking glomerular basement membrane thickening in diabetic nephropathy: adaptive or pathogenic?. Am J Physiol Renal Physiol, 311:F831–F843.
24.
SumerS, Aktug DemirN, KölgelierS, Cagkan InkayaA, ArpaciA, Saltuk DemirL and UralO. (2013). The clinical significance of serum apoptotic cytokeratin 18 neoepitope M30 (CK-18 M30) and matrix metalloproteinase 2 (MMP-2) levels in chronic hepatitis B patients with cirrhosis. Hepat Mon, 13:e10106.
25.
RezvanfarMA, Shojaei SaadiHA, GoosheM, AbdolghaffariAH, BaeeriM and AbdollahiM. (2014). Ovarian aging-like phenotype in the hyperandrogenism-induced murine model of polycystic ovary. Oxid Med Cell Longev, 2014:948951.
26.
WangZY and QiuBF. (2010). Increased expression of matrix metalloproteinase-1 and 3 in remission patients of steroid-dependent ulcerative colitis. Gastroenterology Res, 3:120–124.
27.
OrioliD and DellambraE. (2018). Epigenetic regulation of skin cells in natural aging and premature aging diseases. Cells, 7:268.
28.
ZhouY, ZhaoB, ZhangXL, LuYJ, LuST, ChengJ, FuY, LinL, ZhangNY, et al. (2021). Combined topical and systemic administration with human adipose-derived mesenchymal stem cells (hADSC) and hADSC-derived exosomes markedly promoted cutaneous wound healing and regeneration. Stem Cell Res Ther, 12:257.
29.
KobayashiH, EbisawaK, KambeM, KasaiT, SugaH, NakamuraK, NaritaY, OgataA and KameiY. (2018). Editors' choice effects of exosomes derived from the induced pluripotent stem cells on skin wound healing. Nagoya J Med Sci, 80:141–153.
30.
GriceEA, SnitkinES, YockeyLJ, BermudezDM, LiechtyKW and SegreJA. (2010). Longitudinal shift in diabetic wound microbiota correlates with prolonged skin defense response. Proc Natl Acad Sci U S A, 107:14799–14804.
31.
LiuF, ChenDD, SunX, XieHH, YuanH, JiaW and ChenAF. (2014). Hydrogen sulfide improves wound healing via restoration of endothelial progenitor cell functions and activation of angiopoietin-1 in type 2 diabetes. Diabetes, 63:1763–1778.
32.
McLainJM, AlamiWH, GlovakZT, CooleyCR, BurkeSJ, CollierJJ, BaghdoyanHA, KarlstadMD and LydicR. (2018). Sleep fragmentation delays wound healing in a mouse model of type 2 diabetes. Sleep, 41:zsy156.
33.
ShabbirA, CoxA, Rodriguez-MenocalL, SalgadoM and Van BadiavasE. (2015). Mesenchymal stem cell exosomes induce proliferation and migration of normal and chronic wound fibroblasts, and enhance angiogenesis in vitro. Stem Cells Dev, 24:1635–1647.
34.
ZhangJ, GuanJ, NiuX, HuG, GuoS, LiQ, XieZ, ZhangC and WangY. (2015). Exosomes released from human induced pluripotent stem cells-derived MSCs facilitate cutaneous wound healing by promoting collagen synthesis and angiogenesis. J Transl Med, 13:49.
35.
WuP, ZhangB, ShiH, QianH and XuW. (2018). MSC-exosome: a novel cell-free therapy for cutaneous regeneration. Cytotherapy, 20:291–301.
36.
JingH, HeX and ZhengJ. (2018). Exosomes and regenerative medicine: state of the art and perspectives. Transl Res, 196:1–16.
37.
RillaK, MustonenAM, ArasuUT, HarkonenK, MatilainenJ and NieminenP. (2019). Extracellular vesicles are integral and functional components of the extracellular matrix. Matrix Biol, 75–76:201–219.
38.
BaileyAJM, LiH, KirkhamAM, TieuA, MagantiHB, ShorrR, FergussonDA, LaluMM, ElomazzenH and AllanDS. (2021). MSC-derived extracellular vesicles to heal diabetic wounds: a systematic review and meta-analysis of preclinical animal studies. Stem Cell Rev Rep [Epub ahead of print]; DOI: 10.1007/s12015-021-10164-4.
39.
PanBH, ZhangQ, LamCH, YuenHY, KuangS and ZhaoX. (2021). Petite miracles: insight into the nano-management of scarless wound healing. Drug Discov Today [Epub ahead of print]; DOI: 10.1016/j.drudis.2021.01.025.
40.
FurutaT, MiyakiS, IshitobiH, OguraT, KatoY, KameiN, MiyadoK, HigashiY and OchiM. (2016). Mesenchymal stem cell-derived exosomes promote fracture healing in a mouse model. Stem Cells Transl Med, 5:1620–1630.
41.
HanX, WuP, LiL, SahalHM, JiC, ZhangJ, WangY, WangQ, QianH, ShiH and XuW. (2021). Exosomes derived from autologous dermal fibroblasts promote diabetic cutaneous wound healing through the Akt/beta-catenin pathway. Cell Cycle, 20:616–629.
42.
XiaoS, XiaoC, MiaoY, WangJ, ChenR, FanZ and HuZ. (2021). Human acellular amniotic membrane incorporating exosomes from adipose-derived mesenchymal stem cells promotes diabetic wound healing. Stem Cell Res Ther, 12:255.
43.
LiuW, YuM, XieD, WangL, YeC, ZhuQ, LiuF and YangL. (2020). Melatonin-stimulated MSC-derived exosomes improve diabetic wound healing through regulating macrophage M1 and M2 polarization by targeting the PTEN/AKT pathway. Stem Cell Res Ther, 11:259.
44.
LiX, XieX, LianW, ShiR, HanS, ZhangH, LuL and LiM. (2018). Exosomes from adipose-derived stem cells overexpressing Nrf2 accelerate cutaneous wound healing by promoting vascularization in a diabetic foot ulcer rat model. Exp Mol Med, 50:1–14.
45.
LiH, ZhuH, GeT, WangZ and ZhangC. (2021). Mesenchymal stem cell-based therapy for diabetes mellitus: enhancement strategies and future perspectives. Stem Cell Rev Rep, 10:1–18.
46.
BornLJ, ChangKH, ShoureshiP, LayF, BengaliS, HsuATW, AbadchiSN, HarmonJW and JaySM. (2021). HOTAIR-loaded mesenchymal stem/stromal cell extracellular vesicles enhance angiogenesis and wound healing. Adv Healthc Mater [Epub ahead of print]; DOI: 10.1002/adhm.202002070.
47.
XuY, OuyangL, HeL, QuY, HanY and DuanD. (2020). Inhibition of exosomal miR-24-3p in diabetes restores angiogenesis and facilitates wound repair via targeting PIK3R3. J Cell Mol Med, 24:13789–13803.
48.
MaarofM, ChowdhurySR, SaimA, Bt Hj IdrusR and LokanathanY. (2020). Concentration dependent effect of human dermal fibroblast conditioned medium (DFCM) from three various origins on keratinocytes wound healing. Int J Mol Sci, 21:2929.
49.
CaiY, LiJ, JiaC, HeY and DengC. (2020). Therapeutic applications of adipose cell-free derivatives: a review. Stem Cell Res Ther, 11:312.
50.
BroughtonG, 2nd, JanisJE and AttingerCE. (2006). Wound healing: an overview. Plast Reconstr Surg, 117:1e-S–32e-S.
51.
HanG and CeilleyR. (2017). Chronic wound healing: a review of current management and treatments. Adv Ther, 34:599–610.
52.
DaLC, HuangYZ and XieHQ. (2017). Progress in development of bioderived materials for dermal wound healing. Regen Biomater, 4:325–334.
53.
Alonso-AlonsoML, Garcia-PosadasL and DieboldY. (2021). Extracellular vesicles from human adipose-derived mesenchymal stem cells: a review of common Cargos. Stem Cell Rev Rep [Epub ahead of print]; DOI: 10.1007/s12015-021-10155-5.
54.
ViswanathanS, ShiY, GalipeauJ, KramperaM, LeblancK, MartinI, NoltaJ, PhinneyDG and SensebeL. (2019). Mesenchymal stem versus stromal cells: International Society for Cell & Gene Therapy (ISCT®) Mesenchymal Stromal Cell committee position statement on nomenclature. Cytotherapy, 21:1019–1024.
55.
LötvallJ, HillAF, HochbergF, BuzásEI, Di VizioD, GardinerC, GhoYS, KurochkinIV, MathivananS, et al. (2014). Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles. J Extracell Vesicles, 3:26913.
56.
ZhangC, ZhouL, WangZ, GaoW, ChenW, ZhangH, JingB, ZhuX, ChenL, et al. (2021). Eradication of specific donor-dependent variations of mesenchymal stem cells in immunomodulation to enhance therapeutic values. Cell Death Dis, 12:357.
57.
AnT, ChenY, TuY and LinP. (2021). Mesenchymal stromal cell-derived extracellular vesicles in the treatment of diabetic foot ulcers: application and challenges. Stem Cell Rev Rep, 17:369–378.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
