Umbilical cord mesenchymal stem cells sheets treated by small intestinal submucosa promote wound healing
Objective To explore the effect of extracellular matrix(ECM),especially the small intestinal submucosa(SIS)-treated umbilical cord mesenchymal stem cells(UC-MSCs)sheets on wound healing.Methods Primary UC-MSCs were isolated and cultured from umbilical cord tissues using the tissue block apposition method.Decellularized SIS was obtained using a chemical method,and recombinant human fibronectin(rFN),SIS,and Matrigel were immo-bilized on the surface of polystyrene(PS)cell culture dishes by physical adhesion.The effects of PS surface,rFN-PS surface,SIS-PS surface,and Matrigel-PS surface on the morphology of UC-MSCs were investigated,and the changes in gene expression were detected by RT-PCR,then changes in the secretion of extracellular vesicles(EVs)were detec-ted using nanoparticle tracking analysis.Rat skin total excision wounds were treated with ECM-free,rFN-treated,SIS-treated,and Matrigel-treated UC-MSCs cell sheets,respectively,and the therapeutic effect was further verified by cal-culating the ratio of wound area to the original wound area.Results Compared with the PS surface,the morphology of UC-MSCs cultured on the SIS-PS surface was almost unaltered(P>0.05),and RT-PCR assay showed that gene expres-sion of the SIS-PS surface-cultured UC-MSCs was enhanced,such as Nanog homeobox(NANOG,P=0.041),SRY-box transcription factor 2(SOX-2,P=0.009),octamer-binding transcription factor-4(OCT-4,P<0.001),interleukin-10(IL-10,P=0.049),indoleamine 2,3-dioxygenase(IDO,P=0.007),and transforming growth factor beta(TGF-β,P=0.046),meanwhile the ability of the SIS-PS surface-cultured UC-MSCs to secrete EVs was significantly in-creased(P<0.001).The results of the animal experiments revealed that the lowest percentage of wound to original wound area was 26.9%±6.1%in the group of animals treated with SIS-treated UC-MSCs cell sheets at day 7.Conclu-sion UC-MSCs cultured on the SIS-PS surface significantly improved immunosuppressive function and EVs secretion ability,and the corresponding cell sheets have a stronger ability to promote wound healing.
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