Background Severe injuries of the uterus may trigger uterine scar formation, ultimately leading to infertility or obstetrical complications. Scale bars, 150?m … UC-MSCs communicate specific surface antigens and possess multi-lineage differentiation potential According to the characteristics of MSCs defined from the International Society for Cellular Therapy [43], UC-MSCs were checked for adherence to plastic, specific surface antigen manifestation and multipotent differentiation potential. After culturing human being umbilical cord cells for 14?days, spindle-shaped adherent cells were apparent (Fig.?3a). These cells were positive for buy 24144-92-1 CD29, CD44, CD73, CD90 and CD105, and were negative for CD34, CD45 and HLA-DR (Fig.?3bCk). Moreover, these cells displayed the capacity to differentiate into adipocytes, osteoblasts and neural-like cells after induction in vitro (Fig.?3?lCo), indicating their multi-lineage differentiation potential. Fig. 3 UC-MSCs express specific surface antigens and possess multi-lineage differentiation potential. a Morphology of human being UC-MSCs. Scale bars, 30?m. b-k Flow cytometry analysis of immune-markers in human being UC-MSCs. l-o Differentiation assays … Scaffolds promote the long-term retention of UC-MSCs in uterine scars At day time 30 post-transplantation, labelled UC-MSCs were found to primarily spread in the stroma of the scarred uterine walls. Significantly more labelled UC-MSCs were observed in the stroma of the scaffold/UC-MSCs group than in the UC-MSCs group (Fig.?4a, b). Moreover, the CM-Dil-labelled UC-MSCs were positive for vimentin, a signature marker Rabbit Polyclonal to RBM16 for MSCs (Fig.?4cCf). The number of cells positive for CM-Dil and vimentin in the scaffold/UC-MSCs group (10.67??1.67) was significantly higher than that in the UC-MSCs group (2.83??0.75, phosphate-buffered … To assess the fibrosis in uterine scars, Massons trichrome staining was performed. At day time 30 post-transplantation, uterine scars in the PBS group, the scaffold group and the UC-MSCs group showed abundant collagen deposition and a massive loss of native cells. However, the scaffold/UC-MSCs group experienced obvious collagen degradation and apparent regenerated endometrial glands and muscle mass bundles (Fig.?6a). buy 24144-92-1 At day time 60 post-transplantation, uterine scars in the PBS group, the scaffold group and the UC-MSCs group did not show apparent collagen degradation compared with day time 30 post-transplantation. However, collagen fibres buy 24144-92-1 in the scaffold/UC-MSCs group further decreased; while the endometrium and myometrium regenerated (Fig.?6a). Fig. 6 Scaffold/UC-MSCs transplantation facilitates collagen degradation in uterine scars via upregulation of MMP-9. a Massons trichrome staining of uterine scars at days 30 and 60 post-transplantation in the PBS group, the scaffold group, the UC-MSCs … In the uterus, collagen degradation primarily entails matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) [44, 45]. MMP-9 manifestation was recognized in the endometrial stroma. At day time 30 post-transplantation, the number of MMP-9-positive cells in the scaffold/UC-MSCs group was significantly higher than that in the additional three organizations (Fig.?6b). At day time 60 post-transplantation, although improved in all organizations compared with day time 30 post-transplantation, the number of MMP-9-positive cells in the scaffold/UC-MSCs group (25.96??3.63) remained higher than the PBS group (8.19??1.61, indicate buy 24144-92-1 implanted fetuses … Table 1 Assessment of reproductive results among different treatments at day time 60 post-transplantationa Conversation In the present study, we shown for the first time the scaffold/UC-MSCs system facilitated collagen degradation, full-thickness regeneration and fertility repair in uterine scars; and the underlying mechanism of which might rely on the long-term effect of UC-MSCs in vivo. In uterine scars, collagen deposition is a major clinical problem, which impedes the proliferation, differentiation and migration of uterine native cells. Efficient collagen buy 24144-92-1 degradation in uterine scars treated with scaffold/UC-MSCs compared to uterine scars treated with PBS, scaffolds or UC-MSCs was demonstrated in the present study. In the early stage of postmenstrual repair of the endometrium, which represents the only example of cyclic scar-free repair in adult human tissue, expression of MMPs, including MMP-2 and MMP-9 is elevated both in vivo and in vitro, suggesting the essential role of MMPs in preventing the formation of uterine scars [44]. In our study, the substantial increase in MMP-9 expression induced by the scaffold/UC-MSCs construct in uterine scar tissues, which was validated by immunohistochemistry and immunofluorescence staining, might represent a possible explanation for the reduced collagen deposition. Moreover, we found that the co-culture with degradable collagen fibres promoted the production and secretion of MMP-9 by UC-MSCs in vitro compared with the monolayer culture. This is inconsistent with our previous observation of an increased expression of paracrine factors in adipose-derived mesenchymal stem cells co-cultured with degradable collagen fibres compared with the monolayer culture [41]. When mixed with degradable collagen fibres, UC-MSCs interacted with collagen fibres to form a 3D microenvironment, which provided a suitable niche for UC-MSCs to anchor, migrate and function. Besides increased collagen degradation, facilitated regeneration of endometrial endometrium, glandular epithelium, myometrium and blood vessels was also observed in the scaffold/UC-MSCs-treated uterine scars. In the process of endometrial cyclic regeneration, large quantities of cytokines, such as FGF2.