Menstrual Blood Stem Cells Combined With Estrogen-RGD Gel Scaffold To Prevent Intrauterine Adhesion In Rats And Its Mechanism

Objective: Uterine cavity adhesion refers to the partial or complete occlusion of the uterine cavity due to endometrial injury.Surgical removal of adhesions and barriers are routine clinical treatments,but there are problems with poor prognosis and high recurrence rates.In recent years,a breakthrough has been made in the research of stem cell transplantation for the treatment of intrauterine adhesions.Menstrual blood stem cells are adult stem cells derived from female menstrual blood.Our previous studies have proved that autologous menstrual blood stem cell transplantation can improve the endometrial damage of patients with severe intrauterine adhesions,but the problem of low utilization of transplanted cells during the treatment process remains to be solved.Some studies have shown that presenting cells through three-dimensional cell scaffolds can significantly increase the cell planting rate,prolong the survival time of cells in vivo,and exert their therapeutic effects.RGD gel scaffold is a synthetic tunable hydrogel with good biocompatibility and can provide a suitable microenvironment for cell survival.This experiment intends to use estrogen-RGD gel stent to encapsulate menstrual blood stem cells and implant the rat endometrial injury model by intrauterine perfusion to prevent uterine adhesions and observe the effects of menstrual blood stem cells on rat endometrial injury Therapeutic effect,explore its therapeutic mechanism.Methods: In vitro experiments,using live and dead cell staining,cell counting,acetylcholinesterase activity detection,immunofluorescence and other techniques to clarify the preparation conditions of the estrogen-RGD gel culture system and its biocompatibility with menstrual blood stem cells.In vivo experiment,establish rat endometrial injury model by mechanical curettage,use menstrual blood stem cells combined with estrogen-RGD gel stent transplantation treatment,and take samples 18 days after treatment,through tissue section staining,immunofluorescence,Realtime-PCR and other techniques To evaluate the therapeutic effect of combined transplantation on damaged endometrium in rat models.Results: 1.Menstrual blood stem cells can adhere to the RGD gel scaffold and maintain cell viability;estrogen can promote the proliferation of menstrual blood stem cells(P=0.0140),increase their secretion capacity(P=0.0326),and promote menstrual blood stem cells to the endometrial epithelium Cell Differentiation.2.Intrauterine perfusion of menstrual blood stem cells combined with RGD gel can improve the colonization of cells in the uterus and limit the systemic migration of menstrual blood stem cells.3.Transplantation of menstrual blood stem cells combined with estrogen-RGD gel stent can increase the thickness of the endometrium in rats with endometrial injury and the number of endometrial glands,and reduce the area of endometrial fibrosis,up-regulate the expression of endometrial keratin and proliferating cell nuclear antigen.4.After transplantation of menstrual blood stem cells combined with estrogen-RGD gel scaffold,the expression of IL-2,IL-6,IFN-γ,and TNF-α in the endometrial tissue was significantly reduced.Conclusion: 1.The estrogen-RGD gel system can provide suitable microcircles for the survival of menstrual blood stem cells.2.Intrauterine perfusion of menstrual blood stem cells combined with RGD gel can improve the local action efficiency of menstrual blood stem cells in uterine tissue.3.Transplantation of menstrual blood stem cells combined with estrogen-RGD gel stent can promote rat endometrial cell proliferation,promote gland regeneration,reduce endometrial fibrosis,restore uterine cavity shape,and prevent uterine adhesions.4.Transplantation of menstrual blood stem cells combined with estrogen-RGD gel stent can improve endometrial inflammation and promote endometrial repair.