| ObjectiveTo detect the repair of endometrial damage and the improvement of fibrosis in the endometrium of mice after in situ transplantation of micro-ballooned umbilical cord mesenchymal stem cells(3D-h UC-MSCs),and to explore the role and possible mechanism of 3D-h UC-MSCs in repairing endometrial damage in mice.Methods1: Microencapsulated umbilical cord mesenchymal stem cells(3D-h UC-MSCs)were formed using the laboratory’s own high pressure electrostatic spray device,cultured in medium.2: A mouse endometrial injury model was constructed with 95% alcohol and microencapsulated 3D-h UC-MSCs were transplanted in situ to observe their repairing effect on endometrial injury in mice.3: The mice were divided into four groups: control group,model group,2D-h UC-MSCs group and 3D-h UC-MSCs group.The control group was not treated,the model group was with saline after moulding,the 2D-h UC-MSCs group was transplanted with unencapsulated 2D-h UC-MSCs in situ after moulding,and the3D-h UC-MSCs group was transplanted with 3D-h UC-MSCs after moulding.The uterus of the mice were collected after 7 days of treatment and the endometrial thickness and number of glands were observed by HE staining.4: Detection of endometrial fibrosis in each group of mice.masson Staining experiment was used to observe the area of endometrial fibrosis,α-SMA and type I collagen immunohistochemistry was used to observe the fibrosis of endometrium in mice.5: Detection of proteins related to the TGF-β1/smad pathway: TGF-β1,smad,p-smad3 protein expression using western blotting.6: Observe the ability for pregnancy in each group of mice.Mice in each group were caged together after 14 days of treatment to observe the pregnancy rate and number of embryos in each group.Results1: Microscopically,the 3D-h UC-MSCs formed by the high-pressure electrostatic spray device showed clump-like growth with regular morphology and appropriate size of the microballoons,and the cells were more evenly distributed within the balloons.2:In this experiment,IUA model mice were successfully constructed using 95%alcohol.From the overall appearance,the uterus on the moulded side of the mice became thinner and the overall structure was not homogeneous after moulding.From the results of HE staining,the endometrium of the moulded mice became thinner and partially shed,and the number of glands was reduced.3:3D-h UC-MSCs enhanced the repair of IUA.In terms of appearance,the uterus of3D-h UC-MSCs treated mice had a restored appearance and smooth structure,which was closer to the control group.From the results of HE staining,the endometrial thickness was further thickened and the number of glands was further increased in the3D-h UC-MSCs group,which was closer to that of the control group,and the difference was statistically significant(P < 0.05).4:3D-h UC-MSCs could better improve the fibrosis of the endometrium in IUA mice.From the results of masson staining,the area of fibrosis in the uterus of mice treated with 3D-h UC-MSCs was reduced compared with that treated with2D-h UC-MSCs,and the difference was statistically significant(P < 0.05).From the results of immunohistochemistry,the expression of α-SMA and type I collagen was lower in the 3D-h UC-MSCs group compared with the 2D-h UC-MSCs group,and the difference was statistically significant(P < 0.05).5:3D-h UC-MSCs reduced the expression of TGF-β1/smad proteins in mouse uterus.From the western blotting results,the expression of TGF-β1 and phosphorylated smad3 protein was lower in both 2D-h UC-MSCs and 3D-h UC-MSCs treatment than in the modeling group,and the expression of TGF-β1,phosphorylated smad3 protein was lower in 3D-h UC-MSCs treatment than in 2D-h UC-MSCs treatment,and the difference was statistically significant(P < 0.05).6:3D-h UC-MSCs further improved the function of the mouse uterus.From the results of the number of implanted embryos,the number of embryos increased in both2D-h UC-MSCs and 3D-h UC-MSCs treatment compared to the model group,and more embryos implanted after 3D-h UC-MSCs treatment,and the difference was more statistically significant(P < 0.05).Conclusions: Intrauterine infusion of microencapsulated umbilical cord mesenchymal stem cells can repair damaged endometrium via the TGF-β1/smad pathway. |
PDF Full Text Request