| Background:Non-obstructive azoospermia(non-obstructive azoospermia,NOA)accounts for about 1-2%of the male population and up to 60%of the male azoospermia population.Current treatments for non-obstructive azoospermia include drug therapy and Microdissection Testicular sperm extraction(Micro-TESE).The effect of drug treatment NOA is extremely low,only for a small number of low gonadotropin hypogonadism(Hypogonadotropic hypogonadism,HH)caused by the NOA effect is obvious.Nearly half of the patients treated with Micro-TESE still cannot find sperm.In recent years,stem cell transplantation has become a new research direction for the treatment of non-obstructive azoospermia.However,after stem cell transplantation,the retention time in the tissue is insufficient,and it is easy to be lost to the surrounding organs or tissues,so that the role of stem cells is significantly reduced.Collagen is one of the most abundant and easily available protein in animals.It has the characteristics of low antigenicity,high biocompatibility and easy degradation,and is widely used in tissue engineering research.In stem cell transplantation research,collagen as a biological scaffold plays a fixed role in stem cell transplantation,which can enhance the therapeutic effect of transplanted stem cells to some extent.At present,the treatment method of stem cell combined with collagen stent transplantation has been explored in the fields of myocardial repair,ovarian premature failure treatment and so on,and achieved good results.Objective:To investigate the effect of collagen scaffolds on improving spermatogenic function of mesenchymal stem cells(MSCs)in mice with non-obstructive azoospermia.Methods:1.Mice azoospermia model was established using BusulfanThe Balb/c male mice were intraperitoneally injected with 30 mg/kg、40mg/kg of two doses of 2%solution,and the testis and epididymis were sampled at the 4th,8th and 12th weeks after injection for pathological analysis.Based on the pathological results and the presence of azoospermia in the epididymis,it was proved that the stable mouse azoospermia model could be successfully established,which was used as the basis for the subsequent modeling to find the appropriate white elimination dose.2.To compare the therapeutic effects of MSCs with collagen and MSCs in mouse testis transplantation.Fifty male mice were given 30mg/kg intraperitoneal injection of busulfan to establish the model.After 4 weeks,the 46 surviving mice were divided into 4 groups.In the MSCs group,12 mice were injected with 40ul phosphate solution(PBS)and MSCs mixed suspension in the testis.Twelve mice in the MSCs and collagen group were injected with the mixed suspension of 40ulMSCs and collagen in the testis of each mouse.In the collagen scaffold group,11 mice were injected with a mixture of 40ul phosphate solution(PBS)and collagen suspension in the testis of each mouse.In the control group(PBS),11 mice were injected with 40ulPBS solution into their testicles.Another five mice were treated as blank groups.The transplanted mice were fed for 8 weeks,and then the testis and epididymis were removed for pathological analysis.Results:1.The Johnsen score of testicular pathology in MSCs group was higher than that in PBS group and scaffold group(p<0.05)(p<0.05).The MSCs+collagen group was higher than the MSCs group(p<0.05),and higher than the PBS group and the scaffold group(p<0.05)(p<0.05).The blank group was superior to the MSCs+collagen scaffold group(p<0.05).2.The number of spermatogenic cell layers in the MSCs group was higher than that in the scaffold group and the PBS group(p<0.05)(p<0.05).The MSCs+collagen group was more than the MSCs group(p<0.05).MSCs+collagen group was higher than PBS group and scaffold group(p<0.05)(p<0.05).The blank group was superior to the MSCs+collagen scaffold group(p<0.05).3.Under 400x lens,the number and motility of sperm in epididymis of MSCs+collagen scaffold group were better than that of MSCs group,PBS group and scaffold group.The number of epididymal sperm in the blank group was higher than that in the MSCs+collagen scaffold group.Conclusion:This experiment found that stem cells combined with collagen scaffold transplantation can improve the spermatogenesis function of mice better than MSCs transplantation alone. |
PDF Full Text Request