Effect Of Mild Moxibustion Combined With Bone Marrow Mesenchymal Stem Cell Transplantation On The Structure And Function Of Injured Anal Sphincter In Rat

Objective: The Zutshi anal sphincter complex injury model was used to observe the repair effect of different stem cell injection methods on the wound surface,and then explore the effect of mild moxibustion combined with bone marrow mesenchymal stem cell transplantation on the structure and function of injured anal sphincter in rats.Methods: 96 SD rats were randomly allocated into mild moxibustion + local injection group(group A),mild moxibustion + intravenous injection group(group B),local injection group(group C),intravenous injection group(group D)and mild moxibustion group(group E).Zutshi rat anal sphincter complex injury model was used in all rats.After successful modeling,all groups were treated with mild moxibustion for 15 min or GFPpositive bone marrow mesenchymal stem cell transplantation(local and tail vein)for 7 days.All cats were undergone anal pressure examination and anal sphincter electromyography(EMG)on 7th,14 th,and 28 th day after sphincterectomy.Histological examination of the anal sphincter(immunofluorescence,Masson staining,HE staining)was performed on the 28 th postoperative day.Immunofluorescence(?-SMA or Desmin)was used to locate GFP-positive BMSCs in rat sphincter wounds.Masson staining was used to compare the content of new muscle tissue and collagen fibers between the groups.HE staining was used to observe the content of new blood vessels in the wounds of each group.Results: 1.On the 28 th day after sphincterectomy,new muscle fibers in group A was significantly higher than that in other groups(P<0.05),while collagen fibers in group C and group A were lower than other groups,but there was no significant difference between them(P>0.05).2.On the 28 th day after sphincterectomy,the GFP fluorescence intensity of group A was the strongest,suggesting that more BMSCs were colonized on the wound surface,but no fluorescence colocalization was found.3.On 7th,14 th day after surgery,the resting pressure of group A was significantly higher than that of group B,group D and group E(P<0.05);The difference between group A and group C in resting pressure was statistically significant(P<0.05)until 28 days after surgery.At 14 and 28 days after surgery,the peak contraction pressure of group A was significantly higher than that of the other groups(P<0.05).4.At 7 days postoperatively,there was no significant difference in amplitude and frequency between groups.At 14 days after surgery,the amplitude of group A and group C were significantly higher than that of the other groups(P<0.05),but there was no significant difference between them.The frequency of group A was significantly higher than that of other groups from 14 days to 28 days after surgery(P<0.05).The amplitude of group A was significantly higher than that of group C until 28 days after sphincterectomy(P<0.05).Conclusion: In this study,the combined application of mild moxibustion and different bone marrow mesenchymal stem cell injection methods in the treatment of rat sphincter injury model was studied,and it was found that 15 min mild moxibustion combined with local injection of BMSC can significantly promote sphincter regeneration and angiogenesis,and accelerate the recovery of anal pressure and sphincter electromyography.The combined application of mild moxibustion and local injection of BMSC can accelerate the recovery of the structure and function of the injured anal sphincter in rats,providing a direction for clinical mild moxibustion combined with stem cells in the treatment of myogenous fecal incontinence.