Research of Using Silk-fibroin Loading Rabbit Adipose Stem Cells to Construct Apparatus of Urinary Diversion / 华中科技大学学报(医学版)

ABSTRACT

Objective The aim of this research was to produce apparatus of urinary diversion using silk-fibroin loading rab-bit adipose stem cells,and assess the effect of urinary diversion in a rabbit model.Methods Adipose stem cells were obtained and cultured in vitro,and flow cytometry analysis was performed to determine the adipose stem cells.These cultured adipose stem cells were used to seed on the silk-fibroin scaffolds,and after being incubated in the conditioned medium for 7 days,the a-bove compounds were made into apparatus of urinary diversion.This apparatus of urinary diversion was implanted into 20 rab-bits with radical cystectomy to develop urinary diversion.Five rabbits from each experimental group were euthanized at the spe-cific time points(1,2,3,4 months postoperatively),and the implants were harvested for histological and immunohistochemical a-nalysis.In the control group,silk-fibroins with unseeded cells(only silk-protein scaffolds)were also made into apparatus of uri-nary diversion and then used as urinary diversion on another 5 rabbits with the same process.Results Rabbits adipose stem cells were isolated and cultured successfully,and determined by flow cytometry.The silk-fibroin scaffolds were synthesized suc-cessfully.All rabbits were alive in the experimental group until the time of sacrifice.Histological and immunohistochemical anal-ysis showed multilayer uroepithelium coverage in the luminal surface of apparatus of urinary diversion,and as time went on,epi-thelial layers increased continuously.In the control group,all animals were dead within 3 weeks,and urine leakage,severe in-flammatory reaction and tissue destruction were found by autopsy.Conclusion The present experiment has successfully used silk-fibroin loading rabbit adipose stem cells to construct apparatus of urinary diversion,and demonstrates the feasibility of this kind of apparatus for urinary diversion in a rabbit model,which provides some experimental basis for clinical applications.