| Objective The purpose of this research is to establish the way of HAAM's preparation and preservation,investigate the HAAM'S role and the significance in the rabbit urethral regeneration.Whether it be a good material replacement for urethral regeneration.Methods AM(Amniotic membrane)was separated from umbilical cord of pregnant donors by two methods.Method one was that AM was put in 1% formaldehyde-0.2%glutaraldehyde as link-crossed firstly,then were digested in 0.125%trypsinase-0.05M EDTA,and washed in 0.5%triton X-100 lastly. Method two was that AM was put in 1%formaldehyde-0.2%glutaraldehyde as link-crossed firstly,then were digested in 0.25%trypsinase-0.05M EDTA, and washed in 0.5%tritonX-100 lastly.Histology and bacteriology were performed on samples of HAAM.HAAM stored in 100%glycerol at 4℃as short term preservation.It stored in preservation solution at -80℃as long term preservation.A total of 32 NewZealand rabbits,clear grade,weight in approximately 2kg,were divided into 3 groups:group 1(n=12),which were operated with HAAM to repair the urethral defect;group 2(n=12)which urethras were anastomosised directly;group 3(n=8),which were not given urethral operation.The growth,wound and urination of rabbits were observed after operation.Histological examination was performed at day 10,day 21 and day 42.The examinations including Urethrography,Urodynamic studies and weight of bladder were preformed at day 42。 ResultHAAM samples made from method one were soft diaphanous and composed of many red network looking collagen fibrils(hematoxyli-eosin stain)regularly.The fragment cell was not noticed.Basement membrane of HAAM was integrated.Bacteriological tests was negative.but samples made from method two were obvious oncootic.The fragment cell was not noticed.Rabbit urethral regeneration using HAAM1.Most of rabbits grown well,with increasing weight and good urination. No urinary fistula and infection were found.2.Histological examination indicated that host cell infiltration and generous angiogenesis were observed in group 1 at day 10 after operation.The infiltration of mononuclear cells was observed in HAAM.10,representing an acute inflammatory reaction.there were not acute rejection.Several layers of urothelium covered HAAM At day 21 after operation,There were small blood vessel at day 21.Unorganized musclefiber bundles were found at day 42. urothelium of group 2 was oncootic at day 10 after operation.The infiltration of mononuclear cells was observed,urothelium covered completely at day 21 after operation.There were organized muscle fiber bundles at day 42.Group 3 was normal.3.Urodynamic studies indicated that the volume of bladder,maximum and minimum urethral pressure were not significant difference in these 3 groups (34.40±3.36 VS 34.80±2.59,P＞0.05,34.80±2.59 VS 30.20±3.19,P＞0.05,34.40±3.36 VS 30.20±3.19,P＞0.05,15.60±2.51 VS 16.00±2.35,P＞0.05,16.00±2.35 VS 15.604±1.14,P＞0.05,15.60±2.51 VS 15.60±1.14,P＞0.05,9.80±1.30 VS 9.20±1.92,P＞0.05,9.20±1.92 VS 9.20±1.30,P＞0.05,9.80±1.30 VS 9.20±1.30,P＞0.05).4.There were no diverticulum and exosmosis of urine in Urethrography. There was light stenosis of urethra in group 2,there was no abnormality in group 2 and group 3..5 The weight of bladder in group 2 was significantly high than that in group 3(2.60±0.66 VS 1.66±0.42,P＜0.05)but in group 1,there was not significant difference compared with group 3(2.08±0.35 VS 1.66±0.42,P＞0.05).it is the same in group 1 compared with group 2(2.08±0.35 VS 2.60±0.66,P＞0.05).Conclusionsit is suggested that HAAM is a kind of good biomaterial with well histocompatibility,biocompatibility and low antigenicity,which is suitable for cell attachment and growth as well as reconstruction urethrate.It has perspective clinical application.
|
PDF Full Text Request