| Bladder tissue engineering is a broad term used to describe the development of alternative tissue sources for diseased or dysfunctional native bladder. The techniques involved synthetic and natural biodegradable matrices alone, known as "unseeded" scaffolds, and the latest data on "seeded" scaffolds, which are impregnated with cultured cells from urologic or other organs.In this study, we address the function parameters of tissue engineered bladders with bladder accellular organ specific matrix graft (BAMG) seeded with hepatocyte growth factors or cultured cells from autologous urologic or other organs to replace a subtotal cystectomy. We also try to find a new source of "seed"cells with nuclear transplantation techneques.Aims: To study the HGFs function in bladder smooth muscle wound healing, identify the BAMG application in bladder tissues engineering construction, expand the source of "seed" cell with nuclear transplantation techneques.Methods: (1) A cyst-muscle-ectomy only rabbit model treated withmenstruum only A, 15ug pUDK B or 15ug pUDKH C. Gross, histologic and hypertrophic index (HI) analyses of the wound healing were performed postoperative on day 7 and day 14; (2) The adapted preparation is simplified and used to make the BAMG. The BAMG stained with Masson' or in uranyl acetate and lead citrate, was observed with scanning electron microscopy and light microscopy examination; (3) Autologous MSCs, ASCs, smooth muscle and epthlial were isolated from biopsy respectly. Preparation the tissue engineering bladder with BAMG seeded without or with pUDKH + FB, autologous MSCs, ASCs, smooth muscle and epthlial from bladder biopsy respectly. The cystmetre of rabbits were performed preoperatively and postoperatively at 12 weeks after subsequent replacement of a subtotal cystectomy of rabbits. Graphy and histologic analyses were also performed; (4) Swine's ASCs or MSCs were isolated and cultured to passage 3-5 and used as donor cells in nuclear transfer (NT) to produce the swine-mice reconstruction oocyte with direct intracystplasm microinjection techniques. Sire differences were noted in the ability of both MSCs and ASCs to form blastocysts. The chromosome analysis of hatching cells was also performed.Results: (1) A cyst-muscle-ectomy only rabbit model treated with menstruum only A, 15ug pUDK B or 15ug pUDKH C, the wound aera postoperatively was 44 ±8 mm2, 43 ± 9 mm2 and 28 ± 6 mm2 respectively on day 7, and 23 ±7mm2 , 22 ± 6mm2 和 4 ± 4mm2 respectively on day 14, analysis the data with repeat measure method, the wound aera reduce in C group did significant differ (F=16.236, p =0.00, C vs A and B). The hypertrophic index (HI) postoperatively at day 30 in three groups was 1.76±0.17, 1.83±0.18 and 1.11±0.18 respectively. Analyses of the HI showed that there were significant difference betweean the groups (.P=0.00, C vs A and B; P=0.054, A vs B). C group have little wound aera and HI; (2) Thepreparation time of BAMG, which typically indicates essentially no cell nuclei when stained with Masson' dyes or uranyl acetate and lead citrate, was 26 huors with the adapted simple method; (3) The isolation bladder epithelial were cobblestone shapes under phase contrast microscope and positive staining with pancytokeration AE1/AE3, smooth muscle were shuttles forms and positive staining with anti- a -smooth muscle actin. The isolation rabbit MSCs and ASCs shapes like shuttles under phase contrast microscope. Preparation time of BAMG seeded with pUDKH + FB, MSCs, ASCs, smooth muscle and epthlial from bladder biopsy were 30 minutes, 3 weeks and 4 weeks respectly. Analysis of the the cystmetre data of rabbit performed preoperatively and postoperatively with repeat measure method showed that all parameter did not significant differ in tissue engineering bladder groups. T-test showed that the reduction of bladder volume and capacity did significant different between preoperative and postoperative. Histologically, group A, C, D, E, F retrieved bladders showed a normal cellular organization consisti...
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