title: Preliminary experimental study of urethral reconstruction with tissue engineering and RNA interference techniques.
abstract: This study investigated the feasibility of replacing urinary epithelial cells with oral keratinocytes and transforming growth factor-β1 (TGF-β1) small interfering RNA (siRNA)-transfected fibroblasts seeded on bladder acellular matrix graft (BAMG) in order to reconstruct tissue-engineered urethra. Constructed siRNAs, which expressed plasmids targeting TGF-β1, were transfected into rabbit fibroblasts. The effective siRNA was screened out by RT-PCR and was transfected into rabbit fibroblasts again. Synthesis of type I collagen in culture medium was measured by enzyme-linked immuno sorbent assay (ELISA). Autologous oral keratinocytes and TGF-β1 siRNA-transfected fibroblasts were seeded onto BAMGs to obtain a tissue-engineered mucosa. The tissue-engineered mucosa was assessed morphologically and with the help of scanning electron microscopy. The TGF-β1 siRNA decreased the expression of fibroblasts synthesis type I collagen. Oral keratinocytes and TGF-β1 siRNA-transfected fibroblasts were seeded onto sterilized BAMG to obtain a tissue-engineered mucosa for urethral reconstruction. The compound graft was assessed using scanning electron microscope. Oral keratinocytes and TGF-β1 siRNA-transfected fibroblasts had a good compatibility with BAMG. The downregulation of fibroblasts synthesis type I collagen expression by constructed siRNA interfering TGF-β1 provided a potential basis for genetic therapy of urethral scar. Oral keratinocytes and TGF-β1 siRNA-transfected fibroblasts had good compatibility with BAMG and the compound graft could be a new choice for urethral reconstruction.
authors.affiliation: Department of Urology, Sixth People’s Hospital, Shanghai Jiaotong University, Shanghai 200233, China. chaoli1979@126.com
authors.forename: Chao
authors.initials: C
authors.lastname: Li
authors.forename: Yue-Min
authors.initials: YM
authors.lastname: Xu
authors.forename: Hong-Bin
authors.initials: HB
authors.lastname: Li
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002921:Cicatrix / Q000601:surgery; D024042:Collagen Type I / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure; D015870:Gene Expression; D015603:Keratinocytes / Q000648:ultrastructure; D008297:Male; D009061:Mouth Mucosa; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D011817:Rabbits; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism*; D014521:Urethra / Q000601:surgery*; D001743:Urinary Bladder
chemical_list: D024042:Collagen Type I; D034741:RNA, Small Interfering; D053773:Transforming Growth Factor beta1
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