Pariente Jean-Louis, Kim Byung-Soo, Atala Anthony
Laboratory for Tissue Engineering and Cellular Therapeutics, Children's Hospital and Harvard Medical School, Boston, Massachusetts, USA.
J Urol. 2002 Apr;167(4):1867-71.
Tissue engineering of the urinary tract often requires the use of various biomaterials. Adequate biomaterial biocompatibility is necessary for successful urinary reconstruction. In this study using a primary normal human bladder smooth muscle cell culture system we evaluated the in vitro biocompatibility of a number of naturally derived biomaterials, including bladder submucosa, small intestinal submucosa, collagen and alginate, and polymeric biomaterials, including polyglycolic acid, poly(L-lactic acid) and poly(lactic-co-glycolic acid, which have been used for urinary reconstruction experimentally or clinically.
To determine the cytotoxic and bioactive effects of bladder submucosa, small intestinal submucosa, collagen, alginate, polyglycolic acid, poly(L-lactic acid) and poly(lactic-co-glycolic acid) we measured cell viability, metabolic activity, apoptotic properties and DNA synthesis activity with 4 types of assays, namely Neutral Red (Sigma Chemical Co., St. Louis, Missouri), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (Sigma Chemical Co.), apoptotic activity and tritiated thymidine incorporation (Dupont NEN, Boston, Massachusetts) assays. Normal human bladder smooth muscle cells were cultured with the extracts of the biomaterials or cultured in direct contact with the biomaterials.
All naturally derived and synthetic biomaterials tested in this study except alginate exhibited nontoxic and bioactive effects on human bladder smooth muscle cells (HBSMCs) in vitro, as indicated by the 4 types of biocompatibility assays using the extract and direct contact methods. Cell viability, apoptotic properties, metabolic activity and DNA synthesis activity of HBSMCs cultured with the extracts of the biomaterials or cultured in direct contact with the biomaterials were not significantly different from those of negative controls (fresh medium with no extracts or tissue culture plates without biomaterials).
All naturally derived and synthetic biomaterials tested in this study except alginate exhibited nontoxic and bioactive effects on HBSMCs in vitro. This normal primary human bladder smooth muscle cell culture model is suitable for in vitro biocompatibility assessment. It provides information on cell-biomaterial interactions and on the ability of biomaterials to support bioactive cell functions.
尿路组织工程通常需要使用各种生物材料。足够的生物材料生物相容性对于成功的尿路重建至关重要。在本研究中，我们使用原代正常人膀胱平滑肌细胞培养系统，评估了多种天然来源的生物材料（包括膀胱黏膜下层、小肠黏膜下层、胶原蛋白和藻酸盐）以及聚合物生物材料（包括聚乙醇酸、聚（L-乳酸）和聚（乳酸-共-乙醇酸），这些材料已在实验或临床上用于尿路重建）的体外生物相容性。
为了确定膀胱黏膜下层、小肠黏膜下层、胶原蛋白、藻酸盐、聚乙醇酸、聚（L-乳酸）和聚（乳酸-共-乙醇酸）的细胞毒性和生物活性作用，我们用4种检测方法测量了细胞活力、代谢活性、凋亡特性和DNA合成活性，即中性红（西格玛化学公司，密苏里州圣路易斯）、3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四氮唑（西格玛化学公司）、凋亡活性和氚标记胸腺嘧啶核苷掺入（杜邦新英格兰核公司，马萨诸塞州波士顿）检测。正常人膀胱平滑肌细胞与生物材料提取物一起培养或与生物材料直接接触培养。
如使用提取物和直接接触方法的4种生物相容性检测所示，本研究中测试的所有天然来源和合成生物材料（除藻酸盐外）在体外对人膀胱平滑肌细胞（HBSMCs）均表现出无毒和生物活性作用。与生物材料提取物一起培养或与生物材料直接接触培养的HBSMCs的细胞活力、凋亡特性、代谢活性和DNA合成活性与阴性对照（无提取物的新鲜培养基或无生物材料的组织培养板）相比无显著差异。
本研究中测试的所有天然来源和合成生物材料（除藻酸盐外）在体外对HBSMCs均表现出无毒和生物活性作用。这种原代正常人膀胱平滑肌细胞培养模型适用于体外生物相容性评估。它提供了关于细胞-生物材料相互作用以及生物材料支持生物活性细胞功能能力的信息。
