Phenotypic and functional characterization of in vivo tissue engineered smooth muscle from normal and pathological bladders
Phenotypic and functional characterization of in vivo tissue engineered smooth muscle from normal and pathological bladders
复制标题
DOI：
10.1016/s0022-5347(05)64429-1
复制
发表时间：
2002-10-01
影响因子：
6.6
通讯作者：
Atala, A
中科院分区：
文献类型：
--
作者：
Lai, JY;Yoon, CY;Atala, A
目的膀胱组织工程包括从宿主处获取活检组织，扩增细胞，将其种植到基质上，然后将细胞-基质复合材料植入宿主体内。临床上，用于这些技术的细胞可能是从不正常的膀胱中获取的。目前尚不清楚异常的膀胱细胞是否可以被工程改造成功能正常的组织。材料与方法从功能正常的膀胱、外倾性膀胱和神经源性膀胱中分离培养出人的组织工程膀胱平滑肌细胞，并将其接种到聚合物支架上。对16种细胞种植支架进行了体外分析，并将40种细胞种植支架植入裸鼠体内。分别于2周和2个月取材和分析组织工程化构建物。结果体外培养和体内培养2个月后，无论来源如何，人膀胱细胞均表达α-肌动蛋白和肌球蛋白。所有支架在体内都显示出相似的肌肉形成。细胞接种支架对超大电场和卡巴胆碱刺激表现出典型的“收缩-松弛”反应。结论正常和病变膀胱组织工程化肌肉在体外和体内植入后均保持其表型。这些细胞对电和化学刺激表现出相同程度的收缩能力，而不考虑它们的来源。这些结果表明，从尿动力学正常或病理膀胱获得的肌肉细胞在表型或功能上没有差异，无论其来源如何，膀胱肌肉细胞都有可能被工程化为正常的膀胱组织。
PurposeThe engineering of bladder tissue involves obtaining a biopsy from a host, expanding the cells, seeding them onto a matrix and implanting the cell-matrix composite back into the host. Clinically, cells used for these techniques may be harvested from abnormal bladders. It is not known whether abnormal bladder cells may be engineered into functionally normal tissue. We investigated the phenotypic and functional characteristics of tissue engineered bladder smooth muscle derived from patients with functionally normal bladders and functionally abnormal exstrophic and neuropathic bladders.Materials and MethodsHuman smooth muscle cells derived from functionally normal bladders, exstrophic bladders and neurogenic bladders were grown, expanded and seeded onto polymer scaffolds. Sixteen cell seeded scaffolds were analyzed in vitro and 40 cell seeded scaffolds were implanted in athymic mice. The tissue engineered constructs were retrieved and analyzed at 2 weeks and 2 months. The scaffolds were evaluated immunocytochemically, histologically, with organ bath studies and with Western blot analyses.ResultsHuman bladder cells showed similar expression of smooth muscle marker proteins (α-actin and myosin) in vitro and after 2 months in vivo, regardless of their origin. All scaffolds showed similar muscle formation in vivo. The cell seeded scaffolds demonstrated the typical “contraction-relaxation” response to supramaximal electrical field and carbachol stimulation. There were no statistical differences among the experimental groups (normal, exstrophic, neurogenic).ConclusionsTissue engineered muscle from normal and diseased bladders retain their phenotype in vitro and after implantation in vivo. The cells exhibited the same degree of contractility to electrical and chemical stimulation regardless of their origin. These results suggest that there are no phenotypic or functional differences between muscle cells obtained from urodynamically normal or pathological bladders, and that bladder muscle cells, regardless of their origin, may have the potential to be engineered into normal bladder tissues.