Nakase Yuen, Hagiwara Akeo, Nakamura Tatsuo, Kin Syuichi, Nakashima Susumu, Yoshikawa Tetsuji, Fukuda Ken-Ichirou, Kuriu Yoshiaki, Miyagawa Kouji, Sakakura Chohei, Otsuji Eigo, Shimizu Yasuhiko, Ikada Yoshihito, Yamagishi Hisakazu
Department of Surgery and Regenerative Medicine, Division of Surgery and Physiology of Digestive System, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Tissue Eng. 2006 Feb;12(2):403-12. doi: 10.1089/ten.2006.12.403.
In a previously reported attempt to regenerate small intestine with autologous tissues, collagen scaffolds were used without cell seeding or with autologous mesenchymal stem cell seeding. However the regenerated intestine lacked a smooth muscle layer. To accomplish regeneration of a smooth muscle layer, this present study used collagen scaffolds seeded with the smooth muscle cells (SMC) in a canine model. Autologous SMC were isolated from stomach wall and cultured. Two types of scaffolds were fabricated: in SMC (+), cultured SMCs were mixed with collagen solution and poured into a collagen sponge; and in SMC (-), SMCs were omitted. Both scaffolds were implanted into defects of isolated ileum as a patch graft. Animals were euthanized at 4, 8, and 12 weeks; for the last time point, the ileal loop had been reanastomosed at 8 weeks. At 12 weeks, the SMC (-) group showed a luminal surface covered by a regenerated epithelial cell layer with very short villi; however only a thin smooth muscle layer was observed, representing the muscularis mucosae. In the SMC (+) group, the luminal surface was covered completely by a relatively well-developed epithelial layer with numerous villi. Implanted SMCs were seen in the lamina propria and formed a smooth muscle layer. Thus, we concluded that collagen sponge scaffolds seeded with autologous SMCs have a potential for small intestine regeneration.
在先前报道的一项用自体组织再生小肠的尝试中,使用了未接种细胞或接种了自体间充质干细胞的胶原支架。然而,再生的小肠缺乏平滑肌层。为了实现平滑肌层的再生,本研究在犬模型中使用接种了平滑肌细胞(SMC)的胶原支架。从胃壁分离并培养自体SMC。制备了两种类型的支架:在SMC(+)组中,将培养的SMC与胶原溶液混合并倒入胶原海绵中;在SMC(-)组中,省略了SMC。将两种支架作为补片移植植入离体回肠的缺损处。在4周、8周和12周对动物实施安乐死;对于最后一个时间点,回肠袢在8周时已重新吻合。在12周时,SMC(-)组的管腔表面覆盖着一层再生的上皮细胞层,绒毛很短;然而,仅观察到一层薄的平滑肌层,代表黏膜肌层。在SMC(+)组中,管腔表面完全被一层相对发达的上皮层覆盖,有许多绒毛。植入的SMC可见于固有层并形成平滑肌层。因此,我们得出结论,接种自体SMC的胶原海绵支架具有小肠再生的潜力。