Department of Surgery, University of California, Los Angeles, California, USA.
J Surg Res. 2011 Nov;171(1):e21-6. doi: 10.1016/j.jss.2011.08.001. Epub 2011 Aug 27.
Small intestinal submucosa (SIS) is a porcine-derived, acellular, collagen-based matrix that has been tested without seeded smooth muscle cells (SMCs) for intestinal tissue engineering. We examined the expression patterns of contractile proteins of SIS with SMCs implanted in an in vivo rodent model.
Intestinal SMCs were isolated from Lewis rat pups. Four-ply tubular SMCs-seeded SIS or blank SIS scaffolds were implanted in an adult rat jejunal interposition model. Recipients were sacrificed at 2, 4, and 8 wk following the implantation. The retrieved specimens were examined using antibodies against contractile proteins of SMCs.
Cultured intestinal SMCs expressed α-smooth muscle actin (α-SMA), calponin, and less smooth muscle myosin heavy chain (SM-MHC) in vitro. Cell-seeded SIS scaffolds contracted significantly over 8 wk of implantation but were comparable to SIS scaffolds without cell seeding. Implanted cell-seeded SIS scaffolds at 2 wk expressed extensive α-SMA, some calponin, and minimal SM-MHC. At 4 wk, α-SMA-expressing cells decreased significantly, whereas calponin or SM-MHC expressing cells were rarely detected. A small number of α-SMA-expressing cells were present at 8 wk, whereas more calponin or SM-MHC expressing cells emerged in proximity with the anastomotic interface.
Cell-seeded SIS contracted significantly after implantation, but the expressions of contractile proteins were present at the site of SIS interposition. No organized smooth muscle was formed at the site of implantation. A better scaffold design is needed to produce structured smooth muscle.
小肠黏膜下层(SIS)是一种猪源性、去细胞、基于胶原的基质,已在无平滑肌细胞(SMCs)种植的情况下用于肠道组织工程学研究。我们在体内啮齿动物模型中研究了种植 SMC 的 SIS 的收缩蛋白表达模式。
从小鼠肠中分离出肠平滑肌细胞(SMCs)。将 4 层 SMC 种植的 SIS 或空白 SIS 支架植入成年大鼠空肠间置模型中。植入后 2、4 和 8 周处死接受者。用针对 SMC 收缩蛋白的抗体检查回收标本。
体外培养的肠 SMC 表达α-平滑肌肌动蛋白(α-SMA)、钙调蛋白和较少的平滑肌肌球蛋白重链(SM-MHC)。种植细胞的 SIS 支架在植入后 8 周内显著收缩,但与未种植细胞的 SIS 支架相当。植入的细胞种植 SIS 支架在第 2 周表达广泛的 α-SMA、一些钙调蛋白和最小的 SM-MHC。在第 4 周,α-SMA 表达的细胞显著减少,而很少检测到钙调蛋白或 SM-MHC 表达的细胞。在第 8 周时存在少量表达α-SMA 的细胞,而在吻合口附近出现更多表达钙调蛋白或 SM-MHC 的细胞。
种植细胞的 SIS 在植入后显著收缩,但收缩蛋白的表达位于 SIS 间置部位。在植入部位未形成有组织的平滑肌。需要更好的支架设计来产生有结构的平滑肌。