Marzaro Maurizio, Conconi Maria Teresa, Perin Laura, Giuliani Stefano, Gamba Piergiorgio, De Coppi Paolo, Perrino Gian Pietro, Parnigotto Pier Paolo, Nussdorfer Gastone G
Department of Pediatric Surgery, Treviso Regional Hospital, Section of Anatomy, University of Padua, I-35121 Padua, Italy.
Int J Mol Med. 2002 Aug;10(2):177-82.
Acellular matrix obtained from homologous muscular tissue has been previously used to repair muscular defects. However, the implants, although not rejected, give rise to an intense inflammatory response and are rapidly replaced by fibrous tissue. In this study we examined the possibility that co-culture with autologous satellite cells can improve the efficiency of homologous acellular matrix as skeletal muscle substitute. Satellite cells, isolated from rat dorsal muscle, were cultured in vitro on homologous acellular matrix obtained by detergent-enzymatic treatment of abdominal muscle fragments. Scanning electron microscopy revealed that after 24 h of co-culture satellite cells were attached to the matrix, but still possessed a round shape. After 96 h, seeded cells began to flatten and to differentiate, originating few multinucleated myotubes. Patches of homologous matrix, seeded or not with autologous satellite cells, were implanted in the dorsal muscle of rats. At autopsy, the implants were recovered and processed for light microscopy. Two weeks after surgery, fibrous tissue started to replace the grafts composed only by acellular matrix, which at the 4th week were transformed into a fibrous scar. In contrast, at both times post-surgery the structure of implants containing autologous satellite cells was well preserved. The inflammatory reaction was modest and fibrosis was confined to the periphery of the grafts. It is concluded that the presence of autologous satellite cells is an important factor to preserve the structural integrity and to improve in vivo biocompatibility of homologous muscular acellular matrix implants.
从同源肌肉组织获得的脱细胞基质先前已被用于修复肌肉缺损。然而,这些植入物虽然未被排斥,但会引发强烈的炎症反应,并迅速被纤维组织取代。在本研究中,我们探讨了与自体卫星细胞共培养能否提高同源脱细胞基质作为骨骼肌替代物的效率。从大鼠背部肌肉分离出的卫星细胞,在体外培养于通过对腹部肌肉片段进行去污剂 - 酶处理获得的同源脱细胞基质上。扫描电子显微镜显示,共培养24小时后卫星细胞附着于基质,但仍呈圆形。96小时后,接种的细胞开始变平并分化,形成少量多核肌管。将接种或未接种自体卫星细胞的同源基质贴片植入大鼠背部肌肉。尸检时,取出植入物并进行光学显微镜检查。手术后两周,仅由脱细胞基质组成的移植物开始被纤维组织取代,在第4周时转变为纤维瘢痕。相比之下,在手术后的两个时间点,含有自体卫星细胞的植入物结构均保存良好。炎症反应较轻,纤维化局限于移植物周边。结论是自体卫星细胞的存在是保持同源肌肉脱细胞基质植入物结构完整性并改善其体内生物相容性的重要因素。