Tissue engineering of heart valves by recellularization of glutaraidehyde-fixed porcine valves using bone marrow-derived cells
Tissue engineering of heart valves by recellularization of glutaraidehyde-fixed porcine valves using bone marrow-derived cells
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DOI：
10.1038/emm.2006.33
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发表时间：
2006-06-30
影响因子：
12.8
通讯作者：
Kim, Byung-Soo
中科院分区：
文献类型：
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作者：
Kim, Sang-Soo;Lim, Sang-Hyun;Kim, Byung-Soo
为了提高戊二醛（GA）固定瓣膜的生物相容性和耐用性，人们提出了一种带有活内皮细胞（EC）的生物涂层。然而，由于其无法重新填充，稳定的 EC 层尚未在 GA 固定瓣膜上成功形成。在本研究中，为了改善细胞粘附和增殖，通过用柠檬酸处理去除游离醛基来对 GA 固定假体进行解毒。犬骨髓单核细胞（MNC）在体外分化为EC样细胞和肌成纤维细胞样细胞。将解毒的假体接种并用分化的骨髓来源细胞 (BMC) 进行再细胞化 7 天。未经处理的 GA 固定假体用作对照。研究了细胞附着、增殖、代谢活性和活力，并对接种细胞的小叶进行组织学分析。在解毒的 GA 固定假体上，BMC 接种导致 7 天后细胞增殖不受抑制。相反，在未经处理的 GA 固定假体上，细胞附着很差并且没有观察到活细胞。在解毒瓣膜小叶的腔侧观察到平滑肌α-肌动蛋白、CD31和增殖细胞核抗原的阳性染色，表明接种的BMC的分化和增殖。这些结果表明，用柠檬酸处理 GA 固定瓣膜建立了更适合细胞附着和增殖的表面。通过用 BMC 接种脱毒的 GA 固定生物瓣膜假体来设计心脏瓣膜，可以提高假体的生物相容性和耐用性。该方法可作为治疗终末期心脏瓣膜病恢复心脏瓣膜结构和功能的新途径。
To increase the biocompatibility and durability of glutaraldehyde (GA)-fixed valves, a biological coating with viable endothelial cells (ECs) has been proposed. However, stable EC layers have not been formed successfully on GA-fixed valves due to their inability to repopulate. In this study, to improve cellular adhesion and proliferation, the GA-fixed prostheses were detoxified by treatment with citric acid to remove free alldehyde groups. Canine bone marrow mononuclear cells (MNCs) were differentiated into EC-like cells and myofibroblast-like cells in vitro. Detoxified prostheses were seeded and recellularized with differentiated bone marrow-derived cells (BMCs) for seven days. Untreated GA-fixed prostheses were used as controls. Cell attachment, proliferation, metabolic activity, and viability were investigated and cell-seeded leaflets were histologically analyzed. On detoxified GA-fixed prostheses, BMC seeding resulted in uninhibited cell proliferation after seven days. In contrast, on untreated GA-fixed prostheses, cell attachment was poor and no viable cells were observed. Positive staining for smooth muscle a-actin, CD31, and proliferating cell nuclear antigen was observed on the luminal side of the detoxified valve leaflets, indicating differentiation and proliferation of the seeded BMCs. These results demonstrate that the treatment of GA-fixed valves with citric acid established a surface more suitable for cellular attachment and proliferation. Engineering heart valves by seeding detoxified GA-fixed biological valve prostheses with BMCs may increase biocompatibility and durability of the prostheses. This method could be utilized as a new approach for the restoration of heart valve structure and function in the treatment of end-stage heart valve disease.