Modification of Human Pericardium by Chemical Crosslinking
Modification of Human Pericardium by Chemical Crosslinking
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DOI：
10.33549/physiolres.934335
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发表时间：
2020-02-01
影响因子：
2.1
通讯作者：
Bacakova, Lucie
中科院分区：
文献类型：
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作者：
Filova, Elena;Stankova, Lubica;Bacakova, Lucie
用作心血管手术生物材料的自体和同种异体人心包传统上是用戊二醛交联的。在这项工作中，我们已经评估了抵抗胶原酶消化和细胞毒性的人心包与不同浓度的戊二醛交联的心包京尼平，去甲二氢愈创木酸，单宁酸交联，并与未改性的心包相比。交联保留了二次谐波发生显微镜观察到的天然心包的波浪状形态。采用SDS-PAGE、毛细管电泳和羟脯氨酸测定分析胶原酶消化产物。戊二醛和京尼平交联有效地保护了天然心包免受胶原酶III的消化。其它交联剂仅提供低保护。使用xCELLigence通过监测在交联心包洗脱液中培养的猪瓣膜间质细胞的活力来评价交联心包的细胞毒性。在京尼平洗脱液中观察到最高的细胞指数，反映了监测细胞的数量和形状。因此，用京尼平交联心包移植物似乎是传统戊二醛交联的一种有前景的替代方法，因为它提供了类似的高保护，防止胶原酶降解，而没有细胞毒性作用。
Autologous and allogenic human pericardia used as biomaterials for cardiovascular surgery are traditionally crosslinked with glutaraldehyde. In this work, we have evaluated the resistivity to collagenase digestion and the cytotoxicity of human pericardium crosslinked with various concentrations of glutaraldehyde in comparison with pericardium crosslinked by genipin, nordihydroguaiaretic acid, tannic acid, and in comparison with unmodified pericardium. Crosslinking retained the wavy-like morphology of native pericardium visualized by second harmonic generation microscopy. The collagenase digestion products were analyzed using SDS-PAGE, capillary electrophoresis, and a hydroxyproline assay. Glutaraldehyde and genipin crosslinking protected the native pericardium efficiently against digestion with collagenase III. Only low protection was provided by the other crosslinking agents. The cytotoxicity of crosslinked pericardium was evaluated using xCELLigence by monitoring the viability of porcine valve interstitial cells cultured in eluates from crosslinked pericardium. The highest cell index, reflecting both the number and the shape of the monitored cells was observed in eluates from genipin. Crosslinking pericardium grafts with genipin therefore seems to be a promising alternative procedure to the traditional crosslinking with glutaraldehyde, because it provides similarly high protection against degradation with collagenase, without cytotoxic effects.