De-epithelialization of porcine tracheal allografts as an approach for tracheal tissue engineering
De-epithelialization of porcine tracheal allografts as an approach for tracheal tissue engineering
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DOI：
10.1038/s41598-019-48450-4
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发表时间：
2019-08-19
影响因子：
4.6
通讯作者：
Karoubi, Golnaz
中科院分区：
文献类型：
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作者：
Aoki, Fabio G.;Varma, Ratna;Karoubi, Golnaz
大的气管缺陷的替换仍然是一个未得到满足的临床需求。虽然脱细胞气管移植物的再细胞化似乎是一种可行的途径，但来自临床的证据表明并非如此。事后看来，完全去除软骨细胞和重新植入气管软骨基质以实现气管软骨的功能可能是不现实的。相比之下，通过去除上皮而保留免疫优势软骨的混合移植的概念是一条截然不同的道路，最初的报告表明潜在的临床成功。在这里，我们提出了一种新的方法，使用双腔生物反应器来去除气管移植物上皮化，然后用外源细胞重新填充移植物。经内腔注入十二烷基硫酸钠3h可有效去除大部分气管上皮，而外腔注入生长介质可使大部分(68.6+/-7.3%)软骨细胞存活。去上皮化的移植物支持人支气管上皮细胞(BEAS-2B)附着、存活和生长超过7天。虽然不是没有局限性，但我们的方法表明，嵌合同种异体软骨的最终使用价值在于将完整的供体软骨重新上皮化为受体来源的上皮。通过采用简单和部分脱细胞的方法，特别是去除上皮，我们避免了软骨再生的需要。
Replacement of large tracheal defects remains an unmet clinical need. While recellularization of acellular tracheal grafts appeared to be a viable pathway, evidence from the clinic suggests otherwise. In hindsight, complete removal of chondrocytes and repopulation of the tracheal chondroid matrix to achieve functional tracheal cartilage may have been unrealistic. In contrast, the concept of a hybrid graft whereby the epithelium is removed and the immune-privileged cartilage is preserved is a radically different path with initial reports indicating potential clinical success. Here, we present a novel approach using a double-chamber bioreactor to de-epithelialize tracheal grafts and subsequently repopulate the grafts with exogenous cells. A 3 h treatment with sodium dodecyl sulfate perfused through the inner chamber efficiently removes the majority of the tracheal epithelium while the outer chamber, perfused with growth media, keeps most (68.6 +/- 7.3%) of the chondrocyte population viable. De-epithelialized grafts support human bronchial epithelial cell (BEAS-2B) attachment, viability and growth over 7 days. While not without limitations, our approach suggests value in the ultimate use of a chimeric allograft with intact donor cartilage re-epithelialized with recipient-derived epithelium. By adopting a brief and partial decellularization approach, specifically removing the epithelium, we avoid the need for cartilage regeneration.