Evaluation of small intestine grafts decellularization methods for corneal tissue engineering.
Evaluation of small intestine grafts decellularization methods for corneal tissue engineering.
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DOI：
10.1371/journal.pone.0066538
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Campos A
中科院分区：
文献类型：
--
作者：
Oliveira AC;Garzón I;Ionescu AM;Carriel V;Cardona Jde L;González-Andrades M;Pérez Mdel M;Alaminos M;Campos A
通过组织工程技术开发角膜替代物的进展集中在脱细胞组织支架的使用上。在这项工作中，我们评估了小肠组织的不同化学和物理脱细胞方法，以确定最合适的角膜应用脱细胞方案。我们的研究结果表明，最有效的脱细胞剂是SDS和Triton X-100去污剂，它们能够有效地去除大部分细胞核和残留的DNA。组织学和组织化学分析表明，胶原纤维被保存后脱细胞与Triton X-100，NaCl和超声处理，而网状纤维被妥善保存脱细胞与UV曝光。用SDS、Triton X-100和超声处理脱细胞后，细胞外基质糖蛋白得以保留，而蛋白多糖不受任何脱细胞方案的影响。对于所有方案，组织透明度显著高于对照非脱细胞组织，尽管在用SDS和Triton X-100脱细胞的组织中发现了最好的透光率结果。结论：脱细胞小肠移植物可作为角膜组织工程的生物支架材料。用triton X-100脱细胞能够有效地从组织中去除所有细胞，同时保留组织结构和大多数纤维状和非纤维状细胞外基质组分，表明这种特定的脱细胞剂可以安全地用于角膜TE应用的SI组织的有效脱细胞。
Advances in the development of cornea substitutes by tissue engineering techniques have focused on the use of decellularized tissue scaffolds. In this work, we evaluated different chemical and physical decellularization methods on small intestine tissues to determine the most appropriate decellularization protocols for corneal applications. Our results revealed that the most efficient decellularization agents were the SDS and triton X-100 detergents, which were able to efficiently remove most cell nuclei and residual DNA. Histological and histochemical analyses revealed that collagen fibers were preserved upon decellularization with triton X-100, NaCl and sonication, whereas reticular fibers were properly preserved by decellularization with UV exposure. Extracellular matrix glycoproteins were preserved after decellularization with SDS, triton X-100 and sonication, whereas proteoglycans were not affected by any of the decellularization protocols. Tissue transparency was significantly higher than control non-decellularized tissues for all protocols, although the best light transmittance results were found in tissues decellularized with SDS and triton X-100. In conclusion, our results suggest that decellularized intestinal grafts could be used as biological scaffolds for cornea tissue engineering. Decellularization with triton X-100 was able to efficiently remove all cells from the tissues while preserving tissue structure and most fibrillar and non-fibrillar extracellular matrix components, suggesting that this specific decellularization agent could be safely used for efficient decellularization of SI tissues for cornea TE applications.
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影响因子：
3.2
作者：
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