Corneal Decellularization: A Method of Recycling Unsuitable Donor Tissue for Clinical Translation?
Corneal Decellularization: A Method of Recycling Unsuitable Donor Tissue for Clinical Translation?
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DOI：
10.3109/02713683.2015.1062114
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发表时间：
2016-06
影响因子：
2
通讯作者：
Hopkinson A
中科院分区：
文献类型：
--
作者：
Wilson SL;Sidney LE;Dunphy SE;Dua HS;Hopkinson A
背景：临床上需要与尸体供体组织一样有效，最好是上级的仿生角膜。与合成或半合成的工程化组织相比，脱细胞组织是有利的，因为可以保留天然基质超微结构和内在生物线索，包括生长因子、细胞因子和糖胺聚糖。然而，目前还没有可靠的、标准化的人角膜脱细胞方案。 方法：利用不适合移植的角膜眼库组织，系统比较常用的去细胞方案。高渗氯化钠;研究了离子试剂十二烷基硫酸钠;非离子去污剂叔辛基酚聚氧乙烯（Triton-X）;使用Dispase的酶促解聚;机械搅拌;以及核酸酶的使用。广泛评价了脱细胞功效，特别是对人角膜组织。通过组织学、免疫荧光和生物化学测定证明可检测的细胞物质的去除。评价了宏观组织透明度和透光率的保持情况。通过组织学、免疫荧光和定量分析评估角膜结构、胶原和糖胺聚糖的保留。使用细胞增殖试验评估所得支架的生物相容性。 结果：所研究的脱细胞方案均未成功去除100%的细胞组分。具有最少残留细胞材料的技术在结构上受损最大。糖胺聚糖的生化分析证明了脱细胞过程的剥离效果。 结论：利用、再处理和再生被认为“不适合”移植的组织的能力使我们能够挽救有价值的组织。组织的再处理有可能对解决与尸体供体短缺相关的问题产生相当大的影响。患者将直接受益于获得更多的角膜移植，卫生当局将履行其提供有效角膜重建以减轻角膜失明的责任。然而，为了取得进展，我们可能需要后退一步，建立一个“脱细胞化”的标准;这应该平衡有效去除免疫反应物质与维持组织功能。
Background: There is a clinical need for biomimetic corneas that are as effective, preferably superior, to cadaveric donor tissue. Decellularized tissues are advantageous compared to synthetic or semi-synthetic engineered tissues in that the native matrix ultrastructure and intrinsic biological cues including growth factors, cytokines and glycosaminoglycans may be retained. However, there is currently no reliable, standardized human corneal decellularization protocol. Methods: Corneal eye-bank tissue unsuitable for transplantation was utilized to systematically compare commonly used decellularization protocols. Hypertonic sodium chloride; an ionic reagent, sodium dodecyl sulphate; a non-ionic detergent, tert-octylphenol polyoxyethylene (Triton-X); enzymatic disaggregation using Dispase; mechanical agitation; and the use of nucleases were investigated. Decellularization efficacy, specifically for human corneal tissue, was extensively evaluated. Removal of detectable cellular material was evidenced by histological, immunofluorescence and biochemical assays. Preservation of macroscopic tissue transparency and light transmittance was evaluated. Retention of corneal architecture, collagen and glycosaminoglycans was assessed via histological, immunofluorescence and quantitative analysis. Biocompatibility of the resulting scaffolds was assessed using cell proliferation assays. Results: None of the decellularization protocols investigated successfully removed 100% of cellular components. The techniques with the least residual cellular material were most structurally compromised. Biochemical analysis of glycosaminoglycans demonstrated the stripping effects of the decellularization procedures. Conclusion: The ability to utilize, reprocess and regenerate tissues deemed “unsuitable” for transplantation allows us to salvage valuable tissue. Reprocessing the tissue has the potential to have a considerable impact on addressing the problems associated with cadaveric donor shortage. Patients would directly benefit by accessing greater numbers of corneal grafts and health authorities would fulfill their responsibility for the delivery of effective corneal reconstruction to alleviate corneal blindness. However, in order to progress, we may need to take a step back to establish a “decellularization” criterion; which should balance effective removal of immune reactive material with maintenance of tissue functionality.