Manufacturing considerations for producing and assessing decellularized extracellular matrix hydrogels
Manufacturing considerations for producing and assessing decellularized extracellular matrix hydrogels
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DOI：
10.1016/j.ymeth.2019.09.015
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发表时间：
2020-01-15
期刊：
影响因子：
4.8
通讯作者：
Christman, Karen L.
中科院分区：
文献类型：
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作者：
Hernandez, Melissa J.;Yakutis, Grace E.;Christman, Karen L.
尽管几种脱细胞细胞的细胞外基质（ECM）片或贴片已经商业化用于临床，但只有一种可注射的脱细胞的ECM水凝胶（脱细胞的心肌基质）已经进入临床试验。因此，可用于这些材料的既定制造标准或评估的信息非常少。在这里，我们提出了详细的方法，用于研究与猪源性骨骼肌ECM水凝胶的制造优化相关的三个参数-动物间变异性、生物负载降低和收获条件。表征试验的结果（包括残留dsDNA含量和硫酸化糖胺聚糖含量）在个体动物之间或添加生物负荷降低剂后未产生显著差异。然而，在不同收获条件下收集的组织含有不同量的脂肪，并且脱细胞产物的蛋白质组成不同，这可能最终影响随后的体外或体内功效。随着脱细胞ECM水凝胶继续被评估用于各种应用，应彻底考虑实验室规模和制造规模材料批次之间的差异，以避免在规模扩大期间进行昂贵和及时的优化。
Although several decellularized extracellular matrix (ECM) sheets or patches have been commercialized for use in the clinic, only one injectable decellularized ECM hydrogel, a decellularized myocardial matrix, has reached clinical trials. Consequently, very little information is available for established manufacturing standards or assessments of these materials. Here we present detailed methodology for investigating three parameters related to manufacturing optimization for a porcine derived skeletal muscle ECM hydrogel - animal-to-animal variability, bioburden reduction, and harvesting conditions. Results from characterization assays, including residual dsDNA content and sulfated glycosaminoglycan content, did not yield noteworthy differences amongst individual animals or following the addition of a bioburden reducing agent. However, the tissue collected under different harvesting conditions contained varying amounts of fat, and the protein compositions of the decellularized products differed, which could ultimately impact subsequent efficacy in vitro or in vivo. As decellularized ECM hydrogels continue to be evaluated for various applications, the differences between laboratory-scale and manufacturing-scale material batches should be thoroughly considered to avoid costly and timely optimization during scale-up.