Effects of Chondroitinase ABC-Mediated Proteoglycan Digestion on Decellularization and Recellularization of Articular Cartilage.
Effects of Chondroitinase ABC-Mediated Proteoglycan Digestion on Decellularization and Recellularization of Articular Cartilage.
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DOI：
10.1371/journal.pone.0158976
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Bilgen B
中科院分区：
文献类型：
--
作者：
Bautista CA;Park HJ;Mazur CM;Aaron RK;Bilgen B
关节软骨的自我愈合能力有限，因此局灶性缺陷常常导致骨关节炎的发展。当前的软骨组织工程策略寻求通过创建旨在模仿天然关节软骨的独特结构和组成的支架来再生受损组织。去细胞化是一种新颖的策略，旨在保留天然细胞外基质的生物活性因子和 3D 生物物理环境，同时去除潜在的免疫原性因子。本研究的目的是开发一种能够实现完整关节软骨基质脱细胞和再细胞化的程序。通过一系列冻融循环和 0.1% (w/v) 十二烷基硫酸钠洗涤剂循环对全层猪关节软骨塞进行脱细胞。在洗涤剂循环之前应用软骨素酶 ABC (ChABC) 来消化糖胺聚糖，以增强供体软骨细胞的去除和种子细胞的迁移。将猪滑膜来源的间充质干细胞接种到脱细胞软骨支架上并培养长达 28 天。优化的脱细胞方案去除了每个样品湿重 94% 的天然 DNA，同时保留了胶原蛋白含量和排列。洗涤剂循环之前糖胺聚糖的消耗增加了核材料的去除。培养 28 天后，种子细胞渗入软骨深层达 100 μm。 ChABC 治疗通过降低糖胺聚糖含量来增强相对致密、不可渗透的关节软骨的脱细胞化。在静态体外培养下，ChABC 处理似乎不会影响再细胞化过程中的细胞迁移，这突出表明需要更动态的接种方法。
Articular cartilage has a limited capacity to heal itself and thus focal defects often result in the development of osteoarthritis. Current cartilage tissue engineering strategies seek to regenerate injured tissue by creating scaffolds that aim to mimic the unique structure and composition of native articular cartilage. Decellularization is a novel strategy that aims to preserve the bioactive factors and 3D biophysical environment of the native extracellular matrix while removing potentially immunogenic factors. The purpose of this study was to develop a procedure that can enable decellularization and recellularization of intact articular cartilage matrix. Full-thickness porcine articular cartilage plugs were decellularized with a series of freeze-thaw cycles and 0.1% (w/v) sodium dodecyl sulfate detergent cycles. Chondroitinase ABC (ChABC) was applied before the detergent cycles to digest glycosaminoglycans in order to enhance donor chondrocyte removal and seeded cell migration. Porcine synovium-derived mesenchymal stem cells were seeded onto the decellularized cartilage scaffolds and cultured for up to 28 days. The optimized decellularization protocol removed 94% of native DNA per sample wet weight, while collagen content and alignment were preserved. Glycosaminoglycan depletion prior to the detergent cycles increased removal of nuclear material. Seeded cells infiltrated up to 100 μm into the cartilage deep zone after 28 days in culture. ChABC treatment enhances decellularization of the relatively dense, impermeable articular cartilage by reducing glycosaminoglycan content. ChABC treatment did not appear to affect cell migration during recellularization under static, in vitro culture, highlighting the need for more dynamic seeding methods.