Department of Biomedical and Veterinary Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Tech, Leesburg, Virginia, United States of America.
PLoS One. 2013 May 27;8(5):e64151. doi: 10.1371/journal.pone.0064151. Print 2013.
Natural extracellular matrix provides a number of distinct advantages for engineering replacement orthopedic tissue due to its intrinsic functional properties. The goal of this study was to optimize a biologically derived scaffold for tendon tissue engineering using equine flexor digitorum superficialis tendons. We investigated changes in scaffold composition and ultrastructure in response to several mechanical, detergent and enzymatic decellularization protocols using microscopic techniques and a panel of biochemical assays to evaluate total protein, collagen, glycosaminoglycan, and deoxyribonucleic acid content. Biocompatibility was also assessed with static mesenchymal stem cell (MSC) culture. Implementation of a combination of freeze/thaw cycles, incubation in 2% sodium dodecyl sulfate (SDS), trypsinization, treatment with DNase-I, and ethanol sterilization produced a non-cytotoxic biomaterial free of appreciable residual cellular debris with no significant modification of biomechanical properties. These decellularized tendon scaffolds (DTS) are suitable for complex tissue engineering applications, as they provide a clean slate for cell culture while maintaining native three-dimensional architecture.
天然细胞外基质由于其固有功能特性，为工程替代骨科组织提供了许多明显的优势。本研究的目的是使用马屈肌趾浅屈肌腱优化生物衍生的肌腱组织工程支架。我们使用显微镜技术和一系列生化分析来评估总蛋白、胶原蛋白、糖胺聚糖和脱氧核糖核酸含量，研究了几种机械、去污剂和酶去细胞化方案对支架组成和超微结构的影响。还通过静态间充质干细胞（MSC）培养评估了生物相容性。实施冷冻/解冻循环、2%十二烷基硫酸钠（SDS）孵育、胰蛋白酶消化、DNase-I 处理和乙醇消毒的组合产生了无细胞毒性的生物材料，没有明显的细胞残留，生物力学性能也没有显著改变。这些去细胞化的肌腱支架（DTS）适合用于复杂的组织工程应用，因为它们为细胞培养提供了一个干净的平台，同时保持了天然的三维结构。
