The use of decellularized tissues or organs as cell culture scaffolds has proven to be a promising approach for tissue engineering and regenerative medicine, as these decellularized tissues can provide the instructive niche for cell differentiation and functions. Cartilage is a largely avascular tissue with limited regenerative capacity. Lesions caused by arthritis can lead to severe cartilage degeneration. Previous studies have indicated that decellularized cartilage can be used as scaffolds that support the chondrogenic differentiation of adult stem cells. However, these decellularization protocols all require the use of denaturing agents, such as high salt and detergents, that lead to the artifactual disruption of the chemical and physical integrity of the tissue microenvironment. Here, we established a new decellularization method for cartilage, through a combined effect of freezing-thawing, sectioning, and sonication in water. This protocol achieved the complete removal of cells within minutes, instead of hours or days required by existing procedures, and does not use any detergent. The resulting decellularized cartilage preserved the native ultrastructure and biochemical contents, including glycosaminoglycans, which is typically depleted by traditional decellularization methods. Human mesenchymal stem cells could readily adhere onto the decellularized cartilage. Together, this work unveils a simple new method for decellularizing cartilage, which will be useful in studying how tissue microenvironment supports chondrocyte growth and functions. Impact statement In this study, we develop a simple, fast cartilage decellularization method that does not require any detergent, so that the decellularized cartilage chemistry is preserved. Traditional detergent-based decellularization removes the tissue biochemical contents (i.e., glycosaminoglycans). In this new water decellularization protocol, the biochemical contents of cartilage can be preserved. This allows the study of biochemistry and physical content in extracellular matrix as a whole, and this protocol would definitely be useful for studying the effect of tissue microenvironment in supporting chondrocyte growth and functions.

中文翻译：

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软骨的快速无洗涤剂脱细胞作用。

已证明将脱细胞的组织或器官用作细胞培养支架是组织工程和再生医学的有前途的方法，因为这些脱细胞的组织可以为细胞分化和功能提供指导。软骨是一种基本上无血管的组织，其再生能力有限。关节炎引起的病变可导致严重的软骨变性。先前的研究表明，脱细胞的软骨可以用作支持成体干细胞成软骨分化的支架。然而，这些脱细胞方案均需要使用变性剂，例如高盐和去污剂，其导致人为破坏组织微环境的化学和物理完整性。这里，我们通过在水中冻融，切片和超声处理的综合作用，建立了一种新的软骨脱细胞方法。该方案可在数分钟内完全去除细胞，而不是现有程序所需的数小时或数天，并且不使用任何清洁剂。所得的脱细胞软骨保留了天然的超微结构和生化成分，包括通常通过传统脱细胞方法消耗掉的糖胺聚糖。人间充质干细胞很容易粘附在脱细胞的软骨上。总之，这项工作揭示了一种使软骨脱细胞的简单新方法，该方法将有助于研究组织微环境如何支持软骨细胞的生长和功能。影响陈述在这项研究中，我们开发了一个简单的，快速软骨脱细胞方法，不需要任何去污剂，因此可以保留脱细胞的软骨化学成分。传统的基于去污剂的脱细胞作用去除了组织的生化成分（即糖胺聚糖）。在这种新的水脱细胞方案中，可以保留软骨的生化成分。这允许整体上研究细胞外基质中的生物化学和物理含量，并且该方案对于研究组织微环境在支持软骨细胞生长和功能中的作用肯定是有用的。软骨的生化成分可以保存。这允许整体上研究细胞外基质中的生物化学和物理含量，并且该方案对于研究组织微环境在支持软骨细胞生长和功能中的作用肯定是有用的。软骨的生化成分可以保存。这允许整体上研究细胞外基质中的生物化学和物理含量，并且该方案对于研究组织微环境在支持软骨细胞生长和功能中的作用肯定是有用的。