Influence of Damage-Associated Molecular Patterns from Chondrocytes in Tissue-Engineered Cartilage
Influence of Damage-Associated Molecular Patterns from Chondrocytes in Tissue-Engineered Cartilage
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DOI：
10.1089/ten.tea.2019.0185
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发表时间：
2020-02-19
影响因子：
4.1
通讯作者：
Hoshi, Kazuto
中科院分区：
文献类型：
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作者：
Fujihara, Yuko;Abe, Takahiro;Hoshi, Kazuto
为了在再生医学中获得稳定的结果，移植细胞的质量是非常重要的。细胞应激可能导致损伤相关分子模式（DAMPs）的释放并激活免疫反应，影响移植组织的结果。在本研究中，我们有意制备坏死软骨细胞，使其逐渐死亡并释放DAMPs，并研究如何影响组织工程软骨的成熟。采用常规热处理方法制备坏死软骨细胞，24 h后其活力开始下降。将含坏死软骨细胞的组织工程软骨皮下移植到C57BL/6J小鼠体内，软骨基质的积累较对照组减少。同时，免疫组化染色显示，坏死软骨细胞的巨噬细胞和中性粒细胞的定位更加明显，提示坏死软骨细胞的DAMPs可以促进更多免疫细胞的迁移。双向电泳和质谱分析证实，前纤层蛋白是坏死软骨细胞释放的重要生物分子。RAW264培养基中加入prelamin后，Inos和Il1b随prelamin添加量的增加而增加。这表明，来自死亡软骨细胞的DAMPs可以诱导周围巨噬细胞的炎症特性，从而损害组织工程软骨的成熟。总之，当含有释放DAMPs的活性较低的软骨细胞时，组织工程软骨的成熟受到阻碍。在再生医学中，细胞的质量对保证临床安全至关重要。在培养过程中，细胞损伤可能发生，如果不是严重到足以立即导致细胞死亡，但仍然诱导较低的生存状态。损伤相关分子模式（DAMPs）从坏死细胞中释放出来，但它们在再生医学中的影响尚不清楚。在这项研究中，我们阐明了来自软骨细胞的DAMPs如何影响组织工程软骨的成熟。同时，我们对坏死软骨细胞可能产生的DAMPs进行了综合分析，发现prelamin是一个重要的分子，可能用于检测坏死软骨细胞的存在。
To obtain stable outcomes in regenerative medicine, the quality of cells for transplantation is of great importance. Cellular stress potentially results in the release of damage-associated molecular patterns (DAMPs) and activates immunological responses, affecting the outcome of transplanted tissue. In this study, we intentionally prepared necrotic chondrocytes that would gradually die and release DAMPs and investigated how the maturation of tissue-engineered cartilage was affected. Necrotic chondrocytes were prepared by a conventional heat-treatment method, by which their viability started to decrease after 24 h. When tissue-engineered cartilage containing necrotic chondrocytes was subcutaneously transplanted into C57BL/6J mice, accumulation of cartilage matrix was decreased compared to the control. Meanwhile, immunohistochemical staining demonstrated that localization of macrophages and neutrophils was more apparent in the constructs of necrotic chondrocytes, suggesting that DAMPs from necrotic chondrocytes could prompt migration of more immune cells. Two-dimensional electrophoresis and mass spectrometry identified prelamin as a significant biomolecule released from necrotic chondrocytes. Also, when prelamin was added to a culture of RAW264, Inos and Il1b were increased in accordance with the content of added prelamin. It was suggested that DAMPs from dying chondrocytes could induce inflammatory properties in surrounding macrophages, impairing the maturation of tissue-engineered cartilage. In conclusion, maturation of tissue-engineered cartilage was hampered when less viable chondrocytes releasing DAMPs were included.Impact statementIn regenerative medicine, the quality of cells is of great importance to secure clinical safety. During culture, damage of cells could occur, if not critical enough to cause immediate cell death, but still inducing a less viable status. Damage-associated molecular patterns (DAMPs) are released from necrotic cells, but their influence in regenerative medicine has yet to be clarified. In this study, we elucidated how DAMPs from chondrocytes could affect the maturation of tissue-engineered cartilage. Also, possible DAMPs from necrotic chondrocytes were comprehensively analyzed, and prelamin was identified as a significant molecule, which may serve for detecting the existence of necrotic chondrocytes.