Exosomes derived from mature chondrocytes facilitate subcutaneous stable ectopic chondrogenesis of cartilage progenitor cells.
Exosomes derived from mature chondrocytes facilitate subcutaneous stable ectopic chondrogenesis of cartilage progenitor cells.
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来自成熟软骨细胞的外泌体促进软骨祖细胞皮下稳定异位软骨形成
DOI：
10.1186/s13287-018-1047-2
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发表时间：
2018-11-21
影响因子：
7.5
通讯作者：
Liu K
中科院分区：
文献类型：
--
作者：
Chen Y;Xue K;Zhang X;Zheng Z;Liu K
构建具有合适基质成分且能持续软骨生成的软骨仍然是软骨缺损领域的一个长期挑战。基于软骨祖细胞（CPC）的组织工程因其强大的软骨分化能力近期受到关注。然而，由于缺乏合适的软骨生成微环境，CPC再生软骨在皮下环境中的临床应用仍然是一个挑战。在本研究中，软骨细胞来源的外泌体（CC - Exos）被用于在皮下环境中为CPC构建体提供软骨信号，以实现高效的异位软骨再生。
制备兔CPC - 海藻酸盐构建体，并将其皮下植入裸鼠体内。术后以相同剂量（每次100 μL注射含30 μg外泌体）将CC - Exos注射到构建体中，此后每周注射一次，持续12周。骨髓间充质干细胞来源的外泌体（BMSC - Exos）被用作阳性对照。阴性对照组小鼠给予相同体积的磷酸盐缓冲液（PBS）。在植入后4周和12周，通过组织学、免疫染色和蛋白质分析来分析CC - Exos和BMSC - Exos在皮下环境中促进软骨生成和软骨组织稳定性的潜力。通过与CPC和人脐静脉内皮细胞共培养，评估BMSC - Exos和CC - Exos在体外对软骨生成和血管生成特性的影响。
CC - Exos注射增加了工程构建体中的胶原蛋白沉积，并使血管长入最小化，从而高效且可重复地发育成软骨。生成的软骨在表型上稳定，在长达12周的时间内肥大和血管长入最少，而用BMSC - Exos形成的软骨则以肥大分化并伴有血管长入为特征。体外实验表明，CC - Exos刺激CPC增殖，增加软骨生成标志物的表达，同时抑制血管生成。
这些发现表明，新型的CC - Exos为引导CPC稳定的异位软骨生成提供了更优的微环境。CC - Exos的使用可能代表一种现成的、无细胞的治疗方法，用于促进皮下环境中的软骨再生。
BackgroundDeveloping cartilage constructed with the appropriate matrix composition and persistent chondrogenesis remains an enduring challenge in cartilage defects. Cartilage progenitor cell (CPC)-based tissue engineering has attracted recent attention because of its strong chondrogenic differentiation capacity. However, due to the lack of a suitable chondrogenic niche, the clinical application of CPC-regenerated cartilage in the subcutaneous environment remains a challenge. In this study, exosomes derived from chondrocytes (CC-Exos) were used to provide the CPC constructs with a cartilage signal in subcutaneous environments for efficient ectopic cartilage regeneration.MethodsRabbit CPC-alginate constructs were prepared and implanted subcutaneously in nude mice. CC-Exos were injected into the constructs at the same dose (30 μg exosomes per 100 μL injection) after surgery and thereafter weekly for a period of 12 weeks. Exosomes derived from bone mesenchymal stem cells (BMSC-Exos) were used as the positive control. The mice in the negative control were administered with the same volume of PBS. At 4 and 12 weeks after implantation, the potential of CC-Exos and BMSC-Exos to promote chondrogenesis and stability of cartilage tissue in a subcutaneous environment were analyzed by histology, immunostaining, and protein analysis. The influences of BMSC-Exos and CC-Exos on chondrogenesis and angiogenic characteristics in vitro were assessed via coculturing with CPCs and human umbilical vein endothelial cells.ResultsThe CC-Exos injection increased collagen deposition and minimized vascular ingrowth in engineered constructs, which efficiently and reproducibly developed into cartilage. The generated cartilage was phenotypically stable with minimal hypertrophy and vessel ingrowth up to 12 weeks, while the cartilage formed with BMSC-Exos was characterized by hypertrophic differentiation accompanied by vascular ingrowth. In vitro experiments indicated that CC-Exos stimulated CPCs proliferation and increased expression of chondrogenesis markers while inhibiting angiogenesis.ConclusionsThese findings suggest that the novel CC-Exos provides the preferable niche in directing stable ectopic chondrogenesis of CPCs. The use of CC-Exos may represent an off-the-shelf and cell-free therapeutic approach for promoting cartilage regeneration in the subcutaneous environment.
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