Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo.
Cartilage Derived from Bone Marrow Mesenchymal Stem Cells Expresses Lubricin In Vitro and In Vivo.
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DOI：
10.1371/journal.pone.0148777
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Sekiya I
中科院分区：
文献类型：
--
作者：
Nakagawa Y;Muneta T;Otabe K;Ozeki N;Mizuno M;Udo M;Saito R;Yanagisawa K;Ichinose S;Koga H;Tsuji K;Sekiya I
Lubricin在浅表软骨中的表达将是软骨再生成功的关键因素。间充质干细胞（MSC）是一种有吸引力的细胞来源，使用MSC的聚集体在软骨形成潜力和细胞粘附效率方面具有一些优势。Lubricin在移植的MSC中的表达至今尚未完全阐明。我们的目标是确定（1）人MSC的软骨颗粒在体外软骨形成中是否表达润滑素，（2）人MSC的聚集体是否促进润滑素表达，以及（3）MSC的聚集体在移植到大鼠骨软骨缺损中后是否在浅表软骨中表达润滑素。对于体外分析，人骨髓（BM）MSC通过沉淀培养分化成软骨，并且还使用悬滴技术聚集。对于动物研究，将来自GFP转基因大鼠的BM MSC的聚集体移植到野生型大鼠膝关节的trophilar沟中的骨软骨缺损处。Lubricin主要在分化和再生软骨中表达。在体外分析中，在颗粒和条件培养基的表面区域中检测到润滑素。蛋白聚糖4（Prg4），编码润滑素，在小球中的mRNA表达显着高于未分化的MSC。聚集体的表面和深层之间表现出不同的形态特征，聚集体形成后Prg 4 mRNA的表达增加。在聚集体中也发现了润滑素。在大鼠研究中，如宏观和组织学分析所示，MSC组的关节软骨再生明显优于对照组。透射电镜显示MSC组的浅层软骨形态较对照组更接近完整软骨。GFP阳性细胞保留在修复组织中，并在浅层软骨中表达润滑素。骨髓间充质干细胞来源的软骨在体内外均表达润滑素蛋白。骨髓间充质干细胞体外聚集促进润滑素表达，骨髓间充质干细胞聚集体移植修复大鼠骨软骨缺损模型。我们的研究结果表明，聚集的MSC可能是临床相关的治疗方法，关节软骨再生与适当的浅表区在未来。
Lubricin expression in the superficial cartilage will be a crucial factor in the success of cartilage regeneration. Mesenchymal stem cells (MSCs) are an attractive cell source and the use of aggregates of MSCs has some advantages in terms of chondrogenic potential and efficiency of cell adhesion. Lubricin expression in transplanted MSCs has not been fully elucidated so far. Our goals were to determine (1) whether cartilage pellets of human MSCs expressed lubricin in vitro chondrogenesis, (2) whether aggregates of human MSCs promoted lubricin expression, and (3) whether aggregates of MSCs expressed lubricin in the superficial cartilage after transplantation into osteochondral defects in rats. For in vitro analysis, human bone marrow (BM) MSCs were differentiated into cartilage by pellet culture, and also aggregated using the hanging drop technique. For an animal study, aggregates of BM MSCs derived from GFP transgenic rats were transplanted to the osteochondral defect in the trochlear groove of wild type rat knee joints. Lubricin expression was mainly evaluated in differentiated and regenerated cartilages. In in vitro analysis, lubricin was detected in the superficial zone of the pellets and conditioned medium. mRNA expression of Proteoglycan4 (Prg4), which encodes lubricin, in pellets was significantly higher than that of undifferentiated MSCs. Aggregates showed different morphological features between the superficial and deep zone, and the Prg4 mRNA expression increased after aggregate formation. Lubricin was also found in the aggregate. In a rat study, articular cartilage regeneration was significantly better in the MSC group than in the control group as shown by macroscopical and histological analysis. The transmission electron microscope showed that morphology of the superficial cartilage in the MSC group was closer to that of the intact cartilage than in the control group. GFP positive cells remained in the repaired tissue and expressed lubricin in the superficial cartilage. Cartilage derived from MSCs expressed lubricin protein both in vitro and in vivo. Aggregation promoted lubricin expression of MSCs in vitro and transplantation of aggregates of MSCs regenerated cartilage including the superficial zone in a rat osteochondral defect model. Our results indicate that aggregated MSCs could be clinically relevant for therapeutic approaches to articular cartilage regeneration with an appropriate superficial zone in the future.