Optimal combination of soluble factors for tissue engineering of permanent cartilage from cultured human chondrocytes
Optimal combination of soluble factors for tissue engineering of permanent cartilage from cultured human chondrocytes
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DOI：
10.1074/jbc.m608383200
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发表时间：
2007-07-13
影响因子：
4.8
通讯作者：
Hoshi, Kazuto
中科院分区：
文献类型：
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作者：
Liu, Guangyao;Kawaguchi, Hiroshi;Hoshi, Kazuto
由于永久性软骨自身再生能力差，利用组织工程技术从自体软骨细胞中再生永久性软骨可能对各种骨骼疾病的治疗有很大的益处。然而，常规的自体软骨细胞植入在数量和质量上都是不足的，这是由于两个主要的限制：在长期培养期间用于增殖的去分化和通过刺激用于再分化的肥大分化。为了克服这些局限性，本研究试图从12种推定的软骨细胞调节剂中确定无血清条件下原代人软骨细胞培养物的最佳组合。从2（12）= 4，096种组合中，通过部分析因设计选择256种组合，经统计学确定，骨形态发生蛋白-2和胰岛素（BI）是重复传代后引起去分化细胞再分化的最有效组合。我们进一步发现，添加三碘甲腺原氨酸（T3）通过抑制Akt信号通路阻止BI诱导的再分化软骨细胞的肥大分化。在BI + T3刺激下，由在去端胶原和聚（L-乳酸）支架中培养的人软骨细胞形成的植入物由足够的透明软骨组成，其在裸鼠中移植后具有与天然软骨相当的机械性质，表明BI + T3是从自体软骨细胞再生临床实用的永久性软骨的最佳组合。
Since permanent cartilage has poor self-regenerative capacity, its regeneration from autologous human chondrocytes using a tissue engineering techniquemaygreatly benefit the treatment of various skeletal disorders. However, the conventional autologous chondrocyte implantation is insufficient both in quantity and in quality due to two major limitations: dedifferentiation during a long term culture for multiplication and hypertrophic differentiation by stimulation for the redifferentiation. To overcome the limitations, this study attempted to determine the optimal combination in primary human chondrocyte cultures under a serum-free condition, from among 12 putative chondrocyte regulators. From the exhaustive 2(12) = 4,096 combinations, 256 were selected by fractional factorial design, and bone morphogenetic protein-2 and insulin (BI) were statistically determined to be the most effective combination causing redifferentiation of the dedifferentiated cells after repeated passaging. We further found that the addition of triiodothyronine (T3) prevented the BI-induced hypertrophic differentiation of redifferentiated chondrocytes via the suppression of Akt signaling. The implant formed by the human chondrocytes cultured in atelocollagen and poly(L-latic acid) scaffold under the BI + T3 stimulation consisted of sufficient hyaline cartilage with mechanical properties comparable with native cartilage after transplantation in nude mice, indicating that BI + T3 is the optimal combination to regenerate a clinically practical permanent cartilage from autologous chondrocytes.