Saka Natsumi, Watanabe Yoshinobu, Abe Satoshi, Yajima Ayako, Kawano Hirotaka
Department of Orthopaedics, Teikyo University School of Medicine, Tokyo, Japan, 2-11-1 Kaga, Itabashi, Tokyo 173-8606, Japan.
FUJISOFT Co., Ltd, Tokyo, Japan, 2-19-7, Kotobashi, Sumida, Tokyo 130-0022, Japan.
Regen Med Res. 2019;7:1. doi: 10.1051/rmr/190001. Epub 2019 Aug 5.
There is a growing need for chondrocyte implantation for reconstructing cartilage defect. However, ossification of the implanted cartilage is a challenging problem. Implant-type tissue-engineered cartilage from human auricular chondrocytes is a three-dimensional implant type cartilage using PLLA as a scaffold for chondrocytes. Although there is a study which evaluated the ossification of this cartilage in subcutaneous area, there is no study which clarify the possibility of ossification in osteoinductive surroundings. The purpose of this study was to elucidate the possibility of the ossification of implant-type tissue-engineered cartilage using human auricular chondrocyte in an osteoinductive environment.
Human chondrocytes were harvested from ear cartilage. After dispersion by enzyme digestion, they were put into either a poly-L-lactic acid (PLLA) or poly lactic-co-glycolic acid (PLGA) scaffold, with collagen gel. Implant-type tissue-engineered cartilage was interposed between pieces of human iliac bone harvested from the same donor and implanted subcutaneously in nude rats. Scaffold without chondrocytes was used as a control. After 1, 3, and 6 months, ossification and cartilage formation were evaluated by X-ray, hematoxylin-eosin (HE) stain and toluidine blue (TB) stain.
There was no ossification of implant-type cartilage using human chondrocytes, even under osteoinductive conditions. HE staining showed that perichondrium formed around the constructs and chondrocytes were observed 6months after the implantation. TB staining showed metachromasia in every sample, with the area of metachromasia increasing over time, suggesting maturation of the cartilage.
In conclusion, adjacent iliac bone had no apparent effect on the maturation of cartilage in implant-type tissue-engineered cartilage. Cartilage retention and maturation even in the presence of iliac bone could have been due to a scarcity of mesenchymal stem cells in the bone and surrounding area.
软骨细胞植入用于重建软骨缺损的需求日益增长。然而,植入软骨的骨化是一个具有挑战性的问题。人耳软骨细胞来源的植入型组织工程软骨是一种以聚左旋乳酸(PLLA)为软骨细胞支架的三维植入型软骨。尽管有研究评估了这种软骨在皮下区域的骨化情况,但尚无研究阐明其在骨诱导环境中骨化的可能性。本研究的目的是阐明在骨诱导环境下人耳软骨细胞来源的植入型组织工程软骨骨化的可能性。
从耳软骨中获取人软骨细胞。经酶消化分散后,将其与胶原凝胶一起放入聚左旋乳酸(PLLA)或聚乳酸 - 乙醇酸共聚物(PLGA)支架中。将植入型组织工程软骨置于取自同一供体的人髂骨块之间,并皮下植入裸鼠体内。无软骨细胞的支架用作对照。在1、3和6个月后,通过X射线、苏木精 - 伊红(HE)染色和甲苯胺蓝(TB)染色评估骨化和软骨形成情况。
即使在骨诱导条件下,使用人软骨细胞的植入型软骨也未发生骨化。HE染色显示构建体周围形成了软骨膜,植入6个月后观察到软骨细胞。TB染色显示每个样本均有异染性,且异染面积随时间增加,提示软骨成熟。
总之,相邻髂骨对植入型组织工程软骨中软骨的成熟没有明显影响。即使存在髂骨,软骨仍能保留并成熟,这可能是由于骨和周围区域中间充质干细胞数量不足所致。