Use of bone morphogenetic protein 2 and diffusion chambers to engineer cartilage tissue for the repair of defects in articular cartilage
Use of bone morphogenetic protein 2 and diffusion chambers to engineer cartilage tissue for the repair of defects in articular cartilage
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DOI：
10.1002/art.20713
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发表时间：
2005-01-01
影响因子：
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通讯作者：
Takaoka, K
中科院分区：
文献类型：
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作者：
Nawata, M;Wakitani, S;Takaoka, K
Objective.目的：观察重组人骨形态发生蛋白2（rHuBMP-2）异位诱导的软骨样组织对大鼠软骨缺损的修复作用。通过解剖交配后19天的大鼠胚胎的大腿肌肉制备肌源性间充质细胞。细胞在体外单层培养中增殖10天，并与I型胶原（CI）和0、1或10 μ g rHuBMP-2一起装入扩散室（10（6）/室）中，并植入成年大鼠的腹部筋膜下袋中。在植入后2天至6周从扩散室收获组织团块，并通过组织学、逆转录-聚合酶链反应（PCR）检测聚集蛋白聚糖、CII、CIX、CX和CXI、MyoD 1和核心结合因子a1/runt相关基因2，并通过实时PCR检测CII。植入5周后，将腔室中产生的组织颗粒移植到同一品系成年大鼠髌骨沟的全层软骨缺损中。在10 μ g rHuBMP-2的存在下，肌源性间充质细胞在移植后4天表达CII信使RNA，并且在扩散室中移植后5周形成成熟的软骨块。在存在1 μ g rHuBMP-2或不存在rHuBMP-2的情况下不形成软骨。接受10 μ g rHuBMP-2工程化软骨的缺损在移植后6个月内得到修复并恢复到正常形态条件。这种用于修复关节缺损的组织工程方法可以排除在镶嵌关节成形术或自体软骨细胞植入之前采集软骨组织的需要。需要在大型动物中进行进一步研究，以验证该技术在临床实践中的应用。
Objective. To examine the ability of cartilage-like tissue, generated ectopically in a diffusion chamber using recombinant human bone morphogenetic protein 2 (rHuBMP-2), to repair cartilage defects in rats.Methods. Muscle-derived mesenchymal cells were prepared by dissecting thigh muscles of 19-day postcoital rat embryos. Cells were propagated in vitro in monolayer culture for 10 days and packed within diffusion chambers (10(6)/chamber) together with type I collagen (CI) and 0, 1, or 10 mug rHuBMP-2, and implanted into abdominal subfascial pockets of adult rats. Tissue pellets were harvested from the diffusion chambers at 2 days to 6 weeks after implantation, and examined by histology, by reverse transcription-polymerase chain reaction (PCR) for aggrecan, CII, CIX, CX, and CXI, MyoD1, and core binding factor a1/runt-related gene 2, and by real-time PCR for CII. Tissue pellets generated in the chamber 5 weeks after implantation were transplanted into a full-thickness cartilage defect made in the patellar groove of the same strain of adult rat.Results. In the presence of 10 mug rHuBMP-2, muscle-derived mesenchymal cells expressed CII messenger RNA at 4 days after transplantation, and a mature cartilage mass was formed 5 weeks after transplantation in the diffusion chamber. Cartilage was not formed in the presence of 1 mug rHuBMP-2 or in the absence of rHuBMP-2. Defects receiving cartilage engineered with 10 mug rHuBMP-2 were repaired and restored to normal morphologic condition within 6 months after transplantation.Conclusion. This method of tissue engineering for repair of articular defects may preclude the need to harvest cartilage tissue prior to mosaic arthroplasty or autologous chondrocyte implantation. Further studies in large animals will be necessary to validate this technique for application in clinical practice.