Nishikori T, Ochi M, Uchio Y, Maniwa S, Kataoka H, Kawasaki K, Katsube K, Kuriwaka M
Department of Orthopaedics, Shimane Medical University, 89-1 Enya-cho, Izumo-shi, Shimane-ken 693-8501, Japan.
J Biomed Mater Res. 2002 Feb;59(2):201-6. doi: 10.1002/jbm.1226.
The effects of low-intensity pulsed ultrasound (US) on the proliferation and chondroitin sulfate synthesis of cultured chondrocytes embedded in Atelocollagen gel in vitro were examined. Articular cartilage was harvested from the hip, knee, and shoulder joints of 10-week-old Japanese white rabbits. Chondrocytes isolated by collagenase digestion were embedded in type I collagen gel, Atelocollagen gel, and were cultured in Dulbecco's modified eagle's medium for 3 weeks. The US apparatus, SAFHS, was used to deliver an ultrasound signal with spatial and temporal average intensities of 30 mW/cm(2) (US group). The frequency was 1.5 MHz with a 200-microsecond tone burst repeated at 1.0 kHz. US treatments were administered for 20 min per day under culture dishes, with the medium replaced twice a week. Another group of cells was exposed to sham ultrasound as a control. Cell number, histological findings, synthesis of isomers of chondroitin sulfate, and stiffness of the chondrocyte-collagen gel composites were analyzed. US exposure promoted synthesis of chondroitin sulfate, especially chondroitin 6-sulfate, although it did not significantly enhance cell number and stiffness. In this three-dimensional culture model, these results suggest that US exposure may be clinically useful in improving the quality of chondrocyte-Atelocollagen implants for transplantation into articular cartilage defects.
研究了低强度脉冲超声(US)对体外嵌入脱细胞胶原蛋白凝胶中的培养软骨细胞增殖和硫酸软骨素合成的影响。从10周龄日本白兔的髋关节、膝关节和肩关节获取关节软骨。通过胶原酶消化分离得到的软骨细胞被嵌入I型胶原蛋白凝胶、脱细胞胶原蛋白凝胶中,并在杜氏改良伊格尔培养基中培养3周。使用SAFHS超声设备传递空间和时间平均强度为30 mW/cm²的超声信号(超声组)。频率为1.5 MHz,200微秒的音爆以1.0 kHz重复。超声治疗在培养皿下每天进行20分钟,每周更换两次培养基。另一组细胞接受假超声作为对照。分析了细胞数量、组织学结果、硫酸软骨素异构体的合成以及软骨细胞 - 胶原蛋白凝胶复合材料的硬度。超声暴露促进了硫酸软骨素的合成,尤其是6 - 硫酸软骨素,尽管它没有显著增加细胞数量和硬度。在这个三维培养模型中,这些结果表明超声暴露在临床上可能有助于提高用于移植到关节软骨缺损处的软骨细胞 - 脱细胞胶原蛋白植入物的质量。