Yuan Li-Jen, Niu Chi-Chien, Lin Song-Shu, Chan Yi-Sheng, Yang Chuen-Yung, Chen Wen-Jer, Ueng Steve W N
Department of Orthopaedic Surgery and Hyperbaric Oxygen Therapy Center, Chang Gung Memorial Hospital, Taoyuan, Taiwan.
J Orthop Res. 2009 Nov;27(11):1439-46. doi: 10.1002/jor.20889.
The present study investigated the effects of hyperbaric oxygen (HBO) and platelet-derived growth factor-BB (PDGF-BB) in chondrocyte transplantation. In vitro, chondrocytes were treated with HBO, PDGF-BB, and HBO combined with PDGF-BB (H+P). Cell growth was analyzed using cell counting, MTT assay, and FACS analysis. mRNA expression of the PDGF-alpha receptor (PDGFR-alpha) and beta receptor (PDGFR-beta) was detected by RT-PCR. Protein expression of PDGFR-beta was detected by Western blotting. In vivo, chondrocytes and PDGF-BB were suspended in alginate as a transplantation system. Cartilage defects were grafted with this system and with or without HBO treatment. Released PDGF-BB concentration was quantified by ELISA. After 8 weeks, animals were sacrificed and the repaired tissues were examined. In vitro data suggested that each treatment increased cell growth via the up-regulated mRNA expression of PDGFR-alpha and increased cell accumulation in the S-phase. The H+P treatment was more additive in cell growth and in mRNA and protein expression of PDGFR-beta than HBO or PDGF-BB. In vivo results suggested that PDGF-BB delivery lasted for more than 5 weeks. Scoring results showed that each treatment significantly increased the cartilage repair. Safranin-O and type II collagen staining confirmed the hyaline-like cartilage regeneration in the repaired tissues. In situ up-regulation of PDGFR-beta expression partially explains the additive effect of H+P treatment in cartilage repair. Accordingly, H+P offers a potential treatment method for cartilage repair.
本研究调查了高压氧(HBO)和血小板衍生生长因子-BB(PDGF-BB)在软骨细胞移植中的作用。在体外,用HBO、PDGF-BB以及HBO与PDGF-BB联合(H+P)处理软骨细胞。使用细胞计数、MTT法和流式细胞术分析(FACS分析)来分析细胞生长情况。通过逆转录聚合酶链反应(RT-PCR)检测血小板衍生生长因子α受体(PDGFR-α)和β受体(PDGFR-β)的mRNA表达。通过蛋白质免疫印迹法检测PDGFR-β的蛋白表达。在体内,将软骨细胞和PDGF-BB悬浮于藻酸盐中作为移植系统。用该系统对软骨缺损进行移植,并进行或不进行HBO处理。通过酶联免疫吸附测定(ELISA)对释放的PDGF-BB浓度进行定量。8周后,处死动物并检查修复组织。体外数据表明,每种处理均通过上调PDGFR-α的mRNA表达来促进细胞生长,并增加S期的细胞积聚。与HBO或PDGF-BB相比,H+P处理在细胞生长以及PDGFR-β的mRNA和蛋白表达方面具有更强的累加效应。体内结果表明,PDGF-BB的释放持续超过5周。评分结果显示,每种处理均显著促进了软骨修复。番红O和II型胶原染色证实了修复组织中透明样软骨的再生。PDGFR-β表达的原位上调部分解释了H+P处理在软骨修复中的累加效应。因此,H+P为软骨修复提供了一种潜在的治疗方法。