Chua K H, Aminuddin B S, Fuzina N H, Ruszymah B H I
Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, Kuala Lumpur 50300, Malaysia.
Singapore Med J. 2007 Apr;48(4):324-32.
The objectives of this study were to determine the optimum concentration of basic fibroblast growth factor (bFGF) in foetal bovine serum (FBS) or human serum (HS) supplemented medium for adult human nasal septum chondrocyte culture and to evaluate the potential of cartilage regeneration.
Dose effects of bFGF were evaluated from a range of 0.0 ng/ml to 10.0 ng/ml in the culture medium either supplemented with ten percent HS or ten percent FBS. Chondrocyte growth rate, viability and gene expression were evaluated. Cultured chondrocytes were then suspended in hydrogel for cartilage regeneration. Engineered cartilages were evaluated with standard histological staining and gene expression analysis.
Our results showed that the chondrocyte growth rate increased in a dose dependent manner of bFGF until 5.0 ng/ml. This increment is further enhanced with ten percent HS supplementation. All cultured chondrocytes exhibited the same gene expression profile regardless of bFGF concentration and type of serum used. The histological staining and gene expression analysis of engineered cartilage after implantation showed characteristics similar to native cartilage.
bFGF with ten percent HS was able to accelerate the chondrocyte growth rate, provided more chondrocytes for therapeutic purposes and therefore minimised the amount of nasal septum cartilage needed to be harvested from patients. The combination of 5.0 ng/ml of bFGF and ten percent HS in the culture medium was safer and had less risk compared to FBS. It also demonstrated valuable implications on constructing high quality autologous cartilage for treating cartilage defects, especially in head and neck reconstructive surgery.
本研究的目的是确定在添加胎牛血清(FBS)或人血清(HS)的培养基中用于成人鼻中隔软骨细胞培养的碱性成纤维细胞生长因子(bFGF)的最佳浓度,并评估软骨再生的潜力。
在添加10% HS或10% FBS的培养基中,评估bFGF在0.0 ng/ml至10.0 ng/ml范围内的剂量效应。评估软骨细胞的生长速率、活力和基因表达。然后将培养的软骨细胞悬浮在水凝胶中用于软骨再生。用标准组织学染色和基因表达分析评估工程化软骨。
我们的结果表明,直到5.0 ng/ml,软骨细胞的生长速率以bFGF的剂量依赖性方式增加。添加10% HS可进一步增强这种增加。无论bFGF浓度和所用血清类型如何,所有培养的软骨细胞都表现出相同的基因表达谱。植入后工程化软骨的组织学染色和基因表达分析显示出与天然软骨相似的特征。
添加10% HS的bFGF能够加速软骨细胞的生长速率,为治疗目的提供更多的软骨细胞,因此将需要从患者身上采集的鼻中隔软骨量降至最低。与FBS相比,培养基中5.0 ng/ml的bFGF和10% HS的组合更安全且风险更小。它还对构建用于治疗软骨缺损的高质量自体软骨具有重要意义,尤其是在头颈部重建手术中。