Bone and Joint Research Group, Centre for Human Development, Stem Cells and Regeneration, Human Development and Health, Institute of Developmental Sciences University of Southampton, Tremona Road, Southampton, SO16 6YD, UK.
Cancer Sciences Unit, Somers Cancer Research, University of Southampton, Tremona Road, Southampton, SO16 6YD, UK.
Sci Rep. 2016 Aug 31;6:32168. doi: 10.1038/srep32168.
Biomaterial development for tissue engineering applications is rapidly increasing but necessitates efficacy and safety testing prior to clinical application. Current in vitro and in vivo models hold a number of limitations, including expense, lack of correlation between animal models and human outcomes and the need to perform invasive procedures on animals; hence requiring new predictive screening methods. In the present study we tested the hypothesis that the chick embryo chorioallantoic membrane (CAM) can be used as a bioreactor to culture and study the regeneration of human living bone. We extracted bone cylinders from human femoral heads, simulated an injury using a drill-hole defect, and implanted the bone on CAM or in vitro control-culture. Micro-computed tomography (μCT) was used to quantify the magnitude and location of bone volume changes followed by histological analyses to assess bone repair. CAM blood vessels were observed to infiltrate the human bone cylinder and maintain human cell viability. Histological evaluation revealed extensive extracellular matrix deposition in proximity to endochondral condensations (Sox9+) on the CAM-implanted bone cylinders, correlating with a significant increase in bone volume by μCT analysis (p < 0.01). This human-avian system offers a simple refinement model for animal research and a step towards a humanized in vivo model for tissue engineering.
生物材料的开发在组织工程应用中迅速增加，但在临床应用之前需要进行功效和安全性测试。目前的体外和体内模型存在许多局限性，包括费用高、动物模型与人类结果之间缺乏相关性以及需要对动物进行侵入性手术；因此需要新的预测筛选方法。在本研究中，我们验证了这样一个假设，即鸡胚绒毛尿囊膜（CAM）可用作生物反应器，用于培养和研究人类活体骨的再生。我们从人股骨头中提取骨圆柱体，用钻头缺陷模拟损伤，并将骨植入 CAM 或体外对照培养物中。微计算机断层扫描（μCT）用于定量骨体积变化的幅度和位置，然后进行组织学分析以评估骨修复情况。CAM 血管被观察到渗透到人类骨圆柱体中，并维持人类细胞活力。组织学评估显示，在 CAM 植入的骨圆柱体的软骨内骨化（Sox9+）附近有大量细胞外基质沉积，与 μCT 分析显示的骨体积显著增加相关（p<0.01）。这种人类-禽类系统为动物研究提供了一个简单的改进模型，也是向组织工程的人类化体内模型迈进的一步。
