犬去细胞喉支架的生物力学研究

Biomechanical study on decellularized laryngeal scaffold in dogs

目的 评价犬去细胞喉支架的生物力学性能.方法 随机选5只中国杂种犬通过双侧颅甲动脉顺行持续注入去离子剂作为实验组;另取5只同类犬不灌注喉作为对照组.两组分别行大体观察,HE、阿尔新蓝、Masson染色显微镜观察,扫描电镜观察及生物力学性能测定.结果 实验组喉较对照组苍白透明,HE染色观察肌肉黏膜未见细胞残留.阿尔新蓝染色观察软骨糖胺聚糖含量和Masson染色观察软骨胶原含量,实验组灌注喉与未灌注的对照组比较差异均无统计学意义（t值分别为1.462和1.267,P值均＞0.05）.扫描电镜观察实验组软骨基质与对照组比较未出现明显断裂.压凹试验测得甲状软骨压缩模量实验组为（1.06±0.07）MPa（（-x）±s,下同）,对照组为（1.15±0.11）MPa;环状软骨压缩模量实验组为（1.68±0.11）MPa,对照组为（1.67±0.09）MPa,两组比较差异均无统计学意义（t值分别为1.424和0.185,P值均＞0.05）.拉伸试验测得甲状软骨抗拉强度实验组为（5.74±0.88）MPa,对照组为（6.18±1.33）MPa,差异无统计学意义（t=0.627,P值＞0.05）.结论 灌注法去细胞技术构建的犬去细胞喉支架生物力学性能良好,是一种较理想的支架材料.

Objective To evaluate the biomechanical characteristics of the decellularized laryngeal scaffold. Methods Ten Chinese adult dogs were randomly divided into two groups： perfusion group （n=5） and control group （ n = 5）. The acellular larynx scaffold was obtained from dogs through cranial thyroid arteries perfusion with detergents. Comparative examinations were performed by the macroscopic view,histological view （ hematoxylin and eosin stain, Alcian blue stain and Masson stain）, scanning electron microscope （SEM） and biomechanical properties between perfusion group and control group. Results Macroscopic view showed that the decellularized laryngeal scaffold appeared pale asphyxia.HE stain indicated that there were little acellular traces of muscle and mucosa. Alcian blue stain, Masson stain and scanning electron microscope（SEM）suggested that there were no obvious changes about lycosaminoglycan and collagen. The compressive modulus of thyroid cartilage was （ 1.06 ±0. 07） MPa （（-x）±s） in experimental groups and （1.15±0.11） MPa in control group , showing no significant difference （t=1.424,P＞0.05）,neither in compressive modulus of annular cartilage（1.68±0.11）MPa in experimental groups and （1.67±0. 09）MPa in control group （t = 0.185, P＞0.05）. The tensile strength of thyroid cartilage between experimental （5.74±0.88） MPa and control groups （6.18±1.33） MPa did not have the statistical significance（t =0. 627, P ＞0. 05 ）. Conclusion These results indicate that perfusion method can construct a perfect biomechanical acellular larynx scaffold which could be a better selection for laryngeal reconstruction with tissue engineering method.