改良载体提高大鼠成肌细胞移植效率

Improvement in myoblast-implantation efficiency in rats with modified carrier

目的:观察含1g/L透明质酸钠的F12培养基能否提高成肌细胞移植效率和促进组织修复。方法:实验于2003-9/2004-10在四川大学华西医院修复重建实验室完成。①选取5月龄SD大鼠84只,建立胫前肌的深度冻伤的动物模型。②术后第10天,再次手术暴露冻伤肌组织。随机分为4组:取42只,左侧胫前肌注射以含1g/L透明质酸钠的F12培养基为载体的成肌细胞为左侧成肌细胞组;右侧胫前肌注射以F12培养基为载体的成肌细胞为右侧成肌细胞组;另42只,左侧注射含1g/L透明质酸钠的F12培养基溶液为培养液组;右侧不予注射为空白组。③各组分别于术后第2,5,9天,2,4,8,12周取材观察肌质量变化及组织学改变;于术后第2,5,9天进行磷酸肌酸激酶及同工酶测试。结果:84只大鼠均进入结果分析。①各组胫前肌肌质量变化:术后第2天和第5天,空白组肌质量最轻(P<0.05),其余组间差异无显著性(P>0.05)。术后第9天及第2,4,8,12周,左、右侧成肌细胞组肌质量明显高于培养液组和空白组(P<0.05),而左、右侧成肌细胞组之间、培养液组和空白组之间差异无显著性(P>0.05)。②组织学观察:两组移植细胞均分化形成肌纤维,参与组织修复。透明质酸钠还促进早期小血管的再生。③磷酸肌酸激酶及磷酸肌酸激酶同工酶含量:运用两种载体的成肌细胞移植,冻伤组织中的磷酸肌酸激酶及同工酶活性明显增高(P<0.05),显示移植细胞在冻伤局部增殖、分化参与受损组织的修复,以含透明质酸钠溶液为载体的移植组成肌细胞参与修复的总体活性较高。结论:成肌细胞移植对冻伤的肌肉组织有明显的修复作用,以透明质酸钠的溶液为载体可以提高成肌细胞移植效率。

AIM： To investigate whether the F12 nutrient medium, containing 1 g/L of sodium hyaluronate, can enhance the transplant efficiency of myoblast and promote the tissue repair. METHODS： The experiment was conducted in the Laboratory of Repair and Reconstruction, West China Hospital of Sichuan University between Semptember 2003 and October 2004. （1）A total of 84 5-month-old SD rats were selected to establish the model of severe cryo-damaged tibialis anterior＂ muscle. （2）On the 10^th day of post-operation, the cryo-damaged tissues were exposed through re-operation. Animals were randomly divided into 4 groups： 42 out of which were injected with the myoblasts, the cartier of which was F12 nutrient medium that containing 1 g/L sodium hyaluronate, in the left-side tibialis anterior muscle, and taken as the left-side myoblast group. Rats received injection of myoblasts in the right-side tibialis anterior muscle were taken as the right-side myoblast group. Another 42 rats injected with F12 nutrient medium solution, containing 1 g/L sodium hyaluronate in the left side were taken as the nutrient solution group, and rats received no injection in the right side were taken as the blank group. （3）Materials were drawn from all groups respectively at 2, 5, 9 days and 2, 4, 8 and 12 weeks after operation to observe the changes of muscle mass and histological changes. The phosphocreatine kinase and iseenzyme were inspected at 2,5 and 9 days after operation. RESULTS： A total of 84 rats were involved in the analysis of results. （1） The changes of mass of tibialis anterior muscle in all groups： at 2 and 5 days after implantation, the muscle mass in the blank group was the lightest P 〈 0.05）, and no significant difference was found among other groups（P 〉 0.05）. At 9 days, 2 weeks, 4 weeks, 8 weeks and 12 weeks after implantation, the muscle mass of left and right myoblast groups were significantly higher than the nutrient solution group and blank group（P 〈 0.05 ）, while there were no remarkable differences between left and right myoblast groups, and between the nutrient solution group and the blank group （P 〉 0.05）. （2）Histologieal observation： the implanted myoblasts of both groups all differentiated and formed into myofibers, and participated in the tissue repair. The sodium hyaluronate promoted the regeneration of capillary in early stage. （3）The contents of phesphocreatine kinase and phosphocreatine kinase isoenzyme： the activities of phosphocreatine kinase and isoenzyme in cryo-damaged tissues were significantly enhanced by the implantation of myoblasts in two carriers, （P 〈 0.05）, which showed that implanted myoblasts proliferated, differentiated and participated in the repair of tissues. The total activity of myoblasts that took part in the repair was the highest in the implantation group, which took sodium hyaluronate solution as the carrier. CONCLUSION： Implantation of myoblast plays a significant role in repairing of severe cryo-damaged tibialis anterior muscle. The sodium hyaluronate solution, as a carrier, can promote the efficiency of implantation of myoblast.