乳鼠心室细胞在不同处理的封堵器材料上生长的体外研究

目的 通过在体外观察乳鼠心室细胞在不同处理的室间隔缺损封堵器材料上的生长情况,探讨室间隔缺损介入封堵治疗后影响封堵器表面愈合填补的因素.方法 根据室间隔缺损封堵器材料是否有胶原蛋白涂层,将实验分为涂层组和非涂层组,各组又分为3个亚组,分别给予表皮生长因子(EGF组)、5-溴脱氧尿苷(BrdU组)处理及不额外处理(对照组).乳鼠心室肌组织分别与不同处理的封堵器材料(1 cm×2 cm)进行培养,观察组织块开始搏动时间,细胞开始爬行时间,封堵器材料表面生长的细胞种类、含晕及胰岛素样生长因子的含量.结果 (1)涂层组和非涂层组间组织块开始搏动时间差异无统计学意义.亚组中BrdU组开始搏动时间早于EGF组及对照组(P<0.05).(2)涂层组细胞开始爬行时间早于非涂层组(P<0.05).亚组中EGF组开始爬行时间最早(P<0.05).(3)在封堵器材料上生长的细胞包括内皮细胞、成纤维细胞和心肌细胞.内皮细胞含量在EGF组最高,成纤维细胞含量在BrdU组最高,心肌细胞含昔BrdU组与对照组均高于EGF组(P<0.05).(4)各组胰岛素样生长因子的阳性率均在培养的第7天达到最高值.结论 胶原蛋白涂层促进细胞爬行,EGF促进内皮细胞的生长,BrdU可在不影响心肌细胞生长的状态下促进成纤维细胞的生长.

In vitro growth and function of neonatal cardiomyocytes in various transcatheter closure device patches

Objective To observe the effects of collagen-coating, epidermal growth factor (EGF), Bromodeoxyuridine (BrdU) on growth and function of neonatal ventricular cardiomyocytes in transcatheter closure device patches in vitro. Methods Neonatal ventricular cardiomyocytes were cultured with transcatheter closure device patches (1 cm × 2 cm) coated with or without collagen and treated with 10% FBS (control), EGF (20 ng/ml), BrdU (0.1 mmol/L), respectively. In vitro ventricular cardiomyocytes growth and function as well as IGF-I content were determined. Results (1) The beginning time of ventricular cardiomyocytes beating on patches was similar in collagen-coated and uncoated patches treated with PBS, EGF or BrdU, respectively (P > 0.05). The cell beating time was significantly earlier in Brdu group than in PBS and EGF groups (all P < 0.05). (2) Time of cultured cell covering on patches was significantly earlier in coated patches than those uncoated patches in PBS, EGF and BrdU treated groups (all P < 0.05). The ventricular cardiomyocytes covering time on patches was significantly earlier in EGF group than that in PBS and BrdU groups (all P < 0.05). (3) Ventricular cardiomyocytes types survived on patches included endothelial cells, fibroblasts and myocytes. The highest content of endothelial cells was evidenced in EGF group and the highest content of fibroblasts was found in Brdu group. Myocytes content was similar between PBS and BrdU groups (P > 0.05) and significantly higher than that in EGF group (all P < 0.05). (4) IGF-I peaked at the seventh culture day in all groups (all P < 0.01). Conclusions Ventricular cardiomyocytes covering on patches could be enhanced by collagen coating. EGF could promote endothelial cells growth while Brdu could stimulate fibroblasts growth on patches.