CELL growth and viability analysis in poli membranes (L-lactic acid-CO-glycolic acid): an in vitro study

ABSTRACT Objective: Performing an in vitro evaluation of the biological effects on cell growth and viability of fibroblasts in PLGA membranes with and without simvastatin. Methods: Two groups of resorbable synthetic polymeric membranes were used: PLGA, with and without simvastatin, cut into a suitable format to fit to 24 thermometric wells. Fibroblasts were grown on resorbable membranes and evaluated for proliferation and viability at 24, 48 and 72 hours after the beginning of cultivation, being the tests performed in triplicate. For the cell growth analysis, the Trypan blue exclusion method was applied, while cell viability was observed by the MTT test. The results were statistically analyzed applying the Two-Way ANOVA, followed by the Bonferroni test, with 95% confidence interval and P value smaller than 0.05 was accepted as statistically significant. Results: Statistical difference (p <0.01) was seen between the control group (2.16x104 ± 0.51 cells) and the PLGA group with simvastatin (1.58x104 ± 0.36 cells) in the 48-hour period. After 72 hours, statistical differences (p <0.001) were observed between the PLGA group with simvastatin (1.66x104 ± 0.49 cells) and the PLGA group without simvastatin (2.25x104 ± 0.2 cells) when compared to the control group (2.81x104 ± 0.33 cells) for cell proliferation. Statistical differences (p <0.05) were observed between the control group (0.27 ± 0.05) and the PLGA group with simvastatin (0.21 ± 0.03). Likewise, a statistical difference (p <0.001) was seen between the PLGA group without simvastatin (0.19 ± 0.02) and the control group after 24 hours. In the 48 – 72-hour period, statistical differences (p <0.001) were observed between the control group (0.36 ± 0.09 and 0.55 ± 0.05, after 48 and 72 hours respectively) and the PLGA group without simvastatin (0.26 ± 0.05 and 0.34 ± 0.07, after 48 and 72 hours respectively), as well as in the PLGA group with simvastatin (0.27 ± 0.04 and 0.31 ± 0, 04, after 48 and 72 hours respectively) for the cell viability test. Conclusion: The association of simvastatin to PLGA membranes had an inhibitory effect on fibroblast proliferation, as well as induced a reduction in cell viability. Thus, the use of PLGA along with simvastatin may assist in guided bone regeneration.

摘要 目的：本研究旨在体外评估含与不含辛伐他汀的聚乳酸-羟基乙酸共聚物（PLGA）膜对成纤维细胞生长及活性的生物学效应。方法：本研究设置两组可吸收合成聚合物膜：分别为含辛伐他汀与不含辛伐他汀的PLGA膜，将其裁剪至适配24孔热测板的合适规格。将成纤维细胞接种于可吸收膜表面，于培养开始后24、48及72小时评估其增殖与活性，所有实验均设置三次重复。细胞生长分析采用台盼蓝排斥试验，细胞活性则通过MTT试验检测。结果采用双因素方差分析（Two-Way ANOVA）进行统计学分析，随后采用邦费罗尼（Bonferroni）校正检验，以95%置信区间为判定标准，将P值小于0.05认定为具有统计学显著性。结果：培养48小时时，对照组（2.16×10⁴ ± 0.51 个细胞）与含辛伐他汀的PLGA组（1.58×10⁴ ± 0.36 个细胞）间存在统计学差异（p<0.01）。培养72小时时，相较于对照组（2.81×10⁴ ± 0.33 个细胞），含辛伐他汀的PLGA组（1.66×10⁴ ± 0.49 个细胞）与不含辛伐他汀的PLGA组（2.25×10⁴ ± 0.2 个细胞）的细胞增殖均存在统计学差异（p<0.001）。细胞活性检测方面，对照组（0.27 ± 0.05）与含辛伐他汀的PLGA组（0.21 ± 0.03）间存在统计学差异（p<0.05）；培养24小时时，不含辛伐他汀的PLGA组（0.19 ± 0.02）与对照组间亦存在统计学差异（p<0.001）。培养48至72小时期间，对照组（48小时为0.36 ± 0.09、72小时为0.55 ± 0.05）、不含辛伐他汀的PLGA组（48小时为0.26 ± 0.05、72小时为0.34 ± 0.07）与含辛伐他汀的PLGA组（48小时为0.27 ± 0.04、72小时为0.31 ± 0.04）的细胞活性均存在统计学差异（p<0.001）。结论：辛伐他汀与PLGA膜联合使用可抑制成纤维细胞增殖，并降低细胞活性。据此，PLGA膜联合辛伐他汀可辅助引导性骨再生。