聚乳酸降解终产物对成骨样细胞增殖和成骨表型影响的体外实验研究

发布时间：2018-03-06 22:37

  本文选题：骨组织工程　切入点：支架材料　出处：《中国修复重建外科杂志》2014年12期 　论文类型：期刊论文

【摘要】：目的探讨聚乳酸(polylactic acid,PLA)降解终产物乳酸对成骨样细胞增殖和成骨表型的影响,为骨组织工程支架材料的设计和降解调控提供理论依据。方法实验分为A、B、C 3组,A、B组分别采用含4、8、16、22、27 mmol/L右旋乳酸(L-lactic acid,L-LA)和外消旋乳酸(D,L-lactic acid,D,L-LA)的培养液培养Ros17/2.8成骨样细胞,C组以不含乳酸的培养液(p H7.4)培养细胞。培养1、3、5 d,采用MTT法检测细胞相对增殖率(relative growth ratio,PGR)和细胞毒性评级。另取含4 mmol/L L-LA(A组)及4 mmol/L D,L-LA(B组)的培养液培养Ros17/2.8成骨样细胞,以不含乳酸的培养液(p H7.4)培养细胞作为空白对照组(C组)。培养1、5 d时检测相对ALP活性(relative ALP ratio,RAR),21 d时采用von Kossa染色法检测细胞矿化结节生成并行定量分析。结果乳酸浓度为4 mmol/L时,培养5 d内A、B组RGR均值均80%,细胞毒性评级为0级或Ⅰ级,无明显细胞毒性;浓度增大至8 mmol/L和16 mmol/L时,A、B组表现出毒性作用的时间分别为培养5 d和3 d;浓度≥22 mmol/L时,A、B组乳酸培养液在培养1 d时即对细胞增殖产生明显抑制。相同浓度下,各时间点A组RGR均高于B组,除4 mmol/L浓度培养1 d和3 d外,差异均有统计学意义(P0.05)。培养1 d,A、B组RAR分别为144.1%±3.2%和115.2%±9.8%,5 d时分别为129.6%±9.8%和78.2%±6.9%,A组均显著高于B组(P0.05)。von Kossa染色示,培养21 d,A组黑色团状物明显多于B、C组,定量分析分别为91.2%±8.2%、50.3%±7.9%和54.2%±8.6%,A组显著高于B、C组(P0.05);B、C组间差异无统计学意义(P0.05)。结论 PLA降解产物的浓度和组成对成骨样细胞增殖和成骨表型均有显著影响。
[Abstract]:Objective to investigate the effect of lactic acid, the final product of polylactic lactic acid (PLA) degradation, on the proliferation and osteogenic phenotype of osteoblast like cells. Methods to provide theoretical basis for the design and regulation of bone tissue engineering scaffolds. Methods Ros17/2.8 osteoblast cells were cultured in the culture medium of Agna C 3 groups, Acara B group, containing 4 ~ (8) C ~ (16) ~ (22) mmol/L dextral lactate L ~ (lactic) L ~ (lactic) L ~ (LLA) and D ~ (L-lactic acidacidine) DL-LAA), respectively, in vitro culture of Ros17/2.8 osteoblast-like cells, in which Ros17/2.8 osteoblast-like cells were cultured in vitro. The cells were cultured in lactic acid free medium (pH7.4) for 5 days. The relative proliferation rate and cytotoxicity rating of Ros17/2.8 osteoblasts were measured by MTT assay. The osteoblast cells were cultured in 4 mmol/L L-LAA group and 4 mmol/L DL-LAGA-B group respectively. The cells were cultured in lactic acid free medium (pH7.4) as blank control group. The relative ALP activity and relative ALP activity were detected by von Kossa staining at 21 d after culture. The results showed that the concentration of lactic acid was 4 mmol/L. The mean value of RGR in group A B was 80% within 5 days, and the cytotoxicity rating was 0 grade or I grade, and there was no obvious cytotoxicity. When the concentration was increased to 8 mmol/L and 16 mmol/L, the time of toxicity was 5 days and 3 days, respectively, and the lactic acid culture medium of the group with concentration 鈮，

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