目的通过实时无标记细胞分析法(real time cell analysis,RTCA)与MTT法的比较研究,探讨RTCA法用于医疗器械细胞毒性常规检测的可行性。方法首先,通过筛选细胞接种密度建立基于L929细胞的RTCA细胞毒性检测方法。然后,使用不同浓度的DMSO...目的通过实时无标记细胞分析法(real time cell analysis,RTCA)与MTT法的比较研究,探讨RTCA法用于医疗器械细胞毒性常规检测的可行性。方法首先,通过筛选细胞接种密度建立基于L929细胞的RTCA细胞毒性检测方法。然后,使用不同浓度的DMSO稀释液、标准对照材料浸提液、56批真实世界的医疗器械产品,从简单到复杂分别比较RTCA法与MTT法用于细胞毒性检测结果的一致性,并且汇总多次试验的阴性对照和阳性对照结果,比较两种方法的可重复性。结果RTCA法检测不同浓度的DMSO稀释液没有剂量效应关系,检测标准对照材料浸提液与MTT法结果完全一致并高度相关(R=0.9898),检测56批医疗器械产品与MTT法结果一致率为80.4%,仅中度相关(R=0.6183)。RTCA法的可重复性不比MTT法好。结论RTCA法能有效检测医疗器械细胞毒性,但是部分产品可能因溶出物干扰检测结果而不适用。RTCA法作为一种新型的医疗器械细胞毒性检测方法的适用性还需要进一步研究。展开更多
Objective Several studies have indicated that miR-15a,miR-15b and miR-16 may be the important regulators of apoptosis.Since attenuate apoptosis could protect myocardium and reduce infarction size,the present study was...Objective Several studies have indicated that miR-15a,miR-15b and miR-16 may be the important regulators of apoptosis.Since attenuate apoptosis could protect myocardium and reduce infarction size,the present study was aimed to find out whether these miRNAs participate in regulating myocardial ischemia reperfusion （I/R） injury.Methods Apoptosis in mice hearts subjected to I/R was detected by TUNEL assay in vivo,while flow cytometry analysis followed by Annexin V/PI double stain in vitro was used to detect apoptosis in cultured cardiomyocytes which were subjected to hypoxia/reoxygenation （H/R）.Taqman real-time quantitative PCR was used to confirm whether miR-15a/15b/16 were involved in the regulation of cardiac I/R and H/R.Results Compared to those of the controls,I/R or H/R induced apoptosis of cardiomyocytes was significantly iucreased both in vivo （24.4％ ± 9.4％ vs.2.2％ ± 1.9％,P ＜ 0.01,n =5） and in vitro （14.12％ ±0.92％ vs.2.22％ ± 0.08％）.The expression of miR-15a and miR-15b,but not miR-16,was increased in the mice I/R model,and the results were consistent in the H/R model.Conclusions Our data indicate miR-15 and miR-15b are up-regulated in response to cardiac I/R injury,therefore,down-regulation of miR- 15a/b may be a promising strategy to reduce myocardial apoptosis induced by cardiac I/R injury.展开更多
文摘目的通过实时无标记细胞分析法(real time cell analysis,RTCA)与MTT法的比较研究,探讨RTCA法用于医疗器械细胞毒性常规检测的可行性。方法首先,通过筛选细胞接种密度建立基于L929细胞的RTCA细胞毒性检测方法。然后,使用不同浓度的DMSO稀释液、标准对照材料浸提液、56批真实世界的医疗器械产品,从简单到复杂分别比较RTCA法与MTT法用于细胞毒性检测结果的一致性,并且汇总多次试验的阴性对照和阳性对照结果,比较两种方法的可重复性。结果RTCA法检测不同浓度的DMSO稀释液没有剂量效应关系,检测标准对照材料浸提液与MTT法结果完全一致并高度相关(R=0.9898),检测56批医疗器械产品与MTT法结果一致率为80.4%,仅中度相关(R=0.6183)。RTCA法的可重复性不比MTT法好。结论RTCA法能有效检测医疗器械细胞毒性,但是部分产品可能因溶出物干扰检测结果而不适用。RTCA法作为一种新型的医疗器械细胞毒性检测方法的适用性还需要进一步研究。
文摘Objective Several studies have indicated that miR-15a,miR-15b and miR-16 may be the important regulators of apoptosis.Since attenuate apoptosis could protect myocardium and reduce infarction size,the present study was aimed to find out whether these miRNAs participate in regulating myocardial ischemia reperfusion （I/R） injury.Methods Apoptosis in mice hearts subjected to I/R was detected by TUNEL assay in vivo,while flow cytometry analysis followed by Annexin V/PI double stain in vitro was used to detect apoptosis in cultured cardiomyocytes which were subjected to hypoxia/reoxygenation （H/R）.Taqman real-time quantitative PCR was used to confirm whether miR-15a/15b/16 were involved in the regulation of cardiac I/R and H/R.Results Compared to those of the controls,I/R or H/R induced apoptosis of cardiomyocytes was significantly iucreased both in vivo （24.4％ ± 9.4％ vs.2.2％ ± 1.9％,P ＜ 0.01,n =5） and in vitro （14.12％ ±0.92％ vs.2.22％ ± 0.08％）.The expression of miR-15a and miR-15b,but not miR-16,was increased in the mice I/R model,and the results were consistent in the H/R model.Conclusions Our data indicate miR-15 and miR-15b are up-regulated in response to cardiac I/R injury,therefore,down-regulation of miR- 15a/b may be a promising strategy to reduce myocardial apoptosis induced by cardiac I/R injury.
