本文選題：骨組織工程　切入點(diǎn)：支架材料　出處：《中國修復(fù)重建外科雜志》2014年12期 　論文類型：期刊論文

【摘要】：目的探討聚乳酸(polylactic acid,PLA)降解終產(chǎn)物乳酸對(duì)成骨樣細(xì)胞增殖和成骨表型的影響,為骨組織工程支架材料的設(shè)計(jì)和降解調(diào)控提供理論依據(jù)。方法實(shí)驗(yàn)分為A、B、C 3組,A、B組分別采用含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)的培養(yǎng)液培養(yǎng)Ros17/2.8成骨樣細(xì)胞,C組以不含乳酸的培養(yǎng)液(p H7.4)培養(yǎng)細(xì)胞。培養(yǎng)1、3、5 d,采用MTT法檢測(cè)細(xì)胞相對(duì)增殖率(relative growth ratio,PGR)和細(xì)胞毒性評(píng)級(jí)。另取含4 mmol/L L-LA(A組)及4 mmol/L D,L-LA(B組)的培養(yǎng)液培養(yǎng)Ros17/2.8成骨樣細(xì)胞,以不含乳酸的培養(yǎng)液(p H7.4)培養(yǎng)細(xì)胞作為空白對(duì)照組(C組)。培養(yǎng)1、5 d時(shí)檢測(cè)相對(duì)ALP活性(relative ALP ratio,RAR),21 d時(shí)采用von Kossa染色法檢測(cè)細(xì)胞礦化結(jié)節(jié)生成并行定量分析。結(jié)果乳酸濃度為4 mmol/L時(shí),培養(yǎng)5 d內(nèi)A、B組RGR均值均80%,細(xì)胞毒性評(píng)級(jí)為0級(jí)或Ⅰ級(jí),無明顯細(xì)胞毒性;濃度增大至8 mmol/L和16 mmol/L時(shí),A、B組表現(xiàn)出毒性作用的時(shí)間分別為培養(yǎng)5 d和3 d;濃度≥22 mmol/L時(shí),A、B組乳酸培養(yǎng)液在培養(yǎng)1 d時(shí)即對(duì)細(xì)胞增殖產(chǎn)生明顯抑制。相同濃度下,各時(shí)間點(diǎn)A組RGR均高于B組,除4 mmol/L濃度培養(yǎng)1 d和3 d外,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。培養(yǎng)1 d,A、B組RAR分別為144.1%±3.2%和115.2%±9.8%,5 d時(shí)分別為129.6%±9.8%和78.2%±6.9%,A組均顯著高于B組(P0.05)。von Kossa染色示,培養(yǎng)21 d,A組黑色團(tuán)狀物明顯多于B、C組,定量分析分別為91.2%±8.2%、50.3%±7.9%和54.2%±8.6%,A組顯著高于B、C組(P0.05);B、C組間差異無統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論 PLA降解產(chǎn)物的濃度和組成對(duì)成骨樣細(xì)胞增殖和成骨表型均有顯著影響。
[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，

本文編號(hào)：1576826