中国骨科临床与基础研究杂志
中國骨科臨床與基礎研究雜誌
중국골과림상여기출연구잡지
CHINESE JOURNAL OF CLINICAL AND BASIC ORTHO[AEDIC RESEARCH
2013年
1期
21-27
,共7页
张涛%尹庆水%夏虹%黄华扬%张余%李梅%杨小明%蓝国波
張濤%尹慶水%夏虹%黃華颺%張餘%李梅%楊小明%藍國波
장도%윤경수%하홍%황화양%장여%리매%양소명%람국파
镁%合金%涂层%肌,骨骼%细胞,培养的%生物相容性材料%兔
鎂%閤金%塗層%肌,骨骼%細胞,培養的%生物相容性材料%兔
미%합금%도층%기,골격%세포,배양적%생물상용성재료%토
Magnesium%Alloys%Coating%Muscle,skeletal%Cells,cultured%Biocompatible materials%Rabbits
目的探讨钙磷涂层镁合金对兔骨骼肌细胞生物学行为的影响。方法将兔骨骼肌细胞分别接种于钛合金(钛合金组)、镁合金(AZ31B组)及钙磷涂层镁合金(CaP-AZ31B组)试件上培养2、6、24 h,计算细胞黏附率,扫描电镜观察细胞黏附形态;采用AZ31B及CaP-AZ31B浸提液培养兔骨骼肌细胞1、3、5、7 d,同时以不加合金浸提液的细胞作为对照,倒置显微镜观察细胞形态,CCK-8法检测细胞增殖活力,Calcein-AM和EthD-1双染法观察培养4 d后细胞存活情况,并测定细胞内总蛋白。结果随着培养时间的延长,3组细胞黏附率逐渐增加(P<0.05);培养2、6、24 h时,3组细胞黏附率比较,差异有统计学意义(P<0.05)。浸提液培养1、3、5、7 d,AZ31B组细胞相对增殖率低于对照组(P<0.05),CaP-AZ31B组高于AZ31B组(P<0.05);AZ31B组毒性评级为2级,CaP-AZ31B组毒性评级为1级;对照组、CaP-AZ31B组细胞活性较好,无明显细胞红染,AZ31B组部分细胞红染,但绝大多数细胞仍具有活性;3组细胞内总蛋白量呈时间依赖性增加(P<0.05),各时相点3组细胞内总蛋白量比较,差异有统计学意义(P<0.05),从高到低依次为对照组、CaP-AZ31组、AZ31B组。结论钙磷涂层镁合金对于兔骨骼肌细胞具着良好的生物相容性。
目的探討鈣燐塗層鎂閤金對兔骨骼肌細胞生物學行為的影響。方法將兔骨骼肌細胞分彆接種于鈦閤金(鈦閤金組)、鎂閤金(AZ31B組)及鈣燐塗層鎂閤金(CaP-AZ31B組)試件上培養2、6、24 h,計算細胞黏附率,掃描電鏡觀察細胞黏附形態;採用AZ31B及CaP-AZ31B浸提液培養兔骨骼肌細胞1、3、5、7 d,同時以不加閤金浸提液的細胞作為對照,倒置顯微鏡觀察細胞形態,CCK-8法檢測細胞增殖活力,Calcein-AM和EthD-1雙染法觀察培養4 d後細胞存活情況,併測定細胞內總蛋白。結果隨著培養時間的延長,3組細胞黏附率逐漸增加(P<0.05);培養2、6、24 h時,3組細胞黏附率比較,差異有統計學意義(P<0.05)。浸提液培養1、3、5、7 d,AZ31B組細胞相對增殖率低于對照組(P<0.05),CaP-AZ31B組高于AZ31B組(P<0.05);AZ31B組毒性評級為2級,CaP-AZ31B組毒性評級為1級;對照組、CaP-AZ31B組細胞活性較好,無明顯細胞紅染,AZ31B組部分細胞紅染,但絕大多數細胞仍具有活性;3組細胞內總蛋白量呈時間依賴性增加(P<0.05),各時相點3組細胞內總蛋白量比較,差異有統計學意義(P<0.05),從高到低依次為對照組、CaP-AZ31組、AZ31B組。結論鈣燐塗層鎂閤金對于兔骨骼肌細胞具著良好的生物相容性。
목적탐토개린도층미합금대토골격기세포생물학행위적영향。방법장토골격기세포분별접충우태합금(태합금조)、미합금(AZ31B조)급개린도층미합금(CaP-AZ31B조)시건상배양2、6、24 h,계산세포점부솔,소묘전경관찰세포점부형태;채용AZ31B급CaP-AZ31B침제액배양토골격기세포1、3、5、7 d,동시이불가합금침제액적세포작위대조,도치현미경관찰세포형태,CCK-8법검측세포증식활력,Calcein-AM화EthD-1쌍염법관찰배양4 d후세포존활정황,병측정세포내총단백。결과수착배양시간적연장,3조세포점부솔축점증가(P<0.05);배양2、6、24 h시,3조세포점부솔비교,차이유통계학의의(P<0.05)。침제액배양1、3、5、7 d,AZ31B조세포상대증식솔저우대조조(P<0.05),CaP-AZ31B조고우AZ31B조(P<0.05);AZ31B조독성평급위2급,CaP-AZ31B조독성평급위1급;대조조、CaP-AZ31B조세포활성교호,무명현세포홍염,AZ31B조부분세포홍염,단절대다수세포잉구유활성;3조세포내총단백량정시간의뢰성증가(P<0.05),각시상점3조세포내총단백량비교,차이유통계학의의(P<0.05),종고도저의차위대조조、CaP-AZ31조、AZ31B조。결론개린도층미합금대우토골격기세포구착량호적생물상용성。
Objective To investigate the influence of magnesium alloy with calcium phosphate coating on the biological behavior of rabbit skeletal muscle cells. Methods Rabbit skeletal muscle cells were inoculated on titanium alloy, magnesium alloy (AZ31B) and magnesium alloy with calcium phosphate coating (CaP-AZ31B), and were cultured for 2, 6, 24 h. Cell adhesion rates were calculated, and cell adhesion morphology were observed by scanning electron microscope. In AZ31B and CaP-AZ31B leaching liquids, rabbit skeletal muscle cell were cultured for 1, 3, 5, 7 d, and cultured cells without alloy extracts were used as controls. Cell morphology was observed by inverted microscope, cell proliferation was detected by CCK-8 method, cell survival at 4 d after culture were evaluated by Calcein-AM and EthD-1 double staining, and the total cellular protein were determined in those 3 groups. Results Cell adhesion rates in 3 groups increased time-dependently (P <0.05);At 2, 6, 24 h, the differences of cell adhesion rate among 3 groups had statistically significant (P <0.05). Cell relative growth rate in AZ31B group was lower than that of control group at 1, 3, 5, 7 d after the culture with alloy extracts (P <0.05), while the rate in CaP-AZ31B group was higher than that of AZ31B group at each timepoint (P <0.05); Toxicity in AZ31B group was rated as 2 grade, and in CaP-AZ31B group rated as 1 grade; Cell activities were good in CaP-AZ31B group as well as the control group without obvious red-staining cells, while part of the cells were red-stained in AZ31B group with most of them still active; Intracellular total protein content in 3 groups increased in a time-dependent manner (P <0.05), the differences of total protein content among 3 groups at each timepoint were statistically significant (P <0.05), and the ranking of the total protein content from high to low was control group, CaP-AZ31 group, AZ31B group accordingly. Conclusion Magnesium alloy with calcium phosphate coating has a good biocompatibility to rabbit skeletal muscle cells.