东南国防医药
東南國防醫藥
동남국방의약
JOURNAL OF SOUTHEAST CHINA NATIONAL DEFENCE MEDICAL SCIENCE
2015年
1期
1-4
,共4页
孙国静%杨书丰%郭国栋%樊根涛%吴俊%赵建宁
孫國靜%楊書豐%郭國棟%樊根濤%吳俊%趙建寧
손국정%양서봉%곽국동%번근도%오준%조건저
钛合金微粒%重组成骨蛋白-1%成骨细胞%骨溶解
鈦閤金微粒%重組成骨蛋白-1%成骨細胞%骨溶解
태합금미립%중조성골단백-1%성골세포%골용해
titanium alloy particles%recombinant OP-1%osteoblast%osteolysis
目的:在钛-6铝-4钒(Ti-6Al-4V)合金微粒环境下观察重组成骨蛋白-1(recombinant OP-1,rOP-1)对成骨细胞的影响,为防治关节假体无菌性松动提供新的治疗途径。方法根据小鼠颅顶骨前成骨细胞亚克隆14( MC3 T3-E1)中是否加入Ti-6Al-4V微粒和rOP-1,分为微粒组(5、10、15μg/mL Ti-6Al-4V)、处理组(微粒组加入200 ng/mL rOP-1)、阳性组(加入200 ng/mL rOP-1)和对照组,检测各组24、72、120 h MC3T3-E1细胞增殖能力、72 h碱性磷酸酶( akaline phosphatase ,AKP)、骨钙素(osteocalcin,OCN)和骨桥蛋白( osteopontin,OPN) mRNA的表达,及120 h 成骨细胞的矿化能力。结果①rOP-1无促进Ti-6Al-4V微粒环境下成骨细胞增殖能力,与微粒组比较,差异无统计学意义(P>0.05);②rOP-1可提高成骨细胞分化,与对照组比较差异有统计学意义(P<0.05);同时逆转Ti-6Al-4V微粒抑制成骨细胞分化,与微粒组比较差异有统计学意义(P<0.05);③茜素红S染色后Ti-6Al-4V微粒钙结节数量随着浓度增加逐渐降低,和微粒组比较,加入rOP-1后钙结节数量呈增多趋势。结论 Ti-6Al-4V微粒环境下,rOP-1无提高成骨细胞增殖能力,能提高细胞分化矿化能力,rOP-1可以作为潜在治疗关节假体无菌性松动一种方法。
目的:在鈦-6鋁-4釩(Ti-6Al-4V)閤金微粒環境下觀察重組成骨蛋白-1(recombinant OP-1,rOP-1)對成骨細胞的影響,為防治關節假體無菌性鬆動提供新的治療途徑。方法根據小鼠顱頂骨前成骨細胞亞剋隆14( MC3 T3-E1)中是否加入Ti-6Al-4V微粒和rOP-1,分為微粒組(5、10、15μg/mL Ti-6Al-4V)、處理組(微粒組加入200 ng/mL rOP-1)、暘性組(加入200 ng/mL rOP-1)和對照組,檢測各組24、72、120 h MC3T3-E1細胞增殖能力、72 h堿性燐痠酶( akaline phosphatase ,AKP)、骨鈣素(osteocalcin,OCN)和骨橋蛋白( osteopontin,OPN) mRNA的錶達,及120 h 成骨細胞的礦化能力。結果①rOP-1無促進Ti-6Al-4V微粒環境下成骨細胞增殖能力,與微粒組比較,差異無統計學意義(P>0.05);②rOP-1可提高成骨細胞分化,與對照組比較差異有統計學意義(P<0.05);同時逆轉Ti-6Al-4V微粒抑製成骨細胞分化,與微粒組比較差異有統計學意義(P<0.05);③茜素紅S染色後Ti-6Al-4V微粒鈣結節數量隨著濃度增加逐漸降低,和微粒組比較,加入rOP-1後鈣結節數量呈增多趨勢。結論 Ti-6Al-4V微粒環境下,rOP-1無提高成骨細胞增殖能力,能提高細胞分化礦化能力,rOP-1可以作為潛在治療關節假體無菌性鬆動一種方法。
목적:재태-6려-4범(Ti-6Al-4V)합금미립배경하관찰중조성골단백-1(recombinant OP-1,rOP-1)대성골세포적영향,위방치관절가체무균성송동제공신적치료도경。방법근거소서로정골전성골세포아극륭14( MC3 T3-E1)중시부가입Ti-6Al-4V미립화rOP-1,분위미립조(5、10、15μg/mL Ti-6Al-4V)、처리조(미립조가입200 ng/mL rOP-1)、양성조(가입200 ng/mL rOP-1)화대조조,검측각조24、72、120 h MC3T3-E1세포증식능력、72 h감성린산매( akaline phosphatase ,AKP)、골개소(osteocalcin,OCN)화골교단백( osteopontin,OPN) mRNA적표체,급120 h 성골세포적광화능력。결과①rOP-1무촉진Ti-6Al-4V미립배경하성골세포증식능력,여미립조비교,차이무통계학의의(P>0.05);②rOP-1가제고성골세포분화,여대조조비교차이유통계학의의(P<0.05);동시역전Ti-6Al-4V미립억제성골세포분화,여미립조비교차이유통계학의의(P<0.05);③천소홍S염색후Ti-6Al-4V미립개결절수량수착농도증가축점강저,화미립조비교,가입rOP-1후개결절수량정증다추세。결론 Ti-6Al-4V미립배경하,rOP-1무제고성골세포증식능력,능제고세포분화광화능력,rOP-1가이작위잠재치료관절가체무균성송동일충방법。
Objective Our results has been shown that titanium alloy particles (Ti-6Al-4V) inhibited RUNX2 mRNA and-proteinexpression of osteoblasts .In order to improve the boneformation on osteoblasts induced-Ti-6Al-4V, we observed the effect of rOP-1 on the proliferation, diferentiation, andmineralizatio of Ti-6Al-4V induce osteoblast, to provide a new treatment method of artifi-cial joint aseptic loosening .Methods Cell counting Kit8 test used to assay the cell proliferation after 24 h, 72 h and 120 h-incubation with Ti-6Al-4V ±rOP-1, respectively.Furthermore, alkaline phosphatase (AKP), osteocalcin (OCN) and osteopontin (OPN) mRNA level were detected by reverse transcription-polymerase chain reaction (RT-PCR), respectively, at the time point of 72 h.Cell bio-mineralization was assessed through alizarin red S staining after 120 h.Results ①Compared with Ti-6Al-4V group, after 24 h, 72 h and 120 h-incubation with Ti-6Al-4V+rOP-1, the proliferation of osteoblast did not increase significance with comparison of the Ti -6Al-4V group (P>0.05);②compared with Ti-6Al-4V group, AKP, OCN and OPN mRNA level were increased by 72 h-exposure to Ti-6Al-4V+rOP-1, respectively (P<0.05).rOP-1can stimulate osteoblast differentiation and mineralization , compared with Ti-6Al-4V group, there was statistically significance ( P<0.05);③compared with Ti-6Al-4V group, alizarin red staining demonstrated that the quantity of calcium nodules were markedly increased in the Ti-6Al-4V+rOP-1 group at hours 120.Conclusion Our result indi-cate that rOP-1 can stimulate differentiation and mineralization , while the proliferation remained unchanged on osteoblast with Ti-6Al-4V particles.We suppose that rOP-1 may be a therapeutic candidate for the prevention of artificial joint aseptic loosening .