转化生长因子β1对多孔钽/MG63成骨样细胞复合物细胞增殖及分泌功能的影响

BACKGROUND: Previous studies have demonstrated that the Chinese porous tantalum made in China has non-toxicity and good biocompatibility, which can promote osteogenesis.             

中药川芎对成骨细胞作用最佳浓度的体外实验

BACKGROUND: It has been proved that Rhizoma Chuanxiong can promote osteoblast proliferation and differentiation. OBJECTIVE: To further observe the effects of tetramethylpyrazine (Ligustrazine) extracted from Rhizoma Chuanxiong on osteoblast proliferation and type I collagen expression. METHODS: Mouse osteoblast cell line MC3T3-E1 was cultured in medium containing 1, 5, 10, 15 and   20 mg/L Ligustrazine, respectively. Osteoblast proliferation was detected by cell counting kit-8 assay, and alkaline phosphatase activity and type I collagen level measured using ELISA method. RESULTS AND CONCLUSION: Different concentrations of Ligustrazine all significantly promoted the osteoblast proliferation and type I collagen level compared with the blank control group (P < 0.005). The activity of alkaline phosphatase in all Ligustrazine groups except 15 and 20 mg/L groups was significantly higher than that in the blank control group (P < 0.005). These results show that Ligustrazine isolated from Rhizoma Chuanxiong can effectively promote the osteoblast proliferation and type I collagen expression, with the optimum concentration of 10 mg/L. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

磁性四氧化三铁纳米颗粒作用于前成骨细胞的生物相容性

BACKGROUND: Investigations on toxic mechanism and safety of magnetic ferrosoferric oxide (Fe3O4) nanoparticles are extremely necessary when these nanoparticles as an emerging material for bone tissue engineering are implanted into the living body. OBJECTIVE: To investigate the biocompatibility of magnetic Fe3O4 nanoparticles with preosteoblasts. METHODS: Mouse preosteoblasts were cultured in 0, 200, 400, 800 mg/L magnetic Fe3O4 nanoparticles. After 24 hours, alkaline phosphatase activity, osteocalcin level, cell proliferation rate, cellular morphology, cytoskeleton variation, cell apoptosis and autophagy-related genes, such as Caspase-3, LC3A, LC3B, were detected by alkaline phosphatase assay kit, ELISA kit, cell counting kit-8 kit, inverted microscope, laser confocal microscopy and real-time PCR, respectively. RESULTS AND CONCLUSION: After 24 hours of culture, there ware no significant differences between 200 mg/L group and control group. However, in the groups of 400 and 800 mg/L, the ratio of alkaline phosphatase activity to total protein and osteocalcin level increased, the cell proliferation rate decreased, cellular morphology and cytoskeleton changed remarkably, LC3B expression was up-regulated compared with the control group. Additionally, there were also no significant differences in the expression of Caspase-3 and LC3A between 400 and 800 mg/L groups and control group. Therefore, magnetic Fe3O4 nanoparticles at high level contributes to cytotoxicity and up-regulation of LC3B expression, and affects cellular morphology, cytoskeleton and cell proliferation rate, although these nanoparticles can increase the osteoblastic differentiation.        

益肾蠲痹汤含药血清对大鼠成骨细胞和破骨细胞增殖能力的影响

目的 通过研究益肾蠲痹汤的含药血清对大鼠成骨细胞、破骨细胞增殖的影响,评价益肾蠲痹法对强直性脊柱炎骨代谢异常的治疗作用。 方法 SD正常大鼠(SPF级)经灌胃给药益肾蠲痹汤,收集大鼠含药血清。分离大鼠成骨细胞和破骨细胞,通过Van kossa染色和碱性磷酸酶染色鉴定成骨细胞,TRAP染色、甲苯胺蓝染色鉴定破骨细胞。将体外培养的成骨细胞和破骨细胞,分别分为6组:空白对照组,益肾蠲痹汤大、中、小剂量含药血清组,双氯芬酸对照组和阿仑膦酸盐对照组。使用CCK-8法测定成骨细胞、破骨细胞的生长曲线,收集并测定成骨细胞培养液上清碱性磷酸酶活性。 结果 含有益肾蠲痹汤药物的血清加入体外培养成骨细胞后,成骨细胞的生长曲线呈明显上升趋势,加药后细胞培养上清中碱性磷酸酶浓度增加,提示成骨细胞分泌碱性磷酸酶活性增强。生长曲线上升趋势及分泌碱性磷酸酶活性随含药血清浓度增加而作用增强。体外培养破骨细胞加入益肾蠲痹汤含药血清后,细胞增殖曲线呈下降趋势,曲线下降趋势亦随含药血清浓度升高而幅度增大。 结论 益肾蠲痹汤具有促进成骨细胞增殖及提高成骨细胞活性,同时,对破骨细胞增殖有抑制作用,作用强度随药物浓度增加而增强。因此,预期益肾蠲痹汤对强直性脊柱炎骨代谢异常具有治疗作用。      

骨保护素基因修饰的大鼠骨髓基质细胞系的培养与鉴定

目的培养经骨保护素OPG修饰的大鼠骨髓基质细胞系并检测其表达能力,鉴定表达产物,为基因工程技术治疗牙周病提供物质基础。方法利用已构建的pSecTagOPG真核分泌表达穿梭载体,用脂质体法将目的基因转染至大鼠骨髓基质细胞,应用免疫组织化学和Westernblot方法证实转染的细胞表达骨保护素。结果经免疫组织化学和Westernblot方法检测,在转染OPG的大鼠骨髓基质细胞中有OPG表达。结论本实验成功培养出分泌表达OPG的大鼠骨髓基质细胞系。     

右归饮含药血清对成骨细胞硬骨素表达调控的影响

目的探讨右归饮含药血清对硬骨素（SO）在体外成骨细胞分化过程中表达调控的影响。方法将煎制的右归饮液,对成年sD大鼠进行灌胃,获得右归饮含药血清。体外培养新生sD大鼠颅骨成骨细胞,取第3代细胞分实验组、对照组,分别用右归饮含药血清及0．9％NaCl溶液体外诱导培养,分别在7、14、21天进行形态学观察、ALP活性检测,SO的RT—PCR检测。结果经右归饮含药血清体外诱导培养,成骨细胞中ALP及矿化染色随培养时问延长,均有不同程度加深;实验组中SO表达量随培养时间延长呈增长趋势,同对照组相比,在诱导21天时表达显著升高,差异有统计学意义（P〈0．05）。结论右归饮能调控SO基因在成骨细胞分化过程的表达,提示SO可能参与成骨细胞的分化,影响骨重建过程。     

盐酸四环素缓释微球对成骨细胞增殖作用的研究

目的：观察盐酸四环素缓释微球对体外培养大鼠成骨细胞的增殖活性。方法：体外分离培养SD大鼠成骨细胞,通过形态学观察,茜素红钙结节染色鉴定成骨细胞生物学特征;将盐酸四环素缓释微球与成骨细胞共培养,用细胞计数法和MF法检测成骨细胞的增殖情况。结果：盐酸四环素缓释微球能够在实验的9天内持续促进成骨细胞增殖。结论：盐酸四环素缓释微球能促进成骨细胞增殖。     

CCK-8法检测狗脊不同炮制品对成骨细胞的影响

该文采用胰酶-Ⅱ型胶原酶消化法进行成骨细胞原代培养并接种、传代,碱性磷酸酶染液鉴定,制备狗脊不同炮制品正丁醇提取部位及原儿茶酸、原儿茶醛、曲酸三者混合对照药液等与上述成骨细胞共同培养,以CCK-8法对其进行增殖检测。各炮制品正丁醇提取部位对于成骨细胞的增殖作用显著,酒狗脊>烫狗脊>盐狗脊>砂烫酒制狗脊>醇制狗脊>蒸狗脊>生狗脊,q检验显示砂烫酒制狗脊、醇制狗脊、蒸狗脊无显著差异,烫狗脊、盐狗脊无显著差异;各对照品对于成骨细胞增殖具有一定作用,其中混合对照>原儿茶醛>原儿茶酸>曲酸,q检验显示原儿茶醛、原儿茶酸无显著差异。狗脊各炮制品正丁醇提取部位均显著促进成骨细胞增殖,以酒狗脊为最佳;原儿茶酸、原儿茶醛、曲酸等酚类化合物对成骨细胞均具有一定的促进增殖作用。     

维生素D3衍生物对人成骨细胞生长和碱性磷酸酶活性的影响

目的：使用人胎成骨细胞（ＯＢ）为体外模型，观察了１，２５－双羟维生素Ｄ３〔１，２５（ＯＨ）２Ｄ３〕、２４，２５－双羟维生素Ｄ３［２４，２５（ＯＨ）２Ｄ３）］对ＯＢ生长和代谢的影响。方法：用ＡＬＰ测定法和Ｂｒａｄｆｏｒｄ蛋白含量法。结果：１，２５（ＯＨ）２Ｄ３在浓度为１０８ｍｏｌ／Ｌ时，刺激碱性磷酸酶（ＡＬＰ）的活性。但抑制ＯＢ的生长。２４，２５（ＯＨ）２Ｄ３在１０－８ｍｏｌ／Ｌ时无上述作用。结论：１，２５（ＯＨ）２Ｄ３对ＯＢ的ＡＬＰ有直接的促活作用，其作用与时间相关。２４，２５（ＯＨ）２Ｄ３对ＯＢ的ＡＬＰ活性无关。１，２５（ＯＨ）２Ｄ３对人胎ＯＢ生长有抑制作用。     

雌激素对原代成骨细胞骨形成的影响

了解雌激素对培养的原代成骨细胞骨形成的影响。观察添加雌激素后直接对成骨细胞骨形成的作用，并用VOnKossa免疫染色法定量表示骨形成。结果显示：雌激素对该原代成骨细胞无促进骨形成作用。