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磷酸三钙骨移植材料对成骨细胞原代体外培养的影响。

Effects of tricalcium phosphate bone graft materials on primary cultures of osteoblast cells in vitro.

作者信息

Aybar Buket, Bilir Ayhan, Akçakaya Handan, Ceyhan Tasşkin

机构信息

Department of Oral Surgery, School of Dentistry, Istanbul University, Istanbul, Turkey.

出版信息

Clin Oral Implants Res. 2004 Feb;15(1):119-25. doi: 10.1111/j.1600-0501.2004.01002.x.

Abstract

The aim of this study was to evaluate beta-tricalcium phosphate (beta-TCP Cerasorb Curasan-Germany) graft materials on specific parameters of rat osteoblast activity in vitro. Primary culture osteoblastic cells were isolated from neonatal rat calvaria by sequential collagenase digestion. To analyze the effect of biomaterials on cell proliferation, cell numbers and viability of the cells were cultured on the graft material for 24, 48 or 96 h. Osteoblast cells cultured in DMEF-12 media supplemented with 10% fetal calf serum were used as the control group. [3H]thymidine was added during the last 2 h of the incubation. The cell numbers of each well were counted. Cell viability was estimated by counting the number of cells, which excluded trypan blue solution. Scanning electron microscopy was used to observe for visualizing the interactions between osteoblastic cells and TCP graft material. The proportion of cells undergoing DNA synthesis, estimated by thymidine uptake, was significantly (P<0.05) greater on the control group after the 24- and 48-h incubations. Regarding the cell numbers the difference was not statistically significant for the three time points. The number of viable cells recovered was similar for the two groups. No morphological differences were observed in cell morphology on TCP graft material and the control group. The results demonstrate that TCP graft material has no adverse effect on cell count, viability and morphology, and this material provides a matrix that favors limited cell proliferation.

摘要

本研究的目的是在体外评估β-磷酸三钙(β-TCP Cerasorb Curasan-德国)移植材料对大鼠成骨细胞活性特定参数的影响。通过连续胶原酶消化从新生大鼠颅骨分离原代培养的成骨细胞。为了分析生物材料对细胞增殖的影响,将细胞接种在移植材料上培养24、48或96小时,然后分析细胞数量和活力。在补充有10%胎牛血清的DMEF-12培养基中培养的成骨细胞用作对照组。在孵育的最后2小时加入[3H]胸腺嘧啶核苷。对每个孔的细胞数量进行计数。通过计数排斥台盼蓝溶液的细胞数量来估计细胞活力。使用扫描电子显微镜观察成骨细胞与TCP移植材料之间的相互作用。在24小时和48小时孵育后,通过胸腺嘧啶核苷摄取估计的进行DNA合成的细胞比例在对照组中显著更高(P<0.05)。关于细胞数量,在三个时间点差异无统计学意义。两组回收的活细胞数量相似。在TCP移植材料和对照组上未观察到细胞形态的差异。结果表明,TCP移植材料对细胞计数、活力和形态没有不利影响,并且这种材料提供了有利于有限细胞增殖的基质。

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