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作品数:2 被引量:4H指数:2
相关作者:赵翔蔡迅梓师钟丽严世贵吴浩波更多>>
相关机构:浙江大学医学院附属第二医院更多>>
发文基金:国家自然科学基金浙江省自然科学基金浙江省教育厅科研计划更多>>
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Enhanced integrin-mediated human osteoblastic adhesion to porous amorphous calcium phosphate/poly(L-lactic acid) composite被引量:2
2014年
Background The initial osteoblastic adhesion to materials characterizes the first phase of cell-material interactions and influences all the events leading to the formation of new bone. In a previous work, we developed a novel amorphous calcium phosphate (ACP)/poly(L-lactic acid) (PLLA) material that demonstrated morphologic variations in its microstructure. The aim of this study was to investigate the initial interaction between this material and osteoblastic cells. Cellular attachment and the corresponding signal transduction pathways were investigated. Methods A porous ACP/PLLA composite and PLLA scaffold (as a control) were incubated in fetal bovine serum (FBS) containing phosphate-buffered saline (PBS), and the protein adsorption was determined. Osteoblastic MG63 cells were seeded on the materials and cultured for 1, 4, 8, or 24 hours. Cell attachment was evaluated using the MTS method. Cell morphology was examined using scanning electron microscopy (SEM). The expression levels of the genes encoding integrin subunits αl, α5, αv, β1, focal adhesion kinase (FAK), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were determined using real-time reverse transcription polymerase chain reaction (RT-PCR). Results The ACP/PLLA material significantly increased the protein adsorption by 6.4-fold at 1 hour and 2.4-fold at 24 hours, compared with the pure PLLA scaffold. The attachment of osteoblastic cells to the ACP/PLLA was significantly higher than that on the PLLA scaffold. The SEM observation revealed a polygonal spread shape of cells on the ACP/ PLLA, with the filopodia adhered to the scaffold surface. In contrast, the cells on the PLLA scaffold exhibited a spherical or polygonal morphology. Additionally, real-time RT-PCR showed that the genes encoding the integrin subunits αl, αv, β1, and FAK were expressed at higher levels on the ACP/PLLA composite. Conclusions The ACP/PLLA composite promoted protein adsorption and osteoblastic adhesion. The enhanced cell adhesio
Huang XinQi YiyingLi WeixuShi ZhongliWeng WenjianChen KuiHe Rongxin
关键词:OSTEOBLASTINTEGRIN
纳米羟基磷灰石颗粒对成骨样细胞生物活性的尺寸效应被引量:2
2012年
目的探索不同尺寸的纳米羟基磷灰石颗粒nanohydroxyapatite,nHAP)对成骨细胞增殖和凋亡的生物学效应。方法采用化学沉淀的方法,可控制备不同大小的nHAP,包括20nm球形(20HAP)、40nm球形(40HAP)和80nm球形(80HAP)。通过透射电子显微镜、X-射线衍射仪、红外线光谱仪等工具分析颗粒大小、形状以及化学组分。将nHAP颗粒制备成薄膜,与成骨样细胞MG-63细胞共培养5d,通过甲基噻唑基四唑(methylthiazolyltetrazolium,MTT)计数活细胞数量,采用流式细胞仪检测细胞的凋亡情况,通过透射电子显微镜观察细胞器的变化情况以及nHAP在细胞内的分布。结果可控合成并对20HAP、40HAP和80HAP进行大小组分分析。薄膜细胞共培养结果显示,5天时20HAP组的细胞吸光度值高达1.22±0.13,与空白对照组相比差异无统计学意义(F=6.843,P=0.124)。细胞的凋亡率随nHAP尺寸的增加而增大,与空白对照组相比,差异有统计学意义(q=4.284、5.487、2.933,P〈0.05);各组细胞凋亡率分别为1.12%±0.51%(空白对照组),6.68%±1.04%(20HAP组),10.08%±2.10%(40HAP组)和13.61%±2.45%(80HAP组)。透射电子显微镜结果证实,20HAP能够穿透细胞膜而对细胞超微结构和细胞形态不具有毒性作用。结论成骨细胞增殖和凋亡都与nHAP的尺寸相关,其中20HAP的促进细胞增殖和抑制细胞凋亡作用最佳。
师钟丽严世贵王卫吴浩波赵翔蔡迅梓
关键词:羟基磷灰石类成骨细胞
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