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细胞对生物材料表面化学和形态学方面的反应。II. 骨细胞群体的生物合成和迁移反应。

Cellular responses to chemical and morphologic aspects of biomaterial surfaces. II. The biosynthetic and migratory response of bone cell populations.

作者信息

Chesmel K D, Clark C C, Brighton C T, Black J

机构信息

Department of Orthopaedic Surgery Research, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.

出版信息

J Biomed Mater Res. 1995 Sep;29(9):1101-10. doi: 10.1002/jbm.820290910.

Abstract

The biosynthetic and migratory response of bone cells to changes in both surface composition and morphology of polystyrene (PS) substrates was examined. A system was devised wherein micromachined silicon wafers were used as templates to solvent-cast PS replicas [using 0, 1, or 2 wt % styrene (S) monomer additions] with either 0.5- or 5.0- microns-deep surface grooves. Smooth replicas (0% S) served as the control surfaces. The chemical and morphologic characteristics of the nine unique model biomaterial surfaces (MBSs) produced using this system were documented and were found to be distinct. For the biosynthetic studies, bone cells isolated from neonatal rat calvaria were plated onto the MBSs and labeled at postconfluence with [14C]proline for 24 h. Total DNA per surface, total newly synthesized collagenous (CP), and noncollagenous protein (NCP) (cell associated and secreted) were determined. Cell-associated CP was found to increase significantly for the bone cells cultured on the substrates with 0.5-micron grooves and 2% S (P < .05). Cell-associated NCP was found to be elevated for all 2% S substrates and for the 0.5-micron grooves substrates with 1% S. For the migration studies, bone cells were plated first onto 5-mm nitrocellulose disks that were attached to standard Petri dishes using a plasma clot. At confluence, the disks were removed aseptically and placed on the replicas. The cellular area occupied as a result of the outward migration of the bone cells was measured after 4 days of culture using an image analysis system. An average velocity for the leading edge of bone cell populations on each of the nine MBSs was calculated: Cells on surfaces with either 1% S or 5.0-microns grooves displayed significantly higher velocities than did the control cultures. A significant interaction effect between chemistry and morphology was observed. The biosynthetic and migratory responses of in vitro cultures of bone cells were not predictable from the observations of the cellular responses to the individual features, but appeared to depend on cellular responses to more than one substrate factor.

摘要

研究了骨细胞对聚苯乙烯(PS)底物表面组成和形态变化的生物合成及迁移反应。设计了一个系统,其中使用微加工硅片作为模板,通过溶剂浇铸法制作PS复制品(添加0、1或2 wt%苯乙烯(S)单体),表面有0.5微米或5.0微米深的凹槽。光滑复制品(0% S)作为对照表面。记录了使用该系统制备的九种独特模型生物材料表面(MBS)的化学和形态特征,发现它们各不相同。在生物合成研究中,将从新生大鼠颅骨分离的骨细胞接种到MBS上,并在汇合后用[14C]脯氨酸标记24小时。测定每个表面的总DNA、新合成的总胶原蛋白(CP)和非胶原蛋白(NCP)(细胞相关和分泌的)。发现对于在具有0.5微米凹槽和2% S的底物上培养的骨细胞,细胞相关CP显著增加(P < 0.05)。发现所有含2% S的底物以及含1% S的0.5微米凹槽底物上的细胞相关NCP均升高。在迁移研究中,首先将骨细胞接种到5毫米硝酸纤维素圆盘上,该圆盘使用血浆凝块附着在标准培养皿上。汇合时,无菌取出圆盘并放置在复制品上。使用图像分析系统在培养4天后测量由于骨细胞向外迁移而占据的细胞面积。计算了九个MBS上每个骨细胞群体前沿的平均速度:含1% S或5.0微米凹槽表面上的细胞显示出比对照培养物显著更高的速度。观察到化学和形态之间存在显著的相互作用效应。骨细胞体外培养的生物合成和迁移反应无法从细胞对单个特征的反应观察中预测,而是似乎取决于细胞对多个底物因素的反应。

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