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傅里叶变换红外光谱/衰减全反射法研究蛋白质吸附及透钙磷石向羟基磷灰石的转变

FTIR/ATR study of protein adsorption and brushite transformation to hydroxyapatite.

作者信息

Xie Jing, Riley Clyde, Kumar Mukesh, Chittur Krishnan

机构信息

Chemistry Department, University of Alabama in Huntsville, 35899, USA.

出版信息

Biomaterials. 2002 Sep;23(17):3609-16. doi: 10.1016/s0142-9612(02)00090-x.

Abstract

Previous study has demonstrated that brushite (CaHPO4 x 2H2O), modified by partial potassium substitution for calcium, can transform quickly into hydroxyapatite (HA, Ca5(PO4)3OH) when exposed to aqueous salt solutions at room temperature. Analyses techniques used in those studies required sample retrieval from solution, which may alter the sample surface. In this work FTIR/ ATR was used in analysis, enabling in situ study of the transformation within the aqueous environment. To test the biocompatibility of this brushite, cellular response to the transformation needs to be understood. Cellular response was initiated by bovine serum albumin adsorption on the brushite surface. The response was studied by monitoring the conformation of the adsorbed protein, which is critical to cellular reaction. This required monitoring the brushite transformation and surface adsorbed protein conformation simultaneously which can be realized using FTIR/ATR. Based on band fitting and second derivative results from the spectra it was found that the conformation of the adsorbed BSA changes during the brushite transformation to HA. This study also demonstrated that the deposition of the brushite could be monitored in real time which offers the possibility for studying surface bonding during electrodeposition.

摘要

先前的研究表明,通过部分钾替代钙改性的透钙磷石(CaHPO₄·2H₂O)在室温下暴露于盐水溶液时可迅速转化为羟基磷灰石(HA,Ca₅(PO₄)₃OH)。那些研究中使用的分析技术需要从溶液中取出样品,这可能会改变样品表面。在这项工作中,采用傅里叶变换红外光谱/衰减全反射(FTIR/ATR)进行分析,从而能够在水环境中对转化过程进行原位研究。为了测试这种透钙磷石的生物相容性,需要了解细胞对这种转化的反应。细胞反应是由牛血清白蛋白吸附在透钙磷石表面引发的。通过监测吸附蛋白的构象来研究这种反应,而吸附蛋白的构象对细胞反应至关重要。这需要同时监测透钙磷石的转化和表面吸附蛋白的构象,而这可以通过FTIR/ATR实现。基于光谱的谱带拟合和二阶导数结果发现,在透钙磷石向HA转化的过程中,吸附的牛血清白蛋白的构象发生了变化。该研究还表明,可以实时监测透钙磷石的沉积情况,这为研究电沉积过程中的表面键合提供了可能性。

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