Fourier Transform Infrared Imaging as a Tool to Chemically and Spatially Characterize Matrix-Mineral Deposition in Osteoblasts
Fourier Transform Infrared Imaging as a Tool to Chemically and Spatially Characterize Matrix-Mineral Deposition in Osteoblasts
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DOI：
10.1007/s00223-012-9667-5
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发表时间：
2013-01-01
影响因子：
4.2
通讯作者：
Miller, Lisa M.
中科院分区：
文献类型：
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作者：
Faillace, Meghan E.;Phipps, Roger J.;Miller, Lisa M.
矿化成骨细胞经常被用于研究成骨和体内骨形成模型。因此，重要的是要验证在原位形成的矿物和基质与在体内发现的矿物和基质具有可比性。然而，已有研究表明，仅靠组织化学技术是不足以鉴定含磷酸钙矿物的。本研究的目的是证明使用傅里叶变换红外成像（FTIRI）作为表征成骨细胞原位矿化过程中胶原基质和矿物相的空间分布和共定位的工具。将MC3T3-E1小鼠成骨细胞矿化培养28天，利用FTIRI以空间分解的方式评价胶原含量、胶原交联、矿化水平和物种形成以及矿物结晶度随时间的变化。为了测试FTIRI是否能检测矿化过程中的细微变化，用利塞膦酸盐（RIS）处理细胞。结果表明，胶原沉积和矿化随着时间的推移而进展，磷灰石矿物与胶原基质有关，而不是异位矿物。RIS暂时减缓了这一过程，其中成骨细胞功能的抑制导致胶原蛋白生成和交联减慢，导致矿化减少。本研究表明，FTIRI是组织化学的补充工具，可以在空间上关联成骨细胞矿化过程中胶原基质的分布和所产生矿物质的性质。它可以进一步用于检测类骨和矿物沉积过程中的小扰动。
Mineralizing osteoblasts are regularly used to study osteogenesis and model in vivo bone formation. Thus, it is important to verify that the mineral and matrix being formed in situ are comparable to those found in vivo. However, it has been shown that histochemical techniques alone are not sufficient for identifying calcium phosphate-containing mineral. The goal of the present study was to demonstrate the use of Fourier transform infrared imaging (FTIRI) as a tool for characterizing the spatial distribution and colocalization of the collagen matrix and the mineral phase during the mineralization process of osteoblasts in situ. MC3T3-E1 mouse osteoblasts were mineralized in culture for 28 days and FTIRI was used to evaluate the collagen content, collagen cross-linking, mineralization level and speciation, and mineral crystallinity in a spatially resolved fashion as a function of time. To test whether FTIRI could detect subtle changes in the mineralization process, cells were treated with risedronate (RIS). Results showed that collagen deposition and mineralization progressed over time and that the apatite mineral was associated with a collagenous matrix rather than ectopic mineral. The process was temporarily slowed by RIS, where the inhibition of osteoblast function caused slowed collagen production and cross-linking, leading to decreased mineralization. This study demonstrates that FTIRI is a complementary tool to histochemistry for spatially correlating the collagen matrix distribution and the nature of the resultant mineral during the process of osteoblast mineralization. It can further be used to detect small perturbations in the osteoid and mineral deposition process.