Winkler Thomas, Hoenig Elisa, Huber Gerd, Janssen Rolf, Fritsch Daniel, Gildenhaar Renate, Berger Georg, Morlock Michael M, Schilling Arndt F
Biomechanics Section, Hamburg University of Technology, Hamburg - Germany.
Int J Artif Organs. 2010 Apr;33(4):198-203.
Bioresorbable materials have been developed in the hope that the body will replace them with newly formed tissue. The first step of this remodeling process in bone is the bioresorption of the material by osteoclasts. The aim of this study was to analyze osteoclastic resorption of biomaterials in vitro using the commonly used two-dimensional methods of light-microscopy (LM) and scanning electron microscopy (SEM) in comparison with infinite focus microscopy (IFM), a recently developed imaging method allowing for three-dimensional surface analysis.
Human hematopoietic stem cells were cultivated in the presence of the cytokines M-CSF and RANK-L for 4 weeks directly on dentin and a calcium phosphate cement. Osteoclast development was surveyed with standard techniques. After removal of the cells, resorption was characterized and quantified by LM, SEM and IFM.
Osteoclast cultures on the biomaterials presented the typical osteoclast-specific markers. On dentin samples LM, SEM as well as IFM allowed for discrimination of resorption. Quantification of the resorbed area showed a linear correlation between the results (LM vs. SEM: r=0.996, p=0.004; SEM vs. IFM: r=0.989, p=0.011; IFM vs. LM: r=0.995). It was not possible to demarcate resorption pits on GB14 using LM or SEM. With IFM, resorption on GB14 could be visualized and quantified two- and three-dimensionally.
In this paper we introduce IFM as a technology for three-dimensional visualization and quantification of resorption of biomaterials. Better understanding of the bioresorption of biomaterials may help in the design of better materials and might therefore constitute an important step on the avenue to the development of artificial bone.
生物可吸收材料已被研发出来，期望身体能用新形成的组织替代它们。骨骼重塑过程的第一步是破骨细胞对材料的生物吸收。本研究的目的是使用常用的二维光学显微镜（LM）和扫描电子显微镜（SEM）方法，与无限聚焦显微镜（IFM）（一种最近开发的允许进行三维表面分析的成像方法）相比较，在体外分析生物材料的破骨细胞吸收情况。
将人类造血干细胞在细胞因子M - CSF和RANK - L存在的情况下，直接在牙本质和磷酸钙骨水泥上培养4周。用标准技术监测破骨细胞的发育。去除细胞后，通过LM、SEM和IFM对吸收情况进行表征和量化。
生物材料上的破骨细胞培养呈现出典型的破骨细胞特异性标志物。在牙本质样本上，LM、SEM以及IFM都能区分吸收情况。吸收面积的量化结果显示，各结果之间呈线性相关（LM与SEM：r = 0.996，p = 0.004；SEM与IFM：r = 0.989，p = 0.011；IFM与LM：r = 0.995）。使用LM或SEM无法在GB14上划定吸收凹坑。使用IFM，可以在二维和三维上可视化并量化GB14上的吸收情况。
在本文中，我们介绍了IFM作为一种用于生物材料吸收三维可视化和量化的技术。更好地理解生物材料的生物吸收可能有助于设计出更好的材料，因此可能是人工骨开发道路上的重要一步。
