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Intrafibrillar demineralization study of single human dentin collagen fibrils by AFM

机译:原子力显微镜研究单个人牙本质胶原原纤维的纤维内脱矿质

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In situ atomic force microscopy (AFM) was used to investigate the kinetics of demineralization of human dentin collagen fibrils. Individual dentin collagen fibrils containing intrafibrillar mineral were isolated, transferred onto a glass slide and exposed to water for a day prior to demineralization studies. The fibrils then were exposed to trypsin for removal of non-collagenous proteins and subsequently demineralized in 10 vol % citric acid. Topographic images showed a gradual increase in gap-overlap depth of the fibril. The gap-overlap depth varied linearly with the square root of time before saturation at 7 nm in approximately sixty minutes, suggesting a diffusion process for demineralization of intrafibrillar mineral. Micro Raman studies of partially demineralized dentin revealed the existence of a phosphate peak at wave number close to 960 cm~(-1) . The peak gradually disappeared in 60 minutes as the samples were exposed to 10% citric acid, supporting the notion that AFM topography may be correlated to the degree of intrafibrillar mineralization.
机译:原位原子力显微镜(AFM)用于研究人牙本质胶原原纤维脱矿质的动力学。分离含有原纤维内矿物质的单个牙本质胶原原纤维,转移到载玻片上,并在脱矿质研究前暴露于水中一天。然后将原纤维暴露于胰蛋白酶以去除非胶原蛋白,随后在10体积%柠檬酸中脱矿质。地形图显示原纤维的间隙重叠深度逐渐增加。间隙重叠深度随时间的平方根线性变化,直到在大约60分钟内在7 nm处饱和,才表明原纤维内矿物质的脱矿化过程。对部分脱矿的牙本质进行的显微拉曼研究表明,在波数接近960 cm〜(-1)时存在一个磷酸峰。当样品暴露于10%柠檬酸时,该峰在60分钟内逐渐消失,这支持了AFM形貌可能与原纤维内矿化程度相关的观点。

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