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成骨不全症相关牙本质生成不全症中胶原蛋白的表型特性。

Phenotypic Properties of Collagen in Dentinogenesis Imperfecta Associated with Osteogenesis Imperfecta.

机构信息

Department of Paediatric Dentistry, UCL Eastman Dental Institute, University College London, London, UK.

Department of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, University College London, London, UK.

出版信息

Int J Nanomedicine. 2019 Dec 2;14:9423-9435. doi: 10.2147/IJN.S217420. eCollection 2019.

Abstract

INTRODUCTION

Dentinogenesis imperfecta type 1 (OIDI) is considered a relatively rare genetic disorder (1:5000 to 1:45,000) associated with osteogenesis imperfecta. OIDI impacts the formation of collagen fibrils in dentin, leading to morphological and structural changes that affect the strength and appearance of teeth. However, there is still a lack of understanding regarding the nanoscale characterization of the disease, in terms of collagen ultrastructure and mechanical properties. Therefore, this research presents a qualitative and quantitative report into the phenotype and characterization of OIDI in dentin, by using a combination of imaging, nanomechanical approaches.

METHODS

For this study, 8 primary molars from OIDI patients and 8 primary control molars were collected, embedded in acrylic resin and cut into longitudinal sections. Sections were then demineralized in 37% phosphoric acid using a protocol developed in-house. Initial experiments demonstrated the effectiveness of the demineralization protocol, as the ATR-FTIR spectral fingerprints showed an increase in the amide bands together with a decrease in phosphate content. Structural and mechanical analyses were performed directly on both the mineralized and demineralized samples using a combination of scanning electron microscopy, atomic force microscopy, and Wallace indentation.

RESULTS

Mesoscale imaging showed alterations in dentinal tubule morphology in OIDI patients, with a reduced number of tubules and a decreased tubule diameter compared to healthy controls. Nanoscale collagen ultrastructure presented a similar D-banding periodicity between OIDI and controls. Reduced collagen fibrils diameter was also recorded for the OIDI group. The hardness of the (mineralized) control dentin was found to be significantly higher (p<0.05) than that of the OIDI (mineralized) dentine. Both the exposed peri- and intratubular dentinal collagen presented bimodal elastic behaviors (Young's moduli). The control samples presented a stiffening of the intratubular collagen when compared to the peritubular collagen. In case of the OIDI, this stiffening in the collagen between peri- and intratubular dentinal collagen was not observed and the exposed collagen presented overall a lower elasticity than the control samples.

CONCLUSION

This study presents a systematic approach to the characterization of collagen structure and properties in OIDI as diagnosed in dentin. Structural markers for OIDI at the mesoscale and nanoscale were found and correlated with an observed lack of increased elastic moduli of the collagen fibrils in the intratubular OIDI dentin. These findings offer an explanation of how structural changes in the dentin could be responsible for the failure of some adhesive restorative materials as observed in patients affected by OIDI.

摘要

简介

1 型牙本质生成不全(OIDI)被认为是一种与成骨不全症相关的相对罕见的遗传疾病(1:5000 至 1:45000)。OIDI 影响牙本质中胶原纤维的形成,导致形态和结构发生变化,从而影响牙齿的强度和外观。然而,对于该疾病的纳米级特征,包括胶原超微结构和力学性能,我们仍然缺乏了解。因此,本研究通过结合成像和纳米力学方法,对 OIDI 牙本质的表型和特征进行了定性和定量的报告。

方法

为此研究,收集了 8 例 OIDI 患者的 8 颗乳磨牙和 8 颗乳磨牙对照,将其嵌入丙烯酸树脂中并切成纵切面。然后使用内部开发的方案将切片在 37%的磷酸中脱矿。初步实验表明,脱矿方案是有效的,因为 ATR-FTIR 光谱指纹显示酰胺带增加,同时磷酸盐含量减少。使用扫描电子显微镜、原子力显微镜和 Wallace 压痕法对矿化和脱矿样品直接进行结构和力学分析。

结果

中尺度成像显示 OIDI 患者牙本质小管形态发生改变,与健康对照组相比,小管数量减少,小管直径减小。纳米级胶原超微结构显示 OIDI 和对照组之间存在相似的 D 带周期性。OIDI 组的胶原纤维直径也记录减小。发现(矿化)对照组牙本质的硬度明显高于 OIDI(矿化)牙本质(p<0.05)。暴露的内外根管牙本质胶原均表现出双模态弹性行为(杨氏模量)。与外根管胶原相比,对照组的内根管胶原表现出硬化。在 OIDI 的情况下,在外根管和内根管牙本质胶原之间没有观察到胶原的这种硬化,并且暴露的胶原整体表现出比对照组更低的弹性。

结论

本研究提出了一种系统的方法来对 OIDI 牙本质中的胶原结构和特性进行表征。在中尺度和纳米尺度上发现了 OIDI 的结构标记,并与观察到的内根管 OIDI 牙本质中胶原纤维弹性模量增加不足相关。这些发现解释了牙本质的结构变化如何导致一些在 OIDI 患者中观察到的粘固修复材料失效。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/edfe/6897053/8a325801ebc2/IJN-14-9423-g0001.jpg

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