表征交联壳聚糖纳米颗粒对胶原酶降解的胶原稳定作用。
Characterizing the collagen stabilizing effect of crosslinked chitosan nanoparticles against collagenase degradation.
机构信息
Discipline of Endodontics, Faculty of Dentistry, University of Toronto, 124 Edward Street, Toronto, ON M5G1G6, Canada.
Dental Research Institute, Faculty of Dentistry, University of Toronto, 124 Edward Street, Toronto, ON M5G1G6, Canada.
出版信息
Dent Mater. 2016 Aug;32(8):968-77. doi: 10.1016/j.dental.2016.05.005. Epub 2016 May 31.
UNLABELLED
Antibacterial and chelating properties of chitosan has been widely studied for various dental applications.
OBJECTIVE
To characterize the interaction between chitosan-nanoparticles (CSnp) and collagen, and understand their stabilizing effect against collagenase degradation for dentin matrix stabilization.
METHODS
Phase-1: a single Type I collagen-fibril model was used to study the interaction with CSnp along with carbodiimides crosslinking treatment. Degradation of the crosslinked fibrils was studied with bacterial collagenase enzyme and monitored using Fourier Transform Infrared (FTIR) spectroscopy, turbidity measurement (400nm), ninhydrin assay and Atomic Force Microscopy (AFM). Interaction of CSnp with collagenase and Type I collagen, were evaluated using SDS-PAGE, and proteolytic cleavage potential of a synthetic peptide. Phase-2: degradation of dentin collagen crosslinked with/without CSnp was evaluated using FTIR, ninhydrin assay and Scanning Electron Microscopy (SEM). Glutaraldehyde crosslinking was used as a positive control.
RESULTS
Both native collagen-fibrils and dentin collagen after crosslinking showed higher resistance to collagenase degradation, as observed in turbidity measurements and FTIR spectra. AFM images showed the interaction of CSnp with single collagen-fibril and crosslinked collagen resisted collagenase degradation up to 54h. The collagen and collagenase both formed complexes with CSnp resulting in thickening of bands and reduction in collagen degradation. CSnp treated collagenase showed significantly reduced cleavage of the fluorescent peptides. Dentin collagen was coated with CSnp following crosslinking with significant increase in resistance to collagenase degradation.
SIGNIFICANCE
Crosslinked CSnp on collagen stabilized and enhanced the resistance of dentin matrix against bacterial collagenase degradation due to non-specific interaction with both collagen and collagenase.
未加标签
壳聚糖的抗菌和螯合特性已广泛应用于各种牙科领域。
目的
研究壳聚糖纳米粒子（CSnp）与胶原蛋白的相互作用，并了解其对胶原酶降解的稳定作用，以稳定牙本质基质。
方法
第 1 阶段：使用单一的 I 型胶原蛋白纤维模型来研究与 CSnp 的相互作用，同时进行碳二亚胺交联处理。用细菌胶原酶酶降解交联的原纤维，并通过傅里叶变换红外（FTIR）光谱、浊度测量（400nm）、茚三酮测定和原子力显微镜（AFM）进行监测。通过 SDS-PAGE 和合成肽的蛋白水解切割潜力评估 CSnp 与胶原酶和 I 型胶原蛋白的相互作用。第 2 阶段：使用 FTIR、茚三酮测定和扫描电子显微镜（SEM）评估与/不与 CSnp 交联的牙本质胶原蛋白的降解情况。戊二醛交联用作阳性对照。
结果
天然胶原蛋白纤维和交联后的牙本质胶原蛋白在浊度测量和 FTIR 光谱中均显示出对胶原酶降解更高的抵抗力。AFM 图像显示 CSnp 与单根胶原蛋白纤维的相互作用以及交联胶原蛋白抵抗胶原酶降解的能力长达 54 小时。胶原蛋白和胶原酶都与 CSnp 形成复合物，导致条带变厚，胶原蛋白降解减少。用 CSnp 处理的胶原酶明显减少了荧光肽的切割。交联后牙本质胶原蛋白被 CSnp 包裹，显著提高了对胶原酶降解的抵抗力。
意义
交联的 CSnp 与胶原蛋白结合，稳定并增强了牙本质基质对细菌胶原酶降解的抵抗力，这是由于 CSnp 与胶原蛋白和胶原酶的非特异性相互作用。
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