经羧甲基壳聚糖修饰的具有增强生物活性的脱矿牙本质基质，可通过诱导成骨分化。

Peptide decorated demineralized dentin matrix with enhanced bioactivity, osteogenic differentiation via carboxymethyl chitosan.

机构信息

Stomatological Hospital of Chongqing Medical University, Chongqing Medical University, Chongqing, China; Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing, China; Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing, China.

出版信息

Dent Mater. 2021 Jan;37(1):19-29. doi: 10.1016/j.dental.2020.09.019. Epub 2020 Nov 27.

DOI:10.1016/j.dental.2020.09.019

PMID:33257086

Abstract

OBJECTIVES

To improve the biocompatibility and osteogenic activity of demineralized dentin matrix (DDM) by grafting peptides on its surface.

METHODS

DDM was obtained by pulverizing extracted human teeth that had been systematically demineralized and dried. Four groups of materials were evaluated: DDM, DDM/carboxymethyl chitosan (CMC), DDM/CMC/bone forming peptide-1 (BFP-1), and blank. X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FT-IR) and fluorescence localization were used to characterize the surface of the DDM materials. Cell viability was assessed using a CCK8 assay, scanning electron microscopy (SEM) and in vitro osteogenesis was analyzed using real-time RT-PCR (RT-qPCR) and Alizarin red and alkaline phosphatase staining. Three different materials were implanted into mandibular bone defects in rats. After 8 weeks, bone regeneration was assessed by histomorphometry of HE-stained slides.

RESULTS

FT-IR, XPS, and fluorescence microscopy demonstrated that the DDM surfaces were successfully modified with BFP-1. The CCK8 assay indicated that the proliferation of cells is higher on the DDM/CMC/BFP-1 material than on DDM or DDM/CMC (P < 0.05). Cells were more likely to adhere to DDM/CMC/BFP-1, as observed by SEM. Greater in vitro osteogenesis was observed in the DDM/CMC/BFP-1 group which displayed stronger alkaline phosphatase activity, more alizarin red-stained nodules, and higher target gene expression, as detected by RT-qPCR (P＜0.05). HE staining of in vivo explants indicated that greater quantities of new bone had formed in the DDM/CMC/BFP-1 group.

SIGNIFICANCE

Compared with DDM, DDM/CMC/BFP-1 exhibited superior biocompatibility and osteogenesis, using a method of surface modification that has great potential for future clinical use.

摘要

目的

通过在脱矿牙本质基质（DDM）表面接枝肽来提高其生物相容性和成骨活性。

方法

DDM 通过粉碎经过系统脱矿和干燥的提取人牙获得。评估了四组材料：DDM、DDM/羧甲基壳聚糖（CMC）、DDM/CMC/骨形成肽-1（BFP-1）和空白。X 射线光电子能谱（XPS）、傅里叶变换红外光谱（FT-IR）和荧光定位用于表征 DDM 材料的表面。通过 CCK8 测定法、扫描电子显微镜（SEM）评估细胞活力，并通过实时 RT-PCR（RT-qPCR）和茜素红和碱性磷酸酶染色分析体外成骨作用。将三种不同的材料植入大鼠下颌骨缺损中。8 周后，通过 HE 染色切片的组织形态计量学评估骨再生。

结果

FT-IR、XPS 和荧光显微镜显示，DDM 表面成功地用 BFP-1 修饰。CCK8 测定法表明，与 DDM 或 DDM/CMC 相比，DDM/CMC/BFP-1 材料上细胞的增殖更高（P<0.05）。SEM 观察到细胞更倾向于附着在 DDM/CMC/BFP-1 上。在 DDM/CMC/BFP-1 组中观察到更强的碱性磷酸酶活性、更多的茜素红染色结节和更高的目标基因表达，表明体外成骨作用更强，通过 RT-qPCR 检测（P＜0.05）。体内植入物的 HE 染色表明，DDM/CMC/BFP-1 组形成了更多的新骨。