Pigment epithelium-derived factor modulates periodontal homeostasis in mice and induces osteogenic differentiation of human periodontal ligament fibroblasts.

IF 2.8 4区医学 Q3 CELL BIOLOGY

Connective Tissue Research Pub Date : 2022-09-01 Epub Date: 2022-02-05 DOI:10.1080/03008207.2021.2025224

Cheng Xu, Yu Du, Jun Tian, Chang Liu, Yihua Huang, Ti Zhou, Yang Ning

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引用次数: 1

Abstract

Aim: The aim of this study was to investigate the influence of pigment epithelium-derived factor (PEDF) on periodontal homeostasis in mice and the osteogenic differentiation of human periodontal ligament fibroblasts (PDLFs).

Materials and methods: Micro-computed tomography and histology were performed to compare the alveolar bone volume, density, and bone-related markers between PEDF-deficient (PEDF^-/-) and wild-type (WT) mice. Furthermore, after recombinant human PEDF treatment, the PDLF viability and osteogenic differentiation were examined using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, alkaline phosphatase (ALP) activity assay, Von Kossa staining, Alizarin red staining, real-time quantitative polymerase chain reaction (qRT-PCR), and immunoblotting.

Results: The alveolar bone volume and density of PEDF^-/- mice were significantly lower than those of the WT mice. Higher receptor activator for nuclear factor-κB ligand (RANKL) expression and lower osteoprotegerin (OPG) expression levels were observed in the PEDF^-/- group. Moreover, PEDF treatment did not affect the PDLF proliferation. PEDF dose-dependently improved mineral deposition. Compared with the control group, 250 ng/mL PEDF promoted OPG mRNA expression in PDLFs on Day 3 but inhibited RANKL, Wnt5a, GSK3b mRNA, and non-phosphorylated β-catenin protein expression. However, 250 ng/mL PEDF had no significant effect on the expression of Wnt3a. On Day 7, after culture with 250 ng/mL PEDF in osteogenic medium, the ALP and RUNX2 protein levels were upregulated. VEGF protein expression was reduced in a dose-dependent manner after PEDF stimulation. The PEDF protein expression increased as the osteogenic induction time increased.

Conclusion: PEDF gene knockout suppresses periodontal homeostasis in mice, and PEDF treatment induces PDLF osteogenic differentiation in vitro.

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色素上皮衍生因子调节小鼠牙周稳态及诱导人牙周韧带成纤维细胞成骨分化。

目的:探讨色素上皮衍生因子(PEDF)对小鼠牙周稳态及人牙周韧带成纤维细胞成骨分化的影响。材料和方法:通过显微计算机断层扫描和组织学比较PEDF缺陷(PEDF-/-)和野生型(PEDF-/-)小鼠的牙槽骨体积、密度和骨相关标志物。采用3-[4,5-二甲基噻唑-2-酰基]-2,5二苯基溴化四唑(MTT)法、碱性磷酸酶(ALP)活性法、Von Kossa染色法、茜素红染色法、实时定量聚合酶链反应(qRT-PCR)法和免疫印迹法检测重组人PEDF处理后的pdf活力和成骨分化。结果:PEDF-/-小鼠的牙槽骨体积和骨密度明显低于WT小鼠。PEDF-/-组核因子-κ b配体受体激活因子(RANKL)表达升高，骨保护素(OPG)表达降低。此外，PEDF治疗不影响ppdf的增殖。PEDF剂量依赖性改善矿物沉积。与对照组相比，250 ng/mL PEDF在第3天促进了PDLFs中OPG mRNA的表达，但抑制了RANKL、Wnt5a、GSK3b mRNA和非磷酸化β-catenin蛋白的表达。而250 ng/mL PEDF对Wnt3a的表达无显著影响。第7天，在成骨培养基中加入250 ng/mL PEDF培养后，ALP和RUNX2蛋白水平上调。PEDF刺激后VEGF蛋白表达呈剂量依赖性降低。PEDF蛋白表达随成骨诱导时间的延长而增加。结论:敲除PEDF基因可抑制小鼠牙周稳态，PEDF治疗可诱导pdf体外成骨分化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Connective Tissue Research 生物-细胞生物学

CiteScore

6.60

自引率

3.40%

发文量

审稿时长

2 months

期刊介绍： The aim of Connective Tissue Research is to present original and significant research in all basic areas of connective tissue and matrix biology. The journal also provides topical reviews and, on occasion, the proceedings of conferences in areas of special interest at which original work is presented. The journal supports an interdisciplinary approach; we present a variety of perspectives from different disciplines, including Biochemistry Cell and Molecular Biology Immunology Structural Biology Biophysics Biomechanics Regenerative Medicine The interests of the Editorial Board are to understand, mechanistically, the structure-function relationships in connective tissue extracellular matrix, and its associated cells, through interpretation of sophisticated experimentation using state-of-the-art technologies that include molecular genetics, imaging, immunology, biomechanics and tissue engineering.