SEMA4D修饰钛表面通过巨噬细胞调控内皮细胞功能的研究

doi:10.13591/j.cnki.kqyx.2024.02.001

摘要/Abstract

摘要：

目的探究SEMA4D修饰钛表面对巨噬细胞的直接作用以及通过巨噬细胞对内皮细胞功能的间接调控作用。方法纯钛试件抛光清洗后,通过碱热处理和自组装技术在其表面制备SEMA4D涂层。以光滑钛(Ti)试件为对照组,浓度为25、50和100 ng/mL的SEMA4D溶液制备的SEMA4D修饰钛表面(Ti-4D25、Ti-4D50和Ti-4D100)试件为实验组,采用扫描电镜和接触角测量仪分析各组的表面微形貌和亲水性。在4组试件表面接种巨噬细胞,将获得的上清液制成条件培养基培养内皮细胞,通过CCK-8法检测两种细胞的增殖活性,通过实时荧光定量PCR检测巨噬细胞的炎性表达和VEGF表达,通过荧光染色和划痕实验检测内皮细胞的黏附和迁移能力。结果 SEMA4D修饰钛表面呈现多孔微结构并且被有机薄膜覆盖,无细胞毒性。该表面可降低巨噬细胞炎性表达,上调VEGF表达,促进内皮细胞的黏附和迁移。结论 SEMA4D修饰钛表面可以减轻炎症反应并增强内皮细胞的运动功能,从而获得有利于软组织修复的生理微环境。

关键词: SEMA4D, 钛表面, 巨噬细胞, 内皮细胞

Abstract:

Objective To explore the direct impact of SEMA4D-modified titanium surface on macrophages and the indirect regulatory effect on endothelial cell functions through macrophages. Methods Pure titanium specimens were polished and cleaned, and SEMA4D coatings were prepared using alkali heat treatment and self-assembly techniques. The experimental group consisted of titanium specimens coated with SEMA4D solutions at concentrations of 25, 50, and 100 ng/mL, with smooth titanium(Ti)specimens as the control group. The surface micromorphology and hydrophilicity of titanium in each group were analyzed by scanning electron microscopy and a contact angle meter. Macrophages were introduced onto the surface of each group, and the resulting supernatant was used to culture endothelial cells. The proliferative activity of the cells was measured using CCK-8 assay. The inflammatory expression and VEGF expression of macrophages were evaluated using real-time quantitative PCR. The adhesion and migration abilities of endothelial cells were determined using the fluorescent staining and scratch assay. Results The SEMA4D-modified titanium surface showed a porous microstructure covered by an organic film, which was non-cytotoxic. This surface reduced macrophage inflammatory expression, up-regulated VEGF expression and promoted endothelial cells’ adhesion and migration. Conclusion SEMA4D modified titanium surface can reduce inflammatory response and enhance the motor function of endothelial cells, thereby obtaining a physiological microenvironment conducive to soft tissue repairing.

Key words: SEMA4D, titanium surface, macrophage, endothelial cell

中图分类号:

R783.1

周洁仪, 张建兰, 谢玲玲, 柳姚, 邱憬. SEMA4D修饰钛表面通过巨噬细胞调控内皮细胞功能的研究[J]. 口腔医学, 2024, 44(2): 81-87.

ZHOU Jieyi, ZHANG Jianlan, XIE Lingling, LIU Yao, QIU Jing. Study on SEMA4D modified titanium surface modulating endothelial cell functions via macrophages[J]. Stomatology, 2024, 44(2): 81-87.

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基因名称	引物序列
TNF-α	Forward:5'-GCAAAGGGAGAGTGGTCA-3' Reverse:5'-CTGGCTCTGTGAGGAAGG-3'
IL-1β	Forward:5'-CTCCTGGTATGAAGTGGCAAATC-3' Reverse:5'-ACAAGGCTGCCCCGACTAT-3'
IL-6	Forward:5'-AGGCATAACGCACTAGGTTT-3' Reverse:5'-ACAGAAGGAGTGGCTAAGGA-3'
VEGF	Forward:5'-TAGAGTACATCTTCAAGCCGTC-3' Reverse:5'-CTTTCTTTGGTCTGCATTCACA-3'
GAPDH	Forward:5'-AACTTTGGCATTGGGAAGG-3' Reverse:5'-ACACATTGGGGTAGGAACA-3'