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通过硅烷化将抗菌肽 GL13K 固定在钛上的抗炎和生物相容性评价。

Inflammatory and biocompatibility evaluation of antimicrobial peptide GL13K immobilized onto titanium by silanization.

机构信息

Department of Oral Implantology, Affiliated Stomatological Hospital of Fujian Medical University, No. 246, Yangqiao Road, Gulou District, Fuzhou, Fujian 350002, China.

School of Stomatology, Fujian Medical University, Fuzhou, Fujian 350000, China; Fujian Biological Materials Engineering and Technology Center of Stomatology, Fuzhou, Fujian 350000, China.

出版信息

Colloids Surf B Biointerfaces. 2017 Dec 1;160:581-588. doi: 10.1016/j.colsurfb.2017.10.007. Epub 2017 Oct 4.

Abstract

The inflammatory reaction around the implant after implant placement is important not only for osseointegration but also for long-term implant survivals. In our study, GL13K, an antimicrobial peptide, was immobilized onto titanium surfaces to improve its anti-inflammatory properties. The method of silanization was used to immobilize the GL13K, which was confirmed by X-ray photoelectron spectroscopy, scanning electron microscopy, atomic force microscopy, water contact angle measurement. DAPI fluorescence staining and Cell Counting Kit-8 (CCK-8) were used to measure the cell attachment and cell viability of the RAW264.7, which indicated a good cytocompatibility. Cellular morphology of RAW264.7 on modified surfaces showed less cell pseudopod. ELISA and qRT-PCR were performed to measure the inflammatory activity of the modified titanium surfaces. The secretion levels of pro-inflammatory cytokines interleukin (IL)-1β, tumor necrosis factor-alpha (TNF-α) and inducible nitric oxide synthase (iNOS) were downregulated at 12h, 24h, and 48h, while the anti-inflammatory cytokines IL-10 and arginase were upregulated at 12h, 24h, and 48h. All results indicate that the GL13K-coated titanium surfaces make the inflammatory process towards a less pro-inflammatory, which may promote the process of osseointegration.

摘要

种植体植入后周围的炎症反应不仅对骨整合很重要,而且对长期种植体存活率也很重要。在我们的研究中,GL13K,一种抗菌肽,被固定在钛表面上以提高其抗炎特性。硅烷化方法被用于固定 GL13K,这一点通过 X 射线光电子能谱、扫描电子显微镜、原子力显微镜、水接触角测量得到了证实。DAPI 荧光染色和细胞计数试剂盒(CCK-8)用于测量 RAW264.7 的细胞附着和细胞活力,这表明其具有良好的细胞相容性。在改性表面上 RAW264.7 的细胞形态显示出较少的细胞伪足。通过 ELISA 和 qRT-PCR 测量改性钛表面的炎症活性。促炎细胞因子白细胞介素 (IL)-1β、肿瘤坏死因子-α (TNF-α) 和诱导型一氧化氮合酶 (iNOS) 的分泌水平在 12h、24h 和 48h 时下调,而抗炎细胞因子 IL-10 和精氨酸酶在 12h、24h 和 48h 时上调。所有结果表明,GL13K 涂层钛表面使炎症过程向较少的促炎方向发展,这可能促进骨整合过程。

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