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人成骨细胞(hFOB)在硅-双相磷酸钙材料上的生物活性研究及黏附情况

Bioactivity studies and adhesion of human osteoblast (hFOB) on silicon-biphasic calcium phosphate material.

作者信息

Ibrahim S, Sabudin S, Sahid S, Marzuke M A, Hussin Z H, Kader Bashah N S, Jamuna-Thevi K

机构信息

Advanced Materials Research Centre (AMREC), SIRIM Berhad, Lot 34, Jalan Hi-Tech 2/3, Kulim Hi-Tech Park, 09000 Kulim, Kedah Darul Aman, Malaysia.

出版信息

Saudi J Biol Sci. 2016 Jan;23(1):S56-63. doi: 10.1016/j.sjbs.2015.10.024. Epub 2015 Nov 10.

Abstract

Surface reactivity of bioactive ceramics contributes in accelerating bone healing by anchoring osteoblast cells and the connection of the surrounding bone tissues. The presence of silicon (Si) in many biocompatible and bioactive materials has been shown to improve osteoblast cell adhesion, proliferation and bone regeneration due to its role in the mineralisation process around implants. In this study, the effects of Si-biphasic calcium phosphate (Si-BCP) on bioactivity and adhesion of human osteoblast (hFOB) as an in vitro model have been investigated. Si-BCP was synthesised using calcium hydroxide (Ca(OH)2) and phosphoric acid (H3PO4) via wet synthesis technique at Ca/P ratio 1.60 of material precursors. SiO2 at 3 wt% based on total precursors was added into apatite slurry before proceeding with the spray drying process. Apatite powder derived from the spray drying process was pressed into discs with Ø 10 mm. Finally, the discs were sintered at atmospheric condition to obtain biphasic hydroxyapatite (HA) and tricalcium phosphate (TCP) peaks simultaneously and examined by XRD, AFM and SEM for its bioactivity evaluation. In vitro cell viability of L929 fibroblast and adhesion of hFOB cell were investigated via AlamarBlue® (AB) assay and SEM respectively. All results were compared with BCP without Si substitution. Results showed that the presence of Si affected the material's surface and morphology, cell proliferation and cell adhesion. AFM and SEM of Si-BCP revealed a rougher surface compared to BCP. Bioactivity in simulated body fluid (SBF) was characterised by pH, weight gain and apatite mineralisation on the sample surface whereby the changes in surface morphology were evaluated using SEM. Immersion in SBF up to 21 days indicated significant changes in pH, weight gain and apatite formation. Cell viability has demonstrated no cytotoxic effect and denoted that Si-BCP promoted good initial cell adhesion and proliferation. These results suggest that Si-BCP's surface roughness (164 nm) was significantly higher than BCP (88 nm), thus enhancing the adhesion and proliferation of the osteoblast.

摘要

生物活性陶瓷的表面反应性通过锚定成骨细胞以及连接周围骨组织,有助于加速骨愈合。许多生物相容性和生物活性材料中硅(Si)的存在已被证明可改善成骨细胞的黏附、增殖和骨再生,这是因为它在植入物周围的矿化过程中发挥作用。在本研究中,已对作为体外模型的硅双相磷酸钙(Si-BCP)对人成骨细胞(hFOB)的生物活性和黏附的影响进行了研究。采用氢氧化钙(Ca(OH)₂)和磷酸(H₃PO₄)通过湿合成技术,在材料前驱体的钙磷比为1.60的条件下合成Si-BCP。在进行喷雾干燥过程之前,将基于总前驱体3 wt%的SiO₂添加到磷灰石浆料中。将喷雾干燥过程得到的磷灰石粉末压制成直径为10 mm的圆盘。最后,将圆盘在大气条件下烧结,以同时获得双相羟基磷灰石(HA)和磷酸三钙(TCP)峰,并通过X射线衍射(XRD)、原子力显微镜(AFM)和扫描电子显微镜(SEM)对其生物活性进行评估。分别通过AlamarBlue®(AB)检测法和SEM研究了L929成纤维细胞的体外细胞活力和hFOB细胞的黏附情况。所有结果均与未替代硅的BCP进行比较。结果表明,硅的存在影响了材料的表面和形态、细胞增殖及细胞黏附。与BCP相比,Si-BCP的AFM和SEM显示出更粗糙的表面。模拟体液(SBF)中的生物活性通过pH值、重量增加以及样品表面的磷灰石矿化来表征,其中使用SEM评估表面形态的变化。在SBF中浸泡长达21天表明pH值、重量增加和磷灰石形成有显著变化。细胞活力表明无细胞毒性作用,并且表明Si-BCP促进了良好的初始细胞黏附和增殖。这些结果表明,Si-BCP的表面粗糙度(164 nm)显著高于BCP(88 nm),从而增强了成骨细胞的黏附和增殖。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d84a/4705313/fb5b152e746f/gr1.jpg

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