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具有抗菌和治疗抗炎潜力的羟基磷灰石仿生材料的呈现:RAW 264.7细胞中脂多糖诱导应激的抗炎评估。

Presentation of Antibacterial and Therapeutic Anti-inflammatory Potentials to Hydroxyapatite Biomimetic With : An Anti-inflammatory Assessment in Contradiction of LPS-Induced Stress in RAW 264.7 Cells.

作者信息

Nagaraj Anusuya, Samiappan Suja

机构信息

Department of Biochemistry, Bharathiar University, Coimbatore, India.

出版信息

Front Microbiol. 2019 Aug 7;10:1757. doi: 10.3389/fmicb.2019.01757. eCollection 2019.

Abstract

In the present study, for the first time, biomimetization of hydroxyapatite (HA) with (AI) was proposed and established its antioxidant, antibacterial, and anti-inflammatory potential on lipopolysaccharide (LPS). The ethanolic extract of AI was found rich with phenolics and flavonoids, and determined their concentration as 8.98 ± 1.41 mg gallic acid equivalents/g and 5.46 ± 0.84 mg catechin equivalents/g, respectively. The HA was prepared by sol-gel method from calcium nitrate tetrahydrate and orthophosphoric acid, and successfully biomimetization was performed with ethanolic extract of AI. The FTIR analysis settled that as-synthesized HA-AI composite was comprised of both HA and AI. The XRD pattern and Zeta potential revealed that the HA-AI composite was crystalline and negative in charge (-24.0 mV). The average-size distribution, shape, and size of the HA-AI composite was determined as 238.90 d.nm, spherical, and 117.90 nm from size distribution, SEM, and HR-TEM analysis, respectively. The SEM-EDX concluded that the HA-AI composite was comprised of elements of HA as well as AI. The HA-AI composite presented potential antioxidant activity and its EC50 values (dose required to inhibit about half of the radicals) for ABTS and DPPH assays were determined as 115.72 ± 2.33 and 128.51 ± 1.04 μg/ml, respectively. The HA-AI composite showed potent antibacterial activity, and minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) towards (ATCC 700699) and (ATCC 10536) were correspondingly determined as 266.7 ± 28.87 and 600.0 ± 50.0 μg/ml, and 400.0 ± 86.6 and 816.7 ± 76.38 μg/ml. Most importantly, HA-AI composite presented the potential anti-inflammatory response toward lipopolysaccharide (LPS) in RAW 264.7 cells. The dose of 250 μg/ml of HA-AI composite has shown optimum protection against LPS-induced stress (1 μg/ml) by scavenging oxidants and regulating mitochondrial membrane potential (MMP), inflammatory and apoptotic factors. Thus, this study concluded that the impartation of potential biofunctional features to HA from plant sources through biomimetic approach is much beneficial and could find potential application in dentistry and orthopedic.

摘要

在本研究中,首次提出了用(AI)对羟基磷灰石(HA)进行生物矿化,并确定了其对脂多糖(LPS)的抗氧化、抗菌和抗炎潜力。发现AI的乙醇提取物富含酚类和黄酮类化合物,其浓度分别测定为8.98±1.41毫克没食子酸当量/克和5.46±0.84毫克儿茶素当量/克。通过溶胶-凝胶法由四水合硝酸钙和正磷酸制备HA,并成功地用AI的乙醇提取物进行了生物矿化。傅里叶变换红外光谱(FTIR)分析表明,合成的HA-AI复合材料由HA和AI组成。X射线衍射(XRD)图谱和zeta电位表明,HA-AI复合材料是结晶的且带负电荷(-24.0 mV)。根据尺寸分布、扫描电子显微镜(SEM)和高分辨透射电子显微镜(HR-TEM)分析,HA-AI复合材料的平均尺寸分布、形状和尺寸分别确定为238.90 d.nm、球形和117.90 nm。扫描电子显微镜-能谱分析(SEM-EDX)得出结论,HA-AI复合材料由HA以及AI的元素组成。HA-AI复合材料具有潜在的抗氧化活性,其在ABTS和DPPH测定中的半数有效浓度(EC50)值(抑制约一半自由基所需的剂量)分别测定为115.72±2.33和128.51±1.04微克/毫升。HA-AI复合材料显示出强大的抗菌活性,对(美国典型培养物保藏中心700699)和(美国典型培养物保藏中心10536)的最低抑菌浓度(MIC)和最低杀菌浓度(MBC)相应地测定为266.7±28.87和600.0±50.0微克/毫升,以及400.0±86.6和816.7±76.38微克/毫升。最重要的是,HA-AI复合材料对RAW 264.7细胞中的脂多糖(LPS)呈现出潜在的抗炎反应。250微克/毫升剂量的HA-AI复合材料通过清除氧化剂和调节线粒体膜电位(MMP)、炎症和凋亡因子,对LPS诱导的应激(1微克/毫升)显示出最佳保护作用。因此,本研究得出结论,通过仿生方法从植物来源赋予HA潜在的生物功能特性非常有益,并可能在牙科和骨科领域找到潜在应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5b2/6692563/dfe661eb9a93/fmicb-10-01757-g001.jpg

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