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褐藻酸钠/明胶多孔支架与岩藻依聚糖融合对小鼠小胶质细胞BV2的抗炎作用

Anti-inflammatory effects of sodium alginate/gelatine porous scaffolds merged with fucoidan in murine microglial BV2 cells.

作者信息

Nguyen Van-Tinh, Ko Seok-Chun, Oh Gun-Woo, Heo Seong-Yeong, Jeon You-Jin, Park Won Sun, Choi Il-Whan, Choi Sung-Wook, Jung Won-Kyo

机构信息

Department of Biomedical Engineering, and Center for Marine-Integrated Biomedical Technology (BK21 Plus) Pukyong National University, Busan 48513, Republic of Korea; Marine-Integrated Bionics Research Center, Pukyong National University, Busan 48513, Republic of Korea.

Department of Marine Life Sciences, Jeju National University, Jeju 63243, Republic of Korea.

出版信息

Int J Biol Macromol. 2016 Dec;93(Pt B):1620-1632. doi: 10.1016/j.ijbiomac.2016.05.078. Epub 2016 May 24.

DOI:10.1016/j.ijbiomac.2016.05.078
PMID:27234497
Abstract

Microglia are the immune cells of the central nervous system (CNS). Overexpression of inflammatory mediators by microglia can induce several neurological diseases. Thus, the underlying basic requirement for neural tissue engineering is to develop materials that exhibit little or no neuro-inflammatory effects. In this study, we have developed a method to create porous scaffolds by adding fucoidan (Fu) into porous sodium alginate (Sa)/gelatine (G) (SaGFu). For mechanical characterization, in vitro degradation, stress/strain, swelling, and pore size were measured. Furthermore, the biocompatibility was evaluated by assessing the adhesion and proliferation of BV2 microglial cells on the SaGFu porous scaffolds using scanning electron microscopy (SEM) and lactate dehydrogenase (LDH) assay, respectively. Moreover, we studied the neuro-inflammatory effects of SaGFu on BV2 microglial cells. The effect of gelatine and fucoidan content on the various properties of the scaffold was investigated and the results showed that mechanical properties increased porosity and swelling ratio with an increase in the gelatine and fucoidan, while the in vitro biodegradability decreased. The average SaGFu diameter attained by fabrication of SaGFu ranged from 60 to 120μm with high porosity (74.44%-88.30%). Cell culture using gelatine 2.0% (SaG2Fu) and 4.0% (SaG4Fu), showed good cell proliferation; more than 60-80% that with Sa alone. Following stimulation with 0.5μg/mL LPS, microglia cultured in porous SaGFu decreased their expression of nitric oxide (NO), prostaglandin E2 (PGE2), and reactive oxygen species (ROS). SaG2Fu and SaG4Fu also inhibited the activation and translocation of p65 NF-κB protein levels, resulting in reduction of NO, ROS, and PGE2 production. These results provide insights into the diverse biological effects and opens new avenues for the applications of SaGFu in neuroscience.

摘要

小胶质细胞是中枢神经系统(CNS)的免疫细胞。小胶质细胞炎症介质的过度表达可诱发多种神经疾病。因此,神经组织工程的潜在基本要求是开发出几乎没有或没有神经炎症作用的材料。在本研究中,我们开发了一种通过向多孔海藻酸钠(Sa)/明胶(G)(SaGFu)中添加岩藻依聚糖(Fu)来制备多孔支架的方法。对于力学表征,测量了体外降解、应力/应变、膨胀和孔径。此外,分别使用扫描电子显微镜(SEM)和乳酸脱氢酶(LDH)测定法,通过评估BV2小胶质细胞在SaGFu多孔支架上的粘附和增殖来评价生物相容性。此外,我们研究了SaGFu对BV2小胶质细胞的神经炎症作用。研究了明胶和岩藻依聚糖含量对支架各种性能的影响,结果表明,随着明胶和岩藻依聚糖含量的增加,力学性能、孔隙率和膨胀率增加,而体外生物降解性降低。通过制备SaGFu获得的平均直径范围为60至120μm,孔隙率高(74.44%-88.30%)。使用2.0%(SaG2Fu)和4.0%(SaG4Fu)明胶进行细胞培养,显示出良好的细胞增殖;比单独使用Sa时多60%-80%。在用0.5μg/mL脂多糖刺激后,在多孔SaGFu中培养的小胶质细胞降低了一氧化氮(NO)、前列腺素E2(PGE2)和活性氧(ROS)的表达。SaG2Fu和SaG4Fu还抑制了p65 NF-κB蛋白水平的激活和转位,导致NO、ROS和PGE2产生减少。这些结果为多种生物学效应提供了见解,并为SaGFu在神经科学中的应用开辟了新途径。

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