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使用巨噬细胞研究实心与多孔磷酸盐基玻璃微球的体外免疫调节作用。

In vitro immunomodulatory effect of solid versus porous phosphate-based glass microspheres using macrophages.

作者信息

Sheela Soumya, AlGhalban Fatma Mousa, Ahmed Ifty, Abou Neel Ensanya A

机构信息

Research Institute for Medical and Health Sciences, University of Sharjah, Sharjah, United Arab Emirates.

Advanced Materials Research Group, Faculty of Engineering, University of Nottingham, Nottingham, NG7 2RD, United Kingdom.

出版信息

Heliyon. 2023 Dec 2;9(12):e23059. doi: 10.1016/j.heliyon.2023.e23059. eCollection 2023 Dec.

Abstract

This study aimed to investigate the immunomodulatory effect of two different forms of phosphate-based glass microspheres (solid and porous), on human macrophages. Human THP-1 monocytes were converted to M0 macrophages after being treated with 100 ng/mL phorbol 12-myristate 13-acetate for 48 h. The differentiated cells were analysed for the CD14 marker using flow cytometry. The adhesion, spreading, and viability of M0 macrophages grown directly or indirectly (extracts) at varying concentrations of solid and porous glass microspheres (GMs) were analysed via phase contrast microscopy, confocal microscopy, and XTT assay. The expression of IL-8, IL-1β, IL-6, IL-10, TNF-α, and IL-12p70 cytokines was investigated using flow cytometry. The conversion to M0 macrophages was confirmed by their adherent nature, increased granularity, and CD14 expression. The results showed that both solid and porous GMs or extracts favored the attachment, spreading, and proliferation of macrophages in a comparable manner to cells grown in a normal tissue culture medium. Only the higher concentration of porous GMs (10 mg/mL) changed the morphology of M0 macrophages and increased the expression of IL-1β and IL-8 pro-inflammatory cytokines; this could be related to the fast degradation nature of porous GMs. Of the six cytokines analysed, M0 macrophages grown directly or indirectly with GMs only expressed IL-1β, IL-10, and IL-8. Accordingly, solid microspheres may have advantages as regenerative agents due to their controlled degradation.

摘要

本研究旨在探讨两种不同形式的磷酸盐基玻璃微球(实心和多孔)对人巨噬细胞的免疫调节作用。人THP-1单核细胞在100 ng/mL佛波酯12-肉豆蔻酸酯13-乙酸酯处理48小时后转化为M0巨噬细胞。使用流式细胞术分析分化细胞的CD14标志物。通过相差显微镜、共聚焦显微镜和XTT测定法分析在不同浓度的实心和多孔玻璃微球(GMs)上直接或间接(提取物)生长的M0巨噬细胞的粘附、铺展和活力。使用流式细胞术研究IL-8、IL-1β、IL-6、IL-10、TNF-α和IL-12p70细胞因子的表达。通过其贴壁性质、增加的粒度和CD14表达证实了向M0巨噬细胞的转化。结果表明,实心和多孔GMs或提取物均以与在正常组织培养基中生长的细胞相当的方式促进巨噬细胞的附着、铺展和增殖。只有较高浓度的多孔GMs(10 mg/mL)改变了M0巨噬细胞的形态并增加了IL-1β和IL-8促炎细胞因子的表达;这可能与多孔GMs的快速降解性质有关。在所分析的六种细胞因子中,直接或间接与GMs一起生长的M0巨噬细胞仅表达IL-1β、IL-10和IL-8。因此,实心微球由于其可控的降解性可能具有作为再生剂的优势。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eab6/10750036/27b38dbb5426/ga1.jpg

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