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

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.