In Vitro Effects of Calcium Fructoborate upon Production of Inflammatory Mediators by LPS-stimulated RAW 264.7 Macrophages

Abstract

The present study is supported by our previous findings suggesting that calcium fructoborate (CF) has anti-inflammatory and antioxidant properties. Thus, we investigated the effects of CF on a model for studying inflammatory disorders in vitro represented by lipopolysaccharide (LPS)-stimulated murine macrophage RAW 264.7 cells. This investigation was performed by analyzing the levels of some mediators released during the inflammatory process: cytokines such as tumor necrosis factor-alpha (TNF-alpha), interleukins IL-1beta and IL-6 as well as cyclooxygenase-2 (COX-2), the main enzyme responsible for endotoxin/LPS-induced prostaglandin synthesis by macrophages. We also measured production of nitric oxide (NO) that plays an important role in the cytotoxicity activity of macrophages towards microbial pathogens. After CF treatment of LPS-stimulated macrophages we found an up-regulation of TNF-alpha protein level in culture medium, no significant changes in the level of COX-2 protein expression and a decrease in NO production as well as in IL-1beta and IL-6 release. Collectively, this series of experiments indicate that CF affect macrophage production of inflammatory mediators. However, further research is required in order to establish whether CF treatment can be beneficial in suppression of pro-inflammatory cytokine production and against progression of endotoxin-related diseases.

Calcium Fructoborate—Potential Anti-inflammatory Agent

“…The collected results also show that concentration in serum of CF is of 556 μg/ml of serum, CF being detected as an intact molecule in the mice's blood, which can provide health benefits too [2]. This observation suggests that all biological effects of CF noticed thus far could be due to this unique feature of the compound [5].…”

“…In a recent study, we showed that treatment of LPS-stimulated RAW264.7 macrophage cells with CF induced an inhibition of interleukin-1β (IL-1β), interleukin-6 (IL-6), and nitric oxide (NO) release in the culture media, an increase of the tumor necrosis factor-α (TNF-α) production, and had no effects on lipopolysaccharide (LPS)-induced cyclooxygenase-2 (COX-2) protein expression [5]. The pro-inflammatory cytokine IL-1β is synthesized by activated monocytes and macrophages as a 31-kDa, biologically inactive precursor that is proteolytically processed to the biologically active 17-kDa mature molecule by the IL-1β converting enzyme [71].…”

“…scientific in vitro research on CF has showed some biological activities that might have a therapeutic potential [3], such as anti-inflammatory [4,5], anti-osteoporosis [2], antioxidant [6], and antitumor [3,7]. CF also counteracts the harmful effects of vitamin D deficiency, due to the correlation between the intake of CF and serum vitamin D 3 levels [2].…”

Effects of dietary calcium fructoborate supplementation on joint comfort and flexibility and serum inflammatory markers in dogs with osteoarthritis

“…A second study, whereby human polymorphonuclear neutrophils were subjected to 22,500 µM of CFB for 24 hours, observed a 92.9% decrease in ROS levels (Scorei et al, 2007). Another study measured the production of inflammatory markers by lipopolysaccharide (LPS)-stimulated murine macrophage RAW 264.7 cells exposed to 0.2, 0.45, or 1 mM CFB for 24 hours (Scorei et al, 2010). In the same study by Scorei et al (2010), CFB decreased (P < 0.01) IL-1β concentrations by 27% and IL-6 by 90% compared to control cells exposed to LPS without CFB treatment.…”

Identification and Quantification of Fructoborate Ester Complex Using Liquid Chromatography Coupled with Q Exactive Orbitrap Mass Spectrometry

“…Miljkovic, N. Miljkovic & McCarty, 2004). Studies (Mogosanu et al, 2016;Scorei, Ciofrangeanu & Ion, 2010;Scorei & Rotaru, 2011;Scorei, 2011;Militaru et al, 2013) demonstrated the ability of calcium fructoborate to significantly modulate molecular markers associated with inflammatory mechanisms on the elevated serum levels of C-reactive protein, suggesting possible benefit for cardiovascular conditions. Scorei and some researchers (Scorei & Popa, 2010;Scorei et al, 2007) found negative correlation between borate intake and the incidence of other human health issues.…”