Measurement of Urea in the Saliva of Healthy Mice - a Pilot Study.

Salivary urea is studied as a non-invasive alternative for screening and monitoring of renal diseases. Its high variability prevents a wider clinical use. Animal experiments are needed to identify factors affecting this marker. The aim of this study was to describe the inter-individual variability of salivary urea in healthy mice, establish reference intervals, and analyse the effects of sex, age and body weight. Plasma and saliva samples were obtained from 37 male and 41 female healthy adult CD1 mice aged 13-69 weeks (body weight 22-51 g). The reference interval for salivary urea in heathy mice based on our results is 2.7-8.4 mmol/l (CV = 23 %). Multivariate analysis did not show any significant effect of age, sex, or body weight. In addition, salivary urea did not correlate with its plasma concentrations. The high variability of the promising salivary marker of kidney function in healthy mice requires further research before its use to diagnose or monitor renal failure in animal models of kidney diseases. Other potential confounders should be analysed, including intra-individual and pre-analytical variability. In addition, a normalization factor such as total salivary proteins or salivation rate is likely needed.

Markers of oxidative stress and antioxidant status in the plasma, urine and saliva of healthy mice.

Oxidative stress markers are usually measured in plasma, a stable environment for biomarkers. Blood collection is invasive, but the use of alternative biofluids is limited, due to high variability. In this study, we aimed to establish reference values for oxidative stress markers in plasma, urine and saliva of adult, healthy mice and to identify some sources of variability. Samples were obtained from 41 female and 37 male adult, healthy mice of the CD-1 strain, aged 95-480 days, weighing 21-55 grams. Reference ranges of TBARS (thiobarbituric acid reactive substances), AOPP (advanced oxidation protein products), fructosamine, GSH/GSSG (reduced and oxidized glutathione) ratio, TAC (total antioxidant capacity), and FRAP (ferric reducing antioxidant power) were measured in plasma and urine, and TBARS, GSH/GSSG ratio, TAC and FRAP in saliva, using standard spectrophotometric and fluorometric methods. Salivary GSH/GSSG and urinary AOPP were higher in females. Urinary fructosamine, GSH/GSSG and FRAP were higher in males. Urinary TAC and FRAP negatively correlated with age, and urinary GSH/GSSG positively correlated with weight. We determined that urine and saliva can be obtained non-invasively from mice, in sufficient amounts for reliable oxidative status assessment. Further studies are needed to uncover whether these biofluids reflect systemic oxidative status in diseases.

Phase transformations, microstructure formation and in vitro osteoblast response in calcium silicate/brushite cement composites.

Self-setting simple calcium silicate/brushite (B) biocements with various Ca/P ratios were prepared by mutual mixing of both monocalcium silicate hydrate (CSH) or ß-wollastonite (woll) powders with B and the addition of 2 wt% NaH2PO4 solution as a hardening liquid. The phase composition of the final composites and the texture of the surface calcium phosphate/silica layer were controlled by the starting Ca/P ratio in composites and the pH during setting. It was verified that the presence of continuous bone-like calcium phosphate coating on the surface of the samples was not essential for in vitro osteoblast proliferation. The nanocrystalline calcium deficient hydroxyapatite and amorphous silica were found as the main setting products in composite mixtures with a Ca/P ratio close to the region of the formation of deficient hydroxyapatite-like calcium phosphates. No CSH phase with a lower Ca/Si ratio was identified after transformation. The results confirmed a small effect of the monocalcium silicate addition on the compressive strength (CS) of cements up to 30 wt% (around 20-25 MPa) and a significant rise of the value in 50 woll/B cement (65 MPa). The final setting times of the cement composites varied between 5 and 43 min depending on the P/L ratio and the type of monocalcium silicate phase in the cement mixture. 10CSH/B and 50 woll/B cements with different textures but free of both the needle-like and perpendicularly-oriented hydroxyapatite particles on the surface of the samples had low cytotoxicity.