Effect on adhesion of a nanocapsules-loaded adhesive system

<div><p>Abstract This study aimed to evaluate the in situ degree of conversion, contact angle, and immediate and long-term bond strengths of a commercial primer and an experimental adhesive containing indomethacin- and triclosan-loaded nanocapsules (NCs). The indomethacin- and triclosan-loaded NCs, which promote anti-inflammatory and antibacterial effects through controlled release, were incorporated into the primer at a concentration of 2% and in the adhesive at concentrations of 1, 2, 5, and 10%. The in situ degree of conversion (DC, n=3) was evaluated by micro-Raman spectroscopy. The contact angle of the primer and adhesive on the dentin surface (n = 3) was determined by an optical tensiometer. For the microtensile bond strength µTBS test (12 teeth per group), stick-shaped specimens were tested under tensile stress immediately after preparation and after storage in water for 1 year. The data were analyzed using two-way ANOVA, three-way ANOVA and Tukey’s post hoc tests with α=0.05. The use of the NC-loaded adhesive resulted in a higher in situ degree of conversion. The DC values varied from 75.07 ± 8.83% to 96.18 ± 0.87%. The use of NCs in only the adhesive up to a concentration of 5% had no influence on the bond strength. The contact angle of the primer remained the same with and without NCs. The use of both the primer and adhesive with NCs (for all concentrations) resulted in a higher contact angle of the adhesive. The longitudinal μTBS was inversely proportional to the concentration of NCs in the adhesive system, exhibiting decreasing values for the groups with primer containing NCs and adhesives with increasing concentrations of NCs. Adhesives containing up to 5% of nanocapsules and primer with no NCs maintained the in situ degree of conversion, contact angle, and immediate and long-term bond strengths. Therefore, the NC-loaded adhesive can be an alternative method for combining the bond performance and therapeutic effects. The use of an adhesive with up to 5% nanocapsules containing indomethacin and triclosan and a primer with no nanocapsules maintained the long-term bond performance.</p></div

Effects of Two Types of Melatonin-Loaded Nanocapsules with Distinct Supramolecular Structures: Polymeric (NC) and Lipid-Core Nanocapsules (LNC) on Bovine Embryo Culture Model

<div><p>Melatonin has been used as a supplement in culture medium to improve the efficiency of <i>in vitro</i> produced mammalian embryos. Through its ability to scavenge toxic oxygen derivatives and regulate cellular mRNA levels for antioxidant enzymes, this molecule has been shown to play a protective role against damage by free radicals, to which <i>in vitro</i> cultured embryos are exposed during early development. <i>In vivo</i> and <i>in vitro</i> studies have been performed showing that the use of nanocapsules as active substances carriers increases stability, bioavailability and biodistribution of drugs, such as melatonin, to the cells and tissues, improving their antioxidant properties. These properties can be modulated through the manipulation of formula composition, especially in relation to the supramolecular structures of the nanocapsule core and the surface area that greatly influences drug release mechanisms in biological environments. This study aimed to evaluate the effects of two types of melatonin-loaded nanocapsules with distinct supramolecular structures, polymeric (NC) and lipid-core (LNC) nanocapsules, on <i>in vitro</i> cultured bovine embryos. Embryonic development, apoptosis, reactive oxygen species (ROS) production, and mRNA levels of genes involved in cell apoptosis, ROS and cell pluripotency were evaluated after supplementation of culture medium with non-encapsulated melatonin (Mel), melatonin-loaded polymeric nanocapsules (Mel-NC) and melatonin-loaded lipid-core nanocapsules (Mel-LNC) at 10⁻⁶, 10⁻⁹, and 10⁻¹² M drug concentrations. The highest hatching rate was observed in embryos treated with 10⁻⁹ M Mel-LNC. When compared to Mel and Mel-NC treatments at the same concentration (10⁻⁹ M), Mel-LNC increased embryo cell number, decreased cell apoptosis and ROS levels, down-regulated mRNA levels of <i>BAX</i>, <i>CASP3</i>, and <i>SHC1</i> genes, and up-regulated mRNA levels of <i>CAT</i> and <i>SOD2</i> genes. These findings indicate that nanoencapsulation with LNC increases the protective effects of melatonin against oxidative stress and cell apoptosis during <i>in vitro</i> embryo culture in bovine species.</p></div

Effects of free and nanoencapsulated melatonin (10⁻⁹ M) on the relative mRNA abundance of pluripotency-related genes in bovine blastocysts.

<p>Data represent mean ± S.E.M.</p

ROS levels in 4–8 cell stage embryos cultured in the presence of free and nanoencapsulated melatonin.

<p>Embryos were cultured in the presence of 10⁻⁹ M melatonin non-encapsulated (Mel) or encapsulated in nanocapsules (Mel-NC) or lipid-core nanocapsules (Mel-LNC). In the control group embryos were cultured in SOFaa BSA medium alone. Data represent mean ± S.E.M. Different letters above the error bars indicate significant differences between groups (P< 0.05). Representative fluorescent (B, C, D, E) and corresponding bright field (b, c, d, e) images of control, Mel, Mel-NC, and Mel-LNC embryos, respectively. Scale bar = 50 μm. Magnification = 100X.</p

Effects of free and nanoencapsulated melatonin (10⁻⁹ M) on the relative mRNA abundance of oxidative stress-related genes.

<p>Data represent mean ± S.E.M. Different letters (a, b and c) indicate significant differences between groups (P< 0.05).</p

Hatching rates of bovine embryos cultured in the presence of free or nanoencapsulated melatonin.

<p>Mel = Non-encapsulated melatonin, Mel-NC = melatonin-loaded polymeric nanocapsules, Mel-LNC = melatonin-loaded lipid-core nanocapsules, NC = drug-unloaded nanocapsules, LNC = drug-unloaded lipid-core nanocapsules. Melatonin was used at 10⁻¹² M, 10⁻⁹ M or 10⁻⁶ M concentrations. Control group was cultured in SOFaa BSA alone. The numerators represent the numbers of hatched blastocysts and the denominators represent total blastocysts in each group. Different letters (a-c) above the bars indicate significant differences between groups (<i>P</i> < 0.05).</p