Brazilian red propolis: extracts production, physicochemical characterization, and cytotoxicity profile for antitumor activity

Brazilian red propolis has been proposed as a new source of compounds with cytotoxic activity. Red propolis is a resinous material of vegetal origin, synthesized from the bees of the Appis mellifera family, with recognized biological properties. To obtain actives of low polarity and high cytotoxic profile from red propolis, in this work, we proposed a new solvent accelerated extraction method. A complete 23 factorial design was carried out to evaluate the influence of the independent variables or factors (e.g., temperature, number of cycles, and extraction time) on the dependent variable or response (i.e., yield of production). The extracts were analyzed by gas chromatography coupled with mass spectrometry for the identification of chemical compounds. Gas chromatography analysis revealed the presence of hydrocarbons, alcohols, ketones, ethers, and terpenes, such as lupeol, lupenone, and lupeol acetate, in most of the obtained extracts. To evaluate the cytotoxicity profile of the obtained bioactives, the 3-(4,5-dimethyl-2-thiazole)-2,5-diphenyl-2-H-tetrazolium bromide colorimetric assay was performed in different tumor cell lines (HCT116 and PC3). The results show that the extract obtained from 70 °C and one cycle of extraction of 10 min exhibited the highest cytotoxic activity against the tested cell lines. The highest yield, however, did not indicate the highest cytotoxic activity, but the optimal extraction conditions were indeed dependent on the temperature (i.e., 70 °C).FCT -Fundação para a Ciência e a Tecnologia(UIDB/04469/2020)info:eu-repo/semantics/publishedVersio

Otoliths-composed gelatin/sodium alginate scaffolds for bone regeneration

Evidence that otoliths, mineral-rich limestone concrescences present in the inner ear of bone fishes, can accelerate bone formation in vivo has been previously reported. The goal of this work was the development, characterization, and evaluation of the cytocompatibility of otoliths-incorporated sodium alginate and gelatin scaffolds. Cynoscion acoupaderived otoliths were characterized by X-ray fluorescence spectrometry (FRX), particle size, free lime, and weight loss by calcination. Furthermore, otoliths were incorporated into sodium alginate (ALG/OTL-s) or gelatin (GEL/OTL-s) scaffolds, previously developed by freeze-drying. Then, the scaffolds were characterized by thermogravimetric analysis (TGA/DTG), differential scanning calorimetry (DSC), infrared spectroscopy with Fourier transform (FTIR), swelling tests, and scanning electron microscopy (SEM). Cytotoxicity assays were run against J774.G8 macrophages and MC3T3-E1 osteoblasts. Data obtained from TGA/DTG, DSC, and FTIR analyses confirmed the interaction between otoliths and the polymeric scaffolds. SEM showed the homogeneous porous 3D structure rich in otolith micro-fragments in both scaffolds. Swelling of the GEL/OTL-s (63.54±3.0%) was greater than of ALG/OTL-s (13.36±9.9%) (p0.05) and significantly higher than that treated with Triton-X (p0.05). However, by 48 h, only ALG/OTL-s showed growth similar to control (p>0.05), whereas GEL/OTL showed a significantly lower growth index (p<0.05). In conclusion, the physicochemical profiles suggest proper interaction between the otoliths and the two developed polymeric 3D scaffolds. Moreover, both materials showed cytocompatibility with J774.G8 macrophages but the growth of MC3T3-E1 osteoblasts was higher when exposed to ALG/OTL-s. These data suggest that sodium alginate/otoliths scaffolds are potential biomaterials to be used in bone regeneration applications.We would like to thank the National Council for Scientific and Technological Development (CNPq) and the Foundation for Research and Technological Innovation Support of the State of Sergipe for the financial support in this study. EMBS acknowledges the sponsorship of the projects M-ERA-NET-0004/2015-PAIRED and UIDB/04469/2020 (strategic fund), received support from the Portuguese Science and Technology Foundation, Ministry of Science and Education (FCT/MEC) through national funds, and was co-financed by FEDER, under the Partnership Agreement PT2020.info:eu-repo/semantics/publishedVersio