The main source of commercial chitosan is the extensive deacetylation of its parent polymer chitin. It is present in green algae, the cell walls or fungi and in the exoskeleton of crustaceans. A novel procedure for preparing chitosan from shrimp shells was developed. The procedure involves two 10-minutes bleaching steps with ethanol after the usual demineralization and deproteinization processes. Before deacetylation, chitin was immersed in 12.5 M NaOH, cooled down and kept frozen for 24 h. The obtained chitosan was characterized using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), UV, X-ray diffraction (XRD) and viscosimetry. Samples of white chitosan with acetylation degrees below $9\%$ were obtained, as determined by FTIR and UV-first derivative spectroscopy. The change in the morphology of samples was followed by SEM. The ash content of chitosan samples were all below $0.063\%$ . Chitosan was soluble in $1\%$ acetic acid with insoluble contents of $0.62\%$ or less. XRD patterns exhibited the characteristic peaks of chitosan centered at 10 and 20 degrees in $2\theta$ . The molecular weight of chitosan was between 2.3 and 2.8 $\times {10}^5$ g/mol. It is concluded that the procedure developed in the present work allowed obtaining chitosans with physical and chemical properties suitable for pharmaceutical applications.

Film-forming emulsions and films, prepared by incorporating different concentrations of clove essential oil (CEO) and melaleuca essential oil (MEO) into chitosan (CS) were obtained and their properties were evaluated. Film-forming emulsions were characterized in terms of qualitative assessment, hydrogen potential and in vitro antibacterial activity, that was carried by the agar diffusion method, and the growth inhibition effects were tested on the Gram-positive microorganism of Staphylococcus aureus, Gram-negative microorganisms of Escherichia coli, and against isolated fungi such as Candida albicans. In order to study the impact of the incorporation of CEO and MEO into the CS matrix, the appearance and thickness of the films were evaluated. Furthermore, Fourier transform infrared spectroscopy (FTIR), contact angle measurements, a swelling test, scanning electron microscopy and a tensile test were carried out. Results showed that the film-forming emulsions had translucent aspect with cloudy milky appearance and showed antimicrobial properties. The CEO had the highest inhibition against the three strains studied. As regards the films’ properties, the coloration of the films was affected by the type and concentration of bioactive used. The chitosan/CEO films showed an intense yellowish coloration while the chitosan/MEO films presented a slightly yellowish coloration, but in general, all chitosan/EOs films presented good transparency in visible light besides flexibility, mechanical resistance when touched, smaller thicknesses than the dermis and higher wettability than chitosan films, in both distilled water and phosphate-buffered saline (PBS). The interactions between the chitosan and EOs were confirmed by. The chitosan/EOs films presented morphologies with rough appearance and with EOs droplets in varying shapes and sizes, well distributed along the surface of the films, and the tensile properties were compatible to be applied as wound dressings. These results revealed that the CEO and MEO have a good potential to be incorporated into chitosan to make films for wound-healing applications.

The aim of this paper was to prepare, by the freeze-drying method, ionically crosslinked chitosan membranes with different contents of pentasodium tripolyphosphate (TPP) and loaded with 1,4-naphthoquinone (NQ14) drug, in order to evaluate how the physical crosslinking affects NQ14 release from chitosan membranes for cancer therapy application. The membranes were characterized by Fourier transform infrared spectroscopy (FTIR), wide-angle X-ray diffraction (WAXD), scanning electron microscopy (SEM), swelling degree, and through in vitro drug release and cytotoxicity studies. According to the results, the molecular structure, porosity and hydrophilicity of the chitosan membranes were affected by TPP concentration and, consequently, the NQ14 drug release behavior from the membranes was also affected. The release of NQ14 from crosslinked chitosan membranes decreased when the cross-linker TPP quantity increased. Thus, depending on the TPP amount, the crosslinked chitosan membranes would be a potential delivery system to control the release of NQ14 for cancer therapy application. Lastly, the inhibitory potential of chitosan membranes ionically crosslinked with TPP and loaded with NQ14 against the B16F10 melanoma cell line was confirmed through in vitro cytotoxicity studies assessed via MTT assay. The anti-proliferative effect of prepared membranes was directly related to the amount of cross-linker and among all membranes prepared, such that one crosslinked with 0.3% of TPP may become a potential delivery system for releasing NQ14 drug for cancer therapy.

Nano-/micron-sized bioactive glass (BG) particles are attractive candidates for both soft and hard tissue engineering. They can chemically bond to the host tissues, enhance new tissue formation, activate cell proliferation, stimulate the genetic expression of proteins, and trigger unique anti-bacterial, anti-inflammatory, and anti-cancer functionalities. Recently, composites based on biopolymers and BG particles have been developed with various state-of-the-art techniques for tissue engineering. Gelatin, a semi-synthetic biopolymer, has attracted the attention of researchers because it is derived from the most abundant protein in the body, viz., collagen. It is a polymer that can be dissolved in water and processed to acquire different configurations, such as hydrogels, fibers, films, and scaffolds. Searching “bioactive glass gelatin” in the tile on Scopus renders 80 highly relevant articles published in the last ~10 years, which signifies the importance of such composites. First, this review addresses the basic concepts of soft and hard tissue engineering, including the healing mechanisms and limitations ahead. Then, current knowledge on gelatin/BG composites including composition, processing and properties is summarized and discussed both for soft and hard tissue applications. This review explores physical, chemical and mechanical features and ion-release effects of such composites concerning osteogenic and angiogenic responses in vivo and in vitro. Additionally, recent developments of BG/gelatin composites using 3D/4D printing for tissue engineering are presented. Finally, the perspectives and current challenges in developing desirable composites for the regeneration of different tissues are outlined.

Several studies have established an association between diabetes and alterations in bone metabolism; however, the underlying mechanism is not well established. Although zinc is recognized as a potential preventive agent against diabetes-induced bone loss, there is no evidence demonstrating its effect in chronic diabetic conditions. This study evaluated the effects of zinc supplementation in a chronic (90 days) type 1 diabetes-induced bone-loss model. Male Wistar rats were distributed in three groups: control, type 1 diabetes mellitus (T1DM), and T1DM plus zinc supplementation (T1DMS). Serum biochemical analysis; tibia histomorphometric, biomechanical, and collagen-content analyses; and femur mRNA expression were evaluated. Relative to T1DM, the zinc-supplemented group showed increased histomorphometric parameters such as TbWi and BAr and decreased TbSp, increased biomechanical parameters (maximum load, stiffness, ultimate strain, and Young’s modulus), and increased type I collagen content. Interestingly, similar values for these parameters were observed between the T1DMS and control groups. These results demonstrate the protective effect of zinc on the maintenance of bone strength and flexibility. In addition, downregulation of OPG, COL1A, and MMP-9 genes was observed in T1DMS, and the anabolic effects of zinc were evidenced by increased OC expression and serum ALP activity, both related to osteoblastogenesis, demonstrating a positive effect on bone formation. In contrast, T1DM showed excessive bone loss, observed through reduced histomorphometric and biomechanical parameters, characterizing diabetes-associated bone loss. The bone loss was also observed through upregulation of OPG, COL1A, and MMP-9 genes. In conclusion, zinc showed a positive effect on the maintenance of bone architecture and biomechanical parameters. Indeed, OC upregulation and control of expression of OPG, COL1A, and MMP-9 mRNAs, even in chronic hyperglycemia, support an anabolic and protective effect of zinc under chronic diabetic conditions. Furthermore, these results indicate that zinc supplementation could act as a complementary therapy in chronic T1DM.