Chitosan-collagen biopolymer biofilm derived from cephalopod gladius; Evaluation of osteogenesis, angiogenesis and wound healing for tissue engineering application.

Chitosan (Ch) and acid-soluble collagen (ASC) from Doryteuthis singhalensis gladius were isolated to test their osteogenic, angiogenic, and wound healing capabilities in male Wistar rats. The results of the study showed that the ASC yield was 18.58 %, the total protein content was 86.43 % ± 0.18 %, and the amino acid composition was as follows: glycine, 15.68 %; proline, 13.84 %. A, B, I, II, and III show FT-IR amide regional bands at 3392, 2959, 1652, 1471, and 1237 cm-1 respectively. The electrophoretic pattern of ASC validated its molecular weights of 105 kDa and 96 kDa. The 1H NMR spectra showed pure singles at 1.99 ppm, and the UV-Vis spectrum showed a particular absorbance between 238 and 220 nm. The DSC showed two endothermic peaks: one with an To value of 119.72 °C and TP of 126.28 °C, and the other with 147.42 °C and 148.47 °C. Initially, we fabricated Ch and ASC biofilms at an 8.5:1.5 ratio for tissue engineering applications. A cellular-level study demonstrated good biocompatibility and enhanced osteoblastic differentiation of collagen chitosan films (CChF). Additionally, the biofilm exhibited increased angiogenic potency, as observed in the chick embryo chorioallantoic membrane (CAM) assay. The experimental animal model demonstrated that in wound healing, the CChF treated rats (95.75 ± 2.28 %) had a greater decrease in the diameter of the wound than the control rats (22.25 ± 2.45 %), followed by the CF (collagen film) treated rats (63.25 ± 2.08 %) and ChF (chitosan film) (52.67 ± 1.58 %). Rats treated with CChF had 48.82 ± 1.25 mg/g of hydroxyproline in NFGT and 75.25 ± 1.56 mg/g of overall protein. The higher hydroxyproline levels in the CChF-treated groups corroborated these histopathological findings. These results imply that by promoting the development of scars, inflammation, and proliferation, CChF accelerates the healing process.

Gladius and its derivatives as potential biosorbents for marine diesel oil.

The demand for low cost and effective materials to remove contaminants such as residues of oil spills has encouraged studies on new biosorbents produced from wastes. Considering the overgeneration of fishing residues and the necessity to provide an alternative purpose for such materials, this study aimed to evaluate squid gladius and its derivatives (ß-chitin and chitosan) as sorbents to remove marine diesel oil (MDO) from fresh and artificial seawater. It was also executed an attempted to improve their performances through a high-intensity ultrasound treatment (UT-gladius and UT-ß-chitin). All sorbents removed MDO at both salinities. Contact surface area, salinity, and water retention seemed to play a key role in the outcomes. UT-ß-chitin's performance was significantly superior to ß-chitin's and chitosan's in MDO removal at salinity 0, as well as at salinity 30, where gladius and UT-gladius also excelled. Ultrasound treatment improved the oil removal performance of UT-ß-chitin by increasing its contact surface area. This is the first report on the efficiency of gladius and UT-ß-chitin for such purpose, and brought up huge possibilities and new questions that can lead to the achievement of biosorbents of great efficiency.

ß-Chitin and chitosan from squid gladius: Biological activities of chitosan and its application as clarifying agent for apple juice.

Chitin is the second most abundant polysaccharide in biomass after cellulose and the term chitosan usually refers to a family of polymers obtained after chitin deacetylation. The aim of this work was the preparation and the characterization of chitin and chitosan from the gladius (pen) of the European squid (Loligo vulgaris). A high level of deproteinization (more than 80%) was recorded using Alcalase® with an enzyme/protein ratio of 10U/mg. The demineralization of the gladius was completely achieved within 8h at room temperature in HCl. 13C NMR, FTIR, and XRD diffractograms of prepared chitin and chitosan were taken and then degree of deacetylation of chitosan was calculated using 13C CP/MAS-NMR Spectroscopic. Further, in vitro antioxidant capacity of chitosan was evaluated on 1,1-diphenyl-2-picrylhydrazyl method (IC50=3.2mgmL-1) and the ß-carotene bleaching assay (IC50=3.3mgmL-1). Antimicrobial activity was also investigated and assays indicated that prepared chitosan exhibited marked inhibitory activity against all microbial strains tested. Additionally, chitosan was tested such as clarifying agent for apple juice and showed powerful clarification capability, without affecting nutritional value. Furthermore, the results suggested that prepared chitosan could be used as alternative additive in pharmaceutical preparations and food industry.