Microwave-assisted synthesis of crosslinked ureido chitosan for hemostatic applications.

In this study, we present a facile method for introducing hydrophilic ureido groups (NH2-CO-NH-) into chitosan using a microwave-assisted reaction with molten urea, with the aim of enhancing chitosan's interaction with blood components for improved hemostasis. The formation of the ureido groups through nucleophilic addition reaction between the amine groups in chitosan and in situ generated isocyanic acid was confirmed by FTIR, CP/TOSS 13C NMR, and CP/MAS 15N NMR spectroscopic techniques. However, in stark contrast to the glucans, the said modification introduced extensive crosslinking in chitosan. Spectroscopic studies identified these crosslinks as carbamate bridges (-NH-COO-), which were likely formed by the reaction between the ureido groups and hydroxyl groups of adjacent chains through an isocyanate intermediate. These carbamate bridges improved ureido chitosan's environmental stability, making it particularly resistant to changes in pH and temperature. In comparison to chitosan, the crosslinked ureido chitosan synthesized here exhibited good biocompatibility and cell adhesion, rapidly arrested the bleeding in a punctured artery with minimal hemolysis, and induced early activation and aggregation of platelets. These properties render it an invaluable material for applications in hemostasis, particularly in scenarios that necessitate stability against pH variations and degradation.

Solvent-less carboxymethylation-induced electrostatic crosslinking of chitosan.

The successful N-carboxymethylation and concomitant crosslinking of solid chitosan upon heating its mixture with solid monochloroacetic acid, without the use of solvents or catalysts, is reported. The N-carboxymethylation was confirmed through the analysis of the partially depolymerized product using NMR spectroscopy, as well as a control reaction with lysine. This transformation was facilitated by the nucleophilic nature of the free amine group in the repeating unit of chitosan, which possesses lone pair of electrons capable of attacking the carbon center bearing the leaving group and displacing the leaving group in a concerted manner. The crosslinking, on the other hand, was established by the observed insolubility in aqueous acidic solutions, even when subjected to prolonged heating at 60 °C. This crosslinking occurs due to the electrostatic interactions between the carboxylate groups and the adjacent ammonium groups, as supported by evidence from FTIR spectroscopy and a control reaction involving ethyl chloroacetate. The resulting crosslinked carboxymethyl chitosan demonstrated its usefulness in the adsorption of methyl orange and fluorescein, as well as functioning as an organic catalyst for aza-Michael addition, Hantzsch reaction, and substituted perimidine synthesis.

Valorization of agro-wastes for the biosynthesis and characterization of polyhydroxybutyrate by Bacillus sp. isolated from rice bran dumping yard.

Investigations have been made to determine the usage of inexpensive agro-waste products as an alternative carbon source for the production of degradable bacterial polyester. Among 33 bacterial isolates, a gram-positive bacterium PPECLRB-16 isolated from rice bran dumping yard was found to accumulate a relatively higher quantity of PHB and identified as Bacillus sp. through 16S rRNA gene sequence analysis. The higher PHB producing bacterial isolate was grown with different inexpensive agro-wastes to determine the suitable carbon source for its growth and PHB production. The one-factor-at-a-time approach comparatively enhanced PHB yield (5.64 g/L) when grown for 48 h with 1.5% (w/v) of defatted oil cake at a pH of 7.0. The bacterially accumulated PHB was isolated from the cells, purified, and characterized using solid-state 13C NMR, FT-IR, Powder XRD, TGA, GPC, Tensile and HR-SEM analyses. The hydrophobicity and printing accessibility of recovered PHB were demonstrated using contact angle measurement by coating on different surfaces. The results obtained in the present investigation have thrown light on the potential usage of agro-waste by-products, mainly oil cake, as an appropriate carbon source for the commercial production of PHB by Bacillus sp. in a cost-effective way.

Statistical augmentation of polyhydroxybutyrate production by Isoptericola variabilis: Characterization, moulding, in vitro cytocompatibility and biodegradability evaluation.

This study aimed to explore the production of polyhydroxybutyrate (PHB), a polyhydroxyalkanoate (PHA), which has been widely considered as a potential substitute for the synthetic polymers. Among 53 actinomycete isolates, 11 of them were found to be PHB positive and the quantity of PHB from the positive isolates varied from 10.5 to 29.82 wt% on a dry cell weight basis. A strain designated as PPLAT 012, accumulated relatively higher PHB and has been identified as Isoptericola variabilis by 16S rRNA gene sequence analysis. An effort has also been made to optimize the PHB production by the hyper-producing strain using the conventional, one-factor-at-a-time, and statistical response surface methodologies and the maximum PHB production (46.18%) in DSMZ medium, amended with 12% glucose and 9% potassium nitrate with a pH of 7.0. Further, the characteristic properties such as processability, cytocompatibility and biodegradability of the extracted PHB was also demonstrated. The physical properties of the recovered PHB was further improved by blending with PLA and the resultant blends were characterized. The present investigation has demonstrated that the isolate, Isoptericola variabilis, could be utilized as a potential source for the production of PHB with desirable characteristics, suitable for biomedical applications.

Facile preparation of biocompatible macroporous chitosan hydrogel by hydrothermal reaction of a mixture of chitosan-succinic acid-urea.

The facile preparation of macroporous, super water absorbing, biocompatible hydrogels of chitosan involving the hydrothermal reaction of a mixture of chitosan (CH), succinic acid (SA) and urea (UR), all of which are sustainable materials, is reported. The structure of the dry CHSAUR was ascertained by CP MAS-SS NMR spectroscopy, Fourier transform infrared (FTIR) spectroscopy, powder x-ray diffraction analysis (PXRD), and thermogravimetric analysis (TGA). The principle role of UR in the synthesis was identified as the source of ammonia, which increased the pH of the acidic chitosan solution with reaction time, leading to the formation of the insoluble hydrogel of chitosan accompanied by the formation of pores of different sizes and volumes. In addition, a small fraction of urea participated in chemical reaction with the primary hydroxyl groups in the sixth position of the glucosamine repeat units of chitosan resulting in carbamate linkages. The as-prepared hydrogel, following workup and methanol extraction, was found to be chitosan crosslinked with succinic acid through electrostatic interaction. It was macroporous with percentage porosity varying between 49.4% to 64.2%. It also exhibited different extents of water uptake with the maximum of 760 ±â€¯20 g/g being for the one prepared with the weight ratio of 1: 4: 4 of chitosan: succinic acid: urea. The absorption of water is found to arise out of the porosity as well as presence of water attracting chitosan ammonium cation-succinate electrovalent bonds that are formed by the reaction between SA and ammonium cation of the chitosan backbone. The absorption of saline water was relatively poor suggesting that the saline water absorption might be arising largely due to the presence of micropores and specific interaction. The hydrogels exhibited Herschel-Bulkley rheological behavior. The extraction of CHSAUR with 0.1 N NaOH in methanol resulted in the removal of the physical crosslinks, consisting of succinate anions; the presence of chitosan with porous morphology was confirmed additionally by copper (+2) adsorption. In contrast to the widely reported method of preparing microporous chitosan scaffold of cylindrical shape that takes several days to a week, the present method offers a simple means of preparing macroporous chitosan of any shape and size in very large scale with soft foam-like morphology. With its biocompatibility towards mouse fibroblast cells it could find applications in drug delivery, biodegradable super water absorbency and haemostatic applications.