Injectable hybrid hydrogels physically crosslinked based on carrageenan and green graphene for tissue repair.

Injectable and biocompatible novel hybrid hydrogels based on physically crosslinked natural biopolymers and green graphene for potential use in tissue engineering are reported. Kappa and iota carrageenan, locust bean gum and gelatin are used as biopolymeric matrix. The effect of green graphene content on the swelling behavior, mechanical properties and biocompatibility of the hybrid hydrogels is investigated. The hybrid hydrogels present a porous network with three-dimensionally interconnected microstructures, with lower pore size than that of the hydrogel without graphene. The addition of graphene into the biopolymeric network improves the stability and the mechanical properties of the hydrogels in phosphate buffer saline solution at 37 °C without noticeable change in the injectability. The mechanical properties of the hybrid hydrogels were enhanced by varying the dosage of graphene between 0.025 and 0.075 w/v%. In this range, the hybrid hydrogels preserve their integrity during mechanical test and recover the initial shape after removing the applied stress. Meanwhile, hybrid hydrogels with graphene content of up to 0.05 w/v% exhibit good biocompatibility for 3T3-L1 fibroblasts; the cells proliferate inside the gel structure and show higher spreading after 48 h. These injectable hybrid hydrogels with graphene have promising future as materials for tissue repair.

Starch-based composite foam for chicken meat packaging.

The development of compostable packages that maintain fresh meat quality, is an important achievement for the poultry industry. The objective of this study was to evaluate the feasibility of using a starch-based composite foam (SCF) in the packaging of fresh chicken meat during refrigerated storage. SCF was prepared using extrusion process. Nisin (2%) was added as antimicrobial agent (SCFN). Commercial expanded polystyrene (EPS) was used as control. Physical characterization, antimicrobial analysis and storage of fresh chicken meat were carried out. No differences were observed in SEM images between SFC and SCFN samples. Water uptake of SCF were higher than SCFN (p < 0.05). SCFN exhibited higher Young´s modulus and flexural strength (p < 0.05), and antimicrobial effect against foodborne pathogens. During the storage of chicken meat, the starch-based composite foam showed a higher capacity to retain liquid than EPS. The color of chicken meat had slight variations at day 4 compared with the raw meat. Nisin did not retard lipid oxidation of chicken meat, however, the aerobic plate count was lower. Therefore, the starch-based composite foam is suitable for fresh meat storage, being improved with the incorporation of nisin as antimicrobial agent. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05538-6.

Preparation of Poly(vinyl Alcohol) Microparticles for Freeze Protection of Sensitive Fruit Crops.

Poly(vinyl alcohol) (PVA) displays ice recrystallization inhibition (IRI) properties as many antifreeze proteins found in cold tolerant organisms. The molecular architecture and composition (molecular weight and distribution of pendant OH and acetate groups) have been studied to improve the antifreezing properties of PVA, suggesting that the molecular architecture of PVA plays an important role in IRI activity. The present work deals with the preparation of PVA microparticles using an alkaline treatment. The effect of PVA molecular weight on the morphology and antifreezeing properties of PVA microparticles was investigated. The antifreezeing property of PVA microparticles on the susceptibility of flower bud tissues to freeze damage was also evaluated. The alkaline treatment of an aqueous PVA solution produced stable polymer chain aggregates with spherical shapes. The average size of the PVA microparticles increased significantly with the increasing molecular weight of the PVA macromolecule precursor. The PVA microparticles inhibited the growth of ice crystals and blocked ice growth at concentrations as low as 0.01 % w/v. The effect of impeding ice crystal growth by preventing the joining of adjacent ice crystals is attributed to the larger size of the PVA particles adsorbed on the ice surface compared to the aggregated PVA macromolecules in saline solution. The thermal hysteresis activity of PVA macromolecules and microparticles was not detected by differential scanning calorimetry analysis. The PVA microparticles reduced the incidence of freeze injuries in flower bud tissues by 55% and their application, considering the low toxicity of PVA, has a high potential for freeze protection in fruit crops.

Bioactive Pectin-Murta (Ugni molinae T.) Seed Extract Films Reinforced with Chitin Fibers.

This study investigated the biocomposite pectin films enriched with murta (Ugni molinae T.) seed polyphenolic extract and reinforced by chitin nanofiber. The structural, morphological, mechanical, barrier, colorimetric, and antioxidant activity of films were evaluated. The obtained data clearly demonstrated that the addition of murta seed extract and the high load of chitin nanofibers (50%) provided more cohesive and dense morphology of films and improved the mechanical resistance and water vapor barrier in comparison to the control pectin film. The antioxidant activity ranged between 71% and 86%, depending on the film formulation and concentration of chitin nanofibers. The presented results highlight the potential use of chitin nanofibers and murta seed extract in the pectin matrix to be applied in functional food coatings and packaging, as a sustainable solution.

Improvement of endothelial function by Gunnera tinctoria extract with antioxidant properties.

BACKGROUND: Gunnera tinctoria has been collected by Mapuche-Pewenche people for food and medicinal purposes. The high polyphenol content of methanolic extract from G. tinctoria leaves with chemical constituents such as ellagic acid and quercetin derivatives suggests its application to prevent endothelial dysfunction and oxidative stress. The aim of this study was to provide evidence of the protective effect of this extract on endothelial function by reducing oxidative stress induced by high D-glucose and H2O2, as well as by stimulating nitric oxide (NO) levels in human umbilical vein endothelial cells (HUVECs). RESULTS: A methanolic extract with a high content of polyphenols (520 ± 30 mg gallic acid equivalents/g dry extract) was obtained from G. tinctoria leaves. Its main constituent was ellagic acid. The results of Ferric reducing antioxidant power and 2,2-diphenyl-1-picrylhydrazyl radical scavenging assays of the extract confirmed its antioxidant activity by inhibition pathway of radical species. The incubation of HUVECs with the extract decreased the apoptosis and reactive oxygen species (ROS) synthesis induced by high extracellular concentration of D-glucose or hydrogen peroxide. The extract increased endothelial NO levels and reduced vasoconstriction in human placental vessels. CONCLUSIONS: This study provides evidence about the antioxidant and endothelial protective properties of methanolic G. tinctoria leaf extract. The extract improves the availability of NO in HUVECs, inhibiting the production of ROS and vasoconstriction.

Carrageenan-based physically crosslinked injectable hydrogel for wound healing and tissue repairing applications.

In this study, a novel injectable hydrogel based on iota and kappa carrageenan, locust bean gum and gelatin was prepared for wound healing and tissue repairing applications. This injectable hydrogel was obtained via physical crosslinking. FTIR analysis confirmed the physical interaction between the biopolymeric components of the hydrogel. The prepared injectable hydrogel exhibited shear-thinning characteristics and could be injected for minimally invasive applications. Also, the hydrogel showed a porous structure, physiological and mechanical stability and biocompatibility. The in vitro cell culture studies showed that fibroblasts were able to grow, adhere and spread inside the hydrogel, indicating that hydrogel could support tissue repair. Moreover, hydrogel could be useful for the delivery of biomolecules. Vascular endothelial growth factor was encapsulated within the hydrogel and subsequently released, which accelerated the migration of human umbilical vein endothelial cells and facilitated in vitro wound healing. Overall, the results indicate that hydrogel can be a potential injectable delivery vehicle for wound healing and tissue repair.

A Novel Hydrocolloid Film Based on Pectin, Starch and Gunnera tinctoria and Ugni molinae Plant Extracts for Wound Dressing Applications.

BACKGROUND: The biodegradable and biocompatible nature of pectin-based films is of particular interest in wound dressing applications, due to its non-toxicity, pH-sensitivity and gelling activity. An approach to improve the mechanical properties, the release profile of bioactive compounds as well as the performance in wet environments of pectin-based films is mixing with other biopolymers. OBJECTIVE: To prepare hydrocolloid films based on crosslinked pectin / starch blend loaded with bioactive extracts from leaves of G. tinctoria and U. molinae with controlled release of bioactive compounds and healing property. METHODS: The hydrocolloid films were characterized by FTIR, SEM, and TGA-FTIR techniques and their tensile properties, water uptake, and polyphenolic release profile in aqueous media were evaluated. The dermal anti inflammatory activity of the hydrocolloid films was assessed by the mouse ear inflammation test. The wound healing property of the loaded hydrocolloid films was explored in a rat model and in a clinical trial (sacrum pressure ulcer). RESULTS: The films showed an adequate water-uptake capacity between 100-160%. The release of active compounds from the hydrocolloid films followed the Korsmeyer-Peppas equation. The mechanical properties of hydrocolloid films were not affected by the plant extracts within the concentration range used. The incorporation of the bioactive extracts in the polysaccharide films inhibited the topical edematous response by about 50%. The topical application of the loaded hydrocolloid film on the pressure ulcer is completely closed after 17 days without showing any adverse reaction. CONCLUSION: A novel hydrocolloid matrix was produced from crosslinked starch-pectin, which exhibited suitable chemical-physical properties to be used as a carrier of plant extracts with wound healing properties.

Poly(hydroxybutyrate-co-hydroxyvalerate) microparticles embedded in κ-carrageenan/locust bean gum hydrogel as a dual drug delivery carrier.

A novel composite hydrogel was prepared as a dual drug delivery carrier. Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) microparticles were prepared to encapsulate simultaneously ketoprofen and mupirocin, as hydrophobic drug models. These microparticles were embedded in a physically crosslinked hydrogel of κ-carrageenan/locust bean gum. This composite hydrogel showed for both drugs a slower release than the obtained release from microparticles and hydrogel separately. The release of both drugs was observed during a period of 7 days at 37 °C. Different kinetic models were analyzed and the results indicated the best fitting to a Higuchi model suggesting that the release was mostly controlled by diffusion. Also, the drug loaded microparticles were spherical with average mean particle size of 1.0 µm, mesoporous, and distributed homogeneously in the hydrogel. The composite hydrogel showed a thermosensitive swelling behavior reaching 183% of swelling ratio at 37 °C. The composite hydrogel showed the elastic component to be higher than the viscous component, indicating characteristics of a strong hydrogel. The biocompatibility was evaluated with in vitro cytotoxicity assays and the results indicated that this composite hydrogel could be considered as a potential biomaterial for dual drug delivery, mainly for wound healing applications.

Improvement of endothelial function by Gunnera tinctoria extract with antioxidant properties

BACKGROUND: Gunnera tinctoria has been collected by Mapuche-Pewenche people for food and medicinal purposes. The high polyphenol content of methanolic extract from G. tinctoria leaves with chemical constituents such as ellagic acid and quercetin derivatives suggests its application to prevent endothelial dysfunction and oxidative stress. The aim of this study was to provide evidence of the protective effect of this extract on endothelial function by reducing oxidative stress induced by high D-glucose and H2O2, as well as by stimulating nitric oxide (NO) levels in human umbilical vein endothelial cells (HUVECs). RESULTS: A methanolic extract with a high content of polyphenols (520 ± 30 mg gallic acid equivalents/g dry extract) was obtained from G. tinctoria leaves. Its main constituent was ellagic acid. The results of Ferric reducing antioxidant power and 2,2-diphenyl-1-picrylhydrazyl radical scavenging assays of the extract confirmed its antioxidant activity by inhibition pathway of radical species. The incubation of HUVECs with the extract decreased the apoptosis and reactive oxygen species (ROS) synthesis induced by high extracellular concentration of D-glucose or hydrogen peroxide. The extract increased endothelial NO levels and reduced vasoconstriction in human placental vessels. CONCLUSIONS: This study provides evidence about the antioxidant and endothelial protective properties of methanolic G. tinctoria leaf extract. The extract improves the availability of NO in HUVECs, inhibiting the production of ROS and vasoconstriction.

Entrapment of chitosan, pectin or κ-carrageenan within methacrylate based hydrogels: Effect on swelling and mechanical properties.

Composite hydrogels were obtained by the entrapment of chitosan, pectin or κ-carrageenan within methacrylate-based hydrogels to improve their swelling and the mechanical properties. The results indicated that the water uptake (WU) of κ-carrageenan and chitosan hydrogels were until 3.5 and 2.2 times higher than the WU of the synthetic hydrogel, respectively. The surface morphologies of the hydrogels showed that the pectin and κ-carrageenan favors the formation of larger and more defined pores. The mechanical properties indicated that the pectin increased slightly the mechanical properties and the κ-carrageenan improves the mechanical properties of the synthetic hydrogel reaching up 400 N of compression load. Therefore, the entrapment of κ-carrageenan within synthetic hydrogels improved both the swelling and the mechanical properties. The biocompatibility of the hydrogels was evaluated with in vitro cytotoxicity assays and the results indicated that they could be considered as candidates for biomedical use.

Starch films loaded with donut-shaped starch-quercetin microparticles: Characterization and release kinetics.

Starch films loaded with donut-shaped starch-quercetin microparticles were prepared from two different botanical origins. The quercetin release kinetics through the films were studied. The donut-shaped starch-quercetin microparticles were prepared by thermal aqueous-alcoholic treatment. The quercetin loading percentage and therefore the antioxidant activity were higher for the microparticles from legume than those of cereal origins. The starch-quercetin microparticles also showed higher thermal stability than the starch granules. The starch films were produced using the solution casting method. The films with more microparticles content showed higher thermal stability. In-vitro release studies of the quercetin through the films were performed in aqueous-ethanolic medium. The quercetin released reached the equilibrium in 1 to 4 days for the films of cereal starch and in more than a week for the films of legume origin. The release data were fitted to Peppas-Sahlin model that suggests the release kinetics were controlled mainly by fickian diffusion. The produced biofilms can be utilized mainly for active food packaging applications.

Preparation of starch nanoparticles loaded with quercetin using nanoprecipitation technique.

Nanoparticles of starches from different botanical origin were prepared by nanoprecipitation using 0.1M hydrochloric acid as non-solvent. The morphology and the particle size were analyzed using field emission scanning electron microscopy and dynamic light scattering. The nanoparticles were spherical and their sizes vary depending on the origin and the concentration of the starch solution. Starch nanoparticles loaded with quercetin were prepared. In-vitro release studies of the quercetin from the starch nanoparticles were performed in 35% ethanol as a release medium. The starch origin affects the quercetin loading percentage, the release kinetics and the antioxidant activity of the produced nanoparticles. The starch-quercetin nanoparticles from cereal origin showed the lowest loading percentage and the lowest fraction released of quercetin in comparison with nanoparticles from tuber and legume origin. The release kinetics seem to be controlled mainly by Fickian diffusion which have been revealed fitting the release data to the Peppas-Sahlin model.

Preparation of donut-shaped starch microparticles by aqueous-alcoholic treatment.

A simple method for producing donut-shaped starch microparticles by adding ethanol to a heated aqueous slurry of corn starch is presented. The obtained microparticles were analysed by SEM, XRD and DSC. The average size of microparticles was 14.1 ±â€¯0.3 µm with holes of an average size of 4.6 ±â€¯0.2 µm. The crystalline arrangement of the microparticles was of a V-type single helix. The change in crystallinity from A-type of the starch granules to a more open structure, where water molecules could penetrate easier within the microparticles, substantially increased their solubility and swelling power. The microparticles exhibited a higher gelatinization temperature and a lower gelatinization enthalpy than did the starch granules. The donut-shaped microparticles were stable for more than 18 months and can be used as a carrier of an active compound or as a filler in bioplastics.

Effect of nanocellulose as a filler on biodegradable thermoplastic starch films from tuber, cereal and legume.

Starches from different vegetal sources (tuber, cereal and legume) were plasticized with an invariant glycerol content and reinforced with cellulose nanocrystals by solution casting method. The influence of both, starch nature and filler amount, in the crystallinity and the extension of plasticization have been analyzed by X-ray diffraction. Thermoplastic starches (TPS) morphologies were obtained by scanning electron microscopy. Mechanical properties and thermal stability were analyzed by dynamomechanical and thermogravimetric analysis. Water absorption evolution was studied as well. A major extension in plasticization (high amylopectin starches) led to matrices with large starch-rich domains, a good thermal stability and resistance to water absorption but low stiffness. The incorporation of cellulose nanoparticles favoured plasticization and increased the rigidity in TPS films, as well as the thermal stability and moisture resistance. The aim of this work was to obtain bio-based thermoplastic starch films for replacing petroleum-derived ones in packaging industry, especially for short-life applications.

Ni/Ni oxides nanoparticles with potential biomedical applications obtained by displacement of a nickel-organometallic complex.

A new synthesis and stabilization method was developed for paramagnetic nanoparticles composed of nickel and nickel oxides. Nickel/nickel oxides nanoparticles were synthesized by a method based on ligand displacement of bis(1,5-cyclooctadiene)-nickel(0), zerovalent organometallic precursor and simultaneous formation of a thiourea inclusion compound. Nickel/nickel oxides nanoparticles were stabilized with the amphipathic peptide H2N-Cys-Leu-Pro-Phe-Phe-Asp-NH2 having H2N-Leu-Pro-Phe-Phe-Asp-NH2 a peptide with potential properties for Alzheimer's disease therapy. The inclusion compound formed after displacement was characterized by X-ray powder diffraction, and nickel/nickel oxides nanoparticles were characterized using transmission electron microscopy, atomic force microscopy, UV-Visible spectroscopy, X-ray photoelectron spectroscopy, and superconducting quantum interference device magnetometry. In addition, a cell viability assay in primary rat hippocampal neurons was carried out.