Effect of Xylitol on Inhibition and Eradication of Pseudomonas aeruginosa PAO1 and Methicillin-Resistant Staphylococcus aureus Biofilms in an Alginate Bead Model.

Biofilms formed by Pseudomonas aeruginosa and Staphylococcus aureus, along with their antibiotic tolerance have posed challenges to treatment strategies for lung, wound, and other infections, particularly when co-infecting. In the present study, the inhibitory effect of xylitol on biofilm formation, as well as its eradication potential on pre-established biofilms formed by P. aeruginosa strain PAO1, methicillin-resistant S. aureus, and a mix of both species in an alginate bead model were tested. Xylitol concentrations of 2, 1, and 0.5 M reduced biofilm formation by P. aeruginosa strain PAO1, methicillin-resistant S. aureus, and the mixed-species biofilm in a concentration-dependent manner. Additionally, biofilms formed by these species were subjected to treatment with xylitol. Xylitol was also capable of eradicating biofilms established by P. aeruginosa strain PAO1, methicillin-resistant S. aureus, and the mixed-species biofilm by at least 20%, with the most effective eradication observed for P. aeruginosa strain PAO1. The present study indicates the effectiveness of xylitol as both an inhibitory and eradicating agent against biofilms formed by P. aeruginosa strain PAO1, methicillin-resistant S. aureus, and a mix of both species in an alginate bead model, which mimics the in vivo characteristics of P. aeruginosa aggregates.

Biological evaluation of critical bone defect regeneration using hydroxyapatite/ alginate composite granules.

PURPOSE: to evaluate biocompatibility and osteogenic potential of hydroxyapatite/alginate composite after its implantation on rat calvarian critical bone defect. METHODS: thirty adults male Wistar rats were randomly distributed into two groups: GHA - critical bone defect filled with hydroxyapatite/alginate composite granules (HA/Alg) and CG - critical bone defect without biomaterial; evaluated at biological points of 15, 45 and 120 days. RESULTS: the histomorphometrically analyses for GHA showed osteoid matrix deposition (OM) among the granules and towards the center of the defect in centripetal direction throughout the study, with evident new bone formation at 120 days, resulting in filling 4/5 of the initial bone defect. For CG, this finding was restricted to the edges of the bone margins and formation of connective tissue on the residual area was found in all biological points. Inflammatory response on GHA was chronic granulomatous type, discrete and regressive for all biological points. Throughout the study, the CG presented mononuclear inflammatory infiltrate diffuse and regressive. Histomorphometry analyses showed that OM percentage was evident for GHA group when compared to CG group in all analyzed periods (p > 0.05). CONCLUSIONS: the biomaterial evaluated at this study showed to be biocompatible, bioactive, osteoconductive and biodegradable synchronously with bone formation.

Engineering a microparticle-loaded rough membrane for guided bone regeneration modulating osteoblast response without inducing inflammation.

Guided bone regeneration (GBR) is a widely used procedure that prevents the fast in-growth of soft tissues into bone defect. Among the different types of membranes, the use of collagen membranes is the gold standard. However, these membranes are implanted in tissue location where a severe acute inflammation will occur and can be negatively affected. The aim of this study was to develop a collagen-based membrane for GBR that incorporated alginate-hydroxyapatite microparticles. Membranes were manufactured using collagen type I and gelatin and alginate-hydroxyapatite microparticles. Membranes were assessed in terms of topography by scanning electron microscopy and confocal microscopy; stability by swelling after an overnight incubation in saline and enzymatic degradation against collagenase and mechanical properties by tensile tests. Furthermore, the biological response was assessed with SaOs-2 cells and THP-1 macrophages to determine alkaline phosphatase activity and inflammatory cytokine release. Our results showed that the incorporation of different percentages of these microparticles could induce changes in the surface topography. When the biological response was analyzed, either membranes were not cytotoxic to THP-1 macrophages or to SaOs-2 cells and they did not induce the release of pro-inflammatory cytokines. However, the different surface topographies did not induce changes in the macrophage morphology and the release of pro- and anti-inflammatory cytokines, suggesting that the effect of surface roughness on macrophage behavior could be dependent on other factors such as substrate stiffness and composition. Collagen-gelatin membranes with embedded alginate-hydroxyapatite microparticles increased ALP activity, suggesting a positive effect of them on bone regeneration, remaining unaffected the release of pro- and anti-inflammatory cytokines.

Rational design of PspAlgL to improve its thermostability and anti-biofilm activity against Pseudomonas aeruginosa.

Pseudomonas aeruginosa biofilm enhances tolerance to antimicrobials and immune system defenses. Alginate is an important component of biofilm and a virulence factor of P. aeruginosa. The degradation of alginate by alginate lyases has come to serve as an adjunctive therapeutic strategy against P. aeruginosa biofilm, but poor stability of the enzyme limited this application. Thus, PspAlgL, an alginate lyase, can degrade acetylated alginate but has poor thermostability. The 3D structure of PspAlgL was predicted, and the thermostability of PspAlgL was rationally designed by GRAPE strategy, resulting in two variants with better stability. These variants, PspAlgLS270F/E311P and PspAlgLG291S/E311P, effectively degraded the alginate in biofilm. In addition, compared with PspAlgL, these variants were more efficient in inhibiting biofilm formation and degrading the established biofilm of P. aeruginosa PAO1, and they were also able to destroy the biofilm attached to catheters and to increase the sensitivity of P. aeruginosa to the antibiotic amikacin. This study provides one potential anti-biofilm agent for P. aeruginosa infection.

Electrospray Nano-Micro Composite Sodium Alginate Microspheres with Shape-Adaptive, Antibacterial, and Angiogenic Abilities for Infected Wound Healing.

Nonhealing infectious wounds, characterized by bacterial colonization, wound microenvironment destruction, and shape complexity, present an intractable problem in clinical practice. Inspired by LEGOs, building-block toys that can be assembled into desired shapes, we proposed the use of electrospray nano-micro composite sodium alginate (SA) microspheres with antibacterial and angiogenic properties to fill irregularly shaped wounds instantly. Specifically, porous poly(lactic-co-glycolic acid) (PLGA) microspheres (MSs) encapsulating basic fibroblast growth factor (bFGF) were produced by a water-in-oil-in-water double-emulsion method. Then, bFGF@MSs were blended with the SA solution containing ZIF-8 nanoparticles. The resultant solution was electrosprayed to obtain nano-micro composite microspheres (bFGF@MS/ZIF-8@SAMSs). The composite MSs' size could be regulated by PLGA MS mass proportion and electrospray voltage. Moreover, bFGF, a potent angiogenic agent, and ZIF-8, bactericidal nanoparticles, were found to release from bFGF@MS/ZIF-8@SAMSs in a controlled and sustainable manner, which promoted cell proliferation, migration, and tube formation and killed bacteria. Through experimentation on rat models, bFGF@MS/ZIF-8@SAMSs were revealed to adapt to wound shapes and accelerate infected wound healing because of the synergistic effects of antibacterial and angiogenic abilities. In summation, this study developed a feasible approach to prepare bioactive nano-micro MSs as building blocks that can fill irregularly shaped infected wounds and improve healing.

Establishing a 3D-cultured system based on alginate-hydrogel embedding benefits the in vitro maturation of porcine Oocytes.

The in vitro maturation (IVM) quality of oocytes is directly related to the subsequent developmental potential of embryos and a fundamental of in vitro embryo production. However, conventional IVM methods fail to maintain the gap-junction intercellular communication (GJIC) between cumulus-oocyte complexes (COCs), which leads to insufficient oocyte maturation. Herein, we investigated the effects of three different three-dimensional (3D) culture methods on oocyte development in vitro, optimized of the alginate-hydrogel embedding method, and assessed the effects of the alginate-hydrogel embedding method on subsequent embryonic developmental potential of oocytes after IVM and parthenogenetic activation (PA). The results showed that Matrigel embedding and alginate-hydrogel embedding benefited the embryonic developmental potential of oocytes after IVM and PA. With the further optimization of alginate-hydrogel embedding, including crosslinking and decrosslinking of parameters, we established a 3D culture system that can significantly increase oocyte maturation and the blastocyst rate of embryos after PA (27.2 ± 1.5 vs 36.7 ± 2.8, P < 0.05). This 3D culture system produced oocytes with markedly increased mitochondrial intensity and membrane potential, which reduced the abnormalities of spindle formation and cortical granule distribution. The alginate-hydrogel embedding system can also remarkably enhance the GJIC between COCs. In summary, based on alginate-hydrogel embedding, we established a 3D culture system that can improve the IVM quality of porcine oocytes, possibly by enhancing GJIC.

In Vivo feature of the regenerative potential of chitosan and alginate based osteoplastic composites doped with calcium phosphates, zinc ions, and vitamin D2.

PURPOSE: In vivo comparison of the regenerative potential of two calcium phosphate-biopolymer osteoplastic composites: а) based on alginate (Alg) and hydroxyapatite (HA) - Alg/HA/CS/Zn/D2, b) based on chitosan (CS) and brushite (DCPD) - CS/DCPD/D2. MATERIALS AND METHODS: 36 white male laboratory rats aged six months were used. A defect to the bone marrow canal in the middle of the femur diaphysis was made with a dental bur of 2 mm. The bone defect healed under the blood clot (control) in the different animal groups and was filled with Alg/HA/CS/Zn/D2 and CS/DCPD/D2. The regeneration of the bone defect was studied on the 30th, 90th, and 140th days by computer tomography (CT). RESULTS: On the 30th day, all groups' implantation site optical density (OD) was significantly lower than that of the adjacent maternal bone (MB). Intensity of bone formation for Alg/HA/CS/Zn/D2 exceeds CS/DCPD/D2. On the 90th day, the bone trauma site OD with Alg/HA/CS/Zn/D2 (1725.4 ± 86 HU) and CS/DCPD/D2 (1484.9 ± 69 HU) exceeded the OD of the control (942.5 ± 55 HU). On the 140th day, the OD of Alg/HA/CS/Zn/D2 and CS/DCPD/D2 implantation sites was higher than Control and MB OD. Visually, the area of the past injury with the Alg/HA/CS/Zn/D2 could be detected only by the presence of an endosteal bone callus and in the case of CS/DCPD/D2 - by the shadow of the remaining biomaterial in the bone marrow canal. CONCLUSIONS: According to CT data, Alg/HA/CS/Zn/D2 and CS/DCPD/D2 contribute to the complete healing of the femoral diaphysis defect in 140 days, but the regenerative potential of Alg/HA/CS/Zn/D2 from 30 days to 140 days is higher than CS/DCPD/D2 biomaterial.

Continuous exposure to bisphenol S increases the accumulation of endogenous metabolic toxicants by obstructing the glucuronic acid pathway.

Uridine diphosphate glucuronic acid (UDPGA) is an essential substrate in the glucuronidation of exogenous and endogenous lipophilic compounds via the liver glucuronic acid pathway, and its synthesis depends on glucose and energy in the body. Bisphenol S (BPS), as a lipophilic environmental pollutant, has been widely utilized in the manufacturing of daily necessities. The biological effect of BPS in interference with liver energy metabolism might affect UDPGA synthesis and the excretion of lipophilic compounds, but this was not clearly revealed. Here, female zebrafish that were exposed to BPS for 35 days exhibited a significant decrease in UDPGA in the liver with significant accumulation of exogenous BPS and endogenous bilirubin in the body. One vital reason may be that the exposure to BPS for 35 days promoted the lipid formation through PPARg signaling and reduced energy levels in the liver, resulting in the decreased raw materials for UDPGA production in glucuronic acid pathway. Meanwhile, transcriptome analysis showed that BPS inhibited the mRNA expression levels of genes related to the glucuronic acid pathway. The accumulation of endogenous and exogenous lipophilic compounds can trigger a variety of toxicological effect. Thus, weakened liver detoxification might be the primary cause of the toxicological effects of lipophilic pollutants.

Physical, Chemical, and Biological Properties of Chitosan-Coated Alginate Microparticles Loaded with Porcine Interleukin-1ß: A Potential Protein Adjuvant Delivery System.

We previously developed chicken interleukin-1ß (IL-1ß) mutants as single-dose adjuvants that induce protective immunity when co-administered with an avian vaccine. However, livestock such as pigs may require a vaccine adjuvant delivery system that provides long-lasting protection to reduce the need for successive booster doses. Therefore, we developed chitosan-coated alginate microparticles as a carrier for bovine serum albumin (BSA) or porcine IL-1ß (pIL-1ß) and assessed their physical, chemical, and biological properties. Electrospraying of the BSA-loaded alginate microparticles (BSA/ALG MPs) resulted in an encapsulation efficiency of 50%, and those MPs were then coated with chitosan (BSA/ALG/CHI MPs). Optical and scanning electron microscopy, zeta potential analysis, and Fourier transform infrared spectroscopy were used to characterize these MPs. The BSA encapsulation parameters were applied to ALG/CHI MPs loaded with pIL-1ß, which were not cytotoxic to porcine fibroblasts but had enhanced bio-activity over unencapsulated pIL-1ß. The chitosan layer of the BSA/ALG/CHI MPs prevented burst release and facilitated sustained release of pIL-1ß for at least 28 days. In conclusion, BSA/ALG/CHI MPs prepared as a carrier for pIL-1ß may be used as an adjuvant for the formulation of pig vaccines.

Angucyclines containing ß-á´ -glucuronic acid from Streptomyces sp. KCB15JA151.

Two angucyclines, pseudonocardones D (1) and E (2), were isolated from Streptomyces sp. KCB15JA151. The planar structure was elucidated by comprehensive spectroscopic analysis. The absolute configuration of the sugar unit was determined based on the basis of coupling constants, ROESY, chemical derivatization and HPLC analysis. The biological activities of compounds 1 and 2 were examined by performing a computational target prediction, which led to tests of the antiestrogenic activity. The result suggested that compound 1 might be an ERα antagonist.

A Glucuronic Acid-Palmitoylethanolamide Conjugate (GLUPEA) Is an Innovative Drug Delivery System and a Potential Bioregulator.

Palmitoylethanolamide (PEA) is an endogenous anti-inflammatory lipid mediator and a widely used nutraceutical. In this study, we designed, realized, and tested a drug-carrier conjugate between PEA (the active drug) and glucuronic acid (the carrier). The conjugate, named GLUPEA, was characterized for its capability of increasing PEA levels and exerting anti-inflammatory activity both in vitro and in vivo. GLUPEA treatment, compared to the same concentration of PEA, resulted in higher cellular amounts of PEA and the endocannabinoid 2-arachidonoyl glycerol (2-AG), and increased 2-AG-induced transient receptor potential vanilloid type 1 (TRPV1) channel desensitization to capsaicin. GLUPEA inhibited pro-inflammatory monocyte chemoattractant protein 2 (MCP-2) release from stimulated keratinocytes, and it was almost as efficacious as ultra-micronized PEA at reducing colitis in dinitrobenzene sulfonic acid (DNBS)-injected mice when using the same dose. GLUPEA is a novel pro-drug able to efficiently mimic the anti-inflammatory and endocannabinoid enhancing actions of PEA.

Analysis of Safety and Effectiveness of Sodium Alginate/Poly(γ-glutamic acid) Microspheres for Rapid Hemostasis.

Most preventable deaths after trauma are related to hemorrhage and occur early after injury. Timely hemostatic treatment is essential to minimize blood loss and improve survival. Among the various treatment methods, the most economical and effective is to use a hemostatic agent. A powdered hemostatic agent can be used for wounds of any shape or depth with high compactness and excellent accumulation effect. Herein, we chose the natural, hydrophilic polymer poly(γ-glutamic acid) (γ-PGA) to form composite hemostatic microspheres with sodium alginate (SA), which show good biocompatibility, water absorptivity, and viscosity. The morphology and structure of the hemostatic microspheres were determined using Fourier transform infrared spectroscopy and scanning electron microscopy. The overall safety, hemolysis, pyrogenic, and intradermal irritation tests were examined. The relationship between hemostatic pressure and hemostatic time during microsphere use was also measured. The hemostatic effect was analyzed with a liver, spleen, and femoral artery bleeding model. The composite microspheres were well tolerated in vivo and exhibited better hemostatic effects in animal experiments than a microporous polysaccharide powder compound. Research results showed that SA/γ-PGA microspheres are materials with good hemostatic effect, high safety, and great potential in clinical applications.

Structural characterization of a novel polysaccharide from Pleurotus citrinopileatus and its antitumor activity on H22 tumor-bearing mice.

In this research, a novel polysaccharide (PCP) was extracted from Pleurotus citrinopileatus and purified by Sephadex G-150 gel column, and its antitumor activity was investigated using the model H22 tumor-bearing mice. PCP was found to be composed of arabinose, galactose, glucose, xylose, mannose and glucuronic acid in a proportion of 0.66: 14.59: 10.77: 1: 0.69: 0.23 with average molecular weight of 7.30 × 105 Da. Further analysis suggested that PCP was a pyranose with α-type and ß-type glycosidic residues. The antitumor assays in vivo indicated that PCP could effectively suppress H22 solid tumor growth, protect immune organs and improve inflammation and anemia. Besides, Annexin V-FITC/PI double staining and JC-1 staining demonstrated that PCP could induce apoptosis of H22 hepatoma cells. The PI staining assay revealed that PCP induced H22 hepatoma cells apoptosis by arresting cell cycle in S phase. These results suggest that the polysaccharide from Pleurotus citrinopileatus possesses potential value in the treatment of liver cancer.

Effects of glucuronic acid and N-acetyl-D-glucosamine supplementation on the perivitelline space during the IVM of pig oocytes.

The objective of this study was to minimise polyspermic penetration by increasing the perivitelline space (PVS) thickness through supplementation of the hyaluronic acid components glucuronic acid and N-acetyl-d-glucosamine (GlcNAc). Oocytes (n=4690) were supplemented during the first 24h and/or the remainder of maturation (final 16-18h) with 0.01mM glucuronic acid and 0.01mM GlcNAc and then evaluated for PVS thickness, hyaluronic acid, glutathione and glutathione peroxidase concentrations. Fertilised oocytes were evaluated for polyspermic penetration and embryo development. The PVS thickness and amount of hyaluronic acid was significantly (P<0.05) greater in oocytes supplemented with 0.01mM glucuronic acid and 0.01mM GlcNAc during the second part or all of maturation compared with the other treatments. In addition, polyspermic penetration was significantly (P<0.05) less in oocytes supplemented with 0.01mM glucuronic acid and 0.01mM GlcNAc during the second part or all of maturation compared with the other treatments. Supplementing 0.01mM glucuronic acid and GlcNAc during maturation significantly (P<0.05) increased the percentage of cleaved embryos by 48h after IVF and blastocysts formed by 144h after IVF compared those not supplemented. These results indicate that supplementing PVS components during maturation decreases polyspermic penetration by increasing PVS thickness.

Recent Evidence of the Beneficial Effects Associated with Glucuronic Acid Contained in Kombucha Beverages.

PURPOSE OF THE REVIEW: Glucuronic acid is contained naturally in kombucha beverages due to the associations between bacteria and yeasts during its fermentation. The purpose of this review is to describe the literature related to the hepatoprotective effect associated with glucuronic acid present in different kombucha beverages. RECENT FINDINGS: Although previous research supports beneficial hepatoprotective effects of glucuronic acid consumption from kombucha, these effects are mainly attributed to the tea phytochemicals. However, there are some improvements in methodological deficiencies in some in vivo studies that should be considered. There is no sufficient evidence to generalize the adverse effects of kombucha consumption. Consumption of kombucha could be considered a safe practice in healthy populations due to its hepatoprotective effects. The content of the beneficial or toxic components is very variable because it depends on its manufacturing process. In persons with side sickness, other conditions such as pregnancy, and hypersensitivity to some kombucha components, a restriction in its consumption must be advisable.

Use of Alginate Extracted from Moroccan Brown Algae to Stimulate Natural Defense in Date Palm Roots.

Our study aimed to search for seaweed polysaccharides able to stimulate date palm defense mechanisms. Extraction, purification, characterization, and elicitor activity of sodium alginate (FSSA and BBSA) from Moroccan brown seaweeds Fucus spiralis and Bifurcaria bifurcata were investigated. FSSA and BBSA were characterized by proton nuclear magnetic resonance spectroscopy (1H-NMR) and size exclusion chromatography (HPLC-SEC). The mannuronic acid/guluronic acid (M/G) ratio of FSSA was M/G= 0.92 indicating that FSSA contained 48% and 52% of mannuronic and guluronic acids respectively, and the M/G ratio of BBSA was 0.47 indicating that BBSA contained 32% and 68% of mannuronic and guluronic acids respectively. Elicitor activity of FSSA and BBSA was carried out by developing an innovative study model on the date palm. The elicitor capacities were evaluated by investigating phenolic metabolism including phenylalanine ammonia-lyase (PAL) activity and total polyphenol content in seedling roots of date palm maintained in alginates solution (FSSA and BBSA) at different concentrations. The results obtained show that the PAL activity and the phenolic compound content were significantly stimulated with 1 mg.mL-1 of FSSA and BBSA; after 1 day of treatment with FSSA, and after 12 hours of treatment with BBSA. These results show clearly those alginates extracted from Moroccan brown algae induced in date palm roots the stimulation of natural defense mechanisms.

Evaluation of the Glucuronic Acid Production and Antibacterial Properties of Kombucha Black Tea.

BACKGROUND: Kombucha beverage is considered as a dietary supplement and drinking it strengthens the body's immune system which prevents diseases. OBJECTIVE: The purpose of this study was to determine the amount of glucuronic acid and antibacterial activity of Kombucha black tea drink during its production at different storage temperature. METHODS: The extent of glucuronic acid at temperatures of 20°C and 30°C was explored by the use of the HPLC system for 21 days. To analyse the antibacterial property, the influence of Kombucha black tea supernatant on the growth of Salmonella typhimurium, Staphylococcus aureus, and Lactobacillus rhamnosus bacteria was examined via the two procedures of the disc and agar well diffusion. RESULTS: The production of glucuronic acid underwent a variation at 20°C from 17.0 mg/L on day 1 to roughly 27.2 mg/L on day 21, and the difference was significant. Furthermore, the quantity of this acid at 30°C increased from 42.2 mg/L on day 1 to 48.0 mg/L on day 21. The amount of glucuronic acid produced at 30°C was significantly greater than that at 20°C (p<0.05). This study indicated that the Kombucha black tea has antibacterial activity against Salmonella typhimurium and Staphylococcus aureus, but not against Lactobacillus rhamnosus. However, there are no statistical differences in antibacterial activity of Kombucha between incubation at 20oC and 30oC (P>0.05). CONCLUSION: This study offers a perspective on glucuronic acid production (especially in 30°C rather than 20°C) and antibacterial activity of Kombucha black tea beverage.

Material microenvironmental properties couple to induce distinct transcriptional programs in mammalian stem cells.

Variations in a multitude of material microenvironmental properties have been observed across tissues in vivo, and these have profound effects on cell phenotype. Phenomenological experiments have suggested that certain of these features of the physical microenvironment, such as stiffness, could sensitize cells to other features; meanwhile, mechanistic studies have detailed a number of biophysical mechanisms for this sensing. However, the broad molecular consequences of these potentially complex and nonlinear interactions bridging from biophysical sensing to phenotype have not been systematically characterized, limiting the overall understanding and rational deployment of these biophysical cues. Here, we explore these interactions by employing a 3D cell culture system that allows for the independent control of culture substrate stiffness, stress relaxation, and adhesion ligand density to systematically explore the transcriptional programs affected by distinct combinations of biophysical parameters using RNA-seq. In mouse mesenchymal stem cells and human cortical neuron progenitors, we find dramatic coupling among these substrate properties, and that the relative contribution of each property to changes in gene expression varies with cell type. Motivated by the bioinformatic analysis, the stiffness of hydrogels encapsulating mouse mesenchymal stem cells was found to regulate the secretion of a wide range of cytokines, and to accordingly influence hematopoietic stem cell differentiation in a Transwell coculture model. These results give insights into how biophysical features are integrated by cells across distinct tissues and offer strategies to synthetic biologists and bioengineers for designing responses to a cell's biophysical environment.

Influence of different divalent ions cross-linking sodium alginate-polyacrylamide hydrogels on antibacterial properties and wound healing.

Hydrogels are widely used as debriding agents on account of providing a moist environment for wound healing, however the lack of mechanical strength, angiogenesis and antibacterial property limits their applications. In this study, we synthesized novel divalent ion cross-liking hydrogels (copper, zinc, strontium and calcium) and compared the mechanical performance, swelling ratio, antibacterial properties and biocompatibility in vitro and vivo. Thereinto, among the four divalent ions cross-linked hydrogels, copper ion crosslinking exhibited the maximum breaking strength, while strontium and zinc ion cross-linked hydrogels exhibited an excellent mechanical strength. In addition, the swelling ratio and pore size of no-ion cross-linked hydrogels was larger than ion cross-kinked hydrogels. In vitro, the improvements on wound healing after hydrogel application were evaluated by histological and molecular assays by detecting VEGF and TGF-ß expression. In vitro and in vivo study results showed that zinc cross-kinked hydrogel had a spectrum of antibacterial activities, cell viability, mechanical strength and the ability of wound closure by promoting fibroblasts migration, vascularization, collagen deposition and the formation of granulation tissue.

Injectable polysaccharide hydrogel embedded with hydroxyapatite and calcium carbonate for drug delivery and bone tissue engineering.

To meet the progressive requirements for bone regeneration purpose, injectable hydrogels have attracted increasing attention in tissue regeneration and local drug delivery applications. In this study, we report a facile method to prepare injectable and degradable polysaccharide-based hydrogels doubly integrated with hydroxyapatite (HAp) nanoparticles and calcium carbonate microspheres (CMs) under physiological condition. The mechanism of cross-linking is attributed to the Schiff-base reaction between amino and aldehyde groups of carboxymethyl chitosan (CMCS) and oxidized alginate (OAlg), respectively. Synchronously, tetracycline hydrochloride (TH) loaded CMs were fabricated by the precipitation reaction with an average diameter of 6.62 µm. To enhance bioactive and mechanical properties, nano-HAp and CMs containing TH were encapsulated into the polysaccharide-based hydrogel to form injectable gel scaffolds for imitation of bone niche. The gelation time, morphology, mechanical properties, swelling ratio and in vitro degradation of the gel scaffolds could be controlled by varying HAp and CMs contents. Moreover, the composite gel scaffolds had good sustained drug release and antibacterial properties, as confirmed by drugs release calculation and antibacterial evaluation. In addition, the gel scaffolds were found to be self-healing due to dynamic equilibrium of the Schiff-base linkages. These results suggested that the prepared composite gel scaffolds hold great potential for drug delivery and regeneration of irregular bone defects.