Alginate Hydrogels Coated with Chitosan for Wound Dressing

In this work, a coating of chitosan onto alginate hydrogels was realized using the water-soluble hydrochloride form of chitosan (CH-Cl), with the dual purpose of imparting antibacterial activity and delaying the release of hydrophilic molecules from the alginate matrix. Alginate hydrogels with different calcium contents were prepared by the internal setting method and coated by immersion in a CH-Cl solution. Structural analysis by cryo-scanning electron microscopy was carried out to highlight morphological alterations due to the coating layer. Tests in vitro with human mesenchymal stromal cells (MSC) were assessed to check the absence of toxicity of CH-Cl. Swelling, stability in physiological solution and release characteristics using rhodamine B as the hydrophilic model drug were compared to those of relative uncoated hydrogels. Finally, antibacterial activity against Escherichia coli was tested. Results show that alginate hydrogels coated with chitosan hydrochloride described here can be proposed as a novel medicated dressing by associating intrinsic antimicrobial activity with improved sustained release characteristics.     

Marine Structure Derived Calcium Phosphate–Polymer Biocomposites for Local Antibiotic Delivery

Hydrothermally converted coralline hydroxyapatite (HAp) particles loaded with medically active substances were used to develop polylactic acid (PLA) thin film composites for slow drug delivery systems. The effects of HAp particles within PLA matrix on the gentamicin (GM) release and release kinetics were studied. The gentamicin release kinetics seemed to follow Power law Korsmeyer Peppas model with mainly diffusional process with a number of different drug transport mechanisms. Statistical analysis shows very significant difference on the release of gentamicin between GM containing PLA (PLAGM) and GM containing HAp microspheres within PLA matrix (PLAHApGM) devices, which PLAHApGM displays lower release rates. The use of HAp particles improved drug stabilization and higher drug encapsulation efficiency of the carrier. HAp is also the source of Ca²⁺ for the regeneration and repair of diseased bone tissue. The release profiles, exhibited a steady state release rate with significant antimicrobial activity against Staphylococcus aureus (S. aureus) (SH1000) even at high concentration of bacteria. The devices also indicated significant ability to control the growth of bacterial even after four weeks of drug release. Clinical release profiles can be easily tuned from drug-HAp physicochemical interactions and degradation kinetics of polymer matrix. The developed systems could be applied to prevent microbial adhesion to medical implant surfaces and to treat infections mainly caused by S. aureus in surgery.     

Development and characterization of bamboo gauze fabric coated with polymer and drug for wound healing

The bamboo yarn of Ne 40^s was used for the preparation of the Gauze fabric. The physical properties such as areal density and stiffness of fabrics were measured. The fabric was then scoured and bleached as per the standard procedure using distilled water. Chitosan-sodium alginate, Calcium-sodium alginate polymer and their mixture were coated separately on the gauze structure to improve the antibacterial and wound healing property of the bandage. Scanning electron microscope (SEM) analysis was carried out to observe the uniform distribution of polymers in the samples. The antibiotic drugs were selected based on the antibiotic sensitivity test. The drugs such as Tetracycline hydrochloride (250 mg), Chloramphenicol (250 mg) and Rifampicin (250 mg) were immobilized on the polymer coated fabrics to increase the rate of wound healing and antibacterial activity. The drug loaded samples were subjected to drug release study for about four days in a static condition. The results show that good amount of drug was released during all the four days. Further, the antibacterial activity of the drug loaded and polymer coated samples were evaluated against S. aureus and Proteus bacteria. The results show excellent antibacterial activity.     

The effect of bentonite concentration on the drug delivery efficacy of a pH-sensitive alginate/bentonite hydrogel

We produced a protein loaded, pH-sensitive alginate-bentonite hydrogel for wound dressings. Alginate is a nontoxic polysaccharide with favorable pH-sensitive properties that make it useful for the intestinal delivery of protein drugs. However, the use of alginate for drug delivery is limited by drug leaching and rapid dissolution of alginate at the higher pH, which may result in lower entrapment efficiency and a burst in the release of entrapped protein drugs. To overcome these problems, we created a novel cross-linked alginate-bentonite hydrogel by combining mineral-rich bentonite with the alginate matrix along with an additive to ensure controlled release. We analyzed the gel in the drug loading process in an aqueous environment by looking at the release profiles of a model protein drug (BSA) from the hydrogel at pH values of 4.5, 5.2 (skin area) and 7.4, 9.2 (wound area). The swelling ratio decreased with bentonite concentration, but did not fall below 6. The rate of drug release was slowest at a pH value of 4.5 and fastest at a pH value of 9.2. The rate of drug release decreased with bentonite concentration. The presence of bentonite prevents the rapid dissolution of alginate at the higher pH, ensuring the controlled release of the entrapped drug.     

Formulation,Optimization and In Vivo Evaluation of Fucoidan-Based Cream with Anti-Inflammatory Properties

Fucoidan is a polysaccharide found in brown alga with glorious potential for pharmacological activities, among which its anti-inflammatory properties have gained meaningful attention. Due to several advantages of formulations for topical application, this study aimed to develop and optimize a fucoidan-based cream formulation and to investigate its anti-inflammatory potential after topical application in vivo. Fucoidan from Fucus vesiculosus L. was used. The cream base consisting of olive oil and Kolliphor RH40 was optimized followed by in vitro agar diffusion and drug release studies. The fucoidan-based cream with 13% Kolliphor P 407, 1% Transcutol P, and 5% PEG400 showed good spreadability, washability, and colloidal stability, and it did not irritate the skin. The kinetics of fucoidan release from the optimized cream exhibited the best fit to the Korsmeyer–Peppas and Higuchi models with R² > 0.99. Fucoidan release was controlled by drug diffusion and anomalous transport provided by the optimized cream base. The formulation was stable and provided high fucoidan release after storage for 1 year. Topical application of the fucoidan-based cream dose-dependently inhibited carrageenan-induced edema and ameliorated mechanical allodynia in rats. The efficacy of the fucoidan-based cream at a high dose was comparable with the efficacy of diclofenac gel. The fucoidan-based cream could be considered a promising anti-inflammatory formulation.     

Fabrication and Characterization of Nanocomposite Hydrogel Based on Alginate/Nano-Hydroxyapatite Loaded with Linum usitatissimum Extract as a Bone Tissue Engineering Scaffold

In the current paper, we fabricated, characterized, and applied nanocomposite hydrogel based on alginate (Alg) and nano-hydroxyapatite (nHA) loaded with phenolic purified extracts from the aerial part of Linum usitatissimum (LOH) as the bone tissue engineering scaffold. nHA was synthesized based on the wet chemical technique/precipitation reaction and incorporated into Alg hydrogel as the filler via physical cross-linking. The characterizations (SEM, DLS, and Zeta potential) revealed that the synthesized nHA possess a plate-like shape with nanometric dimensions. The fabricated nanocomposite has a porous architecture with interconnected pores. The average pore size was in the range of 100–200 µm and the porosity range of 80–90%. The LOH release measurement showed that about 90% of the loaded drug was released within 12 h followed by a sustained release over 48 h. The in vitro assessments showed that the nanocomposite possesses significant antioxidant activity promoting bone regeneration. The hemolysis induction measurement showed that the nanocomposites were hemocompatible with negligible hemolysis induction. The cell viability/proliferation confirmed the biocompatibility of the nanocomposites, which induced proliferative effects in a dose-dependent manner. This study revealed the fabricated nanocomposites are bioactive and osteoactive applicable for bone tissue engineering applications.     

Interpenetrating polymer network matrices of sodium alginate and carrageenan for controlled drug delivery application

Interpenetrating polymer network (IPN) matrices of sodium alginate and carrageenan were prepared for controlled release application. The propranolol-resin complex (resinate) loaded matrices were prepared by wet granulation/covalent crosslinking method and subsequently compressed into tablets. The SEM, DSC and XRD studies confirmed the amorphous nature of drug in the IPN matrix and FTIR confirmed the IPN formation and stability of drug within IPN matrix. The pure drug propranolol HCl showed rapid and complete dissolution within 60 min, while drug release from resinate was extended for 2.5 h and that from IPN tablets was still slower and drug release prolonged over 18 h. The crosslinking time of granules affected the release of drug from IPN matrix.     

Chitosan-Genipin Microspheres for the Controlled Release of Drugs: Clarithromycin,Tramadol and Heparin

The aim of this study was to first evaluate whether the chitosan hydrochloride-genipin crosslinking reaction is influenced by factors such as time, and polymer/genipin concentration, and second, to develop crosslinked drug loaded microspheres to improve the control over drug release. Once the crosslinking process was characterized as a function of the factors mentioned above, drug loaded hydrochloride chitosan microspheres with different degrees of crosslinking were obtained. Microspheres were characterized in terms of size, morphology, drug content, surface charge and capacity to control in vitro drug release. Clarithromycin, tramadol hydrochloride, and low molecular weight heparin (LMWH) were used as model drugs. The obtained particles were spherical, positively charged, with a diameter of 1–10 μm. X-Ray diffraction showed that there was an interaction of genipin and each drug with chitosan in the microspheres. In relation to the release profiles, a higher degree of crosslinking led to more control of drug release in the case of clarithromycin and tramadol. For these drugs, optimal release profiles were obtained for microspheres crosslinked with 1 mM genipin at 50 ºC for 5 h and with 5 mM genipin at 50 ºC for 5 h, respectively. In LMWH microspheres, the best release profile corresponded to 0.5 mM genipin, 50 ºC, 5 h. In conclusion, genipin showed to be eligible as a chemical-crosslinking agent delaying the outflow of drugs from the microspheres. However, more studies in vitro and in vivo must be carried out to determine adequate crosslinking conditions for different drugs.     

Fabrication of electrospun antimicrobial nanofibers containing metronidazole using nanospider technology

Nanospider technology as a modified electrospinning technique was used for the fabrication of electrospun nanofibers based on poly(vinyl alcohol) (PVA)/poly(ethylene oxide) (PEO) blend as drug delivery system (DDS) for metronidazole (MTZ) as an antimicrobial drug. Electrospun PVA/PEO/MTZ composite nanofibers were stabilized against disintegration in water by heating in oven at 110°C, or by soaking in isopropyl alcohol for 6 hrs. Incorporation of MTZ into electrospun nanofibers was confirmed by SEM, FT-IR spectra and TGA. The drug release results showed that the burst release was suppressed with stabilized electrospun nanofibers compared with non-stabilized ones. Electrospun PVA/PEO/MTZ composite nanofibers exhibited remarkable antimicrobial activity against Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger, Penicillium notatum and Aspergillus flavus which varies with the species of the tested organisms.     

Influences of Molecular Weights on Physicochemical and Biological Properties of Collagen-Alginate Scaffolds

For tissue engineering applications, biodegradable scaffolds containing high molecular weights (MW) of collagen and sodium alginate have been developed and characterized. However, the properties of low MW collagen-based scaffolds have not been studied in previous research. This work examined the distinctive properties of low MW collagen-based scaffolds with alginate unmodified and modified by subcritical water. Besides, we developed a facile method to cross-link water-soluble scaffolds using glutaraldehyde in an aqueous ethanol solution. The prepared cross-linked scaffolds showed good structural properties with high porosity (~93%) and high cross-linking degree (50–60%). Compared with collagen (6000 Da)-based scaffolds, collagen (25,000 Da)-based scaffolds exhibited higher stability against collagenase degradation and lower weight loss in phosphate buffer pH 7.4. Collagen (25,000 Da)-based scaffolds with modified alginate tended to improve antioxidant capacity compared with scaffolds containing unmodified alginate. Interestingly, in vitro coagulant activity assay demonstrated that collagen (25,000 Da)-based scaffolds with modified alginate (C25-A63 and C25-A21) significantly reduced the clotting time of human plasma compared with scaffolds consisting of unmodified alginate. Although some further investigations need to be done, collagen (25,000 Da)-based scaffolds with modified alginate should be considered as a potential candidate for tissue engineering applications.     

Biochemical Characterization and Cold-Adaption Mechanism of a PL-17 Family Alginate Lyase Aly23 from Marine Bacterium Pseudoalteromonas sp. ASY5 and Its Application for Oligosaccharides Production

As an important enzyme involved in the marine carbon cycle, alginate lyase has received extensive attention because of its excellent degradation ability on brown algae, which is widely utilized for alginate oligosaccharide preparation or bioethanol production. In comparison with endo-type alginate lyases (PL-5, PL-7, and PL-18 families), limited studies have focused on PL-17 family alginate lyases, especially for those with special characteristics. In this study, a novel PL-17 family alginate lyase, Aly23, was identified and cloned from the marine bacterium Pseudoalteromonas carrageenovora ASY5. Aly23 exhibited maximum activity at 35 °C and retained 48.93% of its highest activity at 4 °C, representing an excellent cold-adaptation property. Comparative molecular dynamics analysis was implemented to explore the structural basis for the cold-adaptation property of Aly23. Aly23 had a high substrate preference for poly β-D-mannuronate and exhibited both endolytic and exolytic activities; its hydrolysis reaction mainly produced monosaccharides, disaccharides, and trisaccharides. Furthermore, the enzymatic hydrolyzed oligosaccharides displayed good antioxidant activities to reduce ferric and scavenge radicals, such as hydroxyl, ABTS⁺, and DPPH. Our work demonstrated that Aly23 is a promising cold-adapted biocatalyst for the preparation of natural antioxidants from brown algae.     

Chitosomes-In-Chitosan Hydrogel for Acute Skin Injuries: Prevention and Infection Control

Burns and other skin injuries are growing concerns as well as challenges in an era of antimicrobial resistance. Novel treatment options to improve the prevention and eradication of infectious skin biofilm-producing pathogens, while enhancing wound healing, are urgently needed for the timely treatment of infection-prone injuries. Treatment of acute skin injuries requires tailoring of formulation to assure both proper skin retention and the appropriate release of incorporated antimicrobials. The challenge remains to formulate antimicrobials with low water solubility, which often requires carriers as the primary vehicle, followed by a secondary skin-friendly vehicle. We focused on widely used chlorhexidine formulated in the chitosan-infused nanocarriers, chitosomes, incorporated into chitosan hydrogel for improved treatment of skin injuries. To prove our hypothesis, lipid nanocarriers and chitosan-comprising nanocarriers (≈250 nm) with membrane-active antimicrobial chlorhexidine were optimized and incorporated into chitosan hydrogel. The biological and antibacterial effects of both vesicles and a vesicles-in-hydrogel system were evaluated. The chitosomes-in-chitosan hydrogel formulation demonstrated promising physical properties and were proven safe. Additionally, the chitosan-based systems, both chitosomes and chitosan hydrogel, showed an improved antimicrobial effect against S. aureus and S. epidermidis compared to the formulations without chitosan. The novel formulation could serve as a foundation for infection prevention and bacterial eradication in acute wounds.     

Enzyme pretreatment and negative pressure cavitation extraction of genistein and apigenin from the roots of pigeon pea [Cajanus cajan (L.) Millsp.] and the evaluation of antioxidant activity

An enzyme pretreatment and negative-pressure cavitation extraction (E-NPCE) method was investigated for extraction genistein and apigenin from pigeon pea roots. The important parameters involved in E-NPCE process were optimized by single-factor experiments and then critical parameters were investigated by a 2³ full factorial central composite design (CCD) to optimize extract conditions. Under optimal conditions, the yields of genistein and apigenin were 0.628 mg/g and 0.359 mg/g, which represented an increase of 44.70% and 53.05%, respectively, compared to standard NPCE. Furthermore, from DPPH scavenging activity test the extract of E-NPCE showed better antioxidant activity than these of other methods. The results demonstrated that E-NPCE would have lower energy consumption, higher efficiency and could be an alternative method for natural compounds extraction.     

Controlled release of diclofenac from matrix polymer of chitosan and oxidized konjac glucomannan

The controlled release of diclofenac sodium (DFNa) from a chitosan-oxidized konjac glucomannan (CTS-OKG) polymer film was studied. Konjac glucomannan (KGM) was initially oxidized by sodium periodate and then cross-linked to CTS via imine bonds (-C=N-) to form the new CTS-OKG copolymer. The DFNa loaded CTS-OKG polymers were characterized by Fourier transformed infrared spectroscopy (FT-IR) and X-ray diffractometry (XRD). Finally, the release profiles of DFNa from the CTS-OKG polymer matrices were evaluated in a simulated gastrointestinal fluid system comprised of two hours in simulated gastric fluid (SGF; pH 1.2) followed by 24 h in simulated intestinal fluid (SIF; pH 7.4). A 1:2:1 (w/w/w) ratio of CTS:OKG:DFNa prepared at room temperature for 3 hours gave the highest % encapsulation efficiency (EE) of 95.6 ± 0.6 and resulted in a minimal release of DFNa (<1% over 2 h) in SGF (pH 1.2) and a significantly improved sustained release in SIF (pH 7.4) with ~6% and 19% release over 8 and 24 h, respectively), some 15- and five-fold lower than that of the two commercial DFNa preparations, Diclosian and Voltaren. This formulation may be used for further study as a long term intestine controlled release drug model (at least 3 days).     

Cryopreservation of potato shoot tips by encapsulationdehydration

Summary Potato shoot tips excised from 2-week-old in vitro nodal cuttings were cryopreserved after encapsulation in alginate beads. Encapsulated shoot tips were first precultured in sucroseenriched media, dried over silica gel, and rapidly cooled in liquid nitrogen. After slow rewarming in air at room temperature, alginate beads were transferred to solid culture medium for shoot tip recovery. After cooling in liquid nitrogen, shoot yield depended on preculture duration, sucrose concentration and water content of beads. Survival rates above 60% were obtained for each cultivar tested.     

Removal of reactive orange 16 by immobilized Coprinus plicatilis in the batch shaking bioreactor

Coprinus plicatilis, white rot fungi, capable of removing Reactive Orange 16 (Remazol Brillant Orange 3R) was immobilized in calcium alginate gel beads and used for decolorization of RO16 in aqueous in the Batch Shaking Bioreactor. The optimum conditions for immobilization of the microorganism, such as alginate and calcium ion concentration, initial cell amout, hardening time and bead size, were determined with a view to improving the RO16 removal rate. The characteristics of RO16 decolorization by immobilized fungal cells were investigated. The repeated use of immobilized cells for RO16 decolorization was performed and the results revealed that the bioactivity of immobilized cells was stable over 120 hours in the repeated batch cultivation RO16 removal. At the end of decolorization process, the metabolites of the dye decolorization produced by immobilized C. plicatilis, were analyzed via GC-MS. It was concluded that the removal of the dye by immobilized C. plicatilis was achieved.     

Mucoadhesive Microparticles for Gastroretentive Delivery: Preparation,Biodistribution and Targeting Evaluation

The aim of this research was to prepare and characterize alginate-chitosan mucoadhesive microparticles containing puerarin. The microparticles were prepared by an emulsification-internal gelatin method using a combination of chitosan and Ca²⁺ as cationic components and alginate as anions. Surface morphology, particle size, drug loading, encapsulation efficiency and swelling ratio, in vitro drug released, in vitro evaluation of mucoadhesiveness and Fluorescence imaging of the gastrointestinal tract were determined. After optimization of the formulation, the encapsulation efficiency was dramatically increased from 70.3% to 99.2%, and a highly swelling ratio was achieved with a change in particle size from 50.3 ± 11.2 μm to 124.7 ± 25.6 μm. In ethanol induced gastric ulcers, administration of puerarin mucoadhesive microparticles at doses of 150 mg/kg, 300 mg/kg, 450 mg/kg and 600 mg/kg body weight prior to ethanol ingestion significantly protected the stomach ulceration. Consequently, significant changes were observed in inflammatory cytokines, such as prostaglandin E₂ (PGE₂), tumor necrosis factor (TNF-α), interleukin 6 (IL-6), and interleukin1β (IL-1β), in stomach tissues compared with the ethanol control group. In conclusion, core-shell type pH-sensitive mucoadhesive microparticles loaded with puerarin could enhance puerarin bioavailability and have the potential to alleviate ethanol-mediated gastric ulcers.     

Cloning and Characterization of a Novel Alginate Lyase from Paenibacillus sp. LJ-23

As a low molecular weight alginate, alginate oligosaccharides (AOS) exhibit improved water solubility, better bioavailability, and comprehensive health benefits. In addition, their biocompatibility, biodegradability, non-toxicity, non-immunogenicity, and gelling capability make them an excellent biomaterial with a dual curative effect when applied in a drug delivery system. In this paper, a novel alginate lyase, Algpt, was cloned and characterized from a marine bacterium, Paenibacillus sp. LJ-23. The purified enzyme was composed of 387 amino acid residues, and had a molecular weight of 42.8 kDa. The optimal pH of Algpt was 7.0 and the optimal temperature was 45 °C. The analysis of the conserved domain and the prediction of the three-dimensional structure indicated that Algpt was a novel alginate lyase. The dominant degradation products of Algpt on alginate were AOS dimer to octamer, depending on the incubation time, which demonstrated that Algpt degraded alginate in an endolytic manner. In addition, Algpt was a salt-independent and thermo-tolerant alginate lyase. Its high stability and wide adaptability endow Algpt with great application potential for the efficient preparation of AOS with different sizes and AOS-based products.     

Chemical composition and antioxidant and antimicrobial activity of essential oil of Artemisia annua L. from Bosnia

Hydrodistilled volatile oil obtained from the aerial parts of Artemisia annua L., cultivated near Sarajevo, Bosnia, was analyzed by GC-MS. More than one hundred compounds were identi?ed, representing 95.5% of the total oil. The major constituents of essential oil were oxygenated monoterpenes, artemisia ketone (30.7%) and camphor (15.8%). Isolated essential oil was tested for radical-scavenging ability using the stable DPPH radical, the ABTS radical, for reducing power ability with a test based on the reduction of ferric cations, for reducing ability of hydroxy radical in ORAC assay, and for metal chelating ability using the ferrozine assay. In all tests oil did not show a prominent antioxidant activity, but still comparable with thymol, an already known antioxidant. The screening of antimicrobial activity of oil was individually evaluated against representatives of Gram-positive, Gram-negative bacteria and fungi, using the agar diffusion method. All tested microorganisms were inhibited by essential oil. To the best of our knowledge, this is the first report of antimicrobial activity of essential oil of A. annua against Haemophilus influenzae, Enterococcus faecalis, Streptococcus pneumoniae, Micrococcus luteus and Candida krusei microbial strains. The antioxidant, antibacterial and antifungal activity of essential oil of A. annua from Bosnia is presented here for the first time and extends our knowledge in the range of valuable biological activities and possible roles in therapy associated with this medicinal herb.