Antimicrobial effect of two fluoride-releasing adhesive tapes on Streptococcus mutans biofilm.

Fluoride-releasing adhesive tapes have been developed as a new fluoride delivery agent. However, application as caries prevention agents remains underexplored. This study aimed at evaluating the antimicrobial activity of two fluoride-releasing adhesive tapes against S. mutans biofilm. Two polyvinyl alcohol (PVA) tapes were investigated: (i) a fluoride-PVA (F-PVA) tape, (ii) a pullulan incorporated F-PVA (PF-PVA) tape. S. mutan strains were cultured and treated with the tapes. Antimicrobial effects were evaluated using the agar diffusion test, field-emission scanning electron microscopy (FE-SEM), and confocal laser scanning microscopy (CLSM). F-PVA tapes showed higher inhibition-zone diameters than PF-PVA at 48 h and 72 h. However, there were no significant differences (p > 0.05) between the effects of F-PVA and PF-PVA. The bio-volume of S. mutans and extracellular polymeric substances significantly decreased in the F-PVA tapes than in the PF-PVA tapes (p < 0.05). FE-SEM micrographs revealed less S. mutans colonization in F-PVA. F-PVA exhibited better antimicrobial activity against S. mutans than PF-PVA.

From Tooth Adhesion to Bioadhesion: Development of Bioabsorbable Putty-like Artificial Bone with Adhesive to Bone Based on the New Material "Phosphorylated Pullulan".

Bioabsorbable materials have a wide range of applications, such as scaffolds for regenerative medicine and cell transplantation therapy and carriers for drug delivery systems. Therefore, although many researchers are conducting their research and development, few of them have been used in clinical practice. In addition, existing bioabsorbable materials cannot bind to the body's tissues. If bioabsorbable materials with an adhesive ability to biological tissues can be made, they can ensure the mixture remains fixed to the affected area when mixed with artificial bone or other materials. In addition, if the filling material in the bone defect is soft and uncured, resorption is rapid, which is advantageous for bone regeneration. In this paper, the development and process of a new bioabsorbable material "Phosphorylated pullulan" and its capability as a bone replacement material were demonstrated. Phosphorylated pullulan, which was developed based on the tooth adhesion theory, is the only bioabsorbable material able to adhere to bone and teeth. The phosphorylated pullulan and ß-TCP mixture is a non-hardening putty. It is useful as a new resorbable bone replacement material with an adhesive ability for bone defects around implants.

Potential of Pullulan-Based Polymeric Nanoparticles for Improving Drug Physicochemical Properties and Effectiveness.

Pullulan, a natural polysaccharide with unique biocompatibility and biodegradability, has gained prominence in nanomedicine. Its application in nanoparticle drug delivery systems showcases its potential for precision medicine. AIM OF STUDY: This scientific review aims to comprehensively discuss and summarize recent advancements in pullulan-based polymeric nanoparticles, focusing on their formulation, characterization, evaluation, and efficacy. METHODOLOGY: A search on Scopus, PubMed, and Google Scholar, using "Pullulan and Nanoparticle" as keywords, identified relevant articles in recent years. RESULTS: The literature search highlighted a diverse range of studies on the pullulan-based polymeric nanoparticles, including the success of high-selectivity hybrid pullulan-based nanoparticles for efficient boron delivery in colon cancer as the active targeting nanoparticle, the specific and high-efficiency release profile of the development of hyalgan-coated pullulan-based nanoparticles, and the design of multifunctional microneedle patches that incorporated pullulan-collagen-based nanoparticle-loaded antimicrobials to accelerate wound healing. These studies collectively underscore the versatility and transformative potential of pullulan-based polymeric nanoparticles in addressing biomedical challenges. CONCLUSION: Pullulan-based polymeric nanoparticles are promising candidates for innovative drug delivery systems, with the potential to overcome the limitations associated with traditional delivery methods.

Two novel Pleosporales species isolated from the bark of Acer saccharum.

During a survey of culturable microfungi from the bark of sugar maple (Acer saccharum), Atrocalyx glutinosus and Nigrograna rubescens, two novel species of Pleosporales (Dothideomycetes) were isolated from several locations in eastern Ontario, Canada. Formal species descriptions are presented based on unique colony phenotypes and micromorphological characteristics and supported using multi-locus molecular phylogenetic comparisons with similar species. Both A. glutinosus and N. rubescens produce pycnidial asexual morphs in culture. As their names imply, under specific culture conditions, A. glutinosus excretes large amounts of the glutinous polysaccharide pullulan and N. rubescens produces a dark red naphthoquinone pigment that diffuses in the culture medium. Citation: Mack JN, Sproule A, Shields SW, Seifert KA, Smith M, Overy DP (2024). Two novel Pleosporales species isolated from the bark of Acer saccharum . Fungal Systematics and Evolution 13: 1-14. doi: 10.3114/fuse.2024.13.01.

ARTP mutagenesis of Aureobasidium pullulans RM1603 for high pullulan production and transcriptome analysis of mutants.

Pullulan is a microbial exopolysaccharide produced by Aureobasidium spp. with excellent physical and chemical properties, resulting in great application value. In this study, a novel strain RM1603 of Aureobasidium pullulans with high pullulan production of 51.0 ± 1.0 g·L- 1 isolated from rhizosphere soil was subjected to atmospheric and room temperature plasma (ARTP) mutagenesis, followed by selection of mutants to obtain pullulan high-producing strains. Finally, two mutants Mu0816 and Mu1519 were obtained, with polysaccharide productions of 58.7 ± 0.8 and 60.0 ± 0.8 g∙L- 1 after 72-h fermentation, representing 15.1 and 17.6% increases compared with the original strain, respectively. Transcriptome analysis of the two mutants and the original strain revealed that the high expression of α/ß-hydrolase (ABHD), α-amylase (AMY1), and sugar porter family MFS transporters (SPF-MFS) in the mutants may be related to the synthesis and secretion of pullulan. These results demonstrated the effectiveness of ARTP mutagenesis in A. pullulans, providing a basis for the investigation of genes related to pullulan synthesis and secretion.

Improved thermal stabilization of VSV-vector with enhanced vacuum drying in pullulan and trehalose-based films.

One major limitation of effective vaccine delivery is its dependency on a robust cold chain infrastructure. While Vesicular stomatitis virus (VSV) has been demonstrated to be an effective viral vaccine vector for diseases including Ebola, its -70 °C storage requirement is a significant limitation for accessing disadvantaged locations and populations. Previous work has shown thermal stabilization of viral vaccines with a combination of pullulan and trehalose (PT) dried films. To improve the thermal stability of VSV, we optimized PT formulation concentrations and components, as well as drying methodology with enhanced vacuum drying. When formulated in PT films, VSV can be stored for 32 weeks at 4 °C with less than 2 log PFU loss, at 25 °C with 2.5 log PFU loss, and at 37 °C with 3.1 log PFU loss. These results demonstrate a significant advancement in VSV thermal stabilization, decreasing the cold chain requirements for VSV vectored vaccines.

Sinapic acid-pullulan based inflammation responsive nanomicelles for the local treatment of experimental inflammatory arthritis.

Rheumatoid arthritis (RA) is a chronic inflammatory disorder of joints. RA is one of the major causes of disability and morbidity worldwide. Administration of conventional drugs through the systemic route restricts the bioavailability of drugs, systemic toxicity, and reduced efficacy. We have introduced Rebamipide (Reb)-loaded Sinapic acid (SA)-Pullulan (PL) nanomicelles (Reb@SA-PL NMs), a nanotechnology based drug delivery system for the treatment of inflammatory arthritis. PL is a polysaccharide obtained from the fungus Aureobasidium pullulans, and SA is a bioactive polyphenol found in various plants. Both are classified by US-FDA Generally Recognised as Safe (GRAS) materials. Reb@SA-PL NMs found to be cytocompatible. Subsequently, intra-articular administration of Reb@SA-PL NMs enhances the anti-arthritic potential compared to free Reb drug in collagen-induced experimental inflammatory arthritis rat model. Reb@SA-PL NMs reduced the expression of RANKL receptor and Nf-κB. Reb@SA-PL NMs reverses the breakdown of type II collagen, MMP-13, and inhibits the pro-inflammatory markers. Reb@SA-PL NMs prevented bone erosion, cartilage degradation, joint oedema, and synovial inflammation. The results of the study demonstrated that Reb@SA-PL NMs, an enzyme-responsive drug delivery system, has excellent potential for alleviating inflammatory arthritis by blocking MMP-13 and RANKL.

Draft genome sequence and annotation of the polyextremotolerant polyol lipid-producing fungus aureobasidium pullulans NRRL 62042.

OBJECTIVES: The ascomycotic yeast-like fungus Aureobasidium exhibits the natural ability to synthesize several secondary metabolites, like polymalic acid, pullulan, or polyol lipids, with potential biotechnological applications. Combined with its polyextremotolerance, these properties make Aureobasidium a promising production host candidate. Hence, plenty of genomes of Aureobasidia have been sequenced recently. Here, we provide the annotated draft genome sequence of the polyol lipid-producing strain A. pullulans NRRL 62042. DATA DESCRIPTION: The genome of A. pullulans NRRL 62042 was sequenced using Illumina NovaSeq 6000. Genome assembly revealed a genome size of 24.2 Mb divided into 39 scaffolds with a GC content of 50.1%. Genome annotation using Genemark v4.68 and GenDBE yielded 9,596 genes.

Fabrication of Abelmoschus manihot gum/pullulan/magnesium L-ascorbate green composite packaging film and its excellent performance in preserving fresh-cut carrots.

Pullulan-based composite film can be a potential alternative packing material to non-environmentally friendly plastic wrap (PE) to preserve fresh-cut carrots. However, many developed pullulan-based composites either have high water vapor permeability (WVP) and high mechanical strength or vice versa, which limits the practicality of the developed packaging materials for potential commercialization. Herein, Abelmoschus manihot gum (AMG)/pullulan/magnesium L-ascorbate (MLA) was created as a green composite film (APL) to preserve fresh-cut carrots. The optimal amount of MLA was found to be 10 % (APL10), demonstrating a balance of lower WVP and greater mechanical strength and antioxidant performance than many pullulan-based films. This effectively solved many problems faced by other pullulan-based packaging films. After the fresh-cut carrots were packed with the composite film for 4 days, it was found that APL10 was effective in preserving the quality of carrots, in terms of freshness, weight loss rate, Vitamin C (VC), and malondialdehyde (MDA) content after 4 days of storage, much better than non-biodegradable PE. Thus, based on these findings, it is concluded that APL films have huge potential as a green packaging material for food to replace PE in the future.

ß-Cyclodextrin mediated controlled release of phenothiazine from pH-responsive pectin and pullulan-based hydrogel optimized through experimental design.

Drugs with lower permeability and water solubility provide major challenges for producing safe and efficient formulations. The current work aims to prepare ICs of the drug phenothiazine and ß-cyclodextrin via physical, microwave, freeze-drying, and kneading methods. Many analytical methods, such as 1H NMR, ROESY, FT-IR, DSC, SEM, and XRD, were then used to confirm the formation of inclusion complexes. The natural polysaccharide-based hydrogel comprising pectin and pullulan was synthesized in air and optimized through various parameters. In order to maximize the reaction parameters, Response Surface Methodology design was employed for experimental optimization. We use FT-IR, TGA, SEM, EDX, and XRD to investigate hydrogel formation. At 37 °C, an investigation was carried out on the in vitro controlled release of PN at pH 2, 7, and 7.4. The analysis of drug release data revealed that PM and KM exhibited an initial burst release of drugs, with the MW and FD method proving to be the most suitable approach for achieving precise ICs of PN and ß-CD for sustained drug release. The kinetics of drug release were evaluated using various kinetic models, with the Riteger-Peppas and Peppas-Sahlin models demonstrating the best fit for drug release in all instances.

Polymeric liposomes of emtricitabine employing modified pullulan-an attempt to reduce associated hepatotoxicity.

Emtricitabine (FTC) a BCS class I drug, is used for HIV prevention. The high solubility of the drug is the leading cause of severe hepatotoxicity and lactic acidosis. This research focuses on the use of modified pullulan for the preparation of polymeric liposomes of FTC. Modified pullulan was synthesized using cholesterol, and succinic anhydride in a controlled chemical environment. The formation of the polymer was established through analysis of spectra. Varying the drug-polymer ratio (1:1, 1:2, and 1:3), the drug-polymer composite was loaded in the vesicular system of soya phosphatidylcholine and cholesterol. Formulations were evaluated for drug entrapment, particle size, surface morphology, and in vitro and ex vivo drug release. An in vivo study of the pure drug and the best formulation on mice was conducted for 28 days following daily oral administration to evaluate the effect on liver and hematological parameters. The best formulation was further subjected to cytotoxicity study on hepatic cell lines. Spectral analysis confirmed the formation of modified pullulan. All formulations showed high drug entrapment in the nanovesicles. The in vitro and ex vivo drug release profiles depicted a controlled release of the drug. Hematological parameters were found to be under control in the animals throughout the experimentation. A comparative histopathology study on the livers and cytotoxicity study on hepatic cell lines revealed the safety of the best formulation over the pure drug. Hence it can be concluded that polymeric liposomes of FTC can be a promising mode of delivery to overcome its limitations.

Preparation and characterization of tea tree essential oil microcapsule-coated packaging paper with bacteriostatic effect.

We prepared tea tree essential oil microcapsules, and the microcapsules and pullulan were coated on kraft paper to prepare an antibacterial paper. The antibacterial activity, structural characterization, and thermal stability of the prepared microcapsules and packaging paper were then tested. We found that the retention rate of microcapsules reached 87.1% after a 70 min of high-temperature treatment. The minimum inhibitory concentrations of microcapsules to S. aureus and E. coli were 112 mg/mL and 224 mg/mL, and the bacteriostatic zones of the packaging paper to E. coli and S. aureus were 17.49 mm and 22.75 mm, respectively. The prepared microcapsules were irregular. The paper coating was formed via hydrogen bonding, which filled the pores of paper fibers. When compared with the base paper, the roughness of the paper was reduced to 7.16 nm (Rq) and 5.61 nm (Ra), and no thermal decomposition occurred at <288 °C, which together implies a good application prospect.

Integrated In Vivo and In Vitro Evaluation of a Powder-to-Hydrogel, Film-Forming Polymer Complex Base with Tissue-Protective and Microbiome-Supportive Properties.

The study aimed to perform a comprehensive in vitro and in vivo evaluation of a newly developed, patent-pending, powder-to-hydrogel, film-forming polymer complex base, which possesses tissue-protective and microbiome-supportive properties, and to compare its characteristics with poloxamer 407. The study used a combination of in vitro assays, including tissue viability and cell migration, and in vivo wound healing evaluations in male diabetic mice. Microbiome dynamics at wound sites were also analyzed. The in vitro assays demonstrated that the polymer complex base was non-cytotoxic and that it enhanced cell migration over poloxamer 407. In vivo, the polymer complex base demonstrated superior wound healing capabilities, particularly in combination with misoprostol and phenytoin, as evidenced by the reduced wound area and inflammation scores. Microbiome analysis revealed favorable shifts in bacterial populations associated with the polymer complex base-treated wounds. The polymer complex base demonstrates clinical significance in wound care, potentially offering improved healing, safety and microbiome support. Its transformative properties and efficacy in drug delivery make it a promising candidate for advanced wound care applications, particularly in chronic wound management.

Formulations based on pullulan and a derivative as coating material for the food sector.

Carboxymethylated derivatives of pullulan (PU) were synthesized and evaluated as coating for the postharvest preservation of blueberries. Carboxymethylpullulan was obtained by etherification reaction with the substitution degrees of 0.52, 0.34, and 0.26 for CMP1, CMP2, and CMP3 respectively. Infrared spectroscopy and nuclear magnetic resonance results showed characteristic signals of the carbonyl group belonging to the carboxymethyl group. Thermal analysis showed that CMP1, CMP2, and CMP3 derivatives presented thermal stability values of 209.91 C, 214.73 C, and 225.52 °C, respectively, and were lower with respect to PU with Td of 238.84 °C. Furthermore, an increase in the glass transition temperature due to carboxymethylation was determined. The chemical modification decreased the contact angle with respect to PU (71.34°) with values for CMP1, CMP2, and CMP3 of 39.89°, 53.72° and 60.61°, respectively. The carboxymethylation also increased the water vapor permeability and mechanical properties of the films. In addition, it was found that the CMP molecules affected the optical properties. The application of CMP-based coatings reduced the mass loss and ripening rate of blueberries compared to native pullulan, therefore, packaging from CMP molecules could be used as a coating capable of delaying ripening and extending the shelf life of fruits.

Preparation and characterization of carboxymethylated pullulan/butyric acid-modified chitosan active sustainable bi-layer coatings intended for packaging of cheese slices.

Dependence of the food industry on conventional plastic and the generation of enormous amounts of food waste caused by microbiological spoilage have been imposed as inspiration for this work, to develop active sustainable packaging for sliced cheese using the bi-layer design. Pullulan was modified using a green approach to obtain a polyanionic character in the coating formulation. Chitosan, which has a cationic character in an acidic environment, has been modified using a butyric acid to obtain an amphiphilic character. The formed active bi-layer has demonstrated an improved barrier (decreased permeability for moisture vapor 72.2 and 77.7 times) and mechanical properties (increased tensile strength value up to 3.9 and 9.4 times) compared to the monolayer films. A novel approach to microbiological control of sliced cheese has been established, which implies a synergistic effect of Helichrysum italicum essential oil (EO) and corresponding hydrolate (HY) incorporated in separated layers. This design has ensured avoiding surfactants and preserving cheese's sensory properties, prolonging its shelf-life by 50 % at least. Improvements in cheese storage conditions using this packaging lie in the improved barrier, mechanical and antimicrobial properties, the order of lamination, and a good covering of the cheese surface by spraying.

Composite antibacterial hydrogels based on two natural products pullulan and ε-poly-l-lysine for burn wound healing.

Antibacterial hydrogels as burn wound dressings are capable of efficaciously defending against bacterial infection and accelerating burn wound healing. Thus far, a large plethora of antibacterial hydrogels have adopted numerous components and intricate preparation processes, yet restricting their practical industrialization applications. Simple and effective preparation methods of antibacterial hydrogels are hence urgently needed. Herein, an easy but efficacious strategy with the employment of two natural products pullulan and ε-poly-l-lysine (ε-PL) was designed to fabricate composite antibacterial hydrogels for burn wound healing for the first time. The hydrogel crosslinking networks were formed through amidation reactions between carboxylated pullulan derivative (CP) and ε-poly-l-lysine hydrochloride (ε-PL·HCl). The resulting hydrogels possessed high transparency, porous structures, tunable gelation time and gel content, relatively low swelling ratios, appropriate self-degradability, proper mechanical properties, strong in vitro bacteriostatic activities, non-cytotoxicity, capacities of facilitating cell migration and excellent hemocompatibility. In the infected burn model of mice, the hydrogels were observed to display prominent in vivo antibacterial activities and enable the acceleration of burn wound healing. We opine the simply and effectively prepared antibacterial hydrogels as promising dressings for burn wound recovery have broad industrialization prospects.

Formation of Microcapsules of Pullulan by Emulsion Template Mechanism: Evaluation as Vitamin C Delivery Systems.

Pullulan is a polysaccharide that has attracted the attention of scientists in recent times as a former of edible films. On the other hand, its use for the preparation of hydrogels needs more study, as well as the formation of pullulan microcapsules as active ingredient release systems for the food industry. Due to the slow gelation kinetics of pullulan with sodium trimetaphosphate (STMP), capsules cannot be formed through the conventional method of dropping into a solution of the gelling agent, as with other polysaccharides, since the pullulan chains migrate to the medium before the capsules can form by gelation. Pullulan microcapsules have been obtained by using inverse water-in-oil emulsions as templates. The emulsion that acts as a template has been characterized by monitoring its stability and by optical microscopy, and the size of the emulsion droplets has been correlated with the size of the microcapsules obtained, demonstrating that it is a good technique for their production. Although some flocs of droplets form, these remain dispersed during the gelation process and two capsule size distributions are obtained: those of the non-flocculated droplets and the flocculated droplets. The microcapsules have been evaluated as vitamin C release systems, showing zero-order release kinetics for acidic pH and Fickian mechanism for neutral pH. On the other hand, the microcapsules offer good protection of vitamin C against oxidation during an evaluation period of 14 days.

Draft genome sequence of Vibrio sp. PBL-C16, isolated from Batu Laut Beach in Selangor, Malaysia.

Vibrio sp. PBL-C16 is a bacterium that was isolated from Batu Laut Beach in Selangor, Malaysia. Here, we present a high-quality annotated draft genome of strain PBL-C16 and suggest its potential glycoside hydrolase enzymes for polysaccharide degradation.

Preparation of multi-functional active packaging film of Galla chinensis waste CDs/pullulan.

In this study, multifunctional green carbon dots (CDs) have been synthesized using Galla chinensis waste (GCW) via hydrothermal method for the first time. An active packaging film has been developed in this work by combining CDs and pullulan (PL), using the solution-casting method. The microscopic morphology revealed that the CDs that were prepared using GCW exhibited good compatibility with PL. In addition, it also led to improvement in the toughness of the PL film (14.01 % to 20.26 %), along with its water vapor permeability value [1.31 to 0.53 (g·mm)/(kPa·h·m2)]. The composite films consisting of CDs exhibited good UV blocking rates for the UVA (90.41 %-7.87 %), UVB (87.76 %-0.08 %), and UVC (83.39 %-0 %) spectral ranges. The composite films exhibited strong antioxidant activity, and the clearance of ABTS and DPPH were obtained to be 93.61 % and 86.30 %, respectively. In addition, the composite films showed good antibacterial activity for E. coli and S. aureus, with a high antibacterial rate of up to 99.99 %. Finally, the non-contact preservation of strawberries over a duration of 10 d at room temperature confirmed that the prepared composite film can help preserve the quality of strawberries, as well as extended their shelf-life.

Preparation and functional characterization of pullulan-sodium alginate composite film enhanced with ultrasound-assisted clove essential oil Nanoemulsions for effective preservation of cherries and mushrooms.

Clove essential oil (CEO) exhibited potent antibacterial efficacy and are obtained from Eugenia caryophyllata tree flower buds. Herein, CEO nanoemulsions were prepared using various concentrations of casein protein treated with ultrasound for different time interval. The study demonstrated that CEO nanoemulsions with 5% casein protein subjected to ultrasound for 10 min displayed the most minimal particle size. The pullulan­sodium alginate film incorporated with nanoemulsions treated with ultrasound exhibited enhanced physico-mechanical characteristics. Based on the structural analysis, the application of ultrasonic treatment improved intermolecular compatibility and organized molecular structure by strengthening hydrogen bonds. Furthermore, the composite film displayed remarkable efficacy against E. coli and S. aureus as well as longer retention of essential oils. The use of the developed films to protect cherry fruits and mushrooms produced promising results, emphasizing their potential in food packaging applications.