Formation of amygdalin/ß-cyclodextrin derivatives inclusion complexes for anticancer activity assessment in human cervical carcinoma HeLa cell line.

Nanoencapsulation has gained considerable attention because of its unique features and advantages in anticancer drug delivery. Amygdalin (AMY) is an anticancer compound, showing limitations in its applications by low stability. Herein, the inclusion complexes (ICs) of AMY with ß-cyclodextrin (ßCD), and its derivatives such as 2-hydroxypropyl-ßCD (HPßCD) and methyl-ßCD (MßCD) were fabricated. The fabricated AMY/CD-ICs were thoroughly evaluated using Fourier-transform infrared spectroscopy, powder X-ray diffraction, thermogravimetric/differential thermal analysis, proton nuclear magnetic resonance, ultraviolet-visible diffuse reflectance spectroscopy, and photoluminescence techniques. Double reciprocal profile study of the absorption and fluorescence spectra revealed that the AMY formed the ICs with ßCD derivatives at a guest/host stoichiometric ratio of 1/1. The thermal stability of AMY was enhanced as the IC formation aid observed by the shift of thermal degradation temperature of AMY from the range of âˆ¼ 220-250 °C to > 295 °C. Theoretical analyses of the energetic, electronic, and global reactivity parameters of the AMY/CD-ICs were evaluated using the PM3 method. Further assessment of the dissolution diagrams of AMY/CD-ICs revealed a burst release profile. In addition, cell toxicity was evaluated using the MTT assay, and the results showed that AMY/CD-ICs had significantly more efficacious in inhibiting HeLa cancer cells than AMY. These results proved that the IC formations with CDs significantly enhanced the anticancer activity of AMY.

Recent Advances in Drug Delivery Systems Targeting Insulin Signalling for the Treatment of Alzheimer's Disease.

Alzheimer's disease (AD) is a complex neurodegenerative disorder characterized by the accumulation of neurofibrillary tangles and amyloid-ß plaques. Recent research has unveiled the pivotal role of insulin signaling dysfunction in the pathogenesis of AD. Insulin, once thought to be unrelated to brain function, has emerged as a crucial factor in neuronal survival, synaptic plasticity, and cognitive processes. Insulin and the downstream insulin signaling molecules are found mainly in the hippocampus and cortex. Some molecules responsible for dysfunction in insulin signaling are GSK-3ß, Akt, PI3K, and IRS. Irregularities in insulin signaling or insulin resistance may arise from changes in the phosphorylation levels of key molecules, which can be influenced by both stimulation and inactivity. This, in turn, is believed to be a crucial factor contributing to the development of AD, which is characterized by oxidative stress, neuroinflammation, and other pathological hallmarks. Furthermore, this route is known to be indirectly influenced by Nrf2, NF-κB, and the caspases. This mini-review delves into the intricate relationship between insulin signaling and AD, exploring how disruptions in this pathway contribute to disease progression. Moreover, we examine recent advances in drug delivery systems designed to target insulin signaling for AD treatment. From oral insulin delivery to innovative nanoparticle approaches and intranasal administration, these strategies hold promise in mitigating the impact of insulin resistance on AD. This review consolidates current knowledge to shed light on the potential of these interventions as targeted therapeutic options for AD.

Development of carrageenan-immobilized lytic coliphage vB_Eco2571-YU1 hydrogel for topical delivery of bacteriophages in wound dressing applications.

Bacteriophages are employed as cost-effective and efficient antibacterial agents to counter the emergence of antibiotic-resistant bacteria and other host bacteria in phage therapy. The increasing incidence of skin wounds is a significant concern in clinical practice, especially considering the limitations of antibiotic therapy. Furthermore, the lack of an effective delivery system that preserves the stability of bacteriophages hampers their clinical implementation. In recent years, there has been a growing amount of research on bacteriophage applications in veterinary and biomedical sciences. In our study, lytic coliphage vB_Eco2571-YU1 was isolated against pathogenic Escherichia coli host bacteria, and hydrogel wound dressing materials were fabricated with marine polysaccharide carrageenan (carr-vB_Eco2571-YU1) for their antibacterial activity. Transmission electron microscopy (TEM) morphology identified it as a Myoviridae coliphage with an icosahedral head length and width of approximately 60 and 56.8 nm, respectively, and a tail length of 119.7 nm. The one-step growth curve of coliphage revealed a latent period of 10 min, a rise period of 15 min, and a burst size of 120 virions per cell. The bacteriolytic activity of unimmobilized coliphages was observed within 2 h; however, strain-specific phage resistance was acquired after 9 h. In contrast, carr-vB_Eco2571-YU1 showed a sharp decline in the growth of bacteria in the log phase after 2 h and did not allow for the acquisition of phage resistance by the E. coli strain. The stability of coliphage under different pH, temperature, osmolarity, detergents, and organic solvents was evaluated. We also studied the long-term storage of carr-vB_Eco2571-YU1 hydrogels at 4 °C and found that the titer value decreased during a time-dependent period of 28 days. These hydrogels were also found to be hemocompatible using a hemolysis assay. The addition of plasticizer (0.6 % (w/v)) to the carrageenan (2 % (w/v)) to prepare carr-vB_Eco2571-YU1 hydrogels showed a decrease in compressive strength with enhanced elasticity. This phage therapy using polymeric immobilization of bacteriophages is a promising next-generation wound dressing biomaterial alternative to conventional wound and skin care management.

Autoimmune responses and therapeutic interventions for systemic lupus erythematosus: a comprehensive review.

Systemic Lupus Erythematosus (SLE) or Lupus is a multifactorial autoimmune disease of multiorgan malfunctioning of extremely heterogeneous and unclear etiology that affects multiple organs and physiological systems. Some racial groups and women of childbearing age are more susceptible to SLE pathogenesis. Impressive progress has been made towards a better understanding of different immune components contributing to SLE pathogenesis. Recent investigations have uncovered the detailed mechanisms of inflammatory responses and organ damage. Various environmental factors, pathogens, and toxicants, including ultraviolet light, drugs, viral pathogens, gut microbiome metabolites, and sex hormones trigger the onset of SLE pathogenesis in genetically susceptible individuals and result in the disruption of immune homeostasis of cytokines, macrophages, T cells, and B cells. Diagnosis and clinical investigations of SLE remain challenging due to its clinical heterogeneity and hitherto only a few approved antimalarials, glucocorticoids, immunosuppressants, and some nonsteroidal anti-inflammatory drugs (NSAIDs) are available for treatment. However, the adverse effects of renal and neuropsychiatric lupus and late diagnosis make therapy challenging. Additionally, SLE is also linked to an increased risk of cardiovascular diseases due to inflammatory responses and the risk of infection from immunosuppressive treatment. Due to the diversity of symptoms and treatment-resistant diseases, SLE management remains a challenging issue. Nevertheless, the use of next-generation therapeutics with stem cell and gene therapy may bring better outcomes to SLE treatment in the future. This review highlights the autoimmune responses as well as potential therapeutic interventions for SLE particularly focusing on the recent therapeutic advancements and challenges.

A Comprehensive Review of Emerging Trends and Innovative Therapies in Epilepsy Management.

Epilepsy is a complex neurological disorder affecting millions worldwide, with a substantial number of patients facing drug-resistant epilepsy. This comprehensive review explores innovative therapies for epilepsy management, focusing on their principles, clinical evidence, and potential applications. Traditional antiseizure medications (ASMs) form the cornerstone of epilepsy treatment, but their limitations necessitate alternative approaches. The review delves into cutting-edge therapies such as responsive neurostimulation (RNS), vagus nerve stimulation (VNS), and deep brain stimulation (DBS), highlighting their mechanisms of action and promising clinical outcomes. Additionally, the potential of gene therapies and optogenetics in epilepsy research is discussed, revealing groundbreaking findings that shed light on seizure mechanisms. Insights into cannabidiol (CBD) and the ketogenic diet as adjunctive therapies further broaden the spectrum of epilepsy management. Challenges in achieving seizure control with traditional therapies, including treatment resistance and individual variability, are addressed. The importance of staying updated with emerging trends in epilepsy management is emphasized, along with the hope for improved therapeutic options. Future research directions, such as combining therapies, AI applications, and non-invasive optogenetics, hold promise for personalized and effective epilepsy treatment. As the field advances, collaboration among researchers of natural and synthetic biochemistry, clinicians from different streams and various forms of medicine, and patients will drive progress toward better seizure control and a higher quality of life for individuals living with epilepsy.

Polydopamine-Functionalized Bacterial Cellulose as Hydrogel Scaffolds for Skin Tissue Engineering.

Bacterial cellulose (BC) is a natural polysaccharide polymer hydrogel produced sustainably by the strain Gluconacetobacter hansenii under static conditions. Due to their biocompatibility, easy functionalization, and necessary physicochemical and mechanical properties, BC nanocomposites are attracting interest in therapeutic applications. In this study, we functionalized BC hydrogel with polydopamine (PDA) without toxic crosslinkers and used it in skin tissue engineering. The BC nanofibers in the hydrogel had a thickness of 77.8 ± 20.3 nm, and they could be used to produce hydrophilic, adhesive, and cytocompatible composite biomaterials for skin tissue engineering applications using PDA. Characterization techniques, namely Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), and Raman spectroscopy, were performed to investigate the formation of polydopamine on the BC nanofibers. The XRD peaks for BC occur at 2θ = 14.65°, 16.69°, and 22.39°, which correspond to the planes of (100), (010), and (110) of cellulose type Iα. Raman spectroscopy confirmed the formation of PDA, as indicated by the presence of bands corresponding to the vibration of aromatic rings and aliphatic C-C and C-O stretching at 1336 and 1567 cm-1, respectively. FTIR confirmed the presence of peaks corresponding to PDA and BC in the BC/PDA hydrogel scaffolds at 3673, 3348, 2900, and 1052 cm-1, indicating the successful interaction of PDA with BC nanofibers, which was further corroborated by the SEM images. The tensile strength, swelling ratio, degradation, and surface wettability characteristics of the composite BC biomaterials were also investigated. The BC/PDA hydrogels with PDA-functionalized BC nanofibers demonstrated excellent tensile strength and water-wetting ability while maintaining the stability of the BC fibers. The enhanced cytocompatibility of the BC/PDA hydrogels was studied using the PrestoBlue assay. Culturing murine NIH/3T3 fibroblasts on BC/PDA hydrogels showed higher metabolic activity and enhanced proliferation. Additionally, it improved cell viability when using BC/PDA hydrogels. Thus, these BC/PDA composite biomaterials can be used as biocompatible natural alternatives to synthetic substitutes for skin tissue engineering and wound-dressing applications.

Photocatalytic Degradation, Anticancer, and Antibacterial Studies of Lysinibacillus sphaericus Biosynthesized Hybrid Metal/Semiconductor Nanocomposites.

The biological synthesis of nanocomposites has become cost-effective and environmentally friendly and can achieve sustainability with high efficiency. Recently, the biological synthesis of semiconductor and metal-doped semiconductor nanocomposites with enhanced photocatalytic degradation efficiency, anticancer, and antibacterial properties has attracted considerable attention. To this end, for the first time, we biosynthesized zinc oxide (ZnO) and silver/ZnO nanocomposites (Ag/ZnO NCs) as semiconductor and metal-doped semiconductor nanocomposites, respectively, using the cell-free filtrate (CFF) of the bacterium Lysinibacillus sphaericus. The biosynthesized ZnO and Ag/ZnO NCs were characterized by various techniques, such as ultraviolet-visible spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, field emission scanning electron microscopy, transmission electron microscopy, and photoluminescence spectroscopy. The photocatalytic degradation potential of these semiconductor NPs and metal-semiconductor NCs was evaluated against thiazine dye, methylene blue (MB) degradation, under simulated solar irradiation. Ag/ZnO showed 90.4 ± 0.46% photocatalytic degradation of MB, compared to 38.18 ± 0.15% by ZnO in 120 min. The cytotoxicity of ZnO and Ag/ZnO on human cervical HeLa cancer cells was determined using an MTT assay. Both nanomaterials exhibited cytotoxicity in a concentration- and time-dependent manner on HeLa cells. The antibacterial activity was also determined against Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus). Compared to ZnO, Ag/ZnO NCs showed higher antibacterial activity. Hence, the biosynthesis of semiconductor nanoparticles could be a promising strategy for developing hybrid metal/semiconductor nanomaterials for different biomedical and environmental applications.

Physicochemical, mechanical, dielectric, and biological properties of sintered hydroxyapatite/barium titanate nanocomposites for bone regeneration.

Bone implants fabricated using nanocomposites containing hydroxyapatite (HA) and barium titanate (BT) show osteoconductive, osteoinductive, osteointegration, and piezoelectricity properties for bone regeneration applications. In our present study, HA and BT nanopowders were synthesized using high-energy ball-milling-assisted solid-state reaction with precursors of calcium carbonate and ammonium dihydrogen phosphate, and barium carbonate and titanium oxide powder mixtures, respectively. Hexagonal HA and tetragonal BT phases were formed after calcination at 700 and 1000 °C, respectively. Subsequently, hydroxyapatite/barium titanate (HA/BT) nanocomposites with different weight percentages of HA and BT were prepared by ball-milling, then compacted and sintered at two different temperatures to endow these bioceramics with better mechanical, dielectric, and biological properties for bone regeneration. Microstructure, crystal phases, and molecular structure characterizations of these sintered HA/BT nanocomposite compacts (SHBNCs) were performed using field-emission scanning electron microscopy, x-ray diffraction, and Fourier-transform infrared spectroscopy, respectively. Bulk density was evaluated using the Archimedes method. HA/BT nanocomposites with increased BT content showed enhanced dielectric properties, and the dielectric constant (ϵr) value for 5HA/95BT was ∼182 at 100 Hz. Mechanical properties such as Vicker's hardness, fracture toughness, yield strength, and diametral tensile strength were also investigated. The hemolysis assay of SHBNCs exhibited hemocompatibility. The effect of these SHBNCs as implants on thein vitrocytocompatibility and cell viability of MG-63 osteoblast-like cells was assessed by MTT assay and live/dead staining, respectively. 15HA/85BT showed increased metabolic activity with a higher number of live cells than BT after the culture period. Overall, the SHBNCs can be used as orthopedic implants for bone regeneration applications.

Recent Advances in the Biomedical Applications of Functionalized Nanogels.

Nanomaterials have been extensively used in several applications in the past few decades related to biomedicine and healthcare. Among them, nanogels (NGs) have emerged as an important nanoplatform with the properties of both hydrogels and nanoparticles for the controlled/sustained delivery of chemo drugs, nucleic acids, or other bioactive molecules for therapeutic or diagnostic purposes. In the recent past, significant research efforts have been invested in synthesizing NGs through various synthetic methodologies such as free radical polymerization, reversible addition-fragmentation chain-transfer method (RAFT) and atom transfer radical polymerization (ATRP), as well as emulsion techniques. With further polymeric functionalizations using activated esters, thiol-ene/yne processes, imines/oximes formation, cycloadditions, nucleophilic addition reactions of isocyanates, ring-opening, and multicomponent reactions were used to obtain functionalized NGs for targeted delivery of drug and other compounds. NGs are particularly intriguing for use in the areas of diagnosis, analytics, and biomedicine due to their nanodimensionality, material characteristics, physiological stability, tunable multi-functionality, and biocompatibility. Numerous NGs with a wide range of functionalities and various external/internal stimuli-responsive modalities have been possible with novel synthetic reliable methodologies. Such continuous development of innovative, intelligent materials with novel characteristics is crucial for nanomedicine for next-generation biomedical applications. This paper reviews the synthesis and various functionalization strategies of NGs with a focus on the recent advances in different biomedical applications of these surface modified/functionalized single-/dual-/multi-responsive NGs, with various active targeting moieties, in the fields of cancer theranostics, immunotherapy, antimicrobial/antiviral, antigen presentation for the vaccine, sensing, wound healing, thrombolysis, tissue engineering, and regenerative medicine.

Photocatalytic degradation activity of goji berry extract synthesized silver-loaded mesoporous zinc oxide (Ag@ZnO) nanocomposites under simulated solar light irradiation.

Different approaches have been developed for the synthesis of various nanostructured materials with unique morphologies. This study demonstrated the photocatalytic and antimicrobial abilities of silver-loaded zinc oxide nanocomposites (Ag@ZnO NCs). Initially, ZnO with a unique mesoporous ellipsoidal morphology in the size range of 0.59 ± 0.11 × 0.33 ± 0.09 µm (length × width) was synthesized using aqueous precipitation in a mild hydrothermal condition (80 °C) with the aqueous fruit extract of goji berry (GB) (as an additive) and calcined in air at 200 °C/2 h and 250 °C/3 h. Powder X-ray diffraction (XRD) revealed the formation of a hexagonal phase of the wurtzite (WZ) structure. The average crystallite size of ZnO was 23.74 ± 4.9 nm as calculated using Debye-Scherrer's equation. It also possesses higher thermal stability with the surface area, pore volume, and pore size of 11.77 m2/g, 0.027 cm3/g, and 9.52 nm, respectively. Furthermore, different mesoporous Ag@ZnO NCs loaded with face-centered cubic (fcc) silver nanoparticles (Ag NPs) in the range of 90-160 nm were synthesized by GB extract as a reducing and capping agent on the surface of ZnO after calcination in air. The immobilization of Ag NPs was confirmed by XRD, X-ray photoelectron spectroscopy (XPS), field-emission scanning electron microscopy (FE-SEM), FE-transmission electron microscopy (FE-TEM), and energy-dispersive X-ray spectroscopy (EDS). It was found that Ag0.2@ZnO NC (0.2 wt% of Ag) showed excellent photocatalytic degradation of both methylene blue (MB) (cationic) and congo red (CR) (anionic) dyes under simulated solar irradiation. The photocatalytic degradation of 99.3 ± 0.35% MB and 98.5 ± 1.3% CR occurred in 90 and 55 min, respectively, at room temperature by Ag0.2@ZnO NC. Besides, these NCs also showed broad-spectrum antibacterial activity against both Gram-positive and Gram-negative bacteria. The mechanistic concept of generating reactive oxygen species (ROS) by electron and hole charge (e‾/h+) carriers seems to be responsible for the photocatalytic degradation of commercial dyes and antibacterial activities by Ag@ZnO NCs. Thus, these silver-loaded mesoporous ellipsoidal ZnO NCs are promising candidates as photocatalysts for industrial/wastewater treatment as well as in antimicrobial therapeutics.

Tissue Adhesive, Self-Healing, Biocompatible, Hemostasis, and Antibacterial Properties of Fungal-Derived Carboxymethyl Chitosan-Polydopamine Hydrogels.

In this work, fungal mushroom-derived carboxymethyl chitosan-polydopamine hydrogels (FCMCS-PDA) with multifunctionality (tissue adhesive, hemostasis, self-healing, and antibacterial properties) were developed for wound dressing applications. The hydrogel is obtained through dynamic Schiff base cross-linking and hydrogen bonds between FCMCS-PDA and covalently cross-linked polyacrylamide (PAM) networks. The FCMCS-PDA-PAM hydrogels have a good swelling ratio, biodegradable properties, excellent mechanical properties, and a highly interconnected porous structure with PDA microfibrils. Interestingly, the PDA microfibrils were formed along with FCMCS fibers in the hydrogel networks, which has a high impact on the biological performance of hydrogels. The maximum adhesion strength of the hydrogel to porcine skin was achieved at about 29.6 ± 2.9 kPa. The hydrogel had good self-healing and recoverable properties. The PDA-containing hydrogels show good antibacterial properties on Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) bacteria. Moreover, the adhesive hydrogels depicted good viability and attachment of skin fibroblasts and keratinocyte cells. Importantly, FCMCS and PDA combined resulted in fast blood coagulation within 60 s. Hence, the adhesive hydrogel with multifunctionality has excellent potential as a wound dressing material for infected wounds.

Fabrication strategies and biomedical applications of three-dimensional bacterial cellulose-based scaffolds: A review.

Bacterial cellulose (BC), an extracellular polysaccharide, is a versatile biopolymer due to its intrinsic physicochemical properties, broad-spectrum applications, and remarkable achievements in different fields, especially in the biomedical field. Presently, the focus of BC-related research is on the development of scaffolds containing other materials for in-vitro and in-vivo biomedical applications. To this end, prime research objectives concern the biocompatibility of BC and the development of three-dimensional (3D) BC-based scaffolds. This review summarizes the techniques used to develop 3D BC scaffolds and discusses their potential merits and limitations. In addition, we discuss the various biomedical applications of BC-based scaffolds for which the 3D BC matrix confers desired structural and conformational features. Overall, this review provides comprehensive coverage of the idea, requirements, synthetic strategies, and current and prospective applications of 3D BC scaffolds, and thus, should be useful for researchers working with polysaccharides, biopolymers, or composite materials.

Peptide ligases: A Novel and potential enzyme toolbox for catalytic cross-linking of protein/peptide-based biomaterial scaffolds for tissue engineering.

The fabrication of novel biomaterial scaffolds with improved biological interactions and mechanical properties is an important aspect of tissue engineering. The three-dimensional (3D) protein/peptide-based polymeric scaffolds are promising in vitro biomaterials to replicate the in vivo microenvironment mimicking the extracellular matrix (ECM) for cell differentiation and subsequent tissue formation. Among different strategies in the fabrication of scaffolds, bioorthogonal enzymatic reactions for rapid in situ zero-length cross-linking are advantageous. Peptide ligases as a novel toolbox have the potentiality to enzymatically cross-link natural/synthetic protein/peptide-based polymeric chains for a wide range of biomedical applications. Although natural peptide ligases, such as sortases and butelase 1 are known cysteine proteases with ligase activity, some serine proteases, such as trypsin and subtilisin, are protein engineered to form trypsiligase and subtiligase, respectively, which exhibited efficient ligase activity by linking proteins/peptides with a great variety of molecules. Peptide ligase activity by these engineered proteases is more efficient than the hydrolysis of peptide bonds (peptidase activity). Peptide esters form acyl-enzyme intermediate with serine/cysteine residues of these proteases, with subsequent aminolysis forming covalent peptide bond with N-terminal residue of another polymeric chain. In addition, peptide ligases have the potential to conjugate with cell-adhesive ECM proteins or motifs and growth factors to (bio)polymeric networks to enhance cell attachment, growth, and differentiation. Here, we review the potential and limitations of natural and engineered peptide ligases as an enzyme toolbox with a focus on sortases (classes A-D), butelase 1, trypsiligase, and subtilisin variants, and the mechanisms for their zero-length cross-linking of (bio)polymeric scaffolds for various tissue engineering and regenerative applications.

Bacterial exo-polysaccharides in biofilms: role in antimicrobial resistance and treatments.

BACKGROUND: Bacterial biofilms are aggregation or collection of different bacterial cells which are covered by self-produced extracellular matrix and are attached to a substratum. Generally, under stress or in unfavorable conditions, free planktonic bacteria transform themselves into bacterial biofilms and become sessile. MAIN BODY: Various mechanisms involving interaction between antimicrobial and biofilm matrix components, reduced growth rates, and genes conferring antibiotic resistance have been described to contribute to enhanced resistance. Quorum sensing and multi-drug resistance efflux pumps are known to regulate the internal environment within the biofilm as well as biofilm formation; they also protect cells from antibiotic attack or immune attacks. This review summarizes data supporting the importance of exopolysaccharides during biofilm formation and its role in antibiotic resistance. CONCLUSIONS: Involvement of quorum sensing and efflux pumps in antibiotic resistance in association with exopolysaccharides. Also, strategies to overcome or attack biofilms are provided.

Antibacterial properties of starch-reduced graphene oxide-polyiodide nanocomposite.

Graphene-based nanocomposites with superior antibacterial activity are highly sought after by the food packaging industries. Here, we report for the first time a method that utilizes soluble starch biopolymer as a functionalizing and reducing agent for the preparation of starch-reduced graphene oxide (SRGO), whereby polyiodide binds to the helical structures of amylose units of the starch (chromophore) to form a SRGO-polyiodide nanocomposite (SRGO-PI NC). UV-visible spectroscopy, X-ray diffraction, Raman spectroscopy, scanning electron microscopy, and energy-dispersive spectroscopy confirmed the presence of polyiodide in SRGO. SRGO-PI NC exhibited good antibacterial activities against pathogenic Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) microbes with minimum inhibitory concentrations (MICs) and minimum bactericidal concentration (MBC) values (as determined by a broth-dilution method) of 2.5 and 5 mg/ml, respectively, for both E. coli and S. aureus. PrestoBlue viability assays showed half-maximal inhibitory concentration (IC50) values of 0.45 and 0.41 mg/ml for E. coli and S. aureus, respectively. Time-kill kinetic and live/dead bacterial viability assays revealed the antimicrobial activities of SRGO-PI NC against both E. coli and S. aureus. The study provides new insights regarding the utilization of graphene-polyiodide NCs as high-efficacy antibacterial starch-based nanomaterials for food packaging applications.

Electrospun poly(vinyl alcohol)/reduced graphene oxide nanofibrous scaffolds for skin tissue engineering.

Graphene is composed of a two-dimensional (2D) layer of carbon atoms arranged in a honeycomb lattice configuration. In this paper, we adopted a green synthetic method of producing reduced graphene oxide using glucose as a reducing and stabilizing agent. We also investigated the fabrication of electrospun nanofibers of glucose-reduced graphene oxide (GRGO) (0-1.0 wt%) reinforced with polyvinyl alcohol (PVA) as (PG) scaffolds, and chemically crosslinked with acidic glutaraldehyde (GA) in acetone medium to mimic the extracellular matrix (ECM) for skin tissue engineering applications. These PG scaffolds were evaluated for morphology, mechanical strength, surface wettability, thermal properties, hemocompatibility, and biocompatibility. Field emission-scanning electron microscopy (FE-SEM) revealed an increase in the thickness of nanofibers in PG scaffolds with an increase in the concentration of GRGO. X-ray diffraction and attenuated total reflectance-infrared and Raman spectra showed the GRGO was incorporated in the PVA nanofibrous matrix. As the concentration of GRGO was increased in PG scaffolds, tensile strengths and elongations at break decreased, whereas thermal properties increased. The biological activities of PG scaffolds were evaluated using in vitro hemolysis, using CCD-986Sk (a human skin fibroblast cell line) viability and proliferation assays, and by live/dead cell imaging. Results showed GRGO inclusion in PVA nanofibers caused a slight hydrophilic to hydrophobic shift. PG scaffolds did not cause hemolysis of red blood cells even at a GRGO loading of 1.0 wt%, and PG-1.0 scaffold (with a GRGO loading of 1.0 wt%) exhibited excellent compatibility with fibroblasts and significantly increased metabolic activity after culture for 21 days as compared with PG-0 controls. DAPI staining and live/dead imaging assays showed that all PG scaffolds increased fibroblast proliferation and viability, indicating the potential for skin tissue engineering applications.

Novel biomimetic chitin-glucan polysaccharide nano/microfibrous fungal-scaffolds for tissue engineering applications.

Naturally occurring many biological structures have provided sources of inspiration for the fabrication of many novel nanostructures for various applications. Electrospun nano/microfibrous structures have great potential as scaffolds for cell attachment and proliferation in the field of tissue engineering. Here, for the first time, we report on the preparation of three-dimensional (3D) fungal mycelial mats with chitin-glucan polysaccharide cell walls as nano/microfibrous scaffolds for tissue engineering applications. Treatment of fungal-scaffolds (F-scaffolds) with ß-mercaptoethanol (BME) improved hemocompatibility, and conferred biocompatibility with respect to the adhesion and proliferation of human keratinocytes. Field-emission scanning electron microscopy (FE-SEM) of BME-treated F-scaffolds revealed a meshwork of nano- and micro-fibrous mycelial structures with an average diameter of 2.94 ± 0.96 µm (range 0.92-5.6 µm). Tensile testing showed F-scaffolds had a mean tensile strength of 0.192 ± 0.07 MPa and a mean elongation at break of 10.74 ± 2.53%, respectively. The degradation rate of the F-scaffolds showed ~19.2 ± 1.9% weight loss in 28 days. FE-SEM of BME-treated F-scaffolds seeded with keratinocytes showed deposition of extracellular matrix (ECM) components and the formation of cell sheets in 14 days. In addition, the in vitro cytocompatibility of BME-treated F-scaffolds with keratinocytes was analyzed using resazurin-based assay, which showed a time-dependent increase in metabolic activity up to culture day 21. Overall, this novel investigation shows that filamentous fungal mats with a nano/microfibrous mycelial architecture are potentially useful for tissue engineering applications.

Biocompatibility and hemocompatibility of hydrothermally derived reduced graphene oxide using soluble starch as a reducing agent.

Graphene-based nanomaterials (GBNs) have many applications as biomedical materials in tissue engineering and regenerative medicine. We report on the preparation of starch-(functionalized) reduced graphene oxide nanosheets (SRGO) using soluble starch as a reducing agent in a hydrothermal method, and their in vitro interactions with human skin fibroblasts and red blood cells (RBCs). Our results indicate that soluble SRGO nanosheets were prepared using graphene oxide (GO) as a raw material. SRGO-1 and -10, which were prepared using different concentrations of soluble starch after hydrothermal treatment, were characterized by ultraviolet-visible spectroscopy and showed a peak shift at 260 nm corresponding to the deoxygenation of GO and restoration of the conjugated aromatic structure. Dynamic light scattering and zeta potential measurements were used to determine Z-average sizes and surface charges of GO and SRGOs. X-ray diffractometry, attenuated total reflectance Fourier-transform infrared spectroscopy, and Raman spectroscopy revealed the progressive elimination of labile oxygen functional groups during hydrothermal treatment and restoration of the π-conjugated network. X-ray photoelectron spectroscopy showed de-oxidation of SRGOs, which had high carbon to oxygen ratios (C/O) as compared with GO. Interactions of SRGO-1 and -10 with skin fibroblasts showed excellent biocompatibility even at a concentration of 200 µg/ml with cell viabilities up to 88% and 90%, respectively, whereas notable cytotoxicity was observed for GO at 20 µg/ml. Similarly, SRGO-1 and -10 did not exhibit toxicity to RBCs compared to GO. Biofilm formation and metabolic activities of biofilm by the bacterium Staphylococcus aureus were also evaluated using a crystal violet and a tetrazolium reduction assay, respectively. The described hydrothermal method used to synthesize SRGO provides a cheap, facile, and environmentally friendly means of producing water-dispersible, biocompatible and hemocompatible reduced GOs for the fabrication of novel GBNs for various biomedical applications.

Biofabrication of Lysinibacillus sphaericus-reduced graphene oxide in three-dimensional polyacrylamide/carbon nanocomposite hydrogels for skin tissue engineering.

The biological synthesis of reduced graphene oxide (rGO) from graphene oxide (GO) is an emerging phenomenon for developing biocompatible nanomaterials for its potential applications in nanomedicine. In this study, we demonstrated a simple, green, and non-toxic method for graphene synthesis using the live biomass of Lysinibacillus sphaericus as the reducing and stabilizing agent under ambient conditions. Ultraviolet-visible spectroscopic analysis confirmed the formation of graphene from GO suspension. X-ray diffraction studies showed the disappearance of the GO peak and the appearance of characteristic graphene broad peak at 2θ = 22.8°. Infrared analysis showed the decrease/disappearance of peaks corresponding to the oxygen-containing functionalities, and appearance of a peak at 1620 cm-1 from unoxidized graphitic domains. Scanning electron microscopic images showed that L. sphaericus-reduced graphene oxide (L-rGO) contains aggregated graphene nanoflakes. Evaluation of the in vitro cytotoxicity of L-rGO nanosheets on human skin fibroblasts using the WST-1 assay did not show any significant effects after 24 h of exposure, which is indicative of biocompatibility. Polyacrylamide hydrogels with L-rGO were synthesized and used as scaffolds to support the growth and proliferation of skin fibroblasts. Cell viability assays and DAPI staining showed proliferation of fibroblasts and exhibited 83% of cell viability even after 28 days. Biofilm formation by Pseudomonas aeruginosa and Staphylococcus aureus was enhanced in nanocomposite hydrogels in the presence of 0.25 mg/mL GO and L-rGO in 48 h. Overall, this study showed that microbially-synthesized L-rGO can be used as a dopant in polymeric scaffolds for tissue engineering and highlighted their role in biofilm formation.

Recombinant helical plant virus-based nanoparticles for vaccination and immunotherapy.

Plant virus-based nanoparticles (PVNs) are self-assembled capsid proteins of plant viruses, and can be virus-like nanoparticles (VLPs) or virus nanoparticles (VNPs). Plant viruses showing helical capsid symmetry are used as a versatile platform for the presentation of multiple copies of well-arrayed immunogenic antigens of various disease pathogens. Helical PVNs are non-infectious, biocompatible, and naturally immunogenic, and thus, they are suitable antigen carriers for vaccine production and can trigger humoral and/or cellular immune responses. Furthermore, recombinant PVNs as vaccines and adjuvants can be expressed in prokaryotic and eukaryotic systems, and plant expression systems can be used to produce cost-effective antigenic peptides on the surfaces of recombinant helical PVNs. This review discusses various recombinant helical PVNs based on different plant viral capsid shells that have been developed as prophylactic and/or therapeutic vaccines against bacterial, viral, and protozoal diseases, and cancer.