l-Proline and GelMA hydrogel complex：An efficient antifreeze system for cell cryopreservation.

Cryopreservation of biological samples is an important technology for expanding their applications in the biomedical field. However, the quality and functionality of samples after rewarming are limited by the toxicity of commonly used cryoprotectant agents (CPAs). Here, we developed a novel preservation system by combining the natural amino acid l-proline (L-Pro) with gelatin methacryloyl (GelMA) hydrogels. Compared with dimethyl sulfoxide (DMSO), L-Pro and GelMA demonstrated excellent biocompatibility when co-culturing with cells. Cryopreservation procedures were optimized using 3T3 as model cells. The results showed that rapid cooling was the most suitable cooling procedure for L-Pro and GelMA among the three cooling procedures. Co-culturing with cells for 3 h before cryopreservation, 6 % L-Pro +7 % GelMA had the highest survival rate, reaching up to 80 %. Differential Scanning Calorimetry (DSC) analysis showed that 6 % L-Pro + 7 % GelMA lowered the freezing point of the solution to -4.2 °C and increased the unfrozen water content to 20 %. To the best of our knowledge, this is the first report of cell cryopreservation using a combination of L-Pro and GelMA hydrogels, which provides a new strategy for improving cell cryopreservation.

Trimeric and Tetrameric Cationic Styryl Dyes as Novel Fluorescence and CD Probes for ds-DNA and ds-RNA.

The wide use of mono- or bis-styryl fluorophores in biomedical applications prompted the presented design and study of a series of trimeric and tetrameric homo-analogues, styryl moieties arranged around a central aromatic core. The interactions with the most common biorelevant targets, ds-DNA and ds-RNA, were studied by a set of spectrophotometric methods (UV-VIS, fluorescence, circular dichroism, thermal denaturation). All studied dyes showed strong light absorption in the 350-420 nm range and strongly Stokes-shifted (+100-160 nm) emission with quantum yields (Φf) up to 0.57, whereby the mentioned properties were finely tuned by the type of the terminal cationic substituent and number of styryl components (tetramers being red-shifted in respect to trimers). All studied dyes strongly interacted with ds-DNA and ds-RNA with 1-10 nM-1 affinity, with dye emission being strongly quenched. The tetrameric analogues did not show any particular selectivity between ds-DNA or ds-RNA due to large size and consequent partial, non-selective insertion into DNA/RNA grooves. However, smaller trimeric styryl series showed size-dependent selective stabilization of ds-DNA vs. ds-RNA against thermal denaturation and highly selective or even specific recognition of several particular ds-DNA or ds-RNA structures by induced circular dichroism (ICD) bands. The chiral (ICD) selectivity was controlled by the size of a terminal cationic substituent. All dyes entered efficiently live human cells with negligible cytotoxic activity. Further prospects in the transfer of ICD-based selectivity into fluorescence-chiral methods (FDCD and CPL) is proposed, along with the development of new analogues with red-shifted absorbance properties.

Isolation and Characterization of Antimicrobial Peptides Isolated from Brevibacillus halotolerans 7WMA2 for the Activity Against Multidrug-Resistant Pathogens.

Multidrug resistance to pathogens has posed a severe threat to public health. The threat could be addressed by antimicrobial peptides (AMPs) with broad-spectrum suppression. In this study, Brevibacillus halotolerans 7WMA2, isolated from marine sediment, produced AMPs against Gram-positive and Gram-negative bacteria. The AMPs were precipitated by ammonium sulfate 30% (w/v) from culture broth and dialyzed by a 1 kDa membrane. Tryptone Soy Agar (TSA) was used for the cultivation and resulted in the largest bacteria-inhibiting zones under aerobic conditions at 25 °C, 48 h. An SDS-PAGE gel overlay test revealed that strain 7WMA2 could produce AMPs of 5-10 kDa and showed no degradation when held at 121 °C for 30 min at a wide pH 2-12 range. The AMPs did not cause toxicity to HeLa cells with concentrations up to 500 µg/mL while increasing the arbitrary unit up to eight times. The study showed that the AMPs produced were unique, with broad-spectrum antimicrobial ability.

Efficacy and non-toxicity of ciclopirox olamine-loaded liposomes against Cryptococcus neoformans clinical isolates.

The aim of this study was to evaluate the efficacy and non-toxicity of ciclopirox olamine-loaded liposomes against Cryptococcus neoformans clinical isolates. Initially, 24-1 fractional experimental design was carried out to obtain an optimized formulation of liposomes containing CPO (CPO-LipoC), which were then used to prepare stealth liposomes (CPO-LipoS). Liposomal formulations were characterized by their mean size diameter, polydispersity index (PDI), and drug encapsulation efficiency (EE%). Immunosuppressed mice were exposed to CPO-LipoS at 0.5 mg/kg/day for 14 days to verify possible histopathological alterations in the liver and kidneys. Immunosuppressed mice infected with C. neoformans were treated with CPO-LipoS at 0.5 mg/kg/day for 14 days to quantify the fungal burden in spleen, liver, lungs, and brain. CPO-LipoS presented a mean size diameter, PDI, and EE% of 101.4 ± 0.7 nm, 0.307, and 96.4 ± 0.9%, respectively. CPO-LipoS was non-toxic for the liver and kidneys of immunosuppressed mice. At the survival curve, all infected animals submitted to treatment with CPO-LipoS survived until the end of the experiment. Treatment with CPO-LipoS reduced C. neoformans cells in the spleen (59.3 ± 3.4%), liver (75.0 ± 3.6%), lungs (75.7 ± 6.7%), and brain (54.2 ± 3.2%). CPO-LipoS exhibit antifungal activity against C. neoformans, and the encapsulation of CPO into stealth liposomes allows its use as a systemic drug for treating cryptococcosis.

Facile Synthesis of Benzimidazoles via Oxidative Cyclization of Acyclic Monoterpene Aldehyde with Diamines: Studies on Antimicrobial and in Vivo Evaluation of Zebrafish.

Citral (1a), a bioactive component of Cymbopogon citratus (lemongrass) could be isolated and semi-synthetic analogs synthesized with improved therapeutic properties. Herein we first report describes citral (1a) as a primary material for the synthesis of benzimidazole derivatives between various o-phenylenediamines (2a-l) in the presence of Diisopropylethylamine (DIPEA) as a commercially available environmentally benign base, ethanol as a green solvent and the yield of all benzimidazole derivatives (3a-l) was between 68-76 %; The semi-synthetically prepared benzimidazole derivatives (3a-l) were assessed for their anti-bacterial and anti-fungal properties. The benzimidazole compounds (3a-b, and 3g-j) exhibit good anti-microbial activity. In addition, in silico study was carried out to determine the specific binding affinity of the diamine halogen substituted benzimidazole derivatives to the specific target proteins. In silico analysis revealed a high correlation between docking results and experimental results. Finally, benzimidazole demonstrated significant antibacterial and antifungal activity. Zebrafish embryos were subjected to In vivo toxicological test found that all of the benzimidazole compounds (3a-l) were non-toxic and had low embryotoxicity after 96 h, with an LC50 of 36.425 µg, which could facilitate the design of novel antimicrobial agents using a cost-effective method.

Eco-friendly cubic-ZnS coupled Cu7S4 spines on chitosan matrix: Unravelling defect-engineered nanoplatform for the photodegradation of p-chlorophenol.

Novel ZnS-Cu7S4 nanohybrid supported on chitosan matrix, as an ideal photocatalyst, was fabricated by the sonochemical method wherein high-resolution transmission electron microscopy (HRTEM) and X-ray powder diffraction (XRD) analysis confirmed the co-existence of both ZnS and Cu7S4; presence of vacancy sites in ZnS was verified by electron paramagnetic resonance (EPR) analysis and their introduction could promote two-photon excitation facilitated visible light response and charge transport/separation. The type II interface is formed in the ZnS-Cu7S4/Chitosan heterojunction owing to interstitial states that promote charge separation. The ZnS-Cu7S4/Chitosan was used for the photodegradation of a pharmaceutical pollutant, p-chlorophenol (PCP); over 98.8% of PCP photodegradation was achieved under visible-light irradiation where the ensued ·O2- and ·OH serve a key role in the photodegradation of PCP. In vitro cytotoxicity studies substantiated that the ZnS-Cu7S4/Chitosan is nontoxic to the ecosystem and human beings and endowed with promising photodegradation properties and accessibility via an environmentally friendly design, bodes well for its potential remediation applications.

Optimistic and possible contribution of nanomaterial on biomedical applications: A review.

Nanomaterials have many advantages over bulk materials, including enhanced surface-to-volume proportion as well as magnetic traits. It has been a steady rise in research with using nanomaterials in various biomedical fields in the past few decades. Constructing nanomaterials has emerged as a leading research primary concern in order to discover specialized biomedical applications. Since, their advantageous properties including chemical stability, non-toxicity, bio - compatibility, relatively high magnetization, and strong magnetic vulnerability, nanoparticles of iron oxide had already influenced implementations in different biomedical fields. Nanomaterials can be divided up into four nanomaterials such as metallic nanomaterials, bimetallic or alloy nanomaterials, metal oxide nanomaterials, as well as magnetic nanomaterials. Hence, the purpose of this review is to conduct such in discussion on emerging advancements in nanomaterials for biomedical, with such a special emphasis upon those options of nanomaterials including metallic nanomaterials: Au and Ag, bimetallic nanomaterials: Fe-Co and Fe-Pt, and metal oxides: TiO2 and CeO2. Securing this information gap will result in a better comprehension of the contribution of nanomaterial type and subsequent huge-scale applications in aspects of both their potential and challenges.

Chitosan crosslinking with a vanillin isomer toward self-healing hydrogels with antifungal activity.

The purpose of the study was to develop new antimicrobial hydrogels from natural resources that may promote wound healing and prevent bacterial skin infection. The new hydrogels were synthesized by crosslinking chitosan with a vanillin isomer, 5-methoxysalicylaldehyde, by a friendly and easy method. To characterize these hydrogels, their structural and morphological properties were explored by FTIR, 1H NMR, SEM, POM, and TGA. In view of the targeted application, swelling behavior, biodegradability, antimicrobial activity and biocompatibility were investigated in vitro. Structural and morphological studies confirmed the formation of new hydrogels via the imination reaction concomitant with the supramolecular organization. The hydrogels were highly porous with the average pore diameter around 80 µm, and a swelling rate controlled by the crosslinking density and medium pH. The hydrogels showed a progressive weight loss in the presence of lysozyme up to 35%, during 21 days of testing. They proved non-cytotoxic effect on Normal Human Dermal Fibroblasts using MTS test and powerful antifungal activity against Candida Albicans, as determined by disk diffusion assay. All these properties indicate the new hydrogels as a promising option for the treatment of various skin lesions.

An investigation of the antioxidant potential and bioaccumulated minerals in Tuber borchii and Tuber maculatum mycelia obtained by submerged fermentation.

Submerged fermentation of Tuber borchii and T. maculatum was performed to produce mycelia towards evaluating their bioactive potential and probable toxicity. Truffle mycelia had been mostly explored for biopolymers previously. Methanolic extraction yield, polyphenol content, and flavonoid content varied with species and period of fermentation. In vitro DPPH, FRAP, ABTS and ORAC radical scavenging activity of mycelial extracts of T. maculatum and T. borchii at 7 and 10 days of bioreactor fermentation corroborated with phenolics content. Absence of toxicity and antimutagenicity in lag and log phases in Saccharomyces cerevisiae D7 cells supported possibly safe nutraceutical use of mycelia of both truffles. Mineral content showed variation with species and fermentation time for macro- (calcium, potassium, phosphorus, sodium, iron, magnesium, and manganese) and trace elements (zinc, selenium, copper, chromium, and nickel). Mineral bioaccumulation can be explored for food-fortification applications. The demonstrated activities and detected constituents show potential applications in food, pharmaceutical, cosmetics and nutraceuticals.

Lignin Nanoparticles and Their Nanocomposites.

Lignin nanomaterials have emerged as a promising alternative to fossil-based chemicals and products for some potential added-value applications, which benefits from their structural diversity and biodegradability. This review elucidates a perspective in recent research on nanolignins and their nanocomposites. It summarizes the different nanolignin preparation methods, emphasizing anti-solvent precipitation, self-assembly and interfacial crosslinking. Also described are the preparation of various nanocomposites by the chemical modification of nanolignin and compounds with inorganic materials or polymers. Additionally, advances in numerous potential high-value applications, such as use in food packaging, biomedical, chemical engineering and biorefineries, are described.

Applications of Carbon Dots (CDs) in Latent Fingerprints Imaging.

Carbon dots (CDs), a new member of the carbon-based material family, possess unique properties, such as high fluorescence, non-toxicity, eco-friendliness, stability and cost-effectiveness. These properties helped CDs to receive tremendous attention in various fields, namely, biological, opto-electronic, bio-imaging and energy-related applications. Although CDs are widely explored in bio-imaging and bio-sensing applications, their effectiveness in forensic science and technology is comparatively new. In this review, applications of CDs pertaining to latent FPs recovery since 2015 to 2020 is summarized comprehensively.

Antibacterial, anti-biofilm and anti-quorum sensing activities of Artemisia dracunculus essential oil (EO): a study against Salmonella enterica serovar Typhimurium and Staphylococcus aureus.

The study evaluates the effect of Artemisia dracunculus essential oil (EO) on two pathogenic bacteria Salmonella enterica serovar Typhimurium and Staphylococcus aureus and Vero cell line. To evaluating the anti-biofilm potential of the EO, a microtiter-plate test (MtP) and scanning electron microscopy (SEM) were performed. The quorum-sensing inhibitory properties were examined by QS-related gene expression at sub-MIC concentrations of Artemisia dracunculus EO. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a tetrazole) test was used to determine the cytotoxicity potential of the EO against the Vero cell line and finally, the major components of the EOs were determined using Gas chromatography-mass spectrometry (GC-MS) analysis. The minimum inhibitory concentration (MIC) of the tested EO against S. Typhimurium and S. aureus were 2.5 and 1.25 µl/ml, respectively. In addition, the minimum bactericidal concentration was 5 and 2.5 µl/ml for S. Typhimurium and S. aureus, respectively. Both MtP and SEM showed an acceptable inhibitory and disruption effect of the EO on the biofilm formation at Sub-MIC concentrations. Significant downregulation of luxS, pfs, and hld genes by treatment with MIC/2 concentration of A. dracunculus EO was observed. The IC50 value of A. dracunculus EO against Vero cells was 20 µl/ml. The main detected compound using GC-MS was estragole (methyl chavicol or tarragon) (64.94%). Anti-biofilm, QSI activity, and non-toxicity of A. dracunculus EO reported for the first time in this study propose the use of these plant compounds as alternatives to antibiotics and chemical additives.

Target Activity of Isaria tenuipes (Hypocreales: Clavicipitaceae) Fungal Strains against Dengue Vector Aedes aegypti (Linn.) and Its Non-Target Activity Against Aquatic Predators.

The present investigation aimed to determine the fungal toxicity of Isaria tenuipes (My-It) against the dengue mosquito vector Aedes aegypti L. and its non-target impact against the aquatic predator Toxorhynchitessplendens. Lethal concentrations (LC50 and LC90) of My-It were observed in 2.27 and 2.93 log ppm dosages, respectively. The sub-lethal dosage (My-It-1 × 104 conidia/mL) displayed a significant oviposition deterrence index and also blocked the fecundity rate of dengue mosquitos in a dose-dependent manner. The level of major detoxifying enzymes, such as carboxylesterase (α-and ß-) and SOD, significantly declined in both third and fourth instar larvae at the maximum dosage of My-It 1 × 105 conidia/mL. However, the level of glutathione S-transferase (GST) and cytochrome P-450 (CYP450) declined steadily when the sub-lethal dosage was increased and attained maximum reduction in the enzyme level at the dosage of My-It (1 × 105 conidia/mL). Correspondingly, the gut-histology and photomicrography results made evident that My-It (1 × 105 conidia/mL) heavily damaged the internal gut cells and external physiology of the dengue larvae compared to the control. Moreover, the non-target toxicity against the beneficial predator revealed that My-It at the maximum dosage (1 × 1020 conidia/mL) was found to be less toxic with <45% larval toxicity against Tx.splendens. Thus, the present toxicological research on Isaria tenuipes showed that it is target-specific and a potential agent for managing medically threatening arthropods.

Is GSH Chelated Pt Molecule Inactive in Anti-Cancer Treatment? A Case Study of Pt6 GS4.

Platinum (Pt) drugs are widely used in anti-cancer treatment although many reports advocated that tumor cells could inactivate Pt drugs via glutathione-Pt (GSH-Pt) adducts formation. To date, GSH chelated Pt molecules have not been assessed in cancer treatment because GSH-Pt adducts are not capable of killing cancer cells, which is widely accepted and well followed. In this report, endogenous biothiol is utilized to precisely synthesize a GSH chelated Pt molecule (Pt6 GS4 ). This Pt6 GS4 molecule can be well taken up by aggressive triple negative breast cancer (TNBC) cells. Subsequently, its metabolites could enter nuclei to interact with DNA, finally the DNA-Pt complex triggers TNBC cell apoptosis via the p53 pathway. Impressively, high efficacy for anti-cancer treatment is achieved by Pt6 GS4 both in vitro and in vivo when compared with traditional first-line carboplatin in the same dosage. Compared with carboplatin, Pt6 GS4 keeps tumor bearing mice alive for a longer time and is non-toxic for the liver and kidneys. This work opens a route to explore polynuclear Pt compound with accurate architecture for enhancing therapeutic effects and reducing systemic toxicity.

In situ Fabrication of Nano ZnO/BCM Biocomposite Based on MA Modified Bacterial Cellulose Membrane for Antibacterial and Wound Healing.

BACKGROUND: Developing an ideal wound dressing that meets the multiple demands of safe and practical, good biocompatibility, superior mechanical property and excellent antibacterial activity is highly desirable for wound healing. Bacterial cellulose (BC) is one of such promising class of biopolymers since it can control wound exudates and can provide moist environment to a wound resulting in better wound healing. However, the lack of antibacterial activity has limited its application. METHODS AND RESULTS: We prepared a flexible dressing based on a bacterial cellulose membrane and then modified it by chemical crosslinking to prepare in situ synthesis of nZnO/BCM via a facile and eco-friendly approach. Scanning electron microscopy (SEM) results indicated that nZnO/BCM membranes were characterized by an ideal porous structure (pore size: 30~ 90 µm), forming a unique string-beaded morphology. The average water vapor transmission of nZnO/BCM was 2856.60 g/m2/day, which improved the moist environment of nZnO/BCM. ATR-FITR further confirmed the stepwise deposition of nano-zinc oxide. Tensile testing indicated that our nanocomposites were flexible, comfortable and resilient. Bacterial suspension assay and plate counting methods demonstrated that 5wt. % nZnO/BCM possessed excellent antibacterial activity against S.aureus and E. coli, while MTT assay demonstrated that they had no measurable cytotoxicity toward mammalian cells. Moreover, skin irritation test and histocompatibility examination supported that 5wt. % nZnO/BCM had no stimulation to skin and had acceptable biocompatibility with little infiltration of the inflammatory cells. Finally, by using a bacteria-infected (S. aureus and E. coli) murine wound model, we found that nZnO/BCM could prevent in vivo bacterial infections and promote wound healing via accelerating the re-epithelialization and wound contraction, and these membranes had no obvious toxicity toward normal tissues. CONCLUSION: Therefore, the constructed nZnO/BCM has great potential for biomedical applications as an efficient antibacterial wound dressing.

Kefiran, a branched polysaccharide: Preparation, properties and applications: A review.

Considering harmful effects of the environmental pollution and increased concerns about the synthetic polymers, kefiran has been introduced as a biodegradable polymer due to its nontoxicity. This edible biopolymer and its structural derivations play specific roles in a large number of applications including scaffolds, the reserve materials, and the encapsulation for drug delivery, thanks to its amazing properties such as the protective features, the bifidogenic effects and the antimicrobial activity against some microorganisms. The aim of the present review was to focus on the methods of producing kefiran, its properties and finally its applications, bringing significant changes in various fields.

A novel BiVO4-GO-TiO2-PANI composite for upgraded photocatalytic performance under visible light and its non-toxicity.

A novel non-toxic hybrid BiVO4-GO-TiO2-polyaniline (PANI) (BVGT-PANI) composite with superior photocatalysis was successfully prepared via a one-pot hydrothermal reaction. The structural and morphological characterizations of the synthesized compounds were analyzed by a series of techniques. We found excellent photocatalytic efficiencies for methylene blue (MB) and phenol degradation under visible light irradiation after adhering the PANI to the photocatalyst. The degradation rates of MB and phenol reach up to approximately 85% and 80%, respectively, after 3 h of irradiation. For photodegradation MB, BVGTA exhibit the highest kapp rate constant of about 1.06 × 10-2 min-1, which is about 1.63-fold faster than BVG and 2.94-fold faster than BVGT. For photodegradation of phenol, BVGTA exhibits the highest kapp rate constant, of about 8.86 × 10-3min-1, which is about 1.2-fold faster than BVG and 1.96-fold faster than BVGT. Furthermore, vitro toxicity test against Bacillus subtilis and Staphylococcus aureus demonstrated that the nanophotocatalyst is non-toxic.

Silver nanoparticles coated with dodecanethiol used as fillers in non-cytotoxic and antifungal PBAT surface based on nanocomposites.

In the present study, we report the preparation of antifungal and non-cytotoxic polymer nanocomposites with potential application in biomedical materials. Dodecanethiol-protected silver nanoparticles (AgNPs-DDT) were synthesized by a reduction/precipitation method and dispersed in chloroform to obtain stable colloidal dispersions. PBAT-based nanocomposites containing 0.25, 0.5 and 2 wt% AgNPs-DDT were prepared by casting method. The incorporation of AgNPs-DDT in PBAT matrix resulted in nanocomposites which combine improved mechanical performance and antifungal properties with a non-cytotoxic characteristic.

Liquid Type Nontoxic Photoluminescent Nanomaterials for High Color Quality White-Light-Emitting Diode.

High-brightness white-light-emitting diodes (w-LEDs) with excellent color quality is demonstrated by using nontoxic nanomaterials. Previously, we have reported the high color quality w-LEDs with heavy-metal phosphor and quantum dots (QDs), which may cause environmental hazards. In the present work, liquid-type white LEDs composed of nontoxic materials, named as graphene and porous silicon quantum dots are fabricated with a high color rendering index (CRI) value gain up to 95. The liquid-typed device structure possesses minimized surface temperature and 25% higher value of luminous efficiency as compare to dispensing-typed structure. Further, the as-prepared device is environment friendly and attributed to low toxicity. The low toxicity and high R9 (87) component values were conjectured to produce new or improve current methods toward bioimaging application.

PLGA Microspheres Loaded with ß-Cyclodextrin Complexes of Epigallocatechin-3-Gallate for the Anti-Inflammatory Properties in Activated Microglial Cells.

Although epigallocatechin-3-gallate (EG) is well-known as a potent antioxidant and free radical scavenger for neurodegenerative diseases, it still has disadvantages that reduce its treatment effectiveness due to low bioavailability, slow absorption, and water solubility. Therefore, the aim of this study is to improve the bioavailability of EG and increase the effectiveness of anti-inflammatory properties to microglial cells by using Poly(Lactide-co-Glycolide) (PLGA) microspheres as carriers. In this study, we used UV⁻Vis spectroscopy to show the formation of the complex of ß-cyclodextrin (ß-CD) and EG (CD-EG). The loading efficiency of EG in PLGA microspheres was optimized by the addition of ß-CD. The highest loading efficiency of 16.34% was found among other formulations. The results of Fourier transform infrared spectroscopy indicated the loading of CD-EG in PLGA microspheres. The scanning electron microscopic images demonstrated the spherical PLGA particles with controlled particles size ranging from 1⁻14 µm. Moreover, the in vitro release of EG was conducted to explore the sustained release property of the PLGA formulations. In the in vitro model of mouse microglial cells (BV-2 cells) stimulated by lipopolysaccharide, the cytotoxicity test showed that for up to 1 mg/mL of PLGA microspheres no toxicity to BV-2 cells was found. PLGA microspheres can significantly suppress the nitric oxide production from BV-2 cells, indicating EG loaded in PLGA microspheres can suppress the inflammation of activated microglial cells. Furthermore, the intracellular iNOS in BV-2 cells was also found to be down regulated. In summary, we have successfully shown that the use of ß-CD can increase the loading efficiency of EG in PLGA microspheres and provide neuroprotective effect on the activated microglial cells.