Frontiers in bioscience (Elite edition)最新文献

Background: Regenerative endodontics requires an innovative delivery system to release antibiotics/growth factors in a sequential trend. This study focuses on developing/characterizing a thermoresponsive core-shell hydrogel designed for targeted drug delivery in endodontics.

Methods: The core-shell chitosan-alginate microparticles were prepared by electrospraying to deliver bone morphogenic protein-2 for 14 days and transforming growth factor-beta 1 (TGF-β1) for 7-14 days. Methylcellulose (MC) and gelatin were utilized to create the core-shell hydrogel to load a modified triple antibiotic combination (penicillin G/metronidazole/ciprofloxacin (PMC)) and growth factor-loaded microparticles in the shell and the core compartments, respectively. Morphological assessment, core-shell structural analysis, FTIR analysis, rheological analysis, swelling, and degradation rate studies were conducted for characterization. The viability of dental pulp stem cells (DPSCs) upon antibiotic exposure, antibacterial activity, and release studies of PMC and growth factors were investigated. Cellular studies (cell viability, alkaline phosphatase (ALP) activity, osteo/odontoblast gene expression (using Reverse transcription-polymerase chain reaction (RT-PCR)) and in vivo studies (inflammatory response and differentiation potential of the developed hydrogel by subcutaneous implantation in rats via histological examination) were assessed.

Results: The hydrogel showed a porous microstructure with interconnected pores. Core-shell structure analysis confirmed the successful extrusion of the MC hydrogel to the surface. FTIR analysis revealed interactions between MC and gelatin. Rheological analysis indicated time-dependent gel formation, supporting thermosensitivity at 37 °C. Swelling occurred rapidly, and degradation reached 62.42% on day 45. Further, antibiotics exhibited no cytotoxicity on DPSCs. Sequential release of antibiotics and growth factors was observed for up to 5 and 14 d, respectively. The hydrogel showed antibacterial activity. DPSCs exhibited increased proliferation, ALP activity, and odontoblast gene expression. In vivo studies showed that the biocompatible drug-loaded hydrogel exhibited more mineralization than the control.

Conclusions: The developed core-shell hydrogel containing PMC and growth factor-loaded core-shell microparticles provided a versatile and biocompatible platform for sequential drug delivery in regenerative endodontics. The system demonstrates promising characteristics for dentin regeneration, making it a potential candidate for clinical applications.

Background: Contamination with crude oil and hydrocarbons has become a global threat. Such threats have urged us to invent solutions to deal with this dilemma. However, chemical treatment comes with limited benefits. The use of bioremediation proved to be the optimal approach to face this problem since it is cost-effective, time-conserving and may improve the quality of soil and increase its fertility.

Methods: Soil samples were collected from three sites with the highest degree of pollution in Iraq: Al-Latifia, Al-Begei, and Basra. These were the source of novel Streptomyces isolates that could degrade contaminants and be used as a source of nutrients. The isolation principle was the degradation of petroleum in these soils as a carbon source.

Results: The most efficient isolate was obtained from Basra soil, characterized by the highest degree of contamination. The bacterium grows on minimal medium with crude oil, diesel fuel, aromatic, and non-aromatic hydrocarbons as the only source of carbon and showed the ability to reduce a hydrocarbon mixture containing 23 analytes with C8-C40 and C13-C30 representing total crude hydrocarbons effectively and with a high mineralization capability reaching 84%. This isolate was found to produce biosurfactants and reduce surface tension significantly.

Conclusions: Using chemical treatment of crude oil contaminants, burying the contaminated areas, or sometimes flooding them with water to eliminate these contaminants may not be an effective solution. Thus, using nonpathogenic microorganisms, especially those indigenous to the soil, maybe the most effective solution to achieve this goal.

Flavonoids occur naturally in different types of fruits and vegetables, including tea, cabbage, cauliflower, elderberries, cranberries, red apples, lettuce, pears, spinach, green hot peppers, white and red onions, kale, blueberries, and nuts. Among these flavonoids is quercetin, a potent natural antioxidant and cytotoxic substance with a number of therapeutic functions. Nowadays, quercetin is a common ingredient in many nutraceutical and cosmeceutical products due to its antioxidant properties. Its antibacterial effects and possible action mechanisms have been explored in many studies. From these, it has been established that quercetin stops the activity of numerous Gram-negative and -positive bacteria, fungi, and viruses. This review clarifies the plant sources and extraction methods of quercetin, as well as its medicinal applications as an antibacterial, antifungal, antiviral, and antioxidant agent, with a particular emphasis on the underlying mechanisms of its biological activity. The mechanism of its antimicrobial effect involves damaging the cell membrane-e.g., by changing its permeability, preventing biofilm formation, reducing the mitochondrial expression of virulence factors, and inhibiting protein and nucleic-acid synthesis. Moreover, quercetin has been shown to impede the activity of a variety of drug-resistant bacterial strains, pointing to the possibility of using it as a strong antimicrobial substance against such strains. In addition, it has occasionally been demonstrated that specific structural alterations to quercetin can increase its antibacterial action in comparison to the parent molecule. Overall, this review synthesizes our understanding of the mode of action of quercetin and its prospects for use as a therapeutic material.

Background: Azo pigments are widely used in the textile and leather industry, and they generate diverse contaminants (mainly in wastewater effluents) that affect biological systems, the rhizosphere community, and the natural activities of certain species.

Methods: This review was performed according to the Systematic Reviews and Meta Analyses (PRISMA) methodology.

Results: In the last decade, the use of Streptomyces species as biological azo-degraders has increased, and these bacteria are mainly isolated from mangroves, dye-contaminated soil, and marine sediments. Azo pigments such as acid orange, indigo carmine, Congo red, and Evans blue are the most studied compounds for degradation, and Streptomyces produces extracellular enzymes such as peroxidase, laccase, and azo reductase. These enzymes cleave the molecule through asymmetric cleavage, followed by oxidative cleavage, desulfonation, deamination, and demethylation. Typically, some lignin-derived and phenolic compounds are used as mediators to improve enzyme activity. The degradation process generates diverse compounds, the majority of which are toxic to human cells and, in some cases, can improve the germination process in some horticulture plants.

Conclusions: Future research should include analytical methods to detect all of the molecules that are generated in degradation processes to determine the involved reactions. Moreover, future studies should delve into consortium studies to improve degradation efficiency and observe the relationship between microorganisms to generate scale-up biotechnological applications in the wastewater treatment industry.

While monoclonal antibodies have shown success in cancer immunotherapy, their limitations prompt exploration of alternative approaches such as aptamers and peptides targeting programmed death ligand 1 (PD-L1). Despite the significance of these biotechnological tools, a comprehensive review encompassing both aptamers and peptides for PD-L1 targeting is lacking. Addressing this gap is crucial for consolidating recent advancements and insights in this field. Biotechnological advances leveraging aptamers and peptides represent a cutting-edge approach in refining the targeting proteins. Our review aims to provide valuable guidance for researchers and clinicians, highlighting the biotechnological advances utilizing aptamers and peptides refining PD-L1 targeting.

Background: A pivotal objective in crop production and plant protection lies in developing environmentally friendly insecticidal preparations and biostimulants.

Methods: We employed Bacillus thuringiensis strains with varied insecticidal spectra and engineered melanogenic mutants.

Results: We demonstrated a significant increase in insecticidal activity in the isolated mutants. Meanwhile, there was no observable impact of the enhanced synthesis of water-soluble melanin on the nature and abundance of spore and crystal formation. This heightened efficacy can be attributed to the photoprotective qualities of the synthesized pigment, shielding spores and crystals against the detrimental effects of UV radiation and insolation. We demonstrated the high biological activity of water-soluble bacterial melanin through in vivo experiments involving multiple plant species.

Conclusions: Our findings indicate that bacterial melanin is a potent phytostimulant. This preparation accelerates and amplifies plant growth and development processes, leading to a substantial increase in crop yield by 20-40%. The simultaneous synthesis of two biologically active substance, melanin and insecticidal toxins, ensures an elevated level of effectiveness in utilizing melaninogenic strains.

Background: Monoclonal antibodies (mAbs) are pioneers in the diagnosis and treatment of many diseases, such as cancer, asthma, poisoning, viral infections, etc. As the market value of mAbs increases in the biopharma industry, the demand for high quantities is met by upscaled production using bioreactor systems. Thus, disposable, porous matrices called cryogels have gained the primary focus for adherent support in the proliferation of hybridoma cells.

Methods: In this study, a gelatin-immobilized polyhydroxyethylmethacrylate-based cryogel material (disc-shaped, 9 mL bed volume) was synthesized, and a mini-bioreactor set up developed for culturing hybridoma cells to produce mAbs continuously. The hybridoma clone, 1B4A2D5, secreting anti-human serum albumin monoclonal antibodies, was immobilized in the cryogel matrix (2 discs, 18 mL bed volume).

Results: The hybridoma cells were attached to the matrix within 12 h after inoculation, and the cells were in the lag phase for seven days, where they were secreted mAb into the circulation medium. During the initial exponential phase, the glucose consumption, lactic acid production, and mAb production were 3.36 mM/day, 3.67 mM/day, and 55.61 µg/mL/day, respectively. The medium was refreshed whenever the glucose in the media went below 50% of the initial glucose concentration. The cryogenic reactor was run continuously for 25 days, and the mAb concentration reached a maximum on the 17th day at 310.59 µg/mL.

Conclusion: The cumulative amount of mAbs produced in 25 days of running was 246 µg/mL, 7.7 times higher than the mAbs produced from T-flask batch cultivation. These results demonstrate that the developed polyhydroxyethylmethacrylate-based cryogel reactor can be used efficiently for continuous mAb production.

Background: Despite attempts to control malaria, poor drug bioavailability means malaria still places enormous pressure on health globally. It has been found that the solubility of highly lipophilic compounds can be enhanced through lipid formulations, e.g., self-emulsifying drug delivery systems (SEDDSs). Thus, quality-by-design and characterization were used to justify the development and determine the feasibility of oral oil-in-water SEDDSs comprising a fixed-dose combination (FDC) of artemether-lumefantrine to treat malaria more effectively without the aid of a fatty meal. These formulations were compared to a commercial product containing the same active compounds.

Methods: Excipient compatibility and spontaneous emulsification capacity of different FDC-excipient combinations were identified by employing isothermal microcalorimetry, solubility, and water titration tests. Pseudoternary phase diagrams were constructed, and checkpoint formulations were selected within the self-emulsification region by reviewing formulation properties essential for optimized drug delivery. SEDDSs capable of enduring phase separation within 24 h were subjected to characterization experiments, i.e., drug concentration determination, cloud point, droplet size, size distribution, self-emulsification time, self-emulsification efficacy, viscosity, zeta potential, and thermodynamic stability analysis. SEDDSs with favorable characteristics were identified in the micro or nano range (SNEDDSs) before being subjected to drug release studies.

Results: All final formulations depicted enhanced artemether and lumefantrine release compared to the commercial product, which could not release lumefantrine at a quantifiable concentration in this study. The avocado oil (AVO)4:6 and olive oil (OLV)3:7 SNEDDSs overall portrayed the ideal characteristics and depicted the highest percentage of drug release.

Conclusions: This study offers evidence that SNEDDSs from selected natural oils comprising an artemether-lumefantrine FDC can potentially enhance the bioavailability of these lipophilic drugs.

Background: This research explores the significance of miR-215-5p and vasculogenic mimicry (VM) in forecasting the prognosis for hepatocellular carcinoma (HCC).

Methods: We analyzed HCC-associated miRNA expression profiles using data from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO). Samples included tissue and blood from 80 early-stage HCC patients and serum from 120 healthy individuals. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was employed to measure miR-215-5p and zinc finger E-box binding homeobox 2 (ZEB2) gene expressions. Hematoxylin and eosin (H&E) and CD34/Periodic Acid-Schiff (PAS) double staining assessed VM presence in HCC tissue sections. Bioinformatics tools predicted interactions between miR-215-5p and ZEB2, confirmed through luciferase reporter assays. We also examined the impact of miR-215-5p or ZEB2 overexpression on HCC cell invasion, migration, and VM formation using scratch, Transwell invasion assays, and Matrigel 3D cultures.

Results: Bioinformatics analysis indicated that miR-215-5p was under-expressed in HCC, particularly in cases with vascular invasion, which correlated with worse patient outcomes. In contrast, ZEB2, targeted by miR-215-5p, was overexpressed in HCC. RT-qPCR validated these expression patterns in HCC tissues. Among the HCC patients, 38 were VM positive and 42 VM negative. Logistic regression highlighted a negative correlation between miR-215-5p levels and VM positivity in HCC tissues and a positive correlation for ZEB2 with VM positivity and tumor vascular invasion. Lower miR-215-5p levels were linked to increased HCC recurrence and metastasis. Both bioinformatics analysis and luciferase assays demonstrated a direct interaction between miR-215-5p and ZEB2. Enhancing miR-215-5p levels reduced ZEB2 expression, consequently diminishing invasion, migration, and VM formation of the HCC cells in vitro.

Conclusions: miR-215-5p expression inversely correlates with VM occurrence in HCC tissues, while ZEB2 expression shows a direct correlation. By targeting ZEB2, miR-215-5p may hinder VM in HCC tissues, helping to prevent vascular invasion and HCC recurrence. Thus, miR-215-5p emerges as a vital prognostic indicator for predicting vascular invasion and recurrence in HCC.

Elevated concentrations of toxic organic compounds observed in food products pose serious dangers to human health. Both natural and artificial pollutants can cause food contamination. The stages of food production, packaging, transportation, and storage can also largely cause the appearance of undesirable substances in food products. The health consequences of ingesting food containing toxic contaminants range from mild gastroenteritis to deaths resulting from dysfunctional internal organs and neurological syndromes. The World Health Organization (WHO) sets recommendations for the content of such chemicals in food, including a minimum allowable concentration considered safe for human consumption. However, the control of food products from chemical pollutants is necessary. Moreover, fast, sensitive, and inexpensive methods are needed to detect them at the point of need. Currently, immune analysis methods are most widely used to determine pollutants in food. The development of fluorescence polarization immunoassay (FPIA) methods in a competitive format is a powerful and modern tool for detecting organic molecules in various matrices, thereby making FPIA methods useful for food safety applications. Due to the availability of portable devices for measuring the fluorescence polarization signal, FPIA methods can be used at the point of need. The variety of fluorescent labels and recognizing elements (receptors, monoclonal and polyclonal antibodies, and nanobodies) permits fluorescence polarization (FP) assays to detect significantly lower limits of organic substances. The FP assay is a homogeneous, fast, and quantitative method. The development of various formats of FP assays makes them promising in determining food pollutants. This review summarizes publications on FP analyses for detecting organic contaminants (pesticides, hormones, toxins, antibiotics, and other pharmaceuticals) in food products during 2018-2023. Further, it demonstrates the prospects for using this method to determine pollutants at the point of need and for detecting high molecular weight substances, fungi, and bacterial infections during food safety inspections.