L929 Cell Line Research Articles

Structural, mechanical, and in-vitro characterization of hydroxyapatite loaded PLA composites

This article is intended to present an optimized Biocomposites material to prepare customized implants by using additive manufacturing technology. The most preferred implant material, polylactic acid (PLA) and hydroxyapatite (HAp), is chosen as matrix and reinforcements with different weight percentages of reinforcements. The 3D-printed composites underwent structural, mechanical, thermal stability, and in-vitro characterization. In the composites, the functional groups “OH, CH, CO, CO, and PO4” identified by Fourier transform infrared spectroscopy (FTIR). The thermal stability of the composites was identified by Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC). For the composition of PLA-HAp, a maximum tensile strength of 62 ± 0.5 MPa (30 wt%), a maximum compressive strength of 61 ± 0.5 MPa at 50 wt%), a maximum flexural strength of 46 ± 0.5 MPa (20 wt%), and a Shore-D hardness of 82 ± 0.5 SHN (50 wt%) were obtained. Modeled maximal principal stress in PLA and HAp. The 30 wt% of HAp-PLA composite carried out the in-vitro direct cytotoxicity, and it revealed 97% cell viability and 3% cytotoxicity through the L929 cell line. The results indicate that the addition of reinforcement increased the mechanical strength and cell visibility; therefore, the HAp-PLA composite would be used in biomedical applications.

Characterization and investigation of cytotoxicity and antimicrobial properties of coencapsulated limonene and thymol into the Ferula assafoetida gum microparticles

Thymol (Th) and d-limonene (L) exhibit low stability and are prone to oxidation when exposed to air, light, humidity, and high temperatures. This study examined the coencapsulation of Th and L into Ferula assafoetida gum (AFG) microparticles. Scanning electron microscope (SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and thermogravimetric analyzer (TGA) were done to characterize the obtained complexes. Furthermore, the encapsulation efficiency, antibacterial properties, cytotoxicity, and anticancer properties of both the free and encapsulated forms of L and Th were measured. For all samples, by increasing the percentage of bioactive compound (L, Th, and L-Th) from 2.5 to 5 % w/w, the EE was increased. FTIR and XRD analysis results demonstrated that Th and L were successfully incorporated into the AFG. Additionally, thermogravimetric analysis showed that in the thermal graphs of all samples, the first weight loss occurred between 30 °C and 160 °C, which was due to the evaporation of water. In the free L and Th graph, a sharp reduction peak was observed in which 80 % of compounds were lost. These reduction peaks disappeared in the thermal graphs of L: AFG and Th: AFG revealing that the thermal stability of Th and L was significantly increased upon their incorporation into the AFG. The inclusion of Th into the AFG also led to an increase in its antibacterial activity, while L exhibited acceptable antibacterial activity, albeit not as high as Th. Additionally, according to the MIC results, Th: AFG had the best antibacterial activity among all compounds, especially on gram-positive bacteria. According to the result of the MTT assay, there was a significant difference between the IC50 of free Th (123.4 μg/ml) and Th: AFG (2312 μg/ml), and free L (1762 μg/ml) and L: AFG (2480 μg/ml) showing that encapsulated Th and L into the AFG has decreased the cytotoxicity of free compounds against L929 cell line. Also, Th: AFG had the best anticancer activity against Hella and CT26 cell lines among all compounds. Finally, the flow cytometry analysis demonstrated that the encapsulated particles effectively eliminated cancer cells. The outcomes imply that AFG can be employed as a suitable delivery system to enhance the use of Th and L into the food and pharmaceutical industries.

Development and Validation of the Cell-based Functional Method for Neutralizing Anti-adalimumab Antibodies Detection in Human Serum

Introduction. Adalimumab, a fully humanized monoclonal antibody, is a tumor necrosis factor (TNFα) inactivator that is used against a number of autoimmune diseases such as rheumatoid arthritis and other most common inflammatory arthropathies (ankylosing spondylitis, psoriatic arthritis). Despite the proven efficacy of adalimumab treatment, there is a risk of adverse events, tied up with the formation of anti-drug antibodies, including neutralizing antibodies. Currently, the evaluation and characterization of neutralizing antibodies has become an important part of clinical trials in the development of new drugs and biosimilars.Aim. The aim of this study is to develop and validate the cell-based functional method for neutralizing anti-adalimumab antibodies determination in human serum.Materials and methods. For determination of neutralizing anti-adalimumab antibodies, the cell line L-929 has been employed. L-929 is a cell line sensitive to the TNFα-mediated apoptosis; the neutralizing antibodies interact with adalimumab that leads to TNFα-mediated cytotoxicity. Cytotoxicity was measured using resazurin, an aromatic compound that is a redox indicator.Results and discussion. The developed method was validated for cut point, selectivity, sensitivity, precision, specificity and stability (short- and long-term). An important part of a method development for determining neutralizing antibodies is the selection of concentrations of TNFα (4 ng/ml) and adalimumab (250 ng/ml), as well as determining the minimum required dilution – this parameter is established as 1 : 20. Cut point was chosen as a «floating» cut point, and a correction factor (normalization factor) was determined equal to 0,86. The sensitivity of the developed method was estimated at 108,9 ng/ml of neutralizing anti-adalimumab antibodies.Conclusion. The obtained results can be applied for determining anti-adalimumab neutralizing antibodies in the assessment of the adalimumab immunogenicity, including clinical trials.

Characterization of hFOB 1.19 Cell Line for Studying Zn-Based Degradable Metallic Biomaterials.

In vitro testing is the first important step in the development of new biomaterials. The human fetal osteoblast cell line hFOB 1.19 is a very promising cell model; however, there are vast discrepancies in cultivation protocols, especially in the cultivation temperature and the presence of the selection reagent, geneticin (G418). We intended to use hFOB 1.19 for the testing of Zn-based degradable metallic materials. However, the sensitivity of hFOB 1.19 to zinc ions has not yet been studied. Therefore, we compared the toxicity of zinc towards hFOB 1.19 under different conditions and compared it with that of the L929 mouse fibroblast cell line. We also tested the cytotoxicity of three types of Zn-based biomaterials in two types of media. The presence of G418 used as a selection reagent decreased the sensitivity of hFOB 1.19 to Zn2+. hFOB 1.19 cell line was more sensitive to Zn2+ at elevated (restrictive) temperatures. hFOB 1.19 cell line was less sensitive to Zn2+ than L929 cell line (both as ZnCl2 and extracts of alloys). Therefore, the appropriate cultivation conditions of hFOB 1.19 during biomaterial testing should be chosen with caution.

Biocompatibility and sub-chronic toxicity studies of phlorotannin/polycaprolactone coated trachea tube for advancing medical device applications

The phlorotannin-polycaprolactone-coated endotracheal tube (PP tube) has been developed with the aim of preventing tracheal stenosis that can result from endotracheal intubation, a factor that can lead to a serious airway obstruction. Its preventive efficacy has been assessed through both in vitro and in vivo investigations. However, there is a lack of studies concerning its biocompatibility and sub-chronic toxicity in animal models, a crucial factor to ensure the safety of its usage as a functional endotracheal tube. Thus, this study aimed to evaluate the biocompatibility and sub-chronic (13 weeks) toxicity of the PP tube through L929 cell line and diverse in vivo models. The cytotoxicity testing was performed using the extracts of PP tube on L929 cells for 72 h. Furthermore, other tests conducted on animal models, including ICR mice (acute systemic toxicity), New Zealand white rabbit (intradermal reactivity and pyrogen tests), guinea pig (maximization sensitization), and Sprague Dawley rats (sub-chronic toxicity). In both biocompatibility and sub-chronic toxicity analyses, no significant adverse effects are observed in the groups exposed to the PP tube, when compared to control group. Altogether, the findings suggested that the PP tube exhibits relative non-toxic and safety, supporting its suitability for clinical usage. However, extended periods of intubation may produce mild irritant responses, highlighting the clinical caution of limiting intubation duration to less than 13 weeks.

L-arginine–phosphatidylcholine thermoresponsive gel for enhanced transdermal delivery and L-929 cell cytotoxicity evaluation

The vasodilator effect of the semi-essential amino acid, L-arginine (LA) is crucial in the clinical management of angina pain and erectile dysfunction. Moreover, the limited bioavailability and the drug’s polar nature navigate the development of new formulation strategies for effective transdermal delivery, especially for the penile route. Thus, herein, we prepared the L-arginine–phosphatidylcholine (LA–PC) complex by solvent evaporation technique and converted it into a thermoresponsive gel (LA–PC gel). The optimized complex was selected for the preparation of gel using poloxamer 407, lactic acid, and purified water. Simultaneously we prepared the LA thermoresponsive gel (LA gel). The gels were characterized for their appearance, particle size, pH, osmolality, spreadability, SEM, FTIR, viscosity, TEM, viscoelastic properties, and skin permeability using the Strat-M® membrane at low pH for effective and safe penile and vaginal delivery. We observed that the LA–PC gel showed the smallest droplet size (<100 nm), a zeta potential of −59 mV indicates its stability with good viscoelastic behavior. The in vitro skin permeation reveals that; at 15 min, the permeation of LA for LA–PC gel, was 33.20% with a flux of 2.19 mg/cm2/min, and in the case of LA gel, the permeation of LA was only 18% at 45 minutes with a flux of 1.02 mg/cm2/min. The in vitro cytocompatibility was conducted using an L-929 cell line, and we found that both gels were nontoxic up to 100 µg/mL concentration. In summary, LA–PC gel could have a significant impact on the quick and sustained release, compared to LA gel.

Sprayable hydroxypropyl chitin/collagen extract of Ampelopsis brevipedunculata hydrogel accelerates wound healing.

Keeping a wound moist can allow effective and rapid healing, and it can control the formation of scabs, thereby allowing cell proliferation and epithelial formation. When regularly changing a dressing, thermosensitive hydrogel as a moist dressing does not cause a secondary wound from adhesion. The main aim of this study was to evaluate the effect of a new sprayable thermosensitive hydrogel on wound healing. The hydrophobic N-acetyl group of chitin was removed by microwave reaction with lye until the degree of acetylation was 60%, followed by reaction with propylene oxide to obtain hydroxypropyl chitin (HPCH) with a degree of substitution of 40%. After mixing HPCH with fish scale collagen (FSC), a thermosensitive hydrogel with a gel temperature of 26.5°C was obtained. Ampelopsis brevipedunculata extracts (ABE), which have been found to accelerate wound repair and improve healing, were added. HPCH/FSC is not toxic to the mouse L929 cell line and forms a hydrogel at body surface temperature. It can be easily sprayed on a wound. The HPCH/FSC has a three-dimensional network porous structure with a swelling ratio of 10.95:1 and a water vapour transmission rate of 2386.03±228.87g/m2/day; it can facilitate the penetration of water and air, and promote absorption of wound exudate. Wound repair was performed on five Sprague-Dawley rats. Each rat had three wounds, which were treated with medical gauze, HPCH/FSC and HPCH/FSC/ABE, respectively. The wounds in the HPCH/FSC/ABE group recovered the fastest in vivo, the mature wound site was smoother, the re-epithelialisation was even and thicker, and the angiogenesis developed rapidly to the mature stage. In this study, HPCH/FSC/ABE thermosensitive hydrogel was shown to effectively accelerate wound healing and was convenient for practical application.

Cytotoxicity of Laser-Synthesized Nanoparticles of Elemental Bismuth.

High X-ray absorption combined with photothermal properties make bismuth nanoparticles (Bi NP) a promising agent for multimodal cancer theranostics. However, the synthesis of Bi NP by the "classical" chemical methods has numerous limitations, including potential toxicity of the produced nanomaterials. Here we studied in vitro toxicity of laser-synthesized Bi NP coated with Pluronic F-127 on mouse fibroblast cell line L929. The survival of L929 cells decreased linearly with increasing the concentration of Bi NP in a concentration range of 3-500 μg/ml; the LC50 value was 57 μg/ml. The unique combination of functional properties and moderate toxicity of the laser-synthesized Bi NP makes them a new promising platform for sensitization of multimodal cancer theranostics.

Transcriptional Levels of Intercellular Junction Proteins in an Alveolar Epithelial Cell Line Exposed to Irradiation or Bleomycin.

We performed a comparative study of the effects of X-ray irradiation and bleomycin on the mRNA levels of E-cadherin and tight junction proteins (claudin-3, claudin-4, claudin-18, ZO-2, and occludin) in an alveolar epithelial cell line L2. Irradiation decreased claudin-4 levels and increased occludin levels, while the levels of other mRNAs remained unchanged. Bleomycin increased the expression levels of all proteins examined except claudin-3. Irradiation and bleomycin have different effects on the expression level of intercellular junction proteins, indicating different reactions triggered in alveolar epithelial cells and a great prospects of further comparative studies.

IN VITRO EVALUATION OF THE BIOLOGICAL ACTIVITY POTENTIAL OF ALOE VERA GEL: ANTIOXIDANT ACTIVITY AND CYTOTOXIC EFFECTS IN HEPG2 AND L929

Background: The place of natural and herbal products in alternative therapy is very important. A.vera is also an important plant that can be used in alternative therapy. Our study aimed to determine the antioxidant activity and cytotoxic effects of Aloe vera gel on HepG2 and L929 cell lines. Methods: The effects of A.vera on HepG2 and L929 were evaluated via the MTT analysis. The total antioxidant status and DPPH methods were used to determine the antioxidant capacity. Results: In our study, the mean cytotoxic effect on HepG2 of A.vera was determined to be 60.47 mg/mL at 24 hours and 44.64 mg/mL at 48 hours. In a healthy L929 fibroblast cell line, the best stimulating effect of A. vera was in 48 hours, and the EC50 value was calculated as 21.98±18.87 µg/ml. TAS was determined to be 0.2 mmol/L in A. vera. Free radical scavenging activity was determined at 21.58 mg/mL Conclusions: According to the obtained data, it was determined that A. vera gel has a concentration- and time-dependent cytotoxic effect on HepG2, while it has a proliferative effect on healthy cells. The fact that A. vera gel has antioxidant capacity, together with its positive effects on cells, shows that it can be an important support in treatments.

Metabolite profiling and in-vitro evaluation of cytotoxic efficacy of Psychotria dalzellii Hook.f. on MCF-7 and L929 cell lines

Metabolite profiling and in-vitro evaluation of cytotoxic efficacy of Psychotria dalzellii Hook.f. on MCF-7 and L929 cell lines

EVALUATION OF IN VITRO CYTOTOXIC POTENTIAL OF DIFFERENT CARMUSTINE FORMULATIONS AGAINST U-87 MG HUMAN GLIOBLASTOMA CELL LINE

Glioblastoma (GBM) is a frequent as well as violent type of brain tumor. In this research work, different carmustine formulations were prepared and screened for their cytotoxic activity against U-87 MG glioblastoma and normal human fibroblast L-929 cell lines. The flexible liposomes embedded in situ nasal gel exhibited maximum percentage of growth inhibition against U-87 MG glioblastoma cell line, and in situ nasal gel exhibited lowest percentage of growth inhibition against U-87 MG glioblastoma cell lines. Flexible liposomes embedded in situ nasal gel is observed to be safe and biocompatible against normal human fibroblast L-929 cell line. In cellular uptake study, U-87 MG cell line treated with flexible liposomes embedded in situ thermoreversible intranasal gel emitted stronger and higher intensity fluorescence. It seems that the presence of flexible liposomes embedded in situ thermoreversible nasal gel inside the tumor cells, and would be the best carmustine delivery approach for the management of GBM.

Green tea-mediated synthesis of silver nanoparticles: Enhanced anti-cancer activity and reduced cytotoxicity melanoma and normal murine cell lines

BackgroundGreen tea has been extensively studied as a therapeutic agent in various types of cancer, demonstrating its potential through its antioxidant, anti-aging, anti-collagen production and accumulation, anti-inflammatory, and immune-enhancing properties. Another promising field of research is the biosynthesis of silver nanoparticles (AgNPs), which are utilized for antimicrobial and anticancer therapies. In this study, we explore the green synthesis of AgNPs using an extract derived from Green tea. Additionally, we investigate the cytotoxic effects of these nanoparticles on both malignant murine melanoma cell lines (B16F10) and normal murine cell lines (L929). Material and methodsGreen tea extract-mediated natural synthesis of silver nanoparticles (AgNPs) was verified using UV–vis spectroscopy, DLS, TEM, and AFM techniques. The MTT assay was also used to study the cytotoxic effect of Green tea-AgNPs. The apoptosis rate was determined using anexin/PI and real-time PCR was performed for cellular and molecular levels. ResultsThe obtained results demonstrate the successful synthesis of uniformly distributed silver nanoparticles (AgNPs) using Green tea extract, yielding nanoparticles with an average size of 91 nm. MTT results showed Green tea-AgNPs reduced B16F10 cell viability with an IC50 of 20.2 but had no effect on L929 cell lines. Furthermore, Green tea-AgNPs enhanced the percentage of apoptosis in B16F10 but decreased the rate of apoptosis in L929 when compared to AgNPs. The BAX/BCL2 ratio increased after B16F10 was exposed to Green tea-AgNPs, however it reduced in L929. ConclusionThe findings of the study reveal that the Green tea-synthesized silver nanoparticles (Green tea-AgNPs) exhibit enhanced anti-cancer activity specifically against melanoma cells, while concurrently reducing the cytotoxic effects on normal cells. This suggests that the Green tea-AgNPs produced through this method hold great potential for further exploration and application in anti-cancer research.

In Vitro Release of Glycyrrhiza Glabra Extract by a Gel-Based Microneedle Patch for Psoriasis Treatment.

Microneedle patches are attractive drug delivery systems that give hope for treating skin disorders. In this study, to first fabricate a chitosan-based low-cost microneedle patch (MNP) using a CO2 laser cutter for in vitro purposes was tried and then the delivery and impact of Glycyrrhiza glabra extract (GgE) on the cell population by this microneedle was evaluated. Microscopic analysis, swelling, penetration, degradation, biocompatibility, and drug delivery were carried out to assess the patch's performance. DAPI staining and acridine orange (AO) staining were performed to evaluate cell numbers. Based on the results, the MNs were conical and sharp enough (diameter: 400-500 μm, height: 700-900 μm). They showed notable swelling (2 folds) during 5 min and good degradability during 30 min, which can be considered a burst release. The MNP showed no cytotoxicity against fibroblast cell line L929. It also demonstrated good potential for GgE delivery. The results from AO and DAPI staining approved the reduction in the cell population after GgE delivery. To sum up, the fabricated MNP can be a useful recommendation for lab-scale studies. In addition, a GgE-loaded MNP can be a good remedy for skin disorders in which cell proliferation needs to be controlled.

Anti-Inflammatory Potency of Human Wharton's Jelly Mesenchymal Stem Cell-Derived Exosomes on L2 Cell Line Induced by Lipopolysaccharides.

At present, therapeutic interventions to treat acute lung injury (ALI) remain largely limited to lung-protective strategies, as no real molecular-driven therapeutic intervention has yet become available. The administration of bacterial lipopolysaccharides (LPS) is known as an inflammatory activator, representing a frequently used model of ALI. This study investigated the biological function of normoxic (21% O2 ) vs. hypoxic conditions (5% O2 ) obtained from human Wharton's Jelly mesenchymal stem cells (hWJ-MSCs) and discovered that exosomes have the ability to suppress inflammatory responses by specifically targeting TNF-α, IL-1β, IL-6. and identify the toll-like receptor 4 (TLR4) NF-κβ gene expression. Primer culture hWJ-MSCs characterization with trilineage differentiation and CD markers was conducted. To obtain exosomes, hWJ-MSCs were stimulated with two different oxygen levels: 21% (nor-exo) and 5% (hypo-exo). Then, the L2 cell line was induced with LPS 1 µg/mL. Inflamed-L2 was treated with nor-exo, hypo-exo, and dexamethasone as a positive control. The RNA extracted from treated L2 cells was utilized to examine the gene expression profiles of TLR4 and NF-κβ, and the medium was used to measure tumor necrosis factor α (TNF-α), interleukin (IL)-1β, and IL-6 levels using ELISA. Lastly, proteomic analysis of the exosome using LC/MS-MS was conducted. Nor-exo and hypo-exo can be characterized and can produce higher yields exosomes under hypoxic conditions. The expression of TLR4 and NF-κβ genes and the proinflammatory levels such as IL-6, IL-1β, and TNF-α levels in nor-exo and hypo-exo treatments decreased. Nor-exo and hypo-exo derived from hWJ-MSCs were proven to have anti-inflammatory activities.

Association of Valproic Acid and Its Main Metabolites' Plasma Concentrations with Clinical Outcomes among Epilepsy Patients: A 10-Year Retrospective Study Based on Therapeutic Drug Monitoring.

Valproic acid (VPA) is a first-line antiepileptic drug with broad efficacy. Due to significant individual differences in its metabolism, therapeutic drug monitoring is commonly used. However, the recommended therapeutic range (50-100 μg/mL) is inadequate for predicting clinical outcomes. Additionally, the relationship between VPA metabolites and clinical outcomes remains unclear. In this retrospective study, 485 Chinese Southern Han epilepsy patients receiving VPA monotherapy were analyzed after reaching steady-state levels. Plasma concentrations of VPA and its five main metabolites were determined by liquid chromatography-mass spectrometry (LC-MS). We assessed the relevance of the recommended therapeutic VPA range for clinical outcomes and explored the association between VPA/metabolites levels and treatment efficacy/adverse effects. Vitro experiments were conducted to assess 4-ene-VPA hepatotoxicity. The therapeutic range of VPA exhibited no significant correlation with clinical outcomes, and plasma concentrations of VPA failed to serve as predictive indicators for treatment response/adverse effects. Treatment responders had higher 2-PGA concentrations (median, 26.39 ng/mL versus 13.68 ng/mL), with a threshold of 36.5 ng/mL for optimal epilepsy treatment. Patients with abnormal liver function had a higher 4-ene-VPA median concentration (6.41 μg/mL versus 4.83 μg/mL), and the ratio of 4-ene-VPA to VPA better predicted VPA-induced hepatotoxicity (area under the curve, 0.718) than 4-ene-VPA concentration. Vitro experiments revealed that 4-ene-VPA was more hepatotoxic than VPA in HepaRG and L02 cell lines. Total plasma VPA concentration does not serve as a predictor of clinical outcomes. 2-PGA concentrations may be associated with efficacy, whereas the ratio of 4-ene-VPA to VPA may be considered a better biomarker (threshold 10.03%) for VPA-induced hepatotoxicity. SIGNIFICANCE STATEMENT: This was the first and largest observational cohort in China to explore the relationship between patients' parent and metabolites concentrations of VPA and clinical outcomes during the maintenance of VPA monotherapy in epileptic patients. This study provided feasible references of VPA for epilepsy clinical treatment with a larger sample of patients compared with previous studies for a more definitive conclusion based on real-world situations. We found two potential biomarkers in predicting efficacy and liver injury, respectively. This breakthrough has the potential to assist in the rational use of VPA.

Enhanced antimicrobial efficacy of chlorhexidine-encapsulated halloysite nanotubes incorporated in presurgical orthopedic appliances: an in vitro, controlled study.

Presurgical infant's orthopedic appliances (PSIOs) play an increasingly crucial role in the interdisciplinary management of neonatal CLP, aiming to improve and maintain adequate nasolabial aesthetics, followed by primary lip/nasal surgery in both unilateral and bilateral CLP cases. The use of PSIOs in cleft lip and palate patients can lead to contamination with oral microflora, acting as a potential reservoir for infectious microorganisms. Acrylic surfaces might provide retention niches for microorganisms to adhere, and inhabit, which is difficult to control in immunocompromised patients, thus predisposing them to increased infection risks. The objective of this multi-assay in vitro study was to investigate the effects of incorporating chlorhexidine-loaded halloysite nanotubes (CHX-HNTs) fillers on the morphological, cytotoxic, release, and antimicrobial characteristics of self-cured acrylic polymethyl methacrylate (PMMA) material used in pre-surgical orthopedic appliances. Disk-shaped PMMA specimens were prepared with varying proportions of CHX-HNTs. A control group without any addition served as a reference, and four experimental samples contained a range of different concentrations of CHX-HNTs (1.0, 1.5, 3, and 4.5 wt%). The antimicrobial efficacy was assessed using an agar diffusion test against common reference microorganisms: Candida albicans, Staphylococcus aureus, Streptococcus pneumoniae, and Streptococcus agalactiae. Cytotoxicity was examined using the L929 cell line (mouse fibroblasts) through a (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide, MTT) cell viability assay. The release kinetics of CHX were monitored using UV-spectral measurements. The statistical analysis used a one-way ANOVA followed by Tukey's post hoc test. The integration of CHX-HNTs in PMMA exhibited a substantial dose-dependent antifungal and antibacterial effect against microorganisms at tested mass fractions (1.0 to 4.5 wt%). CHX release was sustained for up to 60 days, supporting prolonged antimicrobial activity. Furthermore, no significant cytotoxicity was determined in the L929 fibroblast cell line (control), indicating the biocompatibility of the CHX-HNTs-enhanced PMMA. Incorporating CHX-HNTs in PMMA successfully enhanced its antimicrobial properties, providing sustained CHX release and superior antimicrobial efficacy. These findings demonstrate the potential of antimicrobial nanoparticles in dental therapies to improve therapeutic outcomes. However, rigorous further clinical trials and observational studies are warranted to validate the practical application, safety, and efficacy. This study has the potential to make a major impact on the health of infants born with cleft lip and palate by helping to reduce the prevalence of infectious illnesses. The incorporation of CHX-HNTs into PMMA-based appliances is a novel promising preventive approach to reduce microbial infections.

Electrospun nanofibers combined with Fe-doped mesoporous bioactive glass nanoparticles induce in vitro melanoma cell death

Iron (Fe) doped-mesoporous bioactive glasses are among the most promising candidates in cancer therapy; the present study reports the successful fabrication and characterization of a series of anticancer and pro-regenerative nanocomposites of polycaprolactone and gelatin nanofibers combined with Fe-doped mesoporous bioactive glasses. The anticancer potential and regenerative capacity of the prepared nanocomposite were studied on melanoma (A375 cell line) and normal fibroblast cells (L929 cell line) by using the MTT assay, annexin V/PI apoptosis assay, intracellular reactive oxygen species (ROS) generation, malondialdehyde assay, as well as adhesion and migration assessments. The characterization of the nanofibers containing 20 % (wt.%) glass particles revealed a suitable tensile strength (5.8 MPa), controlled swelling (355.5 % after 24 hours), and degradability (31.33 % after 28 days). The viability of A375 cells was significantly reduced (up to 77 %) after treatment with the 20 % glass-impregnated nanofibers along with 58 % and 8 % increase in their apoptosis and necrosis, respectively. Interestingly, this nanocomposite showed no significant adverse effects on normal fibroblasts. In conclusion, this study suggests the possibility of the bioactive glass nanoparticles/electrospun nanofiber composites in the treatment of cancers (e.g., melanoma) as well as in providing a suitable microenvironment for tissue repair and regeneration. However, performing in vivo studies is required to provide clear evidence of the anticancer potential of this nanocomposite system.

Effect of cold atmospheric plasma on changing of biomolecular structures involved in apoptosis pathways of melanoma cancer.

Cold atmospheric plasma (CAP), is a technology based on non-thermal ionized gas that is used for cancer therapy in research. We evaluated the effect of CAP on malignant melanoma cancer cell line (B16) in comparison with normal cells (L929). The effect of CAP on the cytotoxicity of B16 and L929 cell lines was assayed by the MTT method and inverted microscopy. The induction of apoptosis in cells was evaluated using a fluorescence microscope. FTIR monitored the CAP effect in biomacromolecules changes in these cell lines. QPCR assayed gene expression of BAX, BCL-2, and Caspase-3 (CASP-3). The results of the MTT test showed CAP has a cytotoxic effect on the B16 cancer cell line more than L929 normal cells (p<0.0001). The results of invert and fluorescence microscopy showed CAP-induced apoptotic morphology on cancerous cells. FTIR spectroscopy indicated CAP changes biomacromolecules structure. Evaluation of gene expression showed CAP increased BAX and CASP-3 gene expression. Also, it decreased BCL-2 gene expression. Taken together, CAP may change biomacromolecule structures involved in apoptosis pathways, decrease proliferation and induce apoptosis in cancer cells.

Design and characterization of β-tricalcium phosphate-based self-passivating coatings on magnesium alloys.

Background: Magnesium alloys degrade rapidly in salt solutions, which limits their use without passivating treatments. AZ31 alloy is particularly promising for implant applications owing to its biodegradability and mechanical properties, necessitating effective corrosion-resistant coatings. Aim: In this study, a self-passivating reactive coating was designed and evaluated for AZ31 magnesium alloy plates using β-tricalcium phosphate (TCP) to enhance corrosion resistance and biocompatibility. Methods: Solutions of TCP, trisodium citrate, magnesium nitrate, hydroxyethyl cellulose (HEC), and sodium chloride were used to dip-coat AZ31 plates. The coated samples were immersed in 3.5 wt% NaCl solution. Phase evolution was analysed using gravimetry, X-ray diffraction (XRD), energy-dispersive X-ray (EDX) spectroscopy, and scanning electron microscopy (SEM). The biological response of the coated samples was evaluated through MTT and resazurin assays. Results: The coating formed a stable TCP/HEC layer that gradually dissolved over two weeks, converting the surface to magnesium hydroxide, magnesium oxychloride, and magnesium phosphate phases. The formation of brucite, responsible for passivation in the long term, was observed. The coating effectively prevented excessive magnesium oxychloride formation and stabilised magnesium hydroxide after one week. Biological characterization indicated that the coating on AZ31 is safe on the Saos-2 and L929 cell lines. Conclusion: The TCP-based coating enhances the corrosion resistance of AZ31 alloy in salt solutions, promoting passivating phases and limiting corrosive products, thereby ameliorating biocompatibility issues. This coating demonstrates substantial potential for extending the longevity and functionality of magnesium alloy implants.