Cell Viability Values Research Articles

GATA1 activates HSD17B6 to improve efficiency of cisplatin in lung adenocarcinoma via DNA damage

BackgroundLung adenocarcinoma (LUAD) is the most common histological type of non-small cell lung cancer (NSCLC). Platinum-based chemotherapy, such as cisplatin chemotherapy, is the cornerstone of treatment for LUAD patients. Nevertheless, cisplatin resistance remains the key obstacle to LUAD treatment, for its mechanism has not been fully elucidated.MethodsHSD17B6 mRNA expression data were accessed from TCGA-LUAD database and differential expression analysis was performed. Enrichment analysis of HSD17B6 was conducted by GSEA, and its upstream transcription factors were predicted by hTFtarget. mRNA and protein expression levels of HSD17B6 and GATA1 were assayed by qRT-PCR and WB, and the binding relationship between them was verified by chromatin immunoprecipitation assay and dual luciferase reporter assay. Cell viability and IC50 value of cisplatin-treated cells were measured by cell counting kit-8 assay. Cell cycle was assayed by flow cytometry. DNA damage level and DNA damage marker γ-H2AX expression were assayed by comet assay and western blot, respectively.ResultsHSD17B6 was lowly expressed in LUAD tissues and cells and mainly enriched in homologous recombination and mismatch repair pathways. As cell function experiments revealed, overexpression of HSD17B suppressed malignant phenotypes and cisplatin resistance in LUAD cells through DNA damage. Bioinformatics analysis revealed that GATA1 is the upstream regulator of HSD17B6, which was markedly reduced in LUAD tissues and cells. ChIP and dual luciferase reporter assays ascertained the binding of GATA1 to HSD17B6. Knockdown of GATA1 attenuated the effect of overexpression of HSD17B6 on LUAD cell behaviors and cisplatin resistance.ConclusionTranscription factor GATA1 could activate HSD17B6 to inhibit cisplatin resistance in LUAD through DNA damage, suggesting that GATA1/HSD17B6 axis may be a potential therapeutic target for chemotherapy resistance in LUAD patients.

Anticancer Potential Of Psoralea Corylifolia L. Ethanol Extract: Cytotoxicity, Apoptosis Mechanisms, And Gene Expression Analysis In Mcf7 Breast Cancer Cells Via qPCR

Aim: The aim of this study was to evaluate the phytochemical profile and anticancer potential of Psoralea corylifolia L. extract, specifically focusing on its effects on breast cancer cells (MCF7) and its underlying molecular mechanisms. Methods: The seeds of P. corylifolia were extracted using ethanol and chloroform through Soxhlet extraction. The extracts were analyzed for bioactive compounds through standard phytochemical tests. The anticancer activity of the ethanol extract was assessed using the MTT assay to determine cell viability and IC50 values. Further molecular analysis of apoptosis was performed by quantifying the expression of the Bcl-2 gene using semi-quantitative PCR and evaluating marker of apoptosis. Results: Phytochemical screening revealed that both ethanol and chloroform extracts contained alkaloids, carbohydrates, and terpenoids, with ethanol extract showing a broader range of bioactive compounds, including flavonoids, phenols, saponins, and proteins. The ethanol extract exhibited significant cytotoxicity against MCF7 cells, with an IC50 value of 10.1 µg/ml. Molecular analysis showed that the extract induced apoptosis through down-regulation of the anti-apoptotic protein Bcl-2 and activation of caspase-3 and PARP, suggesting the involvement of the mitochondrial pathway. Conclusion: The results demonstrate the anticancer potential of P. corylifolia ethanol extract against breast cancer cells, primarily through cell cycle arrest and apoptosis induction. These findings highlight P. corylifolia as a promising candidate for further investigation as a safer, plant-based alternative in cancer treatment, particularly for breast cancer, though clinical studies are needed to confirm its therapeutic efficacy in humans.

Investigation of the efficacy of different cryoprotectants in the freezing of testicular tissue and epididymal sperm: Spermatological parameters, tissue viability and PARP-1 gene expression

The presented study covers testicular tissue and epididymal spermatozoa cryopreservation processes in bulls and aims to investigate the effects of these applications on spermatological parameters, cell viability in testicular tissue, and the expression of the PARP-1 gene, a DNA repair enzyme. Testes of 20 bulls over 2 years old, slaughtered in a slaughterhouse, were used in the study. After spermatological evaluations, the semen obtained from the cauda epididymis was frozen in liquid nitrogen vapor according to the straw method and stored in liquid nitrogen (−196 °C). Testicular tissue pieces obtained from the testicles were frozen by the slow freezing method in cryotubes in diluents containing Dimethylsulfoxide (DMSO) and Ethylene Glycol (EG) cryoprotectants and stored in liquid nitrogen (−196 °C). The total motility (TM) (85.89 ± 12.83 %), progressive motility (PM) (54.02 ± 15.77 %), and kinematic parameter values of fresh sperm were significantly higher compared to the TM (57.62 ± 13.13 %), PM (29.60 ± 10.76 %), and kinematic parameter values after thawing (P < 0.05). Significant decreases in plasma membrane integrity (PMI) and viability and an increase in chromatin condensation and morphological disorders in the head, middle part, and tail regions were observed in post-thaw semen samples (P < 0.05). When the effects of DMSO and EG on cell viability after thaw in frozen testicular tissue were evaluated, it was observed that the cell viability values of testicular tissues frozen with EG (45.70 ± 10.00) were statistically significantly lower than those frozen with DMSO (51.20 ± 7.70) (P < 0.05). When the effects of both cryoprotectants on gene expression in tissue and semen samples were examined, it was determined that gene expression increased on average 0.19 ± 0.27 times in the tissue samples in the DMSO group compared to fresh tissue samples and 0.17 ± 0.19 times in the tissue samples in the EG group. It was determined that gene expression levels increased by an average of 1.20 ± 1.08 times in post-thaw epididymal spermatozoa samples compared to fresh semen samples. The results show that cryopreservation can activate cellular repair mechanisms by stimulating PARP-1 gene expression and affect gene expression by activating specific pathways in tissues and cells.

Improvement of antibacterial activity of AgNPs@PVA-PVP ternary nanocomposite films followed by gamma-ray irradiation treatment for biomedical applications

Our study investigates the influence of several doses of gamma rays on the antibacterial behavior of nanocomposite of silver nanoparticles (AgNPs) doped in a blend of poly (vinyl alcohol) (PVA)-Polyvinyl Pyrrolidone (PVP). AgNPs@PVA-PVP nanocomposite films were fabricated via laser ablation route, and then the synthesized films were subjected to various gamma ray's doses. X-ray diffraction (XRD) data shows a diffraction peak at 2θ = 38° assigned to the existence of AgNPs. Ultraviolet–visible (UV–Vis) results confirm the characteristic peak of silver nanoparticles at 425 nm. The cell viability and antibacterial behavior results confirmed the enhancement in the performance of AgNPs@PVA-PVP composite after irradiated to gamma rays. These values of cell viability have been raised by increasing the dose of gamma rays to 94.5 ± 6.5 % for dose at 70 kGy gamma rays. The values of the inhibition zone of microorganisms were enhanced by raising the doses of gamma rays to 19.5 ± 0.5 and 21.3 ± 0.6 against E. coli and S. aureus respectively specifically for nanocomposite with gamma dose 70 kGy. Thus, the improved antibacterial activity of AgNPs@PVA-PVP nanocomposite could be used in biomedical applications.

Interpenetrated Polymer Network Systems (PEG/PNIPAAm) Using Gamma Irradiation: Biological Evaluation for Potential Biomedical Applications

The potential antimicrobial and antibiofouling properties of previously synthesized PEG/NiPAAm interpenetrated polymer networks (IPNs) were investigated against three of the most common bacteria (E. coli, S. aureus, and S. epidermidis). The main goal was to evaluate the material’s biocompatibility and determine its potential use as an antifouling component in medical devices. This was intended to provide an alternative option that avoids drug usage as the primary treatment, thus contributing to the fight against antimicrobial resistance (AMR). Additionally, characterization and mechanical testing of the IPN were carried out to determine its resistance to manipulation processes in medical/surgical procedures. IPNs with different NiPAAm ratios exhibited excellent cytocompatibility with BALB/3T3 murine fibroblast cells, with cell viability values of between 90 and 98%. In addition, the results regarding the adsorption of albumin as a model protein showed a nearly constant adsorption percentage of almost zero. Furthermore, the bacterial inhibition tests yielded promising results, demonstrating effective pathogen growth inhibition after 48 h. These findings suggest the material’s suitability for use in biomedical applications.

Comparative Study of Single and Coaxial Electrospun Antimicrobial Cross-Linked Scaffolds Enriched with Aloe Vera: Characterization, Antimicrobial Activity, Drug Delivery, Cytotoxicity, and Cell Proliferation on Adipose Stem Cells and Human Skin Fibroblast.

The preparation of materials with application in the biomedical field needs to attend some characteristics such as biocompatibility, nontoxicity, adequate mechanical properties, and the ability to mimic the extracellular matrix. Scaffolds for use in cell culture were prepared based on gelatin, polylactic acid (PLA), aloe vera mucilage, and tetracycline. Fibers were prepared in single and coaxial configuration and then cross-linked with glutaraldehyde saturated vapor. The fibers were obtained with cylindric morphology and changed to ribbon morphology and porous membranes, similarly to the extracellular matrix, when cross-linked. Membranes prepared by coaxial electrospinning showed core-shell structures when observed by transversal images, which is beneficial for controlled drug release. Characterization techniques such as scanning electron microscopy, thermogravimetric analysis, and Fourier transform infrared spectroscopy demonstrated the cross-linking due to the increase in diameter, formation of imine groups, and improvement of thermal stability. Antibiotic release tests showed that the prevalent release mechanism is diffusion and can be controlled considering the encapsulation effect, when fibers are prepared with a coaxial configuration, increasing the drug release time, making it a suitable material for controlled release. The biological evaluation of the scaffolds was carried out in two cell lines: mammalian adipose stem cells (ASCs), used as a primary cell culture, and Detroit 548 human skin fibroblasts as a dermal cell model. Aloe vera enriched scaffolds showed better activity in contact with both cell lines, exhibiting cell viability values greater than 90% and favorable results in live-dead assays when no damaged cells were observed. Cell proliferation was evaluated using Detroit 548 human skin fibroblast on gelatin-based scaffolds by the staining of the adhered cells; the images showed good confluence and morphology of the cells on the aloe vera and antibiotic loaded membranes for both of the studied configurations. Antibiotic loaded membranes presented antimicrobial activity against S. aureus, and this behavior increased when aloe vera is included. According to the results, the scaffolds prepared on single configuration enriched with aloe vera and tetracycline could be used in dermal tissue engineering as burn dressings, diabetic foot apposite, and skin substitutes, and the scaffolds prepared with a coaxial configuration are recommended for controlled release systems of antibiotics as treatments for chronic wounds such as diabetic foot and burn healing.

KLF4 activates LATS2 to promote cisplatin sensitivity in ovarian cancer through DNA damage.

We aimed to investigate the role of large tumor suppressor kinase 2 (LATS2) in cisplatin (DDP) sensitivity in ovarian cancer. Bioinformatic analysis explored LATS2 expression, pathways, and regulators. Quantitative reverse transcription-PCR measured LATS2 and KLF4 mRNA levels. Dual-luciferase and chromatin immunoprecipitation assays confirmed their binding relationship. Cell viability, half maximal inhibitory concentration (IC50) values, cell cycle, and DNA damage were assessed using CCK-8, flow cytometry, and comet assays. Western blot analyzed protein expression. The effect of LATS2 on the sensitivity of ovarian cancer to DDP was verified in vivo. LATS2 and KLF4 were downregulated in ovarian cancer, with LATS2 enriched in cell cycle, DNA replication, and mismatch repair pathways. KLF4, an upstream regulator of LATS2, bound to its promoter. Overexpressing LATS2 increased G1-phase cells, reduced cell viability and IC50 values, and induced DNA damage. Silencing KLF4 alone showed the opposite effect on LATS2 overexpression. Knocking out LATS2 reversed the effects of KLF4 overexpression on cell viability, cell cycle, IC50 values, and DNA damage in ovarian cancer cells. Inhibiting LATS2 inactivated the Hippo-YAP signaling pathway. In vivo experiments showed that overexpression of LATS2 enhanced the sensitivity of ovarian cancer to DDP. KLF4 activates LATS2 via DNA damage to enhance DDP sensitivity in ovarian cancer, providing a potential target for improving treatment outcomes.

The Viability of Osteoblasts against SHED Metabolites and EGCG for Biomaterial Osteogenesis

Background: EGCG, the principal flavonoid found in green tea, exhibits numerous advantageous properties, notably promoting bone regeneration by enhancing the activity of osteoblasts and osteogenic differentiation. Cell-free therapy is an alternative to avoiding the side effects of cell-based therapy. By harnessing the potential of metabolites, SHED combined with EGCG can be a biomaterial to increase osteogenesis. Objectives: This study aims to assess the viability of osteoblast cells when exposed to the combination of SHED metabolites and two concentrations of EGCG, namely 10μM and 50μM. Methods: Osteoblast viability is examined with the 3-(4.5-dimethylthiazole-2-yl)2.5-diphenyl tetrazolium bromide (MTT) assays using an ELISA reader 570nm, and the absorbance value is converted to per cent form. CD50 is a parameter that indicates non-toxicity when the percentage value of living cells is more than 50%. Results: The percentage of living cells exceeded 50%, and statistically significant distinctions were observed among the control media, control cell groups, and the groups exposed to the combination of SHED metabolites and EGCG (p = 0.031). Conclusions: The viability of osteoblast cells exposed to the combination of SHED metabolites and EGCG 10µM, as well as the combination of SHED metabolites and EGCG 50µM, showed no toxicity. The combination of 10µM SHED metabolites and EGCG showed a higher osteoblast cell viability value than the combination of SHED metabolites and EGCG 50 µM.

In vitro digestive system simulation and anticancer activity of soymilk fermented by probiotics and synbiotics immobilised on agro-industrial residues

In this study, a variety of probiotic strains, including Lactiplantibacillus plantarum, Lacticaseibacillus casei, Lactobacillus acidophilus, Streptococcus thermophilus, Bifidobacterium longum, Limosilactobacillus reuteri, Lactobacillus delbrueckii subsp. bulgaricus, Lacticaseibacillus rhamnosus, and Bifidobacterium bifidum, were utilized for soymilk fermentation both as free cells and as synbiotics on agro-industrial residuals such as okara, whey protein, banana peels, apple pomace, sugarcane bagasse, orange peels, and lemon peels. Among these, Lacticaseibacillus rhamnosus emerged as the most significant strain for soymilk fermentation, exhibiting a viability of 10.47 log cfu/mL, a pH of 4.41, total acidity of 1.12%, and organic acid contents (lactic and acetic acid) of 11.20 and 7.50 g/L, respectively. As a synbiotic Lacticaseibacillus rhamnosus immobilised on okara, showed even more impressive results, with a viability of 12.98 log cfu/mL, a pH of 4.31, total acidity of 1.27%, and organic acid contents of 13.90 and 9.30 g/L, respectively. Over a 12-h fermentation period, cell viability values increased by 10.47-fold in free cells and 11.19-fold in synbiotics. Synbiotic supplementation of fermented soymilk proved more beneficial than free cells in terms of viability, acidity, and organic acid content. Furthermore, when synbiotic fermented soymilk was freeze-dried to simulate the digestive system in vitro, synbiotics and freeze-dried cells demonstrated superior gastrointestinal tract survival compared to free cells. Both the probiotic bacteria and the synbiotics exhibited cytotoxicity against colon and liver cancer cell lines, with half-maximal inhibitory concentrations ranging from 41.96 to 61.52 μL/well.

Upregulation of CDC25B by transcription factor TEAD4 drives invasion and inhibits cisplatin sensitivity through cell adhesion in stomach adenocarcinoma.

Cisplatin is crucial in management of advanced stomach adenocarcinoma, whereas development of chemotherapy resistance hinders overall efficacy of cisplatin. This work aims to explore role of CDC25B in cisplatin sensitivity in stomach adenocarcinoma and offer a possible mechanism for explaining its function. By using bioinformatics approaches, CDC25B and TEAD4 expression levels in stomach adenocarcinoma tissues and enriched pathways of CDC25B were analyzed. qRT-PCR of CDC25B and TEAD4 expression in stomach adenocarcinoma cells, CCK-8 detection of cell viability and IC 50 values, and colony formation assay on cell proliferation were performed. Cell adhesion experiment detected cell adhesion ability. Western blot detected expression of proteins related to cell adhesion, specifically Muc-1, ICAM-1, VCAM-1. Dual luciferase assay and ChIP experiment verified binding relationship between TEAD4 and CDC25B. CDC25B was upregulated in stomach adenocarcinoma tissues and cells, enriched in focal adhesion pathway. Treatment with cell adhesion inhibitors revealed that CDC25B overexpression inhibits the sensitivity of stomach adenocarcinoma to cisplatin through the cell adhesion pathway. CDC25B has an upstream transcription factor TEAD4, which targeted and bound to CDC25B and was highly expressed in stomach adenocarcinoma. Rescue experiment revealed that knocking down TEAD4 weakened suppressive impact of CDC25B overexpression on sensitivity of stomach adenocarcinoma cells to cisplatin. Transcription factor TEAD4 could activate the transcription of CDC25B through cell adhesion to drive cell invasion and reduce sensitivity of stomach adenocarcinoma to cisplatin. TEAD4 and CDC25B may become new targets for management of stomach adenocarcinoma.

Synthesis of biocompatible Ti-6Al-4V composite reinforced with ZrO2 and bioceramic produced by powder metallurgy: Morphological, structural, and biocompatibility analysis.

In this experimental study, the initial phase involved preparing composite structures with various mix ratios using the Ti-6Al-4V alloy, widely used in clinical applications, in conjunction with ZrO2 and hydroxyapatite (HA) synthesized via the precipitation method, employing powder metallurgy techniques. Subsequently, the microstructures of the resultant hybrid composite materials were imaged, and x-ray diffraction (XRD) phase analyses were conducted. In the final phase of the experimental work, tests were performed to determine the biocompatibility properties of the hybrid composites. For this purpose, cytotoxicity and genotoxicity assays were carried out. The tests and examinations revealed that structures compatible both morphologically and elementally were obtained with no phase transformations that could disrupt the structure. The incorporation of ZrO2 into the Ti-6Al-4V alloy was observed to enhance cell viability values. The value of 98.25 ± 0.42 obtained by adding 20% ZrO2 gave the highest cell viability result. The addition of HA into the hybrid structures further increased the cell viability values by approximately 10%. All viability values for both HA-added and HA-free groups were obtained above the 70% viability level defined in the standard. According to the genotoxicity test results, the highest cytokinesis-block proliferation index values were obtained as 1.666 and 0.620 in structures containing 20% ZrO2 and 10% ZrO2 + 10% HA, respectively. Remarkably, all fabricated composite and hybrid composite materials surpassed established biocompatibility standards and exhibited nontoxic and nongenotoxic properties. This comprehensive study contributes vital insights for future biomechanical and other in vitro and in vivo experiments, as it meticulously addresses fundamental characterization parameters crucial for medical device development. RESEARCH HIGHLIGHTS: Support of optimum doping rates ions on hybrid composites and concentrations. Development of uniform surface appearance and distributions/orientations of microcrystals on ceramic compounds Improvement of cell viability and desired increase in biocompatibility with the doping of HA.

Synergistic anticancer effects of omega-3 fatty acids (EPA/DHA) and anticancer drug Doxorubicin against human lung adenocarcinoma

Single agent therapy in the treatment of cancer did not provide satisfactory clinical outcome in a long term due to drug resistance, cumulative toxicity and suboptimal efficacy. Therefore, combination therapies are apt to improve the quality of life in patients. In the present study, combinatorial effect of EPA and/or DHA with doxorubicin (Dox) was evaluated for their potential synergistic effect on human lung adenocarcinoma (A549) cells. The respective IC50 values obtained for EPA, DHA and Dox were 170, 140 and 2.5 μmol/L and thus EPA and DHA were separately combined with Dox at constant ratio of 68:1 and 56:1 respectively, depending on the ratio of their IC50 values. Different combinations of drugs revealed dose-dependent inhibition of cell viability and IC50 values were reduced to nearly 3-fold (58.75 μmol/L) and 4.5-fold (31.5 μmol/L) in EPA-Dox and DHA-Dox combination respectively. Combination index (CI) revealed strong synergism (CI < 0.5) in DHA-Dox (0.49) and moderate synergism (CI > 0.5) in EPA-Dox (0.79) at 0.5 Fa (Fraction affected), which is further confirmed by dose reduction index and isobologram. The mechanisms underlying synergistic interactions involve intracellular ROS generation and MMP depolarisation which causes changes in the cell morphology viz. Nuclear fragmentation/condensation, membrane blebbing and early/late apoptotic cells. Moreover, cell cycle arrest was observed at the S and G2/M phase, with more prominent effect in G2/M phase. No toxicity was imposed on non-cancerous HEK-293 T cells. These findings suggest that EPA-Dox and DHA-Dox could be explored further for animal studies and clinical trials for lung adenocarcinoma treatment.

TRIP13 Activates Glycolysis to Promote Cell Stemness and Strengthen Doxorubicin Resistance of Colorectal Cancer Cells.

Chemotherapy resistance is one of the main causes of clinical chemotherapy failure. Current cancer research explores the drug resistance mechanism and new therapeutic targets. This work aims to elucidate the mechanism of thyroid hormone receptor interactor 13 (TRIP13) affecting doxorubicin (DOX) resistance in colorectal cancer (CRC). Bioinformatics analyses were employed to clarify TRIP13 expression in CRC tissues and predict the correlation of the TRIP13 enrichment pathway with glycolysis-related genes and stemness index mRNAsi. Quantitative real-time polymerase chain reaction and western blot were adopted to analyze the expression of TRIP13 and glycolysis- related genes. Cell Counting Kit-8 was utilized to determine the cell viability and IC50 value. Western blot was employed to measure the expression of stemness-related factors. Cell function assays were performed to detect cells' sphere-forming ability and glycolysis level. Animal models were constructed to determine the effects of TRIP13 expression on CRC tumor growth. TRIP13 was significantly overexpressed in CRC, concentrated in the glycolysis signaling pathway, and positively correlated with stemness index mRNAsi. High expression of TRIP13 facilitated DOX resistance in CRC. Further mechanistic studies revealed that overexpression of TRIP13 could promote cell stemness through glycolysis, which was also confirmed in animal experiments. TRIP13 was highly expressed in CRC, which enhanced the DOX resistance of CRC cells by activating glycolysis to promote cell stemness. These findings offer new insights into the pathogenesis of DOX resistance in CRC and suggest that TRIP13 may be a new target for reversing DOX resistance in CRC.

Design and synthesis of chitosan/alginate/zinc oxide nanocomposite hydrogel beads as carrier for metformin hydrochloride

In the current study, nanocomposite hydrogel beads were prepared from chitosan, alginate and zinc oxide nanoparticles using sodium tripolyphosphate as the crosslinking agent by ionotropic gelation method. The resulting nanocomposite hydrogel beads were characterized by FT-IR, XRD, TGA and UV methods and the beads were found to be an effective delivery system for the regulated release of the antidiabetic drug metformin. The swelling properties, drug loading and encapsulation efficiencies of the metformin loaded nanocomposite beads were evaluated and these parameters were found to be influenced by the concentrations of chitosan, alginate and ZnO nanoparticles. The rate of release of the drug from the nanocomposite beads was determined by UV spectroscopic method at λmax 233 nm. An increase in the concentration of ZnO enhances the swelling properties but lowers the release rate. Swelling indices of the prepared formulations were in the range 5–150. Maximum swelling index of 150 was observed for the formulation with 3 % chitosan, 2 % alginate and 15 % zinc oxide NPs. However, a maximum release rate of 83.23 % was observed for the formulation without ZnO and with 3 % chitosan, 2 % alginate. Controlled drug release was observed for the drug loaded chitosan/alginate/ZnO, hydrogel beads. The zinc oxide nanoparticles improved the in vitro α-glucosidase inhibitory activity of the formulations and the formulation with 5 % ZnO exhibited a maximum α-glucosidase inhibition rate of 62.92 %. The incorporation of ZnO nanoparticles enhanced the antioxidant properties of the formulations. The cell viability values of the samples were evaluated using L929 mouse fibroblast cells which confirmed the nontoxic nature of the samples. Dinitrosalicylic acid (DNS) studies proved that the prepared nanocomposites are biodegradable.

Comparative cell viability of dentin-bonding adhesive systems on human dental pulp stem cells: time-dependent analysis

BackgroundRestorative materials are in prolonged contact with living tissues such as oral mucosa, dentin, pulp, periodontal, and periapical tissues. Therefore, the potentially harmful effects of these materials and their components on oral tissues should be evaluated before clinical use. This study aimed to compare the cell viability of different adhesive systems (ASs) on human dental pulp stem cells (hDPSCs).MethodsThree ASs that combining methacryloyloxydecyl dihydrogen phosphate (MDP) monomer with new hydrophilic amide monomers [Clearfil Universal Bond Quick(CUBQ), Kuraray Noritake], self-reinforcing 3D monomer [Bond Force II(BFII), Tokuyama)], and dual-cure property [Futurabond DC(FBDC), VOCO] were used. Three (n = 3) samples were prepared for each group. Dental pulp stem cells were isolated from ten patients’ extracted third molar teeth. Samples were incubated in Dulbecco’s modified Eagle’s medium (DMEM) for 24 h (h), 72 h, and 7 days (d) to obtain extracts. For the control group, cells were cultured without DBA samples. Cell viability of ASs extracts was measured using a cell proliferation detection kit (WST-1, Roche). Statistical analysis was performed using two-way ANOVA and post-hoc (Duncan) tests (p < 0.05).ResultsAt 24 and 72 h statistically significant differences were determined between control and BFII, control and FBDC groups (p < 0.05), while no differences between control and CUBQ groups (p > 0.05). On the 7th d, statistically significant differences were found between the control and experimental groups (p < 0.05), while no differences between experimental groups (p > 0.05). A statistically significant difference was detected for the BFII group over the three-time interval (p < 0.05). The lowest cell viability was observed for the FBDC group at 24 h, and the difference was statistically significant when compared with 72 h and 7th d (p < 0.05).ConclusionAll ASs showed different cell viability values at various exposure times. It should be taken into consideration that pH values, as well as the contents of ASs, have a significant effect on the cell viability.

Anticancer activity of the ethylacetate fraction of Vernonia amygdalina Delile towards overexpression of HER-2 breast cancer cell lines

Vernonia amygdalina Delile, coming from the Asteraceae tribe, contains active compounds that can treat breast cancer. This study examines the anticancer activity of Vernonia amygdalina Delile leaves, an active fraction of MCF-7/HER-2 breast cancer cells. Thin-layer chromatography determines the phytochemical screening. Cytotoxic and proliferation analyses were determined using the MTT method for the MCF-7/HER-2 breast cancer cell line. The cell cycle and apoptosis profiles were examined using flow cytometry. The results can be summarized in this study: Vernonia amygdalina Delile fraction contains a flavonoid with great potential for pharmacological activities, for instance, inhibiting the growth of cancer cells. The ethyl acetate fraction was more potent and cytotoxic on MCF-7/HER-2 cells (IC50 = 66 μg/mL) than extract ethanol (IC50 = 130 μg/mL). The ethylacetate fraction of Vernonia amygdalina Delile has been proven to inhibit cell proliferation by decreasing cell viability with an IC50 of 66 μg/mL concentration incubated for 24, 48, and 72 hrs with cell viability values of 64.46%, 61.67%, and 53.89%, respectively, compared to the ethanol extract IC50 of 130 μg/mL concentration with cell viability values of 56.0%, 50.19%, and 58.67%, respectively, which induced apoptosis and inhibited the cell cycle in the G2-M phase. To conclude, the ethyl acetate fraction of Vernonia amygdalina Delile can be used as an anticancer against the MCF-7/HER-2 cell line with an IC50 of 66 μg/mL, inhibiting cell proliferation and the cell cycle and inducing apoptosis activity in the MCF-7/HER-2 cell.

ZNF384 transcriptionally activated MGST1 to confer TMZ resistance of glioma cells by negatively regulating ferroptosis.

Drug resistance is one of the major reasons of the poor prognosis and recurs frequently in glioma. Ferroptosis is considered to be a new therapeutic strategy for glioma. Microsomal glutathione S-transferase 1 (MGST1) expression in glioma samples was ensured through GAPIA database, qRT-PCR, western blotting assay and immunohistochemistry. The interaction between zinc finger protein 384 (ZNF384) and MGST1 promoter was analyzed through UCSC and JASPAR databases and further verified by ChIP and luciferase reporter assay. Cell viability and IC50 value of temozolomide (TMZ) was measured by CCK-8 assay. The production of MDA, GSH and ROS and the level of Fe2+ were determined using the corresponding kit. MGST1 expression was increased in clinical glioma tissues and glioma cells. MGST1 expression was increased but ferroptosis was suppressed in TMZ-resistant cells when contrasted to parent cells. MGST1 silencing downregulated IC50 value of TMZ and cell viability but facilitated ferroptosis in TMZ-resistant cells and parent glioma cells. Moreover, our data indicated that ZNF384 interacted with MGST1 promoter and facilitated MGST1 expression. ZNF384 was also increased expression in TMZ-resistant cells, and showed a positive correlation with MGST1 expression in clinical level.ZNF384 decreasing enhanced the sensitivity of resistant cells to TMZ, while the effect of ZNF384 could be reversed by overexpression of MGST1. MGST1 transcription is regulated by transcription factor ZNF384 in TMZ-resistant cells. ZNF384 confers the resistance of glioma cells to TMZ through inhibition of ferroptosis by positively regulating MGST1 expression. The current study may provide some new understand to the mechanism of TMZ resistance in glioma.

Antibacterial and cytotoxic effects of fresh bovine amniotic membrane with hydroxyapatite (BAM-HA): a laboratory experiment

Introduction: Bacterial infections, particularly by Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) and Porphyromonas gingivalis (P. gingivalis), can worsen alveolar bone resorption after tooth extraction. The capability of Bovine Amniotic Membrane-Hydroxyapatite (BAM-HA) biocomposite to reduce this resorption has been explored. However, before clinical use, cytotoxicity testing is imperative to ensure its biocompatibility. The aim of the study was to assess both the antibacterial effects and cytotoxicity of the BAM-HA biocomposite to ensure its suitability for clinical use biocompatibility of the BAM-HA biocomposite before its clinical application. Methods: The laboratory-based research involved testing BAM combined with HA powder in 4:1 and 4:2 ratios via freeze-drying and underwent antibacterial tests against A. actinomycetemcomitans and P. gingivalis, using the plate count method. Cytotoxicity tests were performed on HGF cells, including negative control, positive control, BAM-HA (4:1), and BAM-HA (4:2) groups, with statistical analysis conducted using One-Way ANOVA and Post Hoc Bonferroni and Tukey tests. Results: Antibacterial tests against A. actinomycetemcomitans revealed significant reduction in colony count with BAM-HA ratios 4:1 (129.0 ± 12.7 CFU/mL) and 4:2 (77.3 ± 15.5 CFU/mL) compared to the negative control (186.6 ± 27.5 CFU/mL). Similar reductions were observed for P. gingivalis, with BAM-HA ratios 4:1 (51.3 ± 6.6 CFU/mL) and 4:2 (3.1 ± 1.5 CFU/mL) compared to the negative control (117.3 ± 22.0 CFU/mL). Cytotoxicity tests showed no significant differences in HGF cell viability and IC50 values between the negative control and BAM-HA (4:1) or BAM-HA (4:2) groups. Conclusion: The BAM - HA biocomposite shows antibacterial effects against A. actinomycetemcomitans and P. gingivalis. Moreover, BAM - HA ratios of 4:1 and 4:2 do not induce cytotoxic effects on human gingival fibroblasts, suggesting potential biocompatibility for clinical applications.KeywordA. actinomycetemcomitans, antibacterial effects, BAM-HA biocomposite, cytotoxicity, P. gingivalis

Design of Highly Porous Materials Based on Chitosan/Pectin Interpolyelectrolyte Complex for Wound Healing Application

Interpolyelectrolyte complexes (IPECs) of polysaccharides are multifunctional polymer materials that improve the mechanical and physicochemical properties of individual polysaccharides. In this study, highly porous (&gt;90%) materials based on IPECs of versatile natural polysaccharides, chitosan (30 and 1,200 kDa) and pectin, are obtained by freeze-drying technique. To enhance the interaction between chitosan and pectin macromolecules, the latter are chemically functionalized with dialdehyde groups. The chitosan-/aldehyde-functionalized pectin (Chit/AF-Pect) polyelectrolyte complex sponges obtained are characterized using SEM, FTIR spectroscopy, and TGA. The swelling capacity study reveals a higher swelling ratio of IPEC sponges with an increase in both the molecular weight and content of chitosan: for Chit30/AF-Pect, the swelling ratio rises from 327% to 480%, while for Chit1200/AF-Pect, from 681% to 1,066%. Additionally, the in vitro degradation test demonstrates higher stability of Chit1200/AF-Pect sponges in comparison with those of Chit30/AF-Pect: after 4 days of incubation, the weight losses are found to be 9%–16% and 18%–41%, respectively. The cytotoxicity study shows that Chit30/AF-Pect sponges are noncytotoxic, with cell viability values &gt;70%. Furthermore, the Chit30/AF-Pect sponges, obtained at chitosan:pectin weight ratio of 5:1, exhibit bactericidal activity against Escherichia coli BIM B-984 G, Pseudomonas aeruginosa BIM B-807 G, Staphylococcus aureus BIM B-1841, and slightly inhibit the growth of Enterococcus faecalis BIM B-1530 G. These findings indicate that the obtained Chit30/AF-Pect sponges can be used to create wound dressings for wound healing applications.

Patient-derived tumor-like cell clusters for personalized chemo- and immunotherapies in non-small cell lung cancer

Many patient-derived tumor models have emerged recently. However, their potential to guide personalized drug selection remains unclear. Here, we report patient-derived tumor-like cell clusters (PTCs) for non-small cell lung cancer (NSCLC), capable of conducting 100-5,000 drug tests within 10days. We have established 283 PTC models with an 81% success rate. PTCs contain primary tumor epithelium self-assembled with endogenous stromal and immune cells and show a high degree of similarity to the original tumors in phenotypic and genotypic features. Utilizing standardized culture and drug-response assessment protocols, PTC drug-testing assays reveal 89% overall consistency in prospectively predicting clinical outcomes, with 98.1% accuracy distinguishing complete/partial response from progressive disease. Notably, PTCs enable accurate prediction of clinical outcomes for patients undergoing anti-PD1 therapy by combining cell viability and IFN-γ value assessments. These findings suggest that PTCs could serve as a valuable preclinical model for personalized medicine and basic research in NSCLC.