Fibroblast Cell Line L929 Research Articles

Oxygen-Glucose Deprivation Promoted Fibroblast Senescence and Collagen Expression via IL11.

Cell senescence is one of the most important forms of injury induced by cardiovascular and other ischemic diseases. Fibroblasts are important participants in tissue repair after ischemic injury and the main source of IL11 secretion. However, the roles of oxygen–glucose deprivation (OGD) and IL11 in promoting fibroblast senescence and their regulatory mechanisms remain unclear. This study selected the NIH3T3 and L929 fibroblast cell lines as research objects. We found that OGD could induce the expression of p53, P16, p21, and collagen in fibroblasts. In the condition of OGD, when IL11 intervened, fibroblasts’ senescence and collagen expression were changed. Some studies have found that changes in kynurenine (KYN) metabolism are related to aging diseases, and indoleamine 2,3-dioxygenase 1 (IDO1) is a key rate-limiting enzyme in the KYN metabolic pathway. We found that KYN secretion decreased after OGD increased fibroblast senescence, and inhibition of IL11 promoted IDO1 and increased KYN secretion. These results suggest that OGD may promote fibroblast senescence and collagen expression via IL11 inhibition of the IDO1/KYN metabolic pathway. Therefore, the revealed mechanism of OGD-promoted fibroblast senescence could provide an effective theoretical basis for the clinical treatment of aging-related ischemic diseases.

In vitro cytotoxic effects, in silico studies, some metabolic enzymes inhibition, and vibrational spectral analysis of novel β-amino alcohol compounds

In this study, an efficient single-step method for the preparation of β-amino alcohols (1–3) in aqueous media was applied. The aim was to investigate the cytotoxic activity of Compounds 1, 2 and 3 in neuroblastoma SH-SY5Y cell line and mouse fibroblast l-929 cell lines. Cytotoxic activities of compounds 1, 2 and 3 in this cell lines were also determined by MTT method. Cells were incubated with different concentrations of Compound 3 showed the highest cytotoxic activity in SHY5Y cells at an IC50 dose of 13.01±0.87 µM at 72 h compared to other compounds. Compound 3 was determined to have lower cytotoxic activity in l-929 cells. The chemical activities of the molecules against the B3LYP, HF, M062X level 3–21 g, 6–31 g, and SDD basis set with the Gaussian package program and biologically against the adenosine A(2A) receptor (PDB ID: 3PWH and 5NM4) proteins for neuroblastoma tumors cell with the Maestro Molecular modeling platform by Schrödinger were compared. Both experimental and theoretical NMR, UV–vis, and IR spectra of the studied molecules were compared. ADME/T analysis was performed to examine the drug properties of the molecules. Finally, these assayed for their activities against metabolic enzymes acetylcholinesterase and α-glucosidase. The most potent compounds against AChE were order compounds 3, 2 and 1 with Ki values of 35.88±6.61, 43.75±8.28, and 45.34±3.50 µM against AChE, respectively. The results indicated that all the synthesized compounds exhibited excellent inhibitory activities against mentioned enzymes as compared with standard inhibitors. These inhibitors may be candidates for drug design.

Pulsed Laser Photo-Crosslinking of Gelatin Methacryloyl Hydrogels for the Controlled Delivery of Chlorpromazine to Combat Antimicrobial Resistance.

Hydrogels are ideal candidates for the sustained local administration of antimicrobial drugs because they have customizable physicochemical properties that allow drug release kinetics to be controlled and potentially address the issue of systemic side effects. Consequently, the purpose of this study was to use 266 nm-pulsed laser beams to photo-crosslink gelatin methacryloyl hydrogels using Irgacure 2959 as a photo-initiator to reduce the curing time and to have an online method to monitor the process, such as laser-induced fluorescence. Additionally, irradiated chlorpromazine was loaded into the hydrogels to obtain a drug delivery system with antimicrobial activity. These hydrogels were investigated by UV–Vis and FTIR absorption spectroscopy, scanning electron microscopy, and laser-induced fluorescence spectroscopy and their structural and morphological characteristics, swelling behavior, and drug release profile were obtained. As a result the morphology, swelling behavior, and drug release profile were influenced by both the energy of the laser beam and the exposure time. The optimal hydrogel was obtained after 1 min of laser irradiation for Irgacure 2959 at 0.05% w/v concentration and gelatin methacryloyl at 10% w/v concentration. The hydrogels loaded with irradiated chlorpromazine show significant antimicrobial activity against Staphylococcus aureus and MRSA bacteria and a non-cytotoxic effect against L929 fibroblast cell lines.

Ultrasound Assisted Extraction of Saponins from Hedera helix L. and an In Vitro Biocompatibility Evaluation of the Extracts.

The aim of this study was to establish the best ultrasound assisted extraction (UAE) conditions of saponins from Hedera helix L. leaves and to evaluate the in vitro biocompatibility of the extracts richest in saponins. Different parameters, such as extraction time, temperature, ultrasound power, solvent to plant material ratio, and solvent concentration, were investigated. The most efficient extraction conditions were a temperature of 50 °C, an ultrasound amplitude of 40%, an extraction time of 60 min, a plant material to solvent ratio of 1:20 (w:v), and 80% ethanol as solvent. In vitro cytotoxicity of the extracts richest in saponins and their influence on the DNA content of L929 (NCTC) fibroblasts were tested. Until 200 µg/mL, the studied extracts were cytocompatible with L929 fibroblast cell lines at 48 h of treatment. These in vitro cell culture results provide useful information for further applications of Hedera helix extracts in a pharmaceutical field.

Synthesis and Characterization of Silver Nanoparticles from Rhizophora apiculata and Studies on Their Wound Healing, Antioxidant, Anti-Inflammatory, and Cytotoxic Activity.

Silver nanoparticles (AgNPs) have recently gained interest in the medical field because of their biological features. The present study aimed at screening Rhizophora apiculata secondary metabolites, quantifying their flavonoids and total phenolics content, green synthesis and characterization of R. apiculata silver nanoparticles. In addition, an assessment of in vitro cytotoxic, antioxidant, anti-inflammatory and wound healing activity of R. apiculata and its synthesized AgNPs was carried out. The powdered plant material (leaves) was subjected to Soxhlet extraction to obtain R. apiculata aqueous extract. The R. apiculata extract was used as a reducing agent in synthesizing AgNPs from silver nitrate. The synthesized AgNPs were characterized by UV-Vis, SEM-EDX, XRD, FTIR, particle size analyzer and zeta potential. Further aqueous leaf extract of R. apiculata and AgNPs was subjected for in vitro antioxidant, anti-inflammatory, wound healing and cytotoxic activity against A375 (Skin cancer), A549 (Lung cancer), and KB-3-1 (Oral cancer) cell lines. All experiments were repeated three times (n = 3), and the results were given as the mean ± SEM. The flavonoids and total phenolics content in R. apiculata extract were 44.18 ± 0.086 mg/g of quercetin and 53.24 ± 0.028 mg/g of gallic acid, respectively. SEM analysis revealed R. apiculata AgNPs with diameters ranging from 35 to 100 nm. XRD confirmed that the synthesized silver nanoparticles were crystalline in nature. The cytotoxicity cell viability assay revealed that the AgNPs were less toxic (IC50 105.5 µg/mL) compared to the R. apiculata extract (IC50 47.47 µg/mL) against the non-cancerous fibroblast L929 cell line. Antioxidant, anti-inflammatory, and cytotoxicity tests revealed that AgNPs had significantly more activity than the plant extract. The AgNPs inhibited protein denaturation by a mean percentage of 71.65%, which was equivalent to the standard anti-inflammatory medication diclofenac (94.24%). The AgNPs showed considerable cytotoxic effect, and the percentage of cell viability against skin cancer, lung cancer, and oral cancer cell lines was 31.84%, 56.09% and 22.59%, respectively. R. apiculata AgNPs demonstrated stronger cell migration and percentage of wound closure (82.79%) compared to the plant extract (75.23%). The overall results revealed that R. apiculata AgNPs exhibited potential antioxidant, anti-inflammatory, wound healing, and cytotoxic properties. In future, R. apiculata should be further explored to unmask its therapeutic potential and the mechanistic pathways of AgNPs should be studied in detail in in vivo animal models.

A convergent synthetic platform of photocurable silk fibroin-polyvinylpyrrolidone hydrogels for local anaesthesia examination

Investigations may quickly and easily convert a sol-gel into a gel-like substance-using photocurable hydrogel, which has several potential uses in biomedical research. N-Vinyl-2-pyrrolidone (NVP) monomer has been converted into Polyvinylpyrrole (PVP) and interacting matrix with silk fibroin macromolecule hydrogel by photoinitiated polymerization in this study. MTT experiment and microscopic inspections proved the PVP/SF hydrogel's biocompatibility with L929 fibroblast cell lines (96% cell survival and growth compatibility). This hydrogel's release pattern is unique from other hydrogels because of its delayed and low cumulative release. Mice models were used to show treated materials' anaesthetic and toxicological effects. Additionally, PVP/SF@LDC hydrogels dramatically suppress in vivo tumour development in mouse models and efficiently minimize their in vivo toxicity. In vitro and in vivo testing revealed that encapsulating LDC in PVP/SF hydrogels improved the drug's anaesthetic activity while extending its duration. Experimental research involving the controlled release of local anaesthetics might make excellent use of this new approach.

A Study on the Effect of Nanoscale MgO and Hydrogen Bonding in Nanofiber Mats for the Controlled Drug Release along with In Vitro Breast Cancer Cell Line and Antimicrobial Studies.

Nanosized metal oxide-incorporated drug carriers have received significant attention due to their biocompatibility, mechanical strength, controlled drug release, and biodegradability. Herein, an attempt was made to fabricate polycaprolactone-based electrospun nanofiber mats involving the 5-fluorouracil (5Fu) drug, MgO nanoparticle, methyl cellulose, and polyethylene glycol. The chemical interactions, surface wettability, mechanical properties, structural and morphological changes, and thermal stability were studied by the respective analyses. The ionic interaction between 5Fu, MgO, and polymers were found to be responsible for the controlled drug release. Zero-order kinetic and model data also revealed that a controlled drug release pattern was observed in a period of 16 days. Furthermore, the nanofiber mats were subjected to cytotoxicity studies against MDA-MB-231 cancer cell line and the results showed higher cytotoxicity in a short time of 24 h and less toxicity to normal L929 fibroblast cell line. The apoptosis in cancer cell lines was also tested by AO/PI staining assay and confirmed by fluorescence microscopy. In addition, the growth inhibition of several bacterial and fungal strains was tested for the mats and the results exhibited good inhibition activity. Hence, the reported nanofiber drug carrier was found to be an efficient implant for the controlled release of anticancer drug along with other significant properties.

Synthesis, characterization, antioxidant and anticancer activities of a new Schiff base and its M(II) complexes derived from 5-fluorouracil

In this study, Schiff base ligand was obtained from the condensation reaction of benzene-1,2-diamine and 5-fluoropyrimidine-2,4(1H,3H)-dione (5-FU). Metal(II) complexes were synthesized with Fe(II), Co(II) and Ni(II) chloride salts. The synthesized ligand and metal complexes were characterized by FT-IR, UV-vis, 1H-13C NMR, elemental analyses, mass spectroscopy, magnetic moments, molar conductivity and thermogravimetric analysis studies. With the help of different techniques reveal Fe(II), Co(II) and Ni(II) complexes have exhibited tetrahedral and octahedral geometry. Ligand acted as bidentate and it binds metal(II) ions through deprotonated-NH, imine-N atom and carbonyl-O atom, respectively. DPPH, ABTS, FRAP, CUPRAC and total antioxidant activity methods were used to determine the antioxidant properties of ligand and metal complexes. According to the results, the synthesized compounds showed very high antioxidant activity compared to 5-FU. The cytotoxicities of the synthesized compounds were performed on MCF-7 (human breast cancer) and L-929 (fibroblast) cell lines using the MTT assay. In addition, the effect of electroporation (EP) on the cytotoxicity of the compounds was investigated. Our results demonstrated that novel Co(II) and Ni(II) complexes show potential as new anticancer agents and ECT may be a viable treatment option for breast cancer.

In vitro cytocompatibility assessment and antibacterial effects of quercetin encapsulated alginate/chitosan nanoparticle

The present work aims at evaluating the in vitro biocompatibility, antibacterial activity and antioxidant capacity of the fabricated and optimized Alginate/Chitosan nanoparticles (ALG/CSNPs) and quercetin loaded Alginate/Chitosan nanoparticles (Q-ALG/CSNPs) with an improved biological efficacy on the hydrophobic flavonoid.The physicochemical properties were determined by TEM and FTIR analysis. The nanoparticles evaluated for the encapsulation of quercetin exerted % encapsulation efficiency (EE) that varied between 76 and 82.4 % and loading capacity (LC) from 31 to 46.5 %. Potential cytotoxicity of the ALG/CSNPs and Q-ALG/CSNPs upon L929 fibroblast cell line was evaluated by MTT reduction Assay and expressed as % cell viability. The in vitro antibacterial property was studied by well diffusion method against gram-positive bacteria Staphylococcus aureus (ATCC 25925) and gram-negative bacteria Escherichia coli (ATCC 25923). The inhibitory efficacy by scavenging free radical intermediates was evaluated by 1,1, diphenyl 2-picrylhydrazyl (DPPH) assay. The results of in vitro cytotoxicity showed biocompatibility towards L929 cells. Quercetin loaded Alginate/Chitosan nanoparticles inhibited the growth of microorganisms than pure quercetin. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging results have shown a high level of antioxidant property for encapsulated Quercetin in Alginate/Chitosan nanoparticles compared to free Quercetin. The findings of our study suggest that the developed ALG/CSNPs and Q-ALG/CSNPs possess the prerequisites and be proposed as a suitable system for delivering quercetin with enhanced therapeutic effectuality.

A Comparative Study of Quercetin-Loaded Nanocochleates and Liposomes: Formulation, Characterization, Assessment of Degradation and In Vitro Anticancer Potential.

Quercetin, a flavonoid, has antioxidant and anti-inflammatory properties and the potential to inhibit the proliferation of cancer, but its therapeutic efficacy is lowered due to poor solubility and bioavailability. Quercetin-loaded nanocochleates (QN) were developed using a trapping method by the addition of calcium ions into preformed negatively charged liposomes (QL) prepared by a thin-film hydration method. Liposomes were optimized by varying the concentration of Dimyristoyl phosphatidyl glycerol and quercetin by applying D-optimal factorial design using Design-Expert® software. Stable rods were observed using TEM with an average particle size, zeta potential and encapsulation efficiency of 502 nm, −18.52 mV and 88.62%, respectively, for QN which were developed from spherical QL showing 111.06 nm, −40.33 mV and 74.2%, respectively. In vitro release of quercetin from QN and QL was extended to 24 h. Poor bioavailability of quercetin is due to its degradation in the liver, so to mimic in vivo conditions, the degradation of quercetin released from QL and QN was studied in the presence of rat liver homogenate (S9G) and results revealed that QN, due to its unique structure, i.e., series of rolled up solid layers, shielded quercetin from the external environment and protected it. The safety and biocompatibility of QL and QN were provenby performing cytotoxicity studies on fibroblast L929 cell lines. QN showed superior anticancer activity compared to QL, as seen for human mouth cancerKB cell lines. Stability studies proved that nanocochleates were more stable than liposomal formulations. Thus, nanocochleates might serve as pharmaceutical nanocarriers for the improved efficacy of drugs with low aqueous solubility, poor bioavailability, poor targeting ability and stability.

The cross-linked bacterial cellulose impregnated with octenidine dihydrochloride-based antiseptic as an antibacterial dressing material for highly-exuding, infected wounds

The highly absorbent, antibacterial dressings with a sustained release of the antimicrobial are considered necessary measures to counteract chronic wound biofilm-based infections. This study aimed to analyze wet and dry bacterial cellulose (BC) materials, modified by chemical cross-linking, and impregnated with an antiseptic based on octenidine dihydrochloride (OCT) in the context of its antibiofilm/antibacterial activity, exudate absorption, and cytotoxicity. The native BC was obtained from cost-effective, ecological-friendly potato juice (leftover from the starch industry). The ability to absorb and retain OCT, exudate absorption capacity, the kinetics of OCT release as well as antibiofilm/antibacterial activity of modified BC materials against biofilm-forming and planktonic bacteria (Staphylococcus aureus and Pseudomonas aeruginosa) were investigated. The performed analyses revealed that modified BC materials, thanks to their layered structure with numerous air spaces, were characterized by sustained exudate absorption and OCT release profile, which allowed them to exhibit high antimicrobial activity for up to 7 days, with a reduction of planktonic and biofilm cells of 84–100% and 69–93%, respectively. The modified BC materials showed also no cytotoxicity against fibroblast cell line L929 in vitro and were characterized by firm adhesion to the curved surfaces. These results indicate that cross-linked BC impregnated with OCT may be a particularly promising dressing material (obtained using sustainable processes), especially in the treatment of biofilm-infected, highly-exuding wounds.

CYTOTOXICITY AND WOUND HEALING CAPACITY OF BIOLOGICALLY SYNTHESIZED SILVER NANOPARTICLES

Metallic nanoparticles are interesting areas of research due to their unique properties which can be advantageous for producing smart products. Silver nanoparticles (AgNPs) are remarkably used in pharmaceutical industry because of their strong biological activities. The aim of the present study was to investigate the cytotoxicity and wound healing capacity of the biologically prepared silver nanoparticles via green synthesis route. Cytotoxicity of the biogenic AgNPs was determined by MTT (3- [4, 5- dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium-bromide) assay against L929 fibroblast cell line. Wound healing properties of the AgNPs were evaluated using in vitro-scratch wound healing assay using 3T3 fibroblast cell line. Biosynthesized AgNPs inhibited the propagation of fibroblasts at a half maximal inhibitory concentration (IC50) of 23.507 μg/mL after 24 h incubation. In vitro wound healing assay also revealed that the biogenic AgNPs stimulated the 3T3 fibroblasts' cell proliferation. It can be suggested that biologically synthesized AgNPs can be used effectively for biomedical applications such as wound dressing materials.

Novel Graphene Oxide/Quercetin and Graphene Oxide/Juglone Nanostructured Platforms as Effective Drug Delivery Systems with Biomedical Applications.

In this paper, novel drug delivery systems (DDS) were designed based on graphene oxide (GO) as nanocarrier, loaded with two natural substances (quercetin (Qu) and juglone (Ju)) at different concentrations. The chemical structure and morphology of the synthesized GO-based materials were characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and Raman spectroscopy. The antibacterial activity was evaluated against standard strains, Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 8739, and Candida albicans ATCC 10231. Results demonstrated excellent antimicrobial activity, with a 5 log reduction of E. coli and a 1 log to 3.04 log reduction of S. aureus populations. Reduction rates were above 90%. Biocompatibility tests were also performed on GO-based materials, and the results showed biocompatible behavior for both L929 fibroblast cell line and BT474 breast cancer cells at lower concentrations. The identity of Qu and Ju was demonstrated by matrix-assisted laser desorption/ionization (MALDI) analysis, showing the compounds’ mass with high accuracy. In addition, specific properties of GO made it a versatile matrix for the MALDI analysis. The results of this study indicated that GO-based platforms may be suitable for applications in many areas for the effective and beneficial use of hydrophobic compounds such as Ju and Qu.

Farklı Antijen Hazırlama Metotları İle Hazırlanan İnsan Meme Kanseri Hücre Lizatlarının Adjuvan Kombinasyonları İle İmmünostimülan/Sitotoksik Aktivitelerinin İncelenmesi

In last decades, immunotherapies became more and more significant as a cancer treatment method. Many in vivo and in vitro studies have been conducted in the literature in order to determine the appropriate diagnostic and treatment methods for cancer. During the development of therapeutic vaccines, determination of vaccine concentration which is the beginning step of in vivo studies, is critical for the healthy progression of the studies. Breast cancer which occurs by uncontrolled proliferation as a result of exposure of healthy cells in breast tissue to factors such as DNA damage, UV, radiation, carcinogens and, various chemicals has a huge portion among the cancer types. In this study, for the first time, antigens generated by two different antigen preparation methods (autoclaving, freezing-thawing) using AU-565 / MCF-7 cell lines. Then, antigens derived from human breast cancer cell lines were applied alone and with different adjuvants (Polyoxidonium, Freund’s) to the J774 macrophage cell culture system and the L929 fibroblast cell line in order to investigate immunostimulant activity and cytotoxicity via Griess reagent and MTT test respectively. At the end of the 48th hour, the cell viability well will be examined and the most appropriate antigen-adjuvant combination and its concentration for vaccine preparation were determined. Among the antigen preparation methods, it was determined that the freeze-thaw method was more effective than the autoclave method, and the antigens prepared by the freeze-thaw method from the AU-565 and MCF-7 cell lines showed the highest immunostimulatory activity at a concentration of 160 µg/ml. Among the adjuvants, the immunostimulant effect of Freund's adjuvant was found to be higher. In the light of these results, the optimal vaccine concentration against breast cancer and the viability analysis of these concentrations were determined. All in all, it is thought that antibodies obtained from in vivo studies using the most appropriate antigen preparation method and the most appropriate vaccine concentration can be used in the diagnosis and treatment of breast cancer.

Selenium increases cell proliferation after citrate treatment on L929 fibroblast cell line

Selenium increases cell proliferation after citrate treatment on L929 fibroblast cell line

The Evaluation of Safety Property and Apoptotic Efficacy of α-L-Guluronic Acid (G2013), as a Novel NSAID, Under In Vitro Examination on L929 and Hepatocellular Carcinoma Cell Lines.

Many investigations have expanded this concept that liver chronic inflammation has an essential role in persistent cell damages along with altering the liver microenvironment leading to fibrosis, cirrhosis, and finally, hepatocellular carcinoma (HCC). To reduce inflammation and relieve symptoms, Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) are commonly used; however, their long-term usage can lead to severe adverse events on vital organs like the liver. Interestingly, the α-L-Guluronic Acid (G2013), as a novel NSAID with immunomodulatory properties, has shown the inhibitory effects on inflammation and metastasis in experimental models. This study was conducted to determine the effects of G2013 on cytotoxicity and induction of apoptosis, as a new therapeutic target for cancer therapy, in the HepG2 cell line and the mouse fibroblast cell line L929, as a control. MTT assay and flow cytometry method were carried out using the different concentrations of G2013 (5, 15, 25, 50, 100, 200 and 400 μg/ml) in 3 distinct incubation times. Our data showed that treatment of HepG2 cells with high concentration (400μg/mL) of G2013 could effectively cause a decrease in cell viability, so that they were statistically different after 72 hours compared to other concentrations (5 to 200 μg/ml) (p<0.05 and p<0.01, respectively). Moreover, the proportion of apoptosis of HepG2 cells at the dose of 200μg/mL considerably increased, suggesting that the induction of apoptosis by G2013 in HepG2 cells is dose- and time-dependent, which could promote its anticancer properties. The present study revealed that G2013 could induce apoptosis in the liver cancer model. Therefore, based on these findings, G2013 might be considered as a therapeutic option in cancer therapy.

Influence of poly(styrene sodium sulfonate) grafted silicone breast implant’s surface on the biological response and its mechanical properties

Prior to its commercialization, a biomaterial must fulfill the regulation in place. The International Organization for Standardization outlines the requirements needed for a material in order to guarantee safety and quality. In previous work, we have successfully grafted a bioactive polymer known as poly(styrene sodium sulfonate) – (polyNaSS) on silicone breast implants’ surfaces using UV irradiation. This paper intends to study the effect of the polyNaSS grafting parameters. It includes the study of (i) the effect of only UV irradiation and (ii) the presence of the polyNaSS grafted silicone on both the mechanical properties of the material and the biological response using the L929 fibroblast cell line for biocompatibility investigations. PolyNaSS aims to overcome the lack of biocompatibility issues, but the grafting process should have a minimal impact on the surface’s properties. The tensile strength and swelling tests showed no apparent modification before and after grafting in crosslinking densities and elasticity moduli. That confirms the impactless aspect of the grafting protocol on the material’s mechanical properties. Surface roughness was investigated by atomic force microscopy to understand cell behavior on surfaces upon various treatments. Biocompatibility tests showed that the grafting of polyNaSS significantly enhanced cell adhesion and viability compared to a non grafted silicone. Overall, polyNaSS confers a highly suitable surface for fibroblasts, demonstrating their active forms (spindle-shaped). This was assessed by optical microscopy, scanning electron microscopy, and atomic force microscopy.

The Influence of Diamond Nanoparticles on Fibroblast Cell Line L929, Cytotoxicity and Bacteriostaticity of Selected Pathogens

The main problem with using modified allotrophic forms of carbon with nanodiamond particles in the production of food packaging is establishing the boundary between safety, as it affects the human body, and the adequate and effective action of the substances. One vital area of concern is the transmission of pathogens in food into the body. The aim of this study was to evaluate the cytotoxicity and bacteriostatic biological activity of two different modifications of diamond nanoparticles: pure detonation nanodiamond particles (DND) obtained by Danienko and plasma-chemically modified detonation nanodiamond particles obtained by the microwave plasma activated chemical vapor deposition method in a rotary chamber (MDP1) An indirect method was used to evaluate the cytotoxicity effect in accordance with ISO 10993–5. The viability of the L929 fibroblast cell line used as a control was 98.5%, for DND 95.14%, and the lowest level of viability for MDP1 was 88.63%. Escherichia coli and Staphylococcus aureus bacteria were used in bacteriostatic tests and the degree of cytotoxicity of the tested materials was classified as low. The in vitro cytotoxicity results indicate no toxic effect on L929 cells nor any effect on any of the samples tested against the bacterial strains us

Cytocompatible manganese dioxide-based hydrogel nanoreactors for MRI imaging

The application of nanoparticles in magnetic resonance imaging (MRI) has been greatly increasing, due to their advantageous properties such as nanoscale dimension and tuneability. In this context, manganese (Mn2+)-based nanoparticles have been greatly investigated, due to their valuable use as a contrast agent, improving signal intensity and specificity in MRI (manganese-enhanced MRI, MEMRI). Additionally, Mn2+ can act as scavengers of reactive oxygen species (ROS), commonly present in the inflammatory processes of neurodegenerative diseases. The aim of the present study was to develop nanoreactors, which can be used as contrast-agent in MEMRI. Several blends of methacrylated gellan gum (GG-MA) and hyaluronic acid (HA) were embedded with different types of manganese dioxide (MnO2) nanoparticles and further physico-chemically characterized. Dynamic light scattering, scanning electron microscopy, water uptake and degradation studies were performed. In vitro cytotoxicity of the different formulations was also evaluated using an immortalized rat fibroblast cell line L929, up to 72 h of culturing. Synthesized nanoparticles were obtained with an average size of 70 nm and round-shaped morphology. The stability of the different formulations of hydrogels was not affected by nanoparticles' concentration or HA ratio. The presence of synthesized MnO2 (MnO2_S) nanoparticles reduced hydrogels' cytocompatibility, whereas the commercially available type 1 (MnO2_C1) nanoparticles were less toxic to cells. Additionally, cell proliferation and viability were enhanced when a lower content of HA was present. Higher concentrations (75 and 100 ng/mL) of MnO2_S and MnO2_C1 nanoparticles did not negatively affected cell viability, whereas the opposite effect was observed for the commercial type 2 (MnO2_C2) nanoparticles. Further studies are required to evaluate the potential application of the most promising nanoreactors' formulations for combined application in MEMRI and as ROS scavengers.

Megakaryocyte–stromal cell interactions: Effect on megakaryocyte proliferation, proplatelet production, and survival

Bone marrow stromal cells provide a proper environment for the development of hematologic lineages. The incorporation of different stromal cells into in vitro culture systems would be an attractive model to study megakaryopoiesis and thrombopoiesis. Our objective was to evaluate the participation of different types of stromal cells in in vitro megakaryopoiesis, thrombopoiesis, and megakaryocyte (MK) survival. CD34-positive progenitors from umbilical cord blood were differentiated into MK precursors and then cocultured with umbilical vein endothelial cells (HUVECs), bone marrow mesenchymal stem cells (MSCs), skin fibroblasts (SFs) (all human), or the mouse fibroblast cell line L929. The number of MKs (CD61-positive cells) was increased in the presence of HUVECs and SFs, whereas L929 cells decreased total and mature MK counts. With respect to thrombopoiesis, HUVECs increased proplatelet (PP)-producing MKs, while MSCs, L929 cells, and SFs had the opposite effect (immunofluorescence staining and microscopic analysis). MK survival was enhanced in MSC and SF co-cultures, as assessed by evaluation of pyknotic nuclei. However, HUVECs and L929 did not prevent apoptosis of MKs. Reciprocally, the cloning efficiency of MSCs was decreased in the presence of MKs, while the ability of stromal cells (either MSCs or SFs) to produce the extracellular matrix proteins type III collagen, fibronectin, dermatan sulfate, heparan sulfate, and prolyl 4-hydroxylase subunit β was preserved. These data indicate that each stromal cell type performs distinctive functions that differentially modulate MK growth and platelet production and, at the same time, that MKs also modify stromal cell behavior. Overall, our results highlight the relevance of considering the influence of stromal cells in MK research as well as the close interplay of different cell types within the bone marrow milieu.