L929 Mouse Fibroblast Cells Research Articles

Agglomerated serum albumin adsorbed protocatechuic acid coated superparamagnetic iron oxide nanoparticles as a theranostic agent

Iron oxide nanoparticles have been one of the most widely used nanomaterials in biomedical applications. However, the incomplete understanding of the toxicity mechanisms limits their use in diagnosis and treatment processes. Many parameters are associated with their toxicity such as size, surface modification, solubility, concentration and immunogenicity. Further research needs to be done to address toxicity-related concerns and to increase its effectiveness in various applications. Herein, colloidally stable nanoparticles were prepared by coating magnetic iron oxide nanoparticles (MIONPs) with protocatechuic acid (PCA) which served as a stabilizer and a linkage for a further functional layer. A new perfusion agent with magnetic imaging capability was produced by the adsorption of biocompatible passivating agent macro-aggregated albumin (MAA) on the PCA-coated MIONPs. PCA-coated MIONPs were investigated using infrared spectroscopy, thermogravimetric analysis and dynamic light scattering while adsorption of MAA was analysed by transmission electron microscopy, Fourier-transform infrared spectroscopy and x-ray diffraction methods. Magnetic measurements of samples indicated that all samples showed superparamagnetic behaviour. Cytotoxicity results revealed that the adsorption of MAA onto PCA-coated MIONPs provided an advantage by diminishing their toxicity against the L929 mouse fibroblast cell line compared to bare Fe3O4.

Reovirus Type 3 Dearing Variants Do Not Induce Necroptosis in RIPK3-Expressing Human Tumor Cell Lines.

Reoviruses are used as oncolytic viruses to destroy tumor cells. The concomitant induction of anti-tumor immune responses enhances the efficacy of therapy in tumors with low amounts of immune infiltrates before treatment. The reoviruses should provoke immunogenic cell death (ICD) to stimulate a tumor cell-directed immune response. Necroptosis is considered a major form of ICD, and involves receptor-interacting protein kinase 1 (RIPK1), RIPK3 and phosphorylation of mixed-lineage kinase domain-like protein (MLKL). This leads to cell membrane disintegration and the release of damage-associated molecular patterns that can activate immune responses. Reovirus Type 3 Dearing (T3D) can induce necroptosis in mouse L929 fibroblast cells and mouse embryonic fibroblasts. Most human tumor cell lines have a defect in RIPK3 expression and consequently fail to induce necroptosis as measured by MLKL phosphorylation. We used the human colorectal adenocarcinoma HT29 cell line as a model to study necroptosis in human cells since this cell line has frequently been described in necroptosis-related studies. To stimulate MLKL phosphorylation and induce necroptosis, HT29 cells were treated with a cocktail consisting of TNFα, the SMAC mimetic BV6, and the caspase inhibitor Z-VAD-FMK. While this treatment induced necroptosis, three different reovirus T3D variants, i.e., the plasmid-based reverse genetics generated virus (T3DK), the wild-type reovirus T3D isolate R124, and the junction adhesion molecule-A-independent reovirus mutant (jin-1) failed to induce necroptosis in HT29 cells. In contrast, these viruses induced MLKL phosphorylation in murine L929 cells, albeit with varying efficiencies. Our study shows that while reoviruses efficiently induce necroptosis in L929 cells, this is not a common phenotype in human cell lines. This study emphasizes the difficulties of translating the results of ICD studies from murine cells to human cells.

Synthesis and characterization of a novel hydrogel based on carboxymethyl chitosan/sodium alginate with the ability to release simvastatin for chronic wound healing

Since wound dressing has been considered a promising strategy to improve wound healing, recent attention has been focused on the development of modern wound dressings based on synthetic and bioactive polymers. In this study, we prepared a multifunctional wound dressing based on carboxymethyl chitosan (CMC)/sodium alginate (Alg) hydrogel containing a nanostructured lipid carrier (NLC) in which simvastatin (SIM) has been encapsulated. This dressing aimed to act as a barrier against pathogens, eliminate excess exudates, and accelerate wound healing. Among various fabricated composites of dressing, the hydrogel composite with a CMC/sodium Alg ratio of 1:2 had an average pore size of about 98.44 ± 26.9 μm and showed 707 ± 31.9% swelling and a 2116 ± 79.2 g m−2 per day water vapor transfer rate (WVTR), demonstrating appropriate properties for absorbing exudates and maintaining wound moisture. The NLC with optimum composition and properties had a spherical shape and uniform particle size distribution (74.46 ± 7.9 nm). The prepared nanocomposite hydrogel displayed excellent antibacterial activity against Escherichia coli and Staphylococcus aureus as well as high biocompatibility on L929 mouse fibroblast cells. It can release the loaded SIM drug slowly and over a prolonged period of time. The highest drug release occurred (80%) within 14 d. The results showed that this novel nanocomposite could be a promising candidate as a wound dressing for treating various chronic wounds in skin tissues.

Production of Two Novel Polyketide Pigments by Fusarium chlamydosporum

Aim: The aim of the present study was to evaluate the pigment production by the fungus Fusarium chlamydosporum in Glucose Peptone Yeast Extract medium and further to assess its toxicity by in vitro methods. Materials and Methods: The coloured crude extract yielded 4 fractions when subjected to column chromatography, upon which Fraction 2 and 4 were subjected to characterization procedures by UV-Visible spectrophotometry, Fourier Transform Infrared Spectroscopy and Molecular weight determination by Liquid Chromatography Mass Spectroscopy (LCMS). The toxicity of the pigments was evaluated in vitro on L-929 Mouse Fibroblast cell lines by MTT Assay. Results: The fungus produced dark red pigment in Glucose Peptone Yeast Extract medium on extraction with ice cold chloroform. The results summarized Fraction 2 and 4 to be the polyketide derivatives, Rubrofusarin and 8-O-methyl fusarubin respectively wherein the IC50 values of rubrofusarin and 8-O-methyl fusarubin was 183.85 and 82.366 μg/ml respectively. Conclusion: The significance of the present study lies in the characterization of 2 novel polyketides, rubrofusarin and 8-O-methyl fusarubin from F. chlamydosporum and there have been no literature reviews on the production of these pigments from the fungus.

Zinc- and Copper-Doped Mesoporous Borate Bioactive Glasses: Promising Additives for Potential Use in Skin Wound Healing Applications.

In this study, zinc (Zn)- and copper (Cu)-doped 13-93B3 borate mesoporous bioactive glasses (MBGs) were successfully synthesized using nitrate precursors in the presence of Pluronic P123. We benefited from computational approaches for predicting and confirming the experimental findings. The changes in the dynamic surface tension (SFT) of simulated body fluid (SBF) were investigated using the Du Noüy ring method to shed light on the mineralization process of hydroxyapatite (HAp) on the glass surface. The obtained MBGs were in a glassy state before incubation in SBF. The formation of an apatite-like layer on the SBF-incubated borate glasses was investigated by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The incorporation of Zn and Cu into the basic composition of 13-93B3 glass led to changes in the glass transition temperature (Tg) (773 to 556 °C), particle size (373 to 64 nm), zeta potential (-12 to -26 mV), and specific surface area (SBET) (54 to 123 m2/g). Based on the K-means algorithm and chi-square automatic interaction detection (CHAID) tree, we found that the SFT of SBF is an important factor for the prediction and confirmation of the HAp mineralization process on the glasses. Furthermore, we proposed a simple calculation, based on SFT variation, to quantify the bioactivity of MBGs. The doped and dopant-free borate MBGs could enhance the proliferation of mouse fibroblast L929 cells at a concentration of 0.5 mg/mL. These glasses also induced very low hemolysis (<5%), confirming good compatibility with red blood cells. The results of the antibacterial test revealed that all the samples could significantly decrease the viability of Pseudomonas aeruginosa. In summary, we showed that Cu-/Zn-doped borate MBGs can be fabricated using a cost-effective method and also show promise for wound healing/skin tissue engineering applications, as especially supported by the cell test with fibroblasts, good compatibility with blood, and antibacterial properties.

Phytochemical composition of Verbascum stachydifolium Boiss & Heldr. var. stachydifolium growing in Türkiye and in vitro analysis of wound healing activity

This study aimed to investigate the phenolic content, antioxidant activity, cytotoxicity and the in vitro wound healing activity of methanolic and aqueous extracts of Verbascum stachydifolium Boiss & Heldr. var. stachydifolium. Total phenolic and flavonoid contents and antioxidant activity were measured using spectrophotometry-based methods. Quantitative analysis of the selected phenolics was performed by HPLC. The cytotoxic effects of the extracts on L929 mouse fibroblast cells were evaluated by the MTT assay. The migration of treated fibroblast cells was assessed by the cell scratch assay. The expressions of type I collagen, FGF7, TGF-?1 and VEGF were evaluated by qRT-PCR and ELISA. The HPLC-based analysis revealed the presence of different phenolic compounds at varying amounts and high antioxidant activities were detected. The cytotoxicity assay results indicated that the methanolic and aqueous extracts did not exhibit any cytotoxic effect on fibroblast cells when used up to 500 ?g/mL concentration. Fibroblast migration was stimulated to the highest degree by the aqueous extract obtained by maceration as observed in the scratch assay at 60.4% closure. The molecular mechanism of the wound healing activity involves the upregulation of the analyzed genes.

Methanolic leaf extract of Catharanthus roseus reveals wound healing activity on mouse fibroblast L929 cell lines

Methanolic leaf extract of Catharanthus roseus reveals wound healing activity on mouse fibroblast L929 cell lines

Improvement in photodynamic activity by a porphyrin-fullerene composite system in lipid membranes.

Porphyrin-fullerene composite systems are attracting great attention as photodynamic agents; however, water-soluble derivatives are still scarce. Herein, we prepared noncovalently a lipid membrane-incorporated porphyrin-fullerene composite system with relative stability in aqueous solution. As in the case of porphyrin-fullerene composite systems in nonpolar solvents, efficient formation of singlet oxygen occurred via photoinduced energy transfer between porphyrin and fullerene as the predominant pathway in the photodynamic activity under the hydrophobic conditions of the lipid membranes, resulting in enhanced photodynamic activity toward Colon26 and HeLa cells compared with the individual porphyrin and fullerene components. Furthermore, the porphyrin-fullerene composite system exhibited high selectivity toward HeLa cells over normal mouse fibroblast L929 cells.

Does thermotropic liquid crystalline self-assembly control biological activity in amphiphilic amino acids? - tyrosine ILCs as a case study.

Amphiphilic amino acids represent promising scaffolds for biologically active soft matter. In order to understand the bulk self-assembly of amphiphilic amino acids into thermotropic liquid crystalline phases and their biological properties a series of tyrosine ionic liquid crystals (ILCs) was synthesized, carrying a benzoate unit with 0-3 alkoxy chains at the tyrosine unit and a cationic guanidinium head group. Investigation of the mesomorphic properties by polarizing optical microscopy (POM), differential scanning calorimetry (DSC) and X-ray diffraction (WAXS, SAXS) revealed smectic A bilayers (SmAd) for ILCs with 4-alkoxy- and 3,4-dialkoxybenzoates, whereas ILCs with 3,4,5-trisalkoxybenzoates showed hexagonal columnar mesophases (Colh), while different counterions had only a minor influence. Dielectric measurements revealed a slightly higher dipole moment of non-mesomorphic tyrosine-benzoates as compared to their mesomorphic counterparts. The absence of lipophilic side chains on the benzoate unit was important for the biological activity. Thus, non-mesomorphic tyrosine benzoates and crown ether benzoates devoid of additional side chains at the benzoate unit displayed the highest cytotoxicities (against L929 mouse fibroblast cell line) and antimicrobial activity (against Escherichia coli ΔTolC and Staphylococcus aureus) and promising selectivity ratio in favour of antimicrobial activity.

Insights into the remarkable attenuation of hen egg white lysozyme amyloid fibril formation mediated by biogenic gold nanoparticles stabilized by quercetin-functionalized tara gum

Aberrant protein misfolding and/or aggregation and fibrillation has been linked to the pathogenesis of several debilitating chronic diseases including Alzheimer's and Parkinson's disease. Inhibiting protein amyloidogenesis has been proposed as a viable strategy to prevent or ameliorate associated disorders. Herein, we investigated the anti-amyloidogenic properties of biogenic gold nanoparticles (QTG-GNP) prepared via a simple green chemistry route and stabilized by quercetin-functionalized tara gum (QTG). The synthesized QTG-GNP was extensively characterized for its physicochemical attributes via UV–visible spectroscopy, TEM, FESEM, EDX, DLS/Zeta potential, FTIR, RAMAN, XRD, XPS, and TGA analyses, as well as for its biological properties. The results revealed that small-sized (5.01 ± 1.17 nm), well-dispersed, highly stable and round-shaped biogenic gold nanoparticles were successfully synthesized at room temperature with QTG as the sole reductant /stabilizer. Importantly, QTG-GNP demonstrated potent anti-aggregation and fibrillation inhibitory effects against amyloidogenic hen egg white lysozyme (HEWL). Also, QTG-GNP was able to dissociate pre-formed HEWL amyloid fibrils. Furthermore, the constructed nanoparticles exhibited potent anti-radical activities against DPPH and ABTS+ and were cytocompatible with mouse L929 fibroblast cells. On the basis of these findings, it was established that QTG-GNP holds strong prospects for further development as an agent for countering protein aggregation and associated disease conditions.

Comparison of Antibacterial Activity, Cytotoxicity, and Fluoride Release of Glass Ionomer Restorative Dental Cements in Dentistry.

BACKGROUND This study aimed to compare the antibacterial activity, cytotoxicity, and fluoride release of 4 different glass ionomer cements (GIC). MATERIAL AND METHODS A total of 200 samples were prepared: Riva Silver, a silver-reinforced GIC; Equia Forte HT, a glass hybrid GIC; ChemFil Rock, a zinc-added GIC; and Ketac™ Molar Easymix. Using the agar diffusion test for antibacterial activity, 30 samples from each group were analyzed for Streptococcus mutans, Lactobacillus acidophilus, and Actinomyces naeslundii. The areas around the samples where no bacterial growth occurred were digitally measured and recorded. For cytotoxicity analysis, the WST-1 test was performed on 10 samples from each group using the L929 mouse fibroblast cell line. The fluoride release property was evaluated using an ion-selective electrode method on 10 samples from each group. RESULTS The group that used Ketac™ Molar Easymix showed the lowest antibacterial activity against S. mutans, L. acidophilus, and A. naeslundii. In all 3 days of cytotoxicity testing, the group that used Riva Silver was found to be the least toxic material, while the group that used ChemFil Rock did not have viable cells after the 1st day. In all 4 materials, fluoride release values gradually increased since the first day, with Ketac™ Molar Easymix having the highest fluoride release. CONCLUSIONS Of all the GICs tested, Ketac™ Molar Easymix demonstrated the least antibacterial activity despite having the highest fluoride release, while Riva Silver was found to be the least cytotoxic material.

Temperature responsive poly(N-isopropylacrylamide-co-styrene) nanofilms for non-enzymatic cell sheet harvesting

In this study, temperature-responsive cell culture substrates with non-enzymatic cell sheet detachment attributes in response to temperature reduction were prepared. Poly(N-isopropylacrylamide) (PNIPAAm) with a physiologically proximal lower critical solution temperature (LCST) induces a temperature responsivity to the substrate, and polystyrene was used to regulate the hydrophobicity of the substrate. Different compositions of poly(NIPAAm-co-styrene) were synthesized by free radical polymerization. The resulting copolymers were characterized using Fourier transform infrared spectroscopy (FTIR) and proton nuclear magnetic resonance (1HNMR) spectroscopy. The copolymer composition was calculated from 1HNMR results. The molecular weight and molecular weight distribution were measured by gel permeation chromatography (GPC). The nanofilms of the prepared copolymers were physically coated by spin-coating technique on tetramethyl orthosilicate (TMOS) pre-coated glass slides. Attenuated total reflection-FTIR (ATR-FTIR) and field emission scanning electron microscopy (FESEM) analyses showed that the spin coating procedure was performed successfully, and the change in the water contact angle (WCA) in response to temperature alteration indicated the temperature-responsive behavior of the polymer layers. L929 mouse fibroblast cell line was cultured on nanofilm-coated glass slides until a confluent cell sheet covered the entire surface. An almost integrated cell sheet was harvested by temperature reduction from 37 °C to 20 °C. The results revealed that copolymer composition is vital in harvesting contiguous cell sheets. This fabrication method, compared to others such as plasma polymerization and surface-initiated atom transfer radical polymerization (SI-ATRP), is very cost-effective and applicable for large-scale cell culture investigations.

Cytotoxicity and Antibacterial Efficacy of AgCu and AgFe NanoAlloys: A Comparative Study.

Although Ag nanoparticles (NPs) have been widely applied in daily life and in biomedical and industrial fields, there is a demand for Ag-based bimetallic nanoalloys (NAs), such as AgCu and AgFe, due to their enhanced antibacterial efficacy and reduced Ag consumption. In this work, we present a comparison study on the antibacterial efficacy and cytotoxicity rates of Ag NPs and AgCu and AgFe NAs to L929 mouse fibroblast cells using the CCK-8 technique based on the relative cell viability. The concept of the minimum death concentration (MDC) is introduced to estimate the cytotoxicity to the cells. It is found that the minimum inhibitory concentrations (MICs) of the NPs against E. coli and S. aureus decrease with the addition of both Cu and Fe. There is a strong correlation between the MDC and MIC, implying that the mechanisms of both antibacterial efficacy and cytotoxicity are similar. The enhanced antibacterial efficacy to bacteria and cytotoxicity toward the cell are attributed to Ag+ release. The following order is found for both the MIC and MDC: AgFe < AgCu < Ag NPs. However, there is no cytotoxicity to the L929 cells for AgFe and AgCu NAs at their MIC Ag concentrations against S. aureus.

Cytotoxicity and anti-biofilm properties of novel hybrid-glass-based caries infiltrant.

To assess the cytotoxicity of an experimental hybrid-glass-based infiltrant and its effect on biofilm attachment, growth and metabolic activity, and to compare it to the resin-based infiltrant Icon. Cytotoxicity of hybrid-glass-based material (EXP) and resin-based infiltrant Icon (Icon) was tested in direct contact tests on freshly cured (direct_mat) and on materials kept for 24h in cell culture medium (direct_exmat), and extract test with materials 24-h extracts (extract). Cell viability of L929 mouse fibroblast cell line was measured with MTT assay, according to ISO10993-5:2009. Biofilm attachment (5h), growth (24h and 48h) and lactic-acid production (24h and 48h) on glass-disk specimens coated with EXP or Icon, or uncoated (control), were assessed using a microcosm biofilm model and Amsterdam Active Attachment system. At indicated time points, biofilms were harvested, plated, and CFU counts were determined, while lactic-acid production was measured colorimetrically. Cell viability reduction by EXP was below 30%-threshold in direct contact tests, while in extract test an increased cell viability was observed. Icon reduced cell viability substantially in all three tests. Significantly less bacteria attached to the surface of EXP after 5h compared to Icon and control. Biofilm growth was significantly lower on EXP than on Icon and control after 24h, but this difference was smaller and statistically insignificant after 48h. There was no difference in lactic-acid production among groups. Novel hybrid-glass-based infiltrant seems to have a better biocompatibility and accumulates on its surface less bacteria than resin-based infiltrant, which makes it an attractive resin-free alternative.

In vitro evaluation of 2-pyrazoline derivatives as DPP-4 inhibitors

Abstract Objectives In this study, the synthesis of three pyrazoline derivatives and the evaluation of their inhibitory effects on dipeptidyl peptidase (DPP-4) were aimed. Methods Pyrazoline-based compounds (1–3) were obtained via the reaction of 1-(2-furyl)-3-(1,3-benzodioxol-5-yl)-2-propen-1-one with 4-substituted phenylhydrazine hydrochloride. The DPP-4 inhibitory effects of compounds 1–3 were determined with a fluorometric assay using Gly-Pro-aminomethylcoumarin as the fluorogenic substrate. The cytotoxicity of compounds 1–3 on L929 mouse fibroblast (healthy) cell line was evaluated using MTT assay. Results 1-(4-Methylsulfonylphenyl)-3-(2-furyl)-5-(1,3-benzodioxol-5-yl)-2-pyrazoline (2) exhibited the highest DPP-4 inhibitory activity (IC50=5.75 ± 0.35 µM). Moreover, compound 2 exerted no significant cytotoxicity against L929 cells (IC50=34.33 ± 7.09 µM). Conclusions Target compounds exhibited moderate DPP-4 inhibitory activity and compound 2 was identified as the most active compound.

Bacteria-Adhesive Nitric Oxide-Releasing Graphene Oxide Nanoparticles for MRPA-Infected Wound Healing Therapy.

A bacteria-infected wound can lead to being life-threatening and raises a great economic burden on the patient. Here, we developed polyethylenimine 1.8k (PEI1.8k) surface modified NO-releasing polyethylenimine 25k (PEI25k)-functionalized graphene oxide (GO) nanoparticles (GO-PEI25k/NO-PEI1.8k NPs) for enhanced antibacterial activity and infected wound healing via binding to the bacterial surface. In vitro antibacterial activity and in vivo wound healing efficacy in an infected wound model were evaluated compared with NO-releasing NPs (GO-PEI25k/NO NPs). Surface modification with PEI1.8k can enhance the ability of nanoparticles to adhere to bacteria. GO-PEI25k/NO-PEI1.8k NPs released NO in a sustained manner for 48 h and exhibited the highest bactericidal activity (99.99% killing) against methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Pseudomonas aeruginosa (MRPA) without cytotoxicity to L929 mouse fibroblast cells at 0.1 mg/mL. In the MRPA-infected wound model, GO-PEI25k/NO-PEI1.8k NPs showed 87% wound size reduction while GO-PEI25k/NO NPs showed 23% wound size reduction at 9 days postinjury. Masson trichrome and hematoxylin and eosin staining revealed that GO-PEI25k/NO-PEI1.8k NPs enhanced re-epithelialization and collagen deposition, which are comparable to healthy mouse skin tissue. GO-PEI25k/NO-PEI1.8k NPs hold promise as effective antibacterial and wound healing agents.

The Design of 3D-Printed Polylactic Acid-Bioglass Composite Scaffold: A Potential Implant Material for Bone Tissue Engineering.

Bio-based and patient-specific three-dimensional (3D) scaffolds can present next generation strategies for bone tissue engineering (BTE) to treat critical bone size defects. In the present study, a composite filament of poly lactic acid (PLA) and 45S5 bioglass (BG) were used to 3D print scaffolds intended for bone tissue regeneration. The thermally induced phase separation (TIPS) technique was used to produce composite spheres that were extruded into a continuous filament to 3D print a variety of composite scaffolds. These scaffolds were analyzed for their macro- and microstructures, mechanical properties, in vitro cytotoxicity and in vivo biocompatibility. The results show that the BG particles were homogeneously distributed within the PLA matrix and contributed to an 80% increase in the mechanical strength of the scaffolds. The in vitro cytotoxicity analysis of PLA-BG scaffolds using L929 mouse fibroblast cells confirmed their biocompatibility. During the in vivo studies, the population of the cells showed an elevated level of macrophages and active fibroblasts that are involved in collagen extracellular matrix synthesis. This study demonstrates successful processing of PLA-BG 3D-printed composite scaffolds and their potential as an implant material with a tunable pore structure and mechanical properties for regenerative bone tissue engineering.

Effect of Currently Available Nanoparticle Synthesis Routes on Their Biocompatibility with Fibroblast Cell Lines.

Nanotechnology has acquired significance in dental applications, but its safety regarding human health is still questionable due to the chemicals utilized during various synthesis procedures. Titanium nanoparticles were produced by three novel routes, including Bacillus subtilis, Cassia fistula and hydrothermal heating, and then characterized for shape, phase state, size, surface roughness, elemental composition, texture and morphology by SEM, TEM, XRD, AFM, DRS, DLS and FTIR. These novel titanium nanoparticles were tested for cytotoxicity through the MTT assay. L929 mouse fibroblast cells were used to test the cytotoxicity of the prepared titanium nanoparticles. Cell suspension of 10% DMEM with 1 × 104 cells was seeded in a 96-well plate and incubated. Titanium nanoparticles were used in a 1 mg/mL concentration. Control (water) and titanium nanoparticles stock solutions were prepared with 28 microliters of MTT dye and poured into each well, incubated at 37 °C for 2 h. Readings were recorded on day 1, day 15, day 31, day 41 and day 51. The results concluded that titanium nanoparticles produced by Bacillus subtilis remained non-cytotoxic because cell viability was >90%. Titanium nanoparticles produced by Cassia fistula revealed mild cytotoxicity on day 1, day 15 and day 31 because cell viability was 60–90%, while moderate cytotoxicity was found at day 41 and day 51, as cell viability was 30–60%. Titanium nanoparticles produced by hydrothermal heating depicted mild cytotoxicity on day 1 and day 15; moderate cytotoxicity on day 31; and severe cytotoxicity on day 41 and day 51 because cell viability was less than 30% (p < 0.001). The current study concluded that novel titanium nanoparticles prepared by Bacillus subtilis were the safest, more sustainable and most biocompatible for future restorative nano-dentistry purposes.

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.

Immobilization of Gelatin on Fibers for Tissue Engineering Applications: A Comparative Study of Three Aliphatic Polyesters.

Immobilization of cell adhesive proteins on the scaffold surface has become a widely reported method that can improve the interaction between scaffold and cells. In this study, three nanofibrous scaffolds obtained by electrospinning of poly(caprolactone) (PCL), poly(L-lactide-co-caprolactone) (PLCL) 70:30, or poly(L-lactide) (PLLA) were subjected to chemical immobilization of gelatin based on aminolysis and glutaraldehyde cross-linking, as well as physisorption of gelatin. Two sets of aminolysis conditions were applied to evaluate the impact of amine group content. Based on the results of the colorimetric bicinchoninic acid (BCA) assay, it was shown that the concentration of gelatin on the surface is higher for the chemical modification and increases with the concentration of free NH2 groups. XPS (X-ray photoelectron spectroscopy) analysis confirmed this outcome. On the basis of XPS results, the thickness of the gelatin layer was estimated to be less than 10 nm. Initially, hydrophobic scaffolds are completely wettable after coating with gelatin, and the time of waterdrop absorption was correlated with the surface concentration of gelatin. In the case of all physically and mildly chemically modified samples, the decrease in stress and strain at break was relatively low, contrary to strongly aminolyzed PLCL and PLLA samples. Incubation testing performed on the PCL samples showed that a chemically immobilized gelatin layer is more stable than a physisorbed one; however, even after 90 days, more than 60% of the initial gelatin concentration was still present on the surface of physically modified samples. Mouse fibroblast L929 cell culture on modified samples indicates a positive effect of both physical and chemical modification on cell morphology. In the case of PCL and PLCL, the best morphology, characterized by stretched filopodia, was observed after stronger chemical modification, while for PLLA, there was no significant difference between modified samples. Results of metabolic activity indicate the better effect of chemical immobilization than of physisorption of gelatin.