3T3 Fibroblast Cell Line Research Articles

Phenolic-Enriched Collagen Fibrillar Coatings on Titanium Alloy to Promote Osteogenic Differentiation and Reduce Inflammation.

The adsorption of biomolecules on biomaterial surfaces can promote their integration with surrounding tissue without changing their bulk properties. For biomaterials in bone reconstruction, the promotion of osteogenic differentiation and reduction of inflammation are desirable. Fibrillar coatings are interesting because of fibrils’ high surface area-volume ratio, aiding adsorption and adhesion. Fibrils also serve as a matrix for the immobilization of biomolecules with biological activity, such as the phenolic compound phloroglucinol (PG), the subunit of marine polyphenols. The aim of this work was to investigate the influence of PG coatings on fibroblast- and osteoblast-like cells to increase the osseointegration of titanium implants. Collagen fibril coatings, containing PG at low and high concentrations, were produced on titanium alloy (Ti6Al4V) scaffolds generated by additive manufacturing (AM). These coatings, especially PG-enriched coatings, reduced hydrophobicity and modulated the behavior of human osteosarcoma SaOS-2 and mouse embryonic fibroblast 3T3 cell lines. Both osteoblastic and fibroblastic cells spread and adhered well on PG-enriched coatings. Coatings significantly reduced the inflammatory response. Moreover, osteogenic differentiation was promoted by collagen coatings with a high PG concentration. Thus, the enrichment of collagen fibril coatings with PG is a promising strategy to improve Ti6Al4V implants for bone contact in orthopedics and dentistry and is worthy of further investigation.

Free Radical Scavenging and Cytotoxic Activities of Substituted Pyrimidines

A library of substituted pyrimidines was synthesized and evaluated for free radical scavenging, and in vitro cytotoxic activity in 3T3 cells. All compounds showed good free radical scavenging activity with IC50 values in the range of 42.9 + 0.31 to 438.3 3.3 {\mu}M as compared to the standard butylated hydroxytoluene having IC50 value of 128.83 2. 1 {\mu}M. The structure activity-relationship was also established. Selected analogues 1, 2, 3, 5, 6, 7, 8, 9, 10, 12, 13, 15, 19, 20, 21, 24, 25, 26 and 28 were tested for cytotoxicity in mouse fibroblast 3T3 cell line using MTT assay, and most of the analogues showed cytotoxicity. This study has identified a number of cytotoxic novel substituted pyrimidines having free radical scavenging activities that can be used as inhibitory compounds for those cancer cells whose growth is mediated by reactive oxygen species.

Antiglycation Activity of Triazole Schiff's Bases Against Fructosemediated Glycation: In Vitro and In Silico Study.

Advanced glycation end products (AGEs) are known to be involved in the pathophysiology of diabetic complications, neurodegenerative diseases, and aging. Preventing the formation of AGEs can be helpful in the management of these diseases. Two classes of previously synthesized traizole Schiff's bases (4H-1,2,4-triazole-4- Schiff's bases 1-14, and 4H-1,2,4-triazole-3-Schiff's bases 15-23) were evaluated for their in vitro antiglycation activity. In vitro fructose-mediated human serum albumin (HSA) glycation assay was employed to assess the antiglycation activity of triazole Schiff's bases. The active compounds were subjected to cytotoxicity analysis by MTT assay on mouse fibroblast (3T3) cell line. Molecular docking and simulation studies were carried out to evaluate the interactions and stability of compounds with HSA. Anti-hyperglycemic and antioxidant activities of selected non-cytotoxic compounds were evaluated by in vitro α-glucosidase inhibition, and DPPH free radical scavenging assays, respectively. Compound 1 (IC50=47.30±0.38 µM) from 4H-1,2,4-triazole-4-Schiff's bases has exhibited antiglycation activity comparable to standard rutin (IC50=54.5±0.05 µM) along with a stable RMSD profile in MD simulation studies. Compound 1 also exhibited a potent α-glucosidase inhibitory activity, and moderate antioxidant property. Other derivatives showed a weak antiglycation activity with IC50 values between 248.1-637.7 µM. Compounds with potential antiglycation profile were found to be non-cytotoxic in a cellular assay. The study identifies triazole Schiff's bases active against fructose-mediated glycation of HSA, thus indicates their potential against late diabetic complications due to production of advancedend products (AGEs).

Lignins from Agroindustrial by-Products as Natural Ingredients for Cosmetics: Chemical Structure and In Vitro Sunscreen and Cytotoxic Activities.

The growing concern about the environmental impact and human health risk related to the excessive use of synthetic ingredients in cosmetics and topical formulations calls for the exploration of safe and sustainable natural alternatives. Lignin-rich lignocellulosic industrial wastes such as hazelnut and walnut shells were used as a lignin polymer source. Agro-derived lignins were evaluated as a potential natural active ingredient for health care products. Aside from the structural characteristics of isolated lignins, which were identified by GPC, Py-GC–MS, and 2D HSQC NMR techniques, functional properties such as antioxidant power and UV absorption ability were investigated. The SPF values found for creams containing 5% of hazelnut and walnut lignin content were 6.9 and 4.5, respectively. Additionally, both lignin types presented appropriate protection against UVA radiation, highly interesting property to block the full ultraviolet spectrum. The biological activity of isolated lignins assessed at different concentrations (0.01–1 mg/mL) and different times (24, 48, and 72 h) on murine fibroblast cell line 3T3 suggested their suitability for cosmetic applications.

Natural compounds as angiogenic enzyme thymidine phosphorylase inhibitors: In vitro biochemical inhibition, mechanistic, and in silico modeling studies.

Natural flora is the richest source of novel therapeutic agents due to their immense chemical diversity and novel biological properties. In this regard, eighteen natural products belonging to different chemical classes were evaluated for their thymidine phosphorylase (TP) inhibitory activity. TP shares identity with an angiogenic protein platelet derived endothelial cell growth factor (PD-ECGF). It assists tumor angiogenesis and is a key player in cancer progression, thus an ideal target to develop anti-angiogenic drugs. Eleven compounds 1–2, 5–10, 11, 15, and 18 showed a good to weak TP inhibitory activity (IC50 values between 44.0 to 420.3 μM), as compared to standards i.e. tipiracil (IC50 = 0.014 ± 0.002 μM) and 7-deazaxanthine (IC50 = 41.0 ± 1.63 μM). Kinetic studies were also performed on active compounds, in order to deduce the mechanism of ligand binding to enzyme. To get further insight into receptor protein (enzyme) and ligand interaction at atomic level, in- sillico studies were also performed. Active compounds were finally evaluated for cytotoxicity test against mouse fibroblast (3T3) cell line. Compound 18 (Masoprocol) showed a significant TP inhibitory activity (IC50 = 44.0 ± 0.5 μM). Kinetic studies showed that it inhibits the enzyme in a competitive manner (Ki = 25.6 ± 0.008 μM), while it adopts a binding pose different than the substrate thymidine. It is further found to be non-toxic in MTT cytotoxicity assay. This is the first report on TP inhibitory activity of several natural compounds, some of which may serve as leads for further research towards drug the development.

Carbazole‐Linked 1,2,3‐Triazoles: In Vitro β‐Glucuronidase Inhibitory Potential, Kinetics, and Molecular Docking Studies

Abstractβ‐Glucuronidase enzyme is a tetrameric glycoprotein present in microsomes and lysosomes of many organs, and body fluids. Over‐expression of this enzyme is observed in several melanomas and carcinomas. Therefore, it has been identified as a target for the treatment of several pathological conditions. In this regard, carbazole linked 1,2,3‐triazoles (1–27) were synthesized according to our previous report, and evaluated for their in vitro β‐glucuronidase inhibitory activities. Compounds 7–10, 17–18, 20, and 22–27 showed potent activities with IC50 values between 0.55–32.5 μM, as compared to the standard, D‐saccharic acid 1,4‐lactone (IC50=45.75 ± 2.16 μM). All active compounds were found to be non‐cytotoxic against mouse fibroblast 3T3 cell line. Compounds 20, 22, 23, 25, 26, and 27 were subjected to enzyme kinetic studies for the determination of their modes of inhibition, and dissociation constants Ki. Compounds 23, 25, and 26 were also studied for their mode of inhibition by using molecular docking methods.

Libidibia ferrea Fruit Crude Extract and Fractions Show Anti-Inflammatory, Antioxidant, and Antinociceptive Effect In Vivo and Increase Cell Viability In Vitro.

Background Libidibia ferrea (L. ferrea) is found throughout the northeastern region of Brazil, where it has been used in folk medicine with beneficial effects on many inflammatory disorders. Purpose This study investigated the phytochemical composition of the crude extract and fractions of L. ferrea fruit and evaluated its anti-inflammatory and antinociceptive activities in vivo and effect on cell viability in vitro. Methods Characterization of polyphenols present in crude extract (CE), hydroalcoholic fractions of 20-80% ethanol (CE20, CE40, CE60, and CE80), aqueous fraction (AqF), and ethyl acetate (EAF) fractions of L. ferrea fruit was performed by chromatographic analysis. Anti-inflammatory activity was evaluated by using a carrageenan-induced peritonitis model submitted to a leukocyte migration assay and myeloperoxidase activity (MPO) analysis. Total glutathione and malondialdehyde (MDA) levels were assessed to evaluate the oxidative stress level. Antinociceptive activity was evaluated by acetic acid-induced abdominal writhing and hot plate test. In vitro cell viability was determined by using MTT assay in a mouse embryonic fibroblast cell line (3T3 cells). Results Chromatography revealed the presence of ellagic acid content in EAF (3.06), CE (2.96), and CE40 (2.89). Gallic acid was found in EAF (12.03), CE 20 (4.43), and CE (3.99). L. ferrea crude extract and all fractions significantly reduced leukocyte migration and MPO activity (p<0.001). L. ferrea antioxidant effect was observed through high levels of total glutathione and reduction of MDA levels (p<0.001). Acetic acid-induced nociception was significantly inhibited after administration of L. ferrea crude extract and all fractions (p<0.001). Crude extract and all fractions significantly increased the viability of the 3T3 cell line (p<0.05). Conclusions The appropriate extraction procedure preserves the chemical components of L. ferrea fruit, such as gallic acid and ellargic acid. Crude extract and fractions of L. ferrea fruit exhibited anti-inflammatory, antioxidant, antinociceptive activities in vivo and enhanced cell viability in vitro.

Bioinformatics: A rational combine approach used for the identification and in-vitro activity evaluation of potent β-Glucuronidase inhibitors

Identification of hotspot drug-receptor interactions through in-silico prediction methods (Pharmacophore mapping, virtual screening, 3DQSAR, etc), is considered as a key approach in drug designing and development process. In the current design study, advanced in-silico based computational techniques were used for the identification of lead-like molecules against the targeted receptor β-glucuronidase. The binding pattern of a potent inhibitor in the ligand-receptor X-ray co-crystallize complex was used to identify and extract the structure-base Pharmacophore features. Based on these observations; five structure-based pharmacophore models were derived to conduct the virtual screening of ICCBS in-house data-base. Top-ranked identified Hits (33 compounds) were selected to subject for in-vitro biological activity evaluation against β-glucuronidase enzyme; out of them, twenty compounds (61% of screened compounds) evaluated as actives, however eleven compounds were found to have significantly higher inhibitory activity, including compounds 1, 5–8, 10, 12–13, and 17–19 with IC50 values ranging from 1.2 μM to 34.9 μM. Out of the eleven potent inhibitors, seven compounds 1, 5, 6, 7, 8, 13, and 19 were found new, and evaluated first time for the β-glucuronidase inhibitory activity. Compounds 1, 5 and 19 exhibited a highly potent inhibition in uM of β-glucuronidase enzyme with non-cytotoxic behavior against the mouse fibroblast (3T3) cell line. Our combined in-silico and in-vitro results revealed that the binding pattern analysis of the eleven potent inhibitors, showed almost similar non-covalent interactions, as observed in case of our validated pharmacophore model. The obtained results thus demonstrated that the virtual screening minimizes false positives, and provide a template for the identification and development of new and more potent β-glucuronidase inhibitors with non-toxic effects.

Effect of Sideritis leptoclada against HT-144 human malignant melanoma.

Sideritis leptoclada O. Schwarz et P.H. Davis extracts were evaluated for its singlet oxygen production capacity using spectrophotometric method. The extracts producing singlet oxygen were then evaluated for cytotoxicity against malignant melanoma cancer (HT-144) and fibroblast (3T3) cell lines using the 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay. The photocytotoxicity against the HT-144 human melanoma cell line in the presence of illumination (∼≥400 nm) was also evaluated. In the standard MTT assay, the ethanol extract of S. leptoclada (100 µg/ml) showed 83.49±3.33% inhibition of HT-144 cancer cells, whereas in the illuminated MTT assay, it showed 77.46±1.97% inhibition of HT-144 cancer cells. The effects of ethanol extract on reactive oxygen species production, apoptosis, and tumor necrosis factor-α secretion were also evaluated on HT-144 cell lines. The extract triggered an increase in intracellular reactive oxygen species production and tumor necrosis factor-α secretion compared with the respective controls. Thus, the ethanol extract may cause apoptosis. The LC-MS/MS analyses of S. leptoclada ethanolic extract showed that it has quinic acid (137213±11.25 µg/g extract), malic acid (1468±0.16 µg/g extract), chlorogenic acid (881.7±0.06 µg/g extract), and apigetrin (223.2±0.13 µg/g extract) as major constituents. The ethanolic extract of S. leptoclada should be further investigated as a potential treatment for malignant melanoma cancer.

Incorporation of Levodopa into Biopolymer Coatings Based on Carboxylated Carbon Nanotubes for pH-Dependent Sustained Release Drug Delivery.

Four drug delivery systems were formulated by non-covalent functionalization of carboxylated single walled carbon nanotubes using biocompatible polymers as coating agent (i.e., Tween 20, Tween 80, chitosan or polyethylene glycol) for the delivery of levodopa, a drug used in Parkinson’s disease. The chemical interaction between the coating agent and carbon nanotubes-levodopa conjugate was confirmed by Fourier transform infrared (FTIR) and Raman studies. The drug release profiles were revealed to be dependent upon the type of applied coating material and this could be further adjusted to a desired rate to meet different biomedical conditions. In vitro drug release experiments measured using UV-Vis spectrometry demonstrated that the coated conjugates yielded a more prolonged and sustained release pattern compared to the uncoated conjugate. Cytotoxicity of the formulated conjugates was studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay using normal mouse embryonic fibroblast 3T3 cell line. Compared to the non-coated conjugate, the MTT data indicated that the coating procedure improved the biocompatibility of all systems by 34–41% when the concentration used exceeded 100 μg/mL. In conclusion, the comprehensive results of this study suggest that carbon nanotubes-based drug carrier coated with a suitable biomaterial may possibly be a potential nanoparticle system that could facilitate drug delivery to the brain with tunable physicochemical properties.

Graphene Oxide as a Nanocarrier for a Theranostics Delivery System of Protocatechuic Acid and Gadolinium/Gold Nanoparticles.

We have synthesized a graphene oxide (GO)-based theranostic nanodelivery system (GOTS) for magnetic resonance imaging (MRI) using naturally occurring protocatechuic acid (PA) as an anticancer agent and gadolinium (III) nitrate hexahydrate (Gd) as the starting material for a contrast agent,. Gold nanoparticles (AuNPs) were subsequently used as second diagnostic agent. The GO nanosheets were first prepared from graphite via the improved Hummer’s protocol. The conjugation of the GO and the PA was done via hydrogen bonding and π–π stacking interactions, followed by surface adsorption of the AuNPs through electrostatic interactions. GAGPA is the name given to the nanocomposite obtained from Gd and PA conjugation. However, after coating with AuNPs, the name was modified to GAGPAu. The physicochemical properties of the GAGPA and GAGPAu nanohybrids were studied using various characterization techniques. The results from the analyses confirmed the formation of the GOTS. The powder X-ray diffraction (PXRD) results showed the diffractive patterns for pure GO nanolayers, which changed after subsequent conjugation of the Gd and PA. The AuNPs patterns were also recorded after surface adsorption. Cytotoxicity and magnetic resonance imaging (MRI) contrast tests were also carried out on the developed GOTS. The GAGPAu was significantly cytotoxic to the human liver hepatocellular carcinoma cell line (HepG2) but nontoxic to the standard fibroblast cell line (3T3). The GAGPAu also appeared to possess higher T1 contrast compared to the pure Gd and water reference. The GOTS has good prospects of serving as future theranostic platform for cancer chemotherapy and diagnosis.

The effect of urinary Foley catheter substrate material on the antimicrobial potential of calixerene-based molecules.

This study was to investigate the antimicrobial activity of a modified calixarene polymer bound to a silicone substrate in the presence of pathogens associated with catheter infections, Escherichia coli and Proteus mirabilis. The molecule and its constituent parts were studied bound and unbound to silicone substrates to ascertain growth effects. Minimum inhibitory and bactericidal concentrations were determined against E.coli and P. mirabilis. Biofilm growth was studied by immersing silicone discs seeded with either P. mirabilis or E.coli in artificial urine. Biofilms were assessed at 3, 7 and 10days. The coated material reduced bacterial cell density compared to the uncoated samples. Direct and indirect toxicity tests were conducted with a fibroblast cell line (3T3); coated and noncoated silicone samples were seeded with cells (1×104 /cm2 ) and incubated for 72h. Hoechst propidium iodide staining identified delayed toxic effect from the coated and noncoated material leachate in all but the platinum-cured medical-grade silicone, which showed no evident toxicity. The calixerene polymer was determined to be the active part of the coating. Biofilm formation was dramatically reduced in the coated platinum-cured medical-grade silicone samples, but cell viability was reduced on the clinical-grade silicones regardless of coating in contrast to cells seeded on the platinum-cured medical-grade silicone. A delayed toxic response was evident to the extract of the coated and noncoated clinical-grade samples, indicating that the toxic effect is due to the underlying substrate. This study has established that the immobilized molecule enhances the antibacterial and antifouling properties of silicone, without toxicity. It also clearly demonstrates that regardless of coating efficacy, the substrate material has the capacity to disrupt its potency and change the nature of the material coating.

Anti-inflammatory and anti-melanogenic effects of major leaf components of Alpinia zerumbet var. excelsa

Background: The leaves of Alpinia zerumbet var. excelsa (alpinia) are used to prepare traditional food items and folk medicines. Objective: This study was designed with a view to explore the anti-inflammatory and anti-melanogenic potentials of major components of alpinia leaves. Materials and Methods: Anti-inflammatory effects of leaf-derived compounds were primarily assessed with protein denaturation and proteinase assay. Their inhibitory effects on nitrite accumulation and prostaglandin E2 (PGE2) production in lipopolysaccharide-induced RAW 264.7 cells were also evaluated. For anti-melanogenic assays, all the compounds were tested on α-melanocyte-stimulating hormone-stimulated B16F10 cells and on mushroom tyrosinase in vitro. Their cytotoxicity was evaluated using fibroblast cell line 3T3 L-1 and brine shrimps. Results: Five compounds, 5,6-dehydrokawain, dihydro-5,6-dehydrokawain, (E)-5-methoxy-8-(4-methoxy-2-oxo-2H-pyran-6-yl)-7-phenyl-1-styryl-2-oxabicyclo[4.2.0]oct-4-en-3-one (AS-2), kaempferol 3-O-β-D-glucuronide (KOG), and quercetin 3-O-β-D-glucuronide (QOG) were purified from the leaves. Of which, AS-2 and QOG were purified for the first time. All compounds significantly inhibited albumin denaturation and proteinase activity. AS-2, KOG, and QOG remarkably inhibited nitric oxide formation with IC50 values of 8.2, 13.3, and 12.6 μM, respectively, in RAW 264.7 cells. They also inhibited PGE2 production with IC50 values 19.8-23.7 μM. They showed anti-melanogenic effects reducing tyrosinase activity (IC50 values 29.6-112.5 μM) and melanin formation (IC50 values 30.8–164.4 μM) in B16F10 cells, and inhibiting mushroom tyrosinase (IC50 values 61.5-456.4 μg/ml). Conclusion: Taken together, major components of alpinia leaf could be utilized as a potent herbal drug and food supplement with therapeutic prospects against inflammatory disorders and hyperpigmentation. Abbreviations used: HPLC: High performance liquid chromatography; DK: 5,6-dehydrokawain; DDK: Dihydro-5,6-dehydrokawain; AS-2: (E)-5-methoxy-8-(4-methoxy-2-oxo-2H-pyran-6-yl)-7-phenyl-1-styryl-2-oxabicyclo[4.2.0]oct-4-en-3-one; KOG: Kaempferol 3-O-β-D-glucuronide; QOG: Quercetin 3-O-β-D-glucuronide; PGE2: Prostaglandin E2; PAK1: RAC/CDC42-activated kinase 1; UV: Ultra violet; α-MSH: α-Melanocyte-stimulating hormone; COX-2: Cyclooxygenase-2; iNOS: Inducible Nitric Oxide Synthase; PDGF: Platelet Derived Growth Factor; MITF: Microphthalmia-Associated Transcription Factor; TRP: Tyrosinase Related Protein; ATCC: American Type Culture Collection; DMEM: Dulbecco's Modified Eagle Medium; FBS: Fetal bovine serum; CS: Newborn calf serum; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide; DMSO: Dimethyl sulfoxide; NO: Nitric oxide; LPS: Lipopolysaccharide; DMRT: Duncan's multiple range test; SPSS: Statistical Package for the Social Sciences.

Aminoquinoline Schiff Bases as Non-Acidic, Non-Steroidal, Anti-Inflammatory Agents

Inflammation is common localized condition which makes body reddened and swollen due to some injury or any infection. The commonly available drugs i. e. diclofenac, ibuprofen etc. in market against inflammation bear acid group which cause severe stomach disturbance in their prolong use. Thus, such issues required significant attention in term of introducing acid free new anti-inflammatory agents with enhance efficacy. In the present study, we screened a series of eighteen aminoquinoline Schiff bases for their anti-inflammatory activity. These compounds were tested for their inhibitory effect on intracellular reactive oxygen species (ROS) produced from phagocytes isolated from human whole blood. Among the synthetic analogs, (E)-4-((quinolin-5-ylimino)methyl)benzene-1,2,3-triol (5), (E)-1-(2,4-dimethoxyphenyl)-N-(quinolin-5-yl)methanimine (6), (E)-2,6-dimethoxy-4-((quinolin-5-ylimino)methyl)phenol (7), and (E)-4-((quinolin-3-ylimino)methyl)benzene-1,2,3-triol (15) were found to exhibit potent activity with IC50 values of 1.9 ± 0.9, 2.7 ± 0.5, 1.4 ± 0.1, and 2.4 ± 0.1 μg/mL, respectively. Ibuprofen with IC50 value of 2.5 ± 0.6 μg/mL has been used standard in this assay. Cytotoxicity of these compounds was also tested against rat fibroblast cell line (3T3 cell line). The synthetic aminoquinoline Schiff bases were found to be non-toxic in cellular model except trihydroxy derivative 5. The active compounds can serve as potential leads for new anti-inflammatory drugs.

Structure and intracellular antioxidant activity of pectic polysaccharide from acerola (Malpighia emarginata)

Malpighia emarginata is a tropical fruit plant, found naturally in the Caribbean islands and South America that produces an edible fruit known as acerola or Barbados Cherry. Its polysaccharides were obtained by aqueous extraction, subjected to a freezing and thawing process and ultrafiltration. A homogeneous fraction (ACWS-01E) was analyzed by sugar composition, HPSEC, methylation and NMR spectroscopy analyses. The results showed an arabinan-rich pectic polysaccharide, with 6.1×104g/mol and formed mainly by a high methyl esterified (DM=86%) homogalacturonan and branched arabinan. This latter is anchored in type I rhamnogalacturonan regions. The main chain of arabinan consisted of (1→5)-linked α-Araf, branched only at O-3. The potential ACWS-01E intracellular antioxidant activity against H2O2-induced oxidative stress in murine fibroblast cell line (3T3) was determined by DCFH-DA assay. The treatment with ACWS-01E significantly reduced H2O2-induced cytotoxic effect and the levels of reactive oxygen species (ROS). These findings suggested that ACWS-01E protected and improved NIH 3T3 cell viability from H2O2-induced toxicity by decreasing intracellular levels of ROS.

Studies on new urease inhibitors by using biochemical, STD-NMR spectroscopy, and molecular docking methods

Discovery of new urease inhibitors is an important approach towards the treatment of diseases caused by ureolytic bacteria. Urease has an important role in several pathologies, such as urolithiasis, peptic and duodenal ulcers, etc. In this regard, urease inhibitory activity of heterocyclic synthetic compounds 1–14, belonging to different chemical classes was evaluated by employing in vitro biochemical assay, and saturation transfer difference-NMR technique. Compounds 1, 3–5, 7, 10, and 12 have shown urease inhibitory potential in vitro. Among them, compound 1 was found to be more potent (IC50 of 12.90 ± 0.63 µM) than a clinical drug, acetohydroxamic acid (IC50 = 41.5 ± 1.50 µM) (Standard). Compounds 3 (IC50 = 15.0 ± 1.10 µM), 4 (IC50 = 24.67 ± 2.87 µM), and 5 (IC50 = 25.50 ± 0.63 µM) were also identified as potent inhibitors of urease enzyme. These compounds showed strong interactions with the urease enzyme (receptor) in saturation transfer difference-NMR spectra. Specifically, aromatic protons of active compounds received maximum Rf saturation from the receptor protein, thus were inferred to be as in close proximity to the protein. New inhibitors identified through biochemical assay and saturation transfer difference-nuclear magnetic resonance techniques were then subjected to kinetic and molecular docking studies to investigate their mode of inhibition, and production of their interactions with the protein at atomic level, respectively. Active compounds were found to be non-cytotoxic against mouse fibroblast (3T3) cell line MTT assay. Present study thereby identifies new inhibitors of urease enzyme in vitro as leads for further investigation towards the development of novel mechanism-based urease inhibitors.

Tyrosine assisted size controlled synthesis of silver nanoparticles and their catalytic, in-vitro cytotoxicity evaluation.

A simple, green approach for the size controllable preparation of silver nanoparticles (SNPs) using tyrosine as reducing and capping agent is shown here. The size of SNPs is controlled by varying the pH of tyrosine solution. The as synthesized SNPs are characterized by using XRD, UV-Visible, DLS, TEM and SAED. Zeta potential measurements revealed the stability of tyrosine capped silver nanocolloids. Furthermore, catalytic activity studies concluded that the smaller SNPs acts as good catalyst and the catalytic activity depends on size of the nanoparticles. Further, the in-vitro cytotoxicity experiments concluded that the cytotoxicity of the prepared SNPs towards mouse fibroblast (3T3) cell lines is size and dose dependent. Additionally, the present approach is substitute to the traditional methods that are being used now-a-days for size controlled synthesis of SNPs.

Cytotoxicity of Etch-and-Rinse, Self-Etch, and Universal Dental Adhesive Systems in Fibroblast Cell Line 3T3.

The aim of this study was to evaluate in fibroblast cultures the direct cytotoxic effects of etch-and-rinse, self-etch, and universal adhesive systems. The sterile glass cover slips (n = 3) were then immersed in culture medium to obtain the eluates for the experimental groups: (1) Adper™ Single Bond 2; (2) Ambar; (3) Adper™ Scotchbond™ Multi-Purpose; (4) Scotchbond™ Universal; (5) Ambar Universal; and (6) OptiBond All-In-One. As a negative control, sterile glass cover slips were immersed in culture medium only. After 24 h, the eluate obtained was applied on fibroblast culture. Cell viability and cell morphology were evaluated by MTT assay and SEM, respectively. Data were analyzed by Kruskal–Wallis and Mann–Whitney tests (α = 0.05). All adhesive systems except universal reduced cell viability in 3T3 cells to between 26.04% and 56.57%, and Scotchbond Universal and Ambar Universal reduced cell viability to 2.13% and 3.57%, respectively, when compared to the negative control. Cytoplasmic membrane shrinkage and cell-free areas with residual membrane fragments from dead cells were observed. In conclusion, improvements in universal adhesive system formulations and their mechanisms of action are not accompanied by increased toxicity compared with those in other systems, warranting commitment to the use of these dentin-pulp complexes.

One-Pot Aqueous Synthesization of Near-Infrared Quantum Dots for Bioimaging and Photodynamic Therapy of Gliomas.

As the early detection and total destruction of gliomas are essential for longer survival, we attempted to synthesize a quantum dot (QD) that is capable of recognizing glioma cells for imaging and photodynamic therapy. Using a one-pot aqueous approach, near infrared-emitting CdTe was produced. After detection of its physicochemical characteriistics, it was conjugated with RGD. The emission images were observed with confocal microscopy. To test its toxicity, CdTe-RGD at various concentrations was separately added to a human glioma cell line (U251) and a mouse embryo fibroblast cell line (3T3) (control) for incubation in dark conditions. To test its photodynamic effect, the U251 and 3T3 cells were then irradiated for 5-60 min, using a 632.8-nm laser. This QD (Φ = 3.75 nm, photoluminescence (PL) peak wavelength = 700 nm, photoluminescence quantum yield (PLQY) = 20 %), was a spherical crystal with excellent monodispersity. Under a confocal microscope, U251 cells were visualized, but not the 3T3 cells. In dark conditions, the survival rates of both U251 and 3T3 cells were above 85 %. After laser irradiation, the survival rate of U251 cells decreased to 37 ± 1.6 % as the irradiation time and the CdTe-RGD concentration were increased. With good physicochemical characteriistics and low toxicity, this QD-RGD has broad prospects for use in the biomedical imaging and photodynamic therapy of gliomas.

Environment friendly approach for size controllable synthesis of biocompatible Silver nanoparticles using diastase.

A green, facile method for the size selective synthesis of silver nanoparticles (AgNPs) using diastase as green reducing and stabilizing agent is reported. The thiol groups present in the diastase are mainly responsible for the rapid reaction rate of silver nanoparticles synthesis. The variation in the size and morphology of AgNPs were studied by changing the pH of diastase. The prepared silver nanoparticles were characterized by using UV-vis, XRD, FTIR, TEM and SAED. The FTIR analysis revealed the stabilization of diastase molecules on the surface of AgNPs. Additionally, in-vitro cytotoxicity experiments concluded that the cytotoxicity of the as-synthesized AgNPs towards mouse fibroblast (3T3) cell lines is dose and size dependent. Furthermore, the present method is an alternative to the traditional chemical methods of size controlled AgNPs synthesis.