L929 Murine Fibroblasts Research Articles

Optimization of Cationic Nanogel PEGylation to Achieve Mammalian Cytocompatibility with Limited Loss of Gram-Negative Bactericidal Activity.

Tuning the composition of antimicrobial nanogels can significantly alter both nanogel cytotoxicity and antibacterial activity. This project investigated the extent to which PEGylation of cationic, hydrophobic nanogels altered their cytotoxicity and bactericidal activity. These biodegradable, cationic nanogels were synthesized by activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP) emulsion polymerization with up to 13.9 wt % PEG (MW = 2000) MA, as verified by 1H NMR. Nanogel bactericidal activity was assessed against Gram-negative E. coli and P. aeruginosa and Gram-positive S. mutans and S. aureus by measuring membrane lysis with a LIVE/DEAD assay. E. coli and S. mutans viability was further validated by measuring metabolic activity with a PrestoBlue assay and imaging bacteria stained with a LIVE/DEAD probe. All tested nanogels decreased the membrane integrity (0.5 mg/mL dose) for Gram-negative E. coli and P. aeruginosa, irrespective of the extent of PEGylation. PEGylation (13.9 wt %) increased the cytocompatibility of cationic nanogels toward RAW 264.7 murine macrophages and L929 murine fibroblasts by over 100-fold, relative to control nanogels. PEGylation (42.8 wt %) reduced nanogel uptake by 43% for macrophages and 63% for fibroblasts. Therefore, PEGylation reduced nanogel toxicity to mammalian cells without significantly compromising their bactericidal activity. These results facilitate future nanogel design for perturbing the growth of Gram-negative bacteria.

First dinuclear rhodium(II) complexes with triazolopyrimidines and the prospect of their potential biological use

Five novel rhodium(II) complexes of general formula [Rh2(μ-OOCCH3)4L2], where L is a triazolopyrimidine derivative, in particular dimethyl-1,2,4-triazolo[1,5-a]pyrimidine (dmtp) for (1), 5,7-diethyl-1,2,4-triazolo[1,5-a]pyrimidine (detp) for (2), 7-isobutyl-5-methyl-1,2,4-triazolo[1,5-a]pyrimidine (ibmtp) for (3), 7-hydroxy-5-methyl-1,2,4-triazolo[1,5-a]pyrimidine (HmtpO) for (4) and 5,7-ditertbutyl-1,2,4-triazolo[1,5-a]pyrimidine (dbtp) for (5) are reported. These first representatives of paddle-wheel dirhodium complexes with triazolopyrimidines have been characterized by IR and NMR spectroscopy as well as by single-crystal X-ray diffraction studies. Three of the new complexes (1), (2) and (5) were thoroughly screened in vitro for their cytotoxicity against human breast cancer cell line MCF-7 and L929 murine fibroblast cells. Favorably, they show significantly less effective inhibition on the cell growth of L929 than cisplatin under identical conditions. Complexes (1) and (5) display moderate cytotoxic activity (IC50 = 16.3–21.5 μM) against MCF-7 cells which is induced via reactive oxygen species-independent pathways. Extensive studies of rhodium complexes (1), (2) and (5) against microorganisms have shown that the tested compounds exhibit antibacterial activity against Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis) while (5) significantly inhibited the growth of Malassezia furfur. The highest antibacterial, and antifungal activity, was observed for (5).

Neuroprotective Effects of Low Intensity and Low Frequency Electromagnetic �in vitro� Stimulation on Glial Cells

Degenerative neurological diseases (senile dementia, Alzheimer`s disease, glaucoma), post-stroke sequelae are increasing as prevalence nowadays, in context of aging in general population, and there are strong evidences that astrocytes may play a significant role in neuroprotection. The study evaluates the effects of low intensity low frequency electromagnetic field (EMF) �in vitro� stimulation (195 mA, 7-8 Hz) on the glial cells, in different conditions, in order to identify the neuroprotective potential. Three cell lines were used for the study: the Clonetics (Lonza) line of normal human astrocytes, U87 glioblastoma tumor cells (ATCC) and L929 murine fibroblasts. The cell cultures were exposed to EMF stimulation for 2 hours/day, for 5 and 10 day, in absence and in the presence of oxidative stress stimuli. Cell viability, apoptotic potential, changes in cell phenotype, stimulation of cell proliferation for normal and malignant transformed cell lines, resilience to oxidative stress were investigated. The viability of the cells was investigated by MTS assay, and the Griess test was used to detect nitrite production as a marker of oxidation and inflammation. Inverted microscopy examination revealed that following exposure to the tested EMF, no morphological changes of the cells were recorded. In none of the cell lines tested, no cytotoxic or cytopathic effects were observed in the cultures exposed under low density conditions. In case of exposure of cell cultures for 10 days to the action of EMF, a viability of 98.2% of glial cells and 97% of exposed fibroblasts is observed, compared to unexposed cells. The viability of the cells exposed to both EMF and oxidative and inflammatory stimuli is increased by 0.96% in the case of H2O2 stimulation and in 3.76% in the case of LPS stimulation, suggesting a possible neuroprotective effect of EMF exposure. Low intensity low frequency EMF �in vitro� stimulation increased viability of the glial cells exposed to oxidative and inflammatory stimuli. No cytotoxic or cytopathic effects were observed in the exposed cultures under conditions of low density. Further tests are needed in order to elucidate the mechanisms of action of EMF on astrocytes, especially on astrocyte-neuron co-cultures to highlight the interactions between these cell types. Confirmation of the neuroprotective effect of low intensity and low frequency EMF opens the way for the development of an innovative, non-invasive therapy.

Phenotypic high-throughput screening platform identifies novel chemotypes for necroptosis inhibition

Regulated necrosis or necroptosis, mediated by receptor-interacting kinase 1 (RIPK1), RIPK3 and pseudokinase mixed lineage kinase domain-like protein (MLKL), contributes to the pathogenesis of inflammatory, infectious and degenerative diseases. Recently identified necroptosis inhibitors display moderate specificity, suboptimal pharmacokinetics, off-target effects and toxicity, preventing these molecules from reaching the clinic. Here, we developed a cell-based high-throughput screening (HTS) cascade for the identification of small-molecule inhibitors of necroptosis. From the initial library of over 250,000 compounds, the primary screening phase identified 356 compounds that strongly inhibited TNF-α-induced necroptosis, but not apoptosis, in human and murine cell systems, with EC50 < 6.7 μM. From these, 251 compounds were tested for RIPK1 and/or RIPK3 kinase inhibitory activity; some were active and several have novel mechanisms of action. Based on specific chemical descriptors, 110 compounds proceeded into the secondary screening cascade, which then identified seven compounds with maximum ability to reduce MLKL activation, IC50 >100 μM, EC50 2.5–11.5 μM under long-term necroptosis execution in murine fibroblast L929 cells, and full protection from ATP depletion and membrane leakage in human and murine cells. As a proof of concept, compound SN-6109, with binding mode to RIPK1 similar to that of necrostatin-1, confirmed RIPK1 inhibitory activity and appropriate pharmacokinetic properties. SN-6109 was further tested in mice, showing efficacy against TNF-α-induced systemic inflammatory response syndrome. In conclusion, a phenotypic-driven HTS cascade promptly identified robust necroptosis inhibitors with in vivo activity, currently undergoing further medicinal chemistry optimization. Notably, the novel hits highlight the opportunity to identify new molecular mechanisms of action in necroptosis.

TEMPO-Nanocellulose/Ca2+ Hydrogels: Ibuprofen Drug Diffusion and In Vitro Cytocompatibility.

Stable hydrogels with tunable rheological properties were prepared by adding Ca2+ ions to aqueous dispersions of 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO)-oxidized and ultra-sonicated cellulose nanofibers (TOUS-CNFs). The gelation occurred by interaction among polyvalent cations and the carboxylic units introduced on TOUS-CNFs during the oxidation process. Both dynamic viscosity values and pseudoplastic rheological behaviour increased by increasing the Ca2+ concentration, confirming the cross-linking action of the bivalent cation. The hydrogels were proved to be suitable controlled release systems by measuring the diffusion coefficient of a drug model (ibuprofen, IB) by high-resolution magic angle spinning (HR-MAS) nuclear magnetic resonance (NMR) spectroscopy. IB was used both as free molecule and as a 1:1 pre-formed complex with β-cyclodextrin (IB/β-CD), showing in this latter case a lower diffusion coefficient. Finally, the cytocompatibility of the TOUS-CNFs/Ca2+ hydrogels was demonstrated in vitro by indirect and direct tests conducted on a L929 murine fibroblast cell line, achieving a percentage number of viable cells after 7 days higher than 70%.

Preparation and Characterization of Fluoride-Incorporated Plasma Electrolytic Oxidation Coatings on the AZ31 Magnesium Alloy

In this study, films with different fluorine contents were prepared on an AZ31 magnesium alloy by using plasma electrolytic oxidation to study the corrosion resistance and cytocompatibility of the alloy. The morphology of the coating surface, phase, and chemical elements were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), and energy-dispersive X-ray spectrometry (EDS). The changes in the corrosion resistance with different fluorine contents were investigated by electrochemical experiments, hydrogen evolution, and long-term immersion tests. In addition, murine fibroblast L-929 cells were adopted for in vitro cytotoxicity tests using the cell counting kit (CCK)-8 assay, and the morphology of the cells was observed simultaneously by inverted microscopy. The results showed that the main form of the fluorine ions in the plasma electrolytic oxidation coatings was magnesium fluoride (MgF2). In addition, the corrosion resistance and cytocompatibilities of the coatings were improved by the addition of fluoride ions. When the content of potassium fluoride reached 10 g/L, the cell compatibility and corrosion resistance were the best, a finding which provides a basis for the clinical applications of the AZ31 magnesium alloy in the biomedical field.

Synthesis, structure and biological evaluation of ruthenium(III) complexes of triazolopyrimidines with anticancer properties.

Six novel ruthenium(III) complexes of general formula [RuCl3(L)3] (1,3,5) and [RuCl3(H2O)(L)2] (2,4,6), where L stands for three different triazolopyrimidine-derived ligands, are reported. The compounds have been structurally characterized (IR, EPR, SCXRD), and their magnetic moments have been determined. The single-crystal X-ray diffraction study revealed a slightly distorted octahedral geometry of the Ru(III) complexes with mer configuration in 1 and 5, and fac configuration in 3. In 2 and 4, three chloride ions are in mer configuration and the two triazolopyrimidines are oriented trans mutually with the water molecule playing the role of the sixth ligand. All complexes have been thoroughly screened for their in vitro cytotoxicity against human breast cancer cell line MCF-7, human cervical cancer cell line HeLa, and L929 murine fibroblast cells, uncovering among others that the most lipophilic complexes 5 and 6, containing the bulky ligand dptp (5,7-diphenyl-1,2,4-triazolo[1,5-a]pyrimidine), display high cytotoxic activity against MCF-7, and HeLa cells. Moreover, it was also revealed that during the interaction of the complexes 1-6 with the cancer MCF-7 cell line, reactive oxygen species are released intracellularly, which could indicate that they are involved in cell apoptosis. Furthermore, extensive studies have been carried out to reveal the mechanism by which complexes 1-6 interact with DNA, albumin, and apotransferrin. The biological studies were complemented by detailed kinetic studies of the hydrolysis of the complexes in the pH range 5-8, to determine the stability of the complexes in solution. Six novel ruthenium(III) complexes with triazolopyrimidine derivatives demonstrated the potential for use as anticancer agents by maintaining the toxic effect on MCF-7 and HeLa cells.

Pullulan/Poly(Vinyl Alcohol) Composite Hydrogels for Adipose Tissue Engineering.

Composite hydrogels based on pullulan (HP) and poly(vinyl alcohol) (PVA) were both prepared by simple chemical crosslinking with sodium trimethaphosphate (STMP) or by dual crosslinking (simultaneously chemical crosslinking with STMP and physical crosslinking by freeze-thaw technique). The resulting hydrogels and cryogels were designed for tissue engineering applications. PVA, with two different molecular weights (47,000 and 125,000 g/mol; PVA47 and PVA125, respectively), as well as different P/PVA weight ratios were tested. The physico-chemical characterization of the hydrogels was performed by FTIR spectroscopy and scanning electron microscopy (SEM). The swelling kinetics, dissolution behavior, and degradation profiles in simulated physiological conditions (phosphate buffer at pH 7.4) were investigated. Pullulan concentration and the crosslinking method had significant effects on the pore size, swelling ratio, and degradation profiles. Cryogels exhibit lower swelling capacities than the conventional hydrogels but have better stability against hydrolitic degradation. Biocompatibility of the hydrogels was also investigated by both MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and LDH (lactaten dehydrogenase) assay. The MTT and LDH assays proved that dual crosslinked HP/PVA125 (75:25, w/w) scaffolds are more biocompatible and promote to a greater extent the adhesion and proliferation of L929 murine fibroblast cells than chemically crosslinked HP/PVA47 (50/50, w/w) scaffolds. Moreover, the HP/PVA125 cryogel had the best ability for the adipogenic differentiation of cells. The overall results demonstrated that the HP/PVA composite hydrogels or cryogels are suitable biomaterials for tissue engineering applications.

Viral persistence of mammalian reovirus in cell culture: a model of virus-cell coevolution.

Although mammalian reovirus exhibits only limited pathogenicity in humans, it has been, and still remains, instrumental in studies of viral replication and pathogenesis. Generally considered as cytolytic, this virus can sometimes establish long-term persistent infections in tissue culture. In fact, in this context, it constitutes one widely used model to demonstrate coevolution between virus and host cells. Initially limited to the murine L929 fibroblasts model, further studies in different cell types appeared in the last few years. Establishment of viral persistence could also become a preferred approach to isolate new viruses that are better adapted to their applications in virotherapy, for example as oncolytic agents against human or animal cancers. A better understanding of the persistence phenomenon, especially of viral genes involved, is thus essential. The development of new tools, such as reverse genetics, appears very promising to achieve these objectives. Actually, this last approach allows us to establish the biological significance of mutations found on viruses selected during viral persistence.

Viral persistence of mammalian reovirus in cell culture: a model of virus-cell coevolution

Although mammalian reovirus exhibits only limited pathogenicity in humans, it has been, and still remains, instrumental in studies of viral replication and pathogenesis. Generally considered as cytolytic, this virus can sometimes establish long-term persistent infections in tissue culture. In fact, in this context, it constitutes one widely used model to demonstrate coevolution between virus and host cells. Initially limited to the murine L929fibroblasts model, further studies in different cell types appeared in the last few years. Establishment of viral persistence could also become a preferred approach to isolate new viruses that are better adapted to their applications in virotherapy, for example as oncolytic agents against human or animal cancers. A better understanding of the persistence phenomenon, especially of viral genes involved, is thus essential. The development of new tools, such as reverse genetics, appears very promising to achieve these objectives. Actually, this last approach allows us to establish the biological significance of mutations found on viruses selected during viral persistence.

Functional Polyimide-Based Electrospun Fibers for Biomedical Application.

The current study focuses on the application of cytotoxicity tests upon one membrane matrix based on electrospun polyimide fibers, appealing for biomedical application, such as scaffolds for cell growth, patches or meshes for wound healing, etc. Assays were performed in order to determine the viability and proliferation of L929 murine fibroblasts after they were kept in direct contact with the studied electrospun polyimide fibers. Increased cell viability and proliferation were detected for cells seeded on electrospun polyimide fibers membrane, in comparison with the control system, either after two or six days of evaluation. The number of live cells was higher on the studied material compared to the control, after two and six days of cell seeding. The tendency of the cells to proliferate on the electrospun polyimide fibers was revealed by confocal microscopy. The morphological stability of electrospun polyimide membrane was evaluated by SEM observation, after immersion of the samples in phosphate buffer saline solution (PBS, 7.4 at 37 °C) at various time intervals. Additionally, the easy production of electrospun polyimide fibers can facilitate the development of these types of matrices into specific biomedical applications in the future.

Synthesis of dehydro-α-lapachones, α- and β-lapachones, and screening against cancer cell lines

14 new naphthoquinones were prepared and tested against human cancer cell lines PC-3 (prostate), HCT-116 (colon carcinoma), SNB-19 (glioblastoma), HL-60 (leukemia) and MCF-7 (breast), and a nontumor cell line L929 (murine fibroblasts) to determine cytotoxicity with the MTT assay. 8-OH-β-lapachones (14a, 14c, 14d) presented best results, showing low IC50 values and high selectivity for HCT-116 and HL-60 tumor cells.

Titanium coated with poly(lactic-co-glycolic) acid incorporating simvastatin: Biofunctionalization of dental prosthetic abutments.

To propose a biofunctionalized prosthetic abutment by analyzing physico-chemical and morphological properties, simvastatin (SIM) release, and biocompatibility of titanium (Ti) disks coated with poly(lactic-co-glycolic) acid (PLGA) incorporating SIM. Titanium disks (8×3mm) were distributed into four groups: Ti: pure Ti; Ti+PLGA: Ti coated with PLGA; Ti+PLGA+SIM6%: Ti+PLGA with 6% SIM; and Ti+PLGA+SIM0.6%: Ti+PLGA incorporating 0.6% SIM. PLGA was prepared through chloroform evaporation technique. After complete dissolution of PLGA, SIM was diluted in the solution. Ti+PLGA, Ti+PLGA+SIM6%, and Ti+PLGA+SIM0.6% were dip coated with PLGA and PLGA+SIM, respectively. Samples were sterilized by ethylene oxide. For SIM release assay, disks were submerged in PBS, pH 7.4, 37°C, 30rpm up to 600hours. At different time intervals, SIM was quantified by spectrophotometry (238nm). For characterization of the biomaterial components, it was performed Fourier-transform infrared spectroscopy, differential scanning calorimetry, scanning electron microscopy (SEM), optical profilometry, and atomic force microscopy. Biocompatibility analyses were performed by MTS colorimetric assay on murine fibroblasts L929, human gingival fibroblasts (HGFs), and stem cells from human exfoliated deciduous teeth (SHEDs). Absorbance was measured at 490nm, and percentages of viable cells were calculated in relation to positive control (Ti). SEM images were obtained to verify cell adhesion and morphology. One-way ANOVA followed by Tukey's post hoc test was applied (P<0.05) for statistical analyses. SIM release was slow and continuous, reaching about 21% of the incorporated SIM after 600hours. Topographical analyses revealed success in coating Ti disks with PLGA incorporating SIM. Regarding biocompatibility test, Ti+PLGA+SIM0.6% showed the highest percentage of L929 viability at days 3 and 7. There was no significant difference for Ti, Ti+PLGA, and Ti+PLGA+SIM0.6% groups on cell viability of both SHEDs and HGFs at days 3 and 7. SEM corroborates that SHEDs and HGFs were able to adhere and proliferate on Ti, Ti+PLGA, and Ti+PLGA+SIM0.6% surfaces. A slow and controlled release of SIM was achieved, attributed to a diffusional mass transfer mechanism. Moreover, a homogenous coating topography was obtained. Additionally, 0.6% SIM incorporated into PLGA coating improved fibroblasts L929 viability compared to titanium or PLGA. Also, 0.6% SIM incorporated into PLGA promoted cell viability of about 100% for HGFs and approximately 150% for human mesenchymal stem cells. Therefore, this study allows to consider the use of PLGA-coated titanium incorporating SIM as a biofunctionalized abutment for dental implants.

Protective effects of Clematichinenoside AR against inflammation and cytotoxicity induced by human tumor necrosis factor-α

Clematichinenoside AR (AR), a major active ingredient extracted from traditional Chinese herb Clematis chinensis Osbeck, has been demonstrated to possess anti-inflammatory and immune-modulatory activities in the treatment of experimental rheumatoid arthritis (RA). The therapeutic potential of AR was supposed to be closely correlated to its ability against tumor necrosis factor-α (TNF-α). Therefore, we aimed to explore the protective effects of Clematichinenoside AR against inflammation and cytotoxicity induced by human TNF-α. AR treatment significantly decreased IL-6 and IL-8 secretion, and attenuated MMP-1 production in human RA-derived fibroblast-like synoviocyte MH7A cells stimulated by recombinant human TNF-α (rhTNF-α). AR might antagonize rhTNF-α-induced responses in MH7A cells through inhibiting p38 and ERK MAPKs signal activation. In TNF-α-sensitive murine fibroblast L929 cells, AR treatment attenuated the proliferation inhibition ratio induced by rhTNF-α/ActD and antagonized rhTNF-α-induced cytotoxicity. The cellular and nuclear morphological alterations in apoptotic characteristics induced by rhTNF-α/ActD in L929 cells were observed to be attenuated by the pretreatment with AR under a phase-contrast and fluorescence microscopy, respectively. The Annexin V-FITC/PI double-staining assay was performed to confirm that AR pretreatment obviously decreased the cell death. The antagonistic effects of AR against rhTNF-α-induced cytotoxicity might be potentially attributed to the degeneration of reactive oxygen species and the increasing of mitochondrial membrane potential, along with the suppression of durative phosphorylation of c-Jun N-terminal kinase (JNK). Collectively, our results indicated that AR antagonizes the inflammatory and cytotoxic activities induced by human TNF-α effectively in vitro, which provided further evidence for a novel mechanism underlying AR for treating RA correlating with excessive TNF-α production.

In vitro biosafety of pro-ecological chitosan-based hydrogels modified with natural substances.

Hydrogels belong to the group of materials with growing interest on the market of polymers. In this article, hydrogels based on Beetosan were obtained using ultraviolet (UV) radiation. Main component of hydrogel matrix-Beetosan-is chitosan obtained from naturally died honeybees. Such hydrogels were modified with active substances, that is, caffeine, bee pollen, Salvia officinalis (sage), and Aloe vera juice. Next, the analysis of cytotoxicity of hydrogels in relation to murine fibroblasts by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide and neutral red uptake assays were conducted. Furthermore, surface morphology, tensile strength, geometry, and roughness of hydrogels were characterized. Hydrogels did not show cytotoxicity to recommended L929 murine fibroblasts. These polymers did not affect adversely the growth and viability of these cells. Moreover, Beetosan hydrogels were characterized by flexibility as well as by diversified surface morphology that could indicate their high absorbency. Therefore these materials may be considered as useful for biomedical purposes with special emphasis on application as modern wound dressings that not only absorb wound exudate but also contain natural substances with therapeutic properties that is beneficial from the point of view of wound healing process.

Mitochondrial Heat Shock Response Induced by Ectromelia Virus is Accompanied by Reduced Apoptotic Potential in Murine L929 Fibroblasts

Poxviruses utilize multiple strategies to prevent activation of extrinsic and intrinsic apoptotic pathways for successful replication. Mitochondrial heat shock proteins (mtHsps), especially Hsp60 and its cofactor Hsp10, are engaged in apoptosis regulation; however, until now, the influence of poxviruses on mtHsps has never been studied. We used highly infectious Moscow strain of ectromelia virus (ECTV) to investigate the mitochondrial heat shock response and apoptotic potential in permissive L929 fibroblasts. Our results show that ECTV-infected cells exhibit mostly mitochondrial localization of Hsp60 and Hsp10, and show overexpression of both proteins during later stages of infection. ECTV infection has only moderate effect on the electron transport chain subunit expression. Moreover, increase of mtHsp amounts is accompanied by lack of apoptosis, and confirmed by reduced level of pro-apoptotic Bax protein and elevated levels of anti-apoptotic Bcl-2 and Bcl-xL proteins. Taken together, we show a positive relationship between increased levels of Hsp60 and Hsp10 and decreased apoptotic potential of L929 fibroblasts, and further hypothesize that Hsp60 and/or its cofactor play important roles in maintaining protein homeostasis in mitochondria for promotion of cell survival allowing efficient replication of ECTV.

A Self-Adhesive Elastomeric Wound Scaffold for Sensitive Adhesion to Tissue.

Pressure sensitive adhesives based on silicone materials are used particularly for skin adhesion, e.g., the fixation of electrocardiogram (ECG) electrodes or wound dressings. However, adhesion to sensitive tissue structures is not sufficiently addressed due to the risk of damage or rupture. We propose an approach in which a poly-(dimethylsiloxane) (PDMS)-based soft skin adhesive (SSA) acts as cellular scaffold for wound healing. Due to the intrinsically low surface free energy of silicone elastomers, functionalization strategies are needed to promote the attachment and spreading of eukaryotic cells. In the present work, the effect of physical adsorption of three different proteins on the adhesive properties of the soft skin adhesive was investigated. Fibronectin adsorption slightly affects adhesion but significantly improves the cellular interaction of L929 murine fibroblasts with the polymeric surface. Composite films were successfully attached to explanted tympanic membranes. This demonstrates the potential of protein functionalized SSA to act as an adhesive scaffold in delicate biomedical applications.

Magnetic assisted curcumin drug delivery using folate receptor targeted hybrid casein-calcium ferrite nanocarrier

Receptor targeted nanocarriers in the multimodal delivery of cancer therapeutics with minimal side effects are being intensively researched. In this study, folate receptor targeted hybrid protein inorganic nanocarrier was investigated for magnetic-assisted curcumin drug delivery. The protein carrier, casein was hybridized with superparamagnetic calcium ferrite nanoparticles (CFNP) in which the anti-cancer drug, curcumin (Cur) was encapsulated. Finally, folic acid (FA) was conjugated to enable receptor-mediated endocytosis. The synthesized materials were characterized using XRD, FTIR, SEM, VSM and DLS analysis confirming their formulation. The drug loading parameters were optimized by Taguchi technique. Curcumin release from the carrier was studied under the influence of pH, initial drug concentrations and magnetic field. Drug release rate was higher in acidic conditions, increased drug concentrations and under the influence of magnetic field. The drug release mechanism from the carrier were proposed using different kinetic models. Biocompatibility and cytotoxicity tests were performed for the synthesized drug carrier systems using L929 murine fibroblast and MCF-7 breast cancer cells, respectively. The IC50 value reduced nearly six fold for MCF-7 cells, for casein-Cur with FA conjugation in comparison to the carrier without FA. The study clearly demonstrated casein-CFNP-Cur-FA as a novel potential formulation for targeted drug delivery.

Synergic formulation of onion peel quercetin loaded chitosan-cellulose hydrogel with green zinc oxide nanoparticles towards controlled release, biocompatibility, antimicrobial and anticancer activity.

The therapeutic prospective of a novel carrier based delivery of phyto-derived quercetin (QE) extracted from onion peel waste was studied in the present work. Dialdehyde cellulose (DAC) developed from sugarcane bagasse (SCB) cellulose effectively crosslinked chitosan hydrogel. The hydrogel matrices were embedded with green zinc oxide nanoparticles (ZnO NPs), phyto-synthesized using musk melon seeds. The hybrid carrier formulation was characterised using analytical techniques such as XRD, FTIR and SEM analysis. The onion peel drug (OPD) analysed for its bioactive components using HPLC was reported as a potential source of QE. Taguchi optimization for the QE drug loading in the nanohybrid hydrogel indicated a remarkable improvement by 39% in comparison to drug loading in hydrogel without ZnO NPs. The maximum QE release was obtained at an optimal drug loading condition with pH 5.0, which favoured the anticancer applications. The drug release revealed a Fickian diffusion mechanism by adopting various kinetic models. The commercial QE and QE in OPD (QE/OPD) loaded nanohybrid hydrogels were found to inhibit the growth of Staphylococcus aureus and Trichophyton rubrum strains. The biocompatibility and anticancer properties of the QE/OPD loaded nanohybrid hydrogels were established against normal L929 murine fibroblast cells and A431 human skin carcinoma cell lines respectively.

Assessment of Manganese-Zinc Ferrite Nanoparticles as a Novel Magnetic Resonance Imaging Contrast Agent for the Detection of 4T1 Breast Cancer Cells.

Background:The aim of the study was to evaluate the potential of manganese-zinc ferrite nanoparticles (MZF NPs) as a novel negative magnetic resonance imaging (MRI) contrast agents for 4T1 (mouse mammary carcinoma) and L929 (murine fibroblast) cell lines.Methods:MZF NPs and its suitable coating, polyethylene glycol (PEG) via covalent bonding, were investigated under in vitro condition. The cytotoxicity of MZF NPs was tested by 3-(4,5-dimethyl thiazolyl-2)-2,5-diphenyltetrazolium bromide assay after 12 and 24 h of incubation. To evaluate the potential of MZF NPs as T2 MRI nanocontrast agent, images were obtained from phantom containing different Fe concentrations and T2 relaxivity (r2) was measured. The viability of both 4T1 breast cancer and L929 murine fibroblast cell lines incubated with different Fe concentrations.Results:In vitro T2-weighted MRI showed that signal intensity of 4T1 cells was lower than that of L929 as control cells. T2-weighted MRI showed that signal intensity of MZF NPs enhanced with increasing concentration of NPs. The values of 1/T2 relaxivity (r2) for coated MZF NPs with PEG found to be 85.5 mM−1 s−1 which is higher than that of commercially clinical used (Sinerem) MRI contrast agent.Conclusion:The results showed that MZF NPs have potential to detect breast cancer cells (4T1) and also have high contrast resolution between normal (L929) and cancerous cells (4T1) which is a suitable nanoprobe for T2-weighted MR imaging contrast agents.