Rajendra H. Patil

Quantitative Characterization of the Influence of the Nanoscale Morphology of Nanostructured Surfaces on Bacterial Adhesion and Biofilm Formation

Bacterial infection of implants and prosthetic devices is one of the most common causes of implant failure. The nanostructured surface of biocompatible materials strongly influences the adhesion and proliferation of mammalian cells on solid substrates. The observation of this phenomenon has led to an increased effort to develop new strategies to prevent bacterial adhesion and biofilm formation, primarily through nanoengineering the topology of the materials used in implantable devices. While several studies have demonstrated the influence of nanoscale surface morphology on prokaryotic cell attachment, none have provided a quantitative understanding of this phenomenon. Using supersonic cluster beam deposition, we produced nanostructured titania thin films with controlled and reproducible nanoscale morphology respectively. We characterized the surface morphology; composition and wettability by means of atomic force microscopy, X-ray photoemission spectroscopy and contact angle measurements. We studied how protein adsorption is influenced by the physico-chemical surface parameters. Lastly, we characterized Escherichia coli and Staphylococcus aureus adhesion on nanostructured titania surfaces. Our results show that the increase in surface pore aspect ratio and volume, related to the increase of surface roughness, improves protein adsorption, which in turn downplays bacterial adhesion and biofilm formation. As roughness increases up to about 20 nm, bacterial adhesion and biofilm formation are enhanced; the further increase of roughness causes a significant decrease of bacterial adhesion and inhibits biofilm formation. We interpret the observed trend in bacterial adhesion as the combined effect of passivation and flattening effects induced by morphology-dependent protein adsorption. Our findings demonstrate that bacterial adhesion and biofilm formation on nanostructured titanium oxide surfaces are significantly influenced by nanoscale morphological features. The quantitative information, provided by this study about the relation between surface nanoscale morphology and bacterial adhesion points towards the rational design of implant surfaces that control or inhibit bacterial adhesion and biofilm formation.

Cost effective technologies and renewable substrates for biosurfactants’ production

Diverse types of microbial surface active amphiphilic molecules are produced by a range of microbial communities. The extraordinary properties of biosurfactant/bioemulsifier (BS/BE) as surface active products allows them to have key roles in various field of applications such as bioremediation, biodegradation, enhanced oil recovery, pharmaceutics, food processing among many others. This leads to a vast number of potential applications of these BS/BE in different industrial sectors. Despite the huge number of reports and patents describing BS and BE applications and advantages, commercialization of these compounds remain difficult, costly and to a large extent irregular. This is mainly due to the usage of chemically synthesized media for growing producing microorganism and in turn the production of preferred quality products. It is important to note that although a number of developments have taken place in the field of BS industries, large scale production remains economically challenging for many types of these products. This is mainly due to the huge monetary difference between the investment and achievable productivity from the commercial point of view. This review discusses low cost, renewable raw substrates, and fermentation technology in BS/BE production processes and their role in reducing the production cost.

Nanowires of silver-polyaniline nanocomposite synthesized via in situ polymerization and its novel functionality as an antibacterial agent.

Silver-polyaniline (Ag-PANI) nanocomposite was synthesized by in situ polymerization method using ammonium persulfate (APS) as an oxidizing agent in the presence of dodecylbenzene sulfonic acid (DBSA) and silver nitrate (AgNO(3)). The as synthesized Ag-PANI nanocomposite was characterized by using different analytical techniques such as UV-visible (UV-vis) and Fourier transform Infrared spectroscopy (FT-IR), field emission scanning electron microscopy (FE-SEM), thermo gravimetric analysis (TGA), X-ray diffraction (XRD), and transmission electron microscopy (TEM). UV-visible spectra of the synthesized nanocomposite showed a sharp peak at ~420 nm corresponding to the surface plasmon resonance (SPR) of the silver nanoparticles (AgNPs) embedded in the polymer matrix which is overlapped by the polaronic peak of polyaniline appearing at that wavelength. Nanowires of Ag-PANI nanocomposite with diameter 50-70 nm were observed in FE-SEM and TEM. TGA has indicated an enhanced thermal stability of nanocomposite as compared to that of pure polymer. The Ag-PANI nanocomposite has shown an antibacterial activity against model organisms, a gram positive Bacillus subtilis NCIM 6633 in Mueller-Hinton (MH) medium, which is hitherto unattempted. The Ag-PANI nanocomposite with monodispersed AgNPs is considered to have potential applications in sensors, catalysis, batteries and electronic devices.

Biological Synthesis of Copper Oxide Nano Particles Using Escherichia coli

In this paper, we report a facile biological method for extracellular synthesis of copper oxide nanoparticles (CONPs) using Es- cherichia coli (E. coli). We report that trichloroacetic acid (TCA) precipitated protein fraction of E. coli has synthesized copper oxide nanoparticles (CONPs), under simple experimental conditions like aerobic environment, neutral pH and room temperature. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) results have shown that proteins of molecular weight ranging from 22 KDa, 52 KDa, and 25 KDa are not only involved in reduction of Cu (II) into CONPs, but also play a significant role in stabilization of formed nanoparticles at room temperature. Further, transmission electron microscopy (TEM), scanning electron microscopy (SEM), X- ray diffraction measurements (XRD) and fourier transform infrared (FTIR) analysis have confirmed the synthesis of nanoparticles through microbial route. CONPs formed were of variable size and shapes.

Synthesis of Ag-Pt alloy nanoparticles in aqueous bovine serum albumin foam and their cytocompatibility against human gingival fibroblasts.

Foams of bovine serum albumin (BSA) have been utilized for synthesizing in situ protein capped Ag-Pt alloy nanoparticles. The in vitro cytotoxicity and the rate of proliferation of human gingival fibroblasts (HGFs) in presence of the above synthesized alloy nanoparticles is investigated. Expression profile of protein involved in detoxification, i.e. metallotheonein (MT) were assayed by ELISA and expression of mRNA transcripts by reverse transcription polymerase chain reaction (RT-PCR). Cytotoxicity results suggested that protein capped nano-alloys might be promising candidates for implants and prosthetic material. RT-PCR and ELISA confirmed the expression of MT, in cells treated with the alloy nanoparticles. Morphology variation studied by SEM also confirms that cells treated with alloy nanoparticles present an intact morphology.

Antileishmanial drug discovery: comprehensive review of the last 10 years

Leishmaniasis, a group of diseases caused by hemoflagellate obligate intracellular protozoa (trypanosomatids) from the genus Leishmania, has not received the attention it deserves and has developed into a major health problem in developing countries. No effective vaccine is available against leishmaniasis, so chemotherapy is the only effective way to treat all forms of the disease. However, the drugs currently used for treatment of human cutaneous and visceral leishmaniasis are toxic, having severe adverse reactions which limit their use. Therefore, development of novel, effective, and safe antileishmanial agents, with reduced side effects, is a major priority for health researchers, and large numbers of research reports have been published on antileishmanial agents in the last 10 years. Herein, we comprehensively review the developments of the last decade, covering all aspects of leishmaniasis including clinically used drugs, various new classes of antileishmanial agents (synthetic as well as natural), patented antileishmanial agents, and possible drug targets.

Micro-nanopatterning as tool to study the role of physicochemical properties on cell–surface interactions

The current nano-biotechnologies interfacing synthetic materials and cell biology requires a better understanding of cell-surface interactions on the micro-to-nanometer scale. Cell-substrate interactions are mediated by the presence of proteins adsorbed from biological fluids to the substrate. The effect of nanotopography and surface chemistry on protein adsorption as well as the mediation effect on subsequent cellular communication with substratum is not well documented. This review discusses the role of physicochemical properties of cell-surface interactions and state-of-the-art methods currently available for micro-nanoscale surface fabrication and patterning. We also briefly discuss the current surface patterning techniques that allow the combination of a fine and independent control on nanotopography and chemistry to understand the effect of surface nanoscale substrate morphology on cell-surface interactions which has never been realized in previous reports. In addition, we discuss the influence of various chemical patterning and modulation of the nano-topography of surfaces on cell functionality and phenotype.

Biosurfactant/s from Lactobacilli species: Properties, challenges and potential biomedical applications.

Lactic acid bacteria are generally believed to have positive roles in maintaining good health and immune system in humans. A number of Lactobacilli spp. are known to produce important metabolites, among which biosurfactants in particular have shown antimicrobial activity against several pathogens in the intestinal tract and female urogenital tract partly through interfering with biofilm formation and adhesion to the epithelial cells surfaces. Around 46 reports are documented on biosurfactant production from Lactobacillus spp. of which six can be broadly classified as cell free biosurfactant and 40 as cell associated biosurfactants and only approximately 50% of those have reported on the structural composition which, in order of occurrence were mainly proteinaceous, glycolipidic, glycoproteins, or glycolipopeptides in nature. Due to the proteinaceous nature, most biosurfactant produced by strains of Lactobacillus are generally believed to be surlactin type with high potential toward impeding pathogens adherence. Researchers have recently focused on the anti‐adhesive and antibiofilm properties of Lactobacilli‐derived biosurfactants. This review briefly discusses the significance of Lactobacilli‐derived biosurfactants and their potential applications in various fields. In addition, we highlight the exceptional prospects and challenges in fermentation economics of Lactobacillus spp.‐derived biosurfactants’ production processes.

Antileishmanial activity of novel indolyl–coumarin hybrids: Design, synthesis, biological evaluation, molecular docking study and in silico ADME prediction

In present work we have designed and synthesized total twelve novel 3-(3-(1H-indol-3-yl)-3-phenylpropanoyl)-4-hydroxy-2H-chromen-2-one derivatives 13(a-l) using Ho(3+) doped CoFe2O4 nanoparticles as catalyst and evaluated for their potential antileishmanial and antioxidant activities. The compounds 13a, 13d and 13h were found to possess significant antileishmanial activity (IC50 value=95.50, 95.00 and 99.00μg/mL, respectively) when compared to the standard sodium stibogluconate (IC50=490.00 μg/mL). The compounds 13a (IC50=12.40 μg/mL), 13d (IC50=13.49 μg/mL), 13g (IC50=13.24 μg/mL) and 13l (IC50=13.74 μg/mL) had shown good antioxidant activity when compared with standards butylated hydroxy toluene (IC50=16.5 μg/mL) and ascorbic acid (IC50=12.8 μg/mL). After performing molecular docking studies, it was found that compounds 13a and 13d had potential to inhibit pteridine reductase 1 enzyme. In silico ADME pharmacokinetic parameters had shown promising results and none of the synthesized compounds had violated Lipinskis rule of five. Thus, suggesting that compounds from the present series can serve as important gateway for the design and development of new antileishmanial as well as antioxidant agent.

Biofilm inhibition of linezolid-like Schiff bases: synthesis, biological activity, molecular docking and in silico ADME prediction.

Herein, we report the synthesis and screening of linezolid-like Schiff bases as inhibitors of biofilm formation. The result of biofilm inhibition of Pseudomonas aeruginosa suggested that compounds 5h (IC50 value=12.97±0.33μM) and 5i (IC50 value=15.63±0.20μM) had more inhibitory activity when compared with standard linezolid (IC50=15.93±0.18μM) without affecting the growth of cells (and thus behave as anti-quorum sensing agents). The compounds 5h (MIC range=2.5-10μg/mL) and 5i (MIC range=3.5-10μg/mL) with 2-chloroquinolinyl and 2-chloro-8-methylquinolinyl motif, respectively, showed antibacterial activity in comparable range of linezolid (MIC range=2-3μg/mL) and were more potent when compared with ciprofloxacin (MIC range=25-50μg/mL). Thus, the active derivatives were not only potent inhibitors of P. aeruginosa biofilm growth but also efficient antibacterial agents. The docking study of most active compounds 5h and 5i against PqsD enzyme of P. aeruginosa exhibited good binding properties. In silico ADME properties of synthesized compounds were also analyzed and showed potential to develop as good oral drug candidates.