Graphene Oxide Based Electrochemical Genosensor for Label Free Detection of Mycobacterium tuberculosis from Raw Clinical Samples.

BACKGROUND: Mycobacterium tuberculosis' rapid detection is still a formidable challenge to have control over the lethal disease. New diagnostic methods such as LED fluorescence microscopy, Genexpert, Interferon Gamma Release Assay (IGRA) are limited on efficacy spectrum owing to their high cost, time-intensive and laborious nature, in addition their low sensitivity hinders their robustness and portability. Electroanalytical methods are now being considered as an excellent alternative, being currently employed for efficient detection of the analytes with the potential of being portable. This report suggests label-free electrochemical detection of Mycobacterium tuberculosis (Mtb) via its marker, insertion sequence (IS6110). METHODS: In this pursuit, graphene oxide-chitosan nanocomposite (GO-CHI), a biocompatible matrix, having a large electroactive area with an overall positively charged surface, is fabricated and characterized. The obtained GO-CHI nanocomposite is then immobilized on the ITO surface to form a positively functionalized electrochemical sensor for the detection of Mtb. DNA probe, specific for the IS6110, was electrostatically anchored on a positively charged electrode surface and the resistance of charge transfer was investigated for the sensitive and specific (complementary vs non-complementary) detection of Mtb by cyclic voltammetry and differential pulse voltammetry techniques. RESULTS: The cyclic voltammetry was found to be diffusion controlled facilitating the absorption of analyte on the electrode surface. The label-free "genosensor" was found to detect a hybridization efficiency with a limit of detection of 3.4 pM, and correlation coefficient R2=0.99 when analysed over a range of concentrations of DNA from 7.86 pM to 94.3pM. The genosensor was also able to detect target DNA from raw sputum samples of clinical isolates without DNA purification. CONCLUSION: This electrochemical genosensor provides high sensitivity and specificity; thus offering a promising platform for clinical diagnosis of TB and other infectious diseases in general.

Synthesis of Silver-Coated Bioactive Nanocomposite Scaffolds Based on Grafted Beta-Glucan/Hydroxyapatite via Freeze-Drying Method: Anti-Microbial and Biocompatibility Evaluation for Bone Tissue Engineering.

Advancement and development in bone tissue engineering, particularly that of composite scaffolds, are of great importance for bone tissue engineering. We have synthesized polymeric matrix using biopolymer (ß-glucan), acrylic acid, and nano-hydroxyapatite through free radical polymerization method. Bioactive nanocomposite scaffolds (BNSs) were fabricated using the freeze-drying method and Ag was coated by the dip-coating method. The scaffolds have been characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and X-ray diffraction analysis (XRD) to investigate their functional groups, surface morphology, and phase analysis, respectively. The pore size and porosity of all BNS samples were found to be dependent on silver concentration. Mechanical testing of all BNS samples have substantial compressive strength in dry form that is closer to cancellous bone. The samples of BNS showed substantial antibacterial effect against DH5 alpha E. coli. The biological studies conducted using the MC3T3-E1 cell line via neutral red dye assay on the scaffolds have found to be biocompatible and non-cytotoxic. These bioactive scaffolds can bring numerous applications for bone tissue repairs and regenerations.

Comparative safety analysis of bactericidal nano-colloids: Assessment of potential functional toxicity and radical scavenging action.

Extensive utilization of silver nanoparticles (AgNP) has raised concerns of their safety profile upon interaction with biological system. In past decade, various nanoparticles (NPs) with excellent antimicrobial potential have been synthesized, a majority of which have struggled with the established toxicity in biological systems. The NPs safety is still a hot debate and various strategies are being adopted to overcome this giant limitation. This paper successfully reports comparative toxicity profiles of previously synthesized antimicrobial NPs in our lab and concludes the effectiveness of biologically synthesized NPs for its safe usage in biological systems. In this study, five of our previously synthesized NPs that showed excellent antimicrobial potential were compared for their in vivo toxicity and corresponding radical scavenging activities. Based on lowest morbidity, mortality, weight loss, toxicity and agglomeration profile, best NPs with highest antimicrobial potentials were screened out and used for further biomedical applications. The previously reported NPs used in this study included Aerva javanica synthesized nanoparticles (AjNPs), Heliotropium crispium synthesized nanoparticles (HcNPs), and violacein capped nanoparticles (VNPs), these showed least toxicity upon in vivo histological analysis. AjNPs among them showed maximum safety and efficacy profile and consistently showed least production of reactive oxygen species, least mortality and morbidity rate as compared to other groups. Present study establishes that all these biologically synthesized NPs and specifically AjNPs can be efficiently employed as antimicrobial agents as they have not exhibited toxic profile and have shown least accumulation into the organs such as liver spleen and kidney.

Formulation of active packaging system using Artemisia scoparia for enhancing shelf life of fresh fruits.

An improved active packaging system was developed for fresh fruits using silver nanoparticles (AgNPs) coupled with calcium alginate (Ca-ALG). For the synthesis of AgNPs aqueous, ethanol and methanol extracts of Artemisia scoparia (AS) were used. These AgNP's were characterized using UV-Vis, SEM, EDS, AFM, FTIR and gel electrophoresis. Ethanol extract of AS (ASE) produced AgNPs with smallest size in comparison to aqueous AS (ASA) and methanol extract of AS (ASM). AgNPs synthesized from ASE had a size range of 12.0-23.3 nm and were tested on Human Corneal Epithelial Cells to evaluate their cytotoxicity. At 0.05 ng/mL of AgNP's concentration, no toxic effects were observed on the evaluated cell line. Therefore, 0.05 ng/mL of AgNPs mixed with edible coating of Ca-ALG were applied on strawberries and loquats as active coating to increase their shelf life. Significant improvement was observed in the quality parameters of strawberries and loquats such as microbial analysis, acidity loss, soluble solid content loss, weight loss and quality decay. Ca-ALG coating incorporated with AgNPs enhanced the shelf life of strawberries and loquats in comparison to no treatment and simple Ca-ALG coatings. This study provides an insight to food industry to extend the shelf life of fresh fruits using AgNP's formulated coatings.