Elango Jeevithan

Type II Collagen and Gelatin from Silvertip Shark (Carcharhinus albimarginatus) Cartilage: Isolation, Purification, Physicochemical and Antioxidant Properties

Type II acid soluble collagen (CIIA), pepsin soluble collagen (CIIP) and type II gelatin (GII) were isolated from silvertip shark (Carcharhinus albimarginatus) cartilage and examined for their physicochemical and antioxidant properties. GII had a higher hydroxyproline content (173 mg/g) than the collagens and cartilage. CIIA, CIIP and GII were composed of two identical α1 and β chains and were characterized as type II. Amino acid analysis of CIIA, CIIP and GII indicated imino acid contents of 150, 156 and 153 amino acid residues per 1000 residues, respectively. Differing Fourier transform infrared (FTIR) spectra of CIIA, CIIP and GII were observed, which suggested that the isolation process affected the secondary structure and molecular order of collagen, particularly the triple-helical structure. The denaturation temperature of GII (32.5 °C) was higher than that of CIIA and CIIP. The antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl radicals and the reducing power of CIIP was greater than that of CIIA and GII. SEM microstructure of the collagens depicted a porous, fibrillary and multi-layered structure. Accordingly, the physicochemical and antioxidant properties of type II collagens (CIIA, CIIP) and GII isolated from shark cartilage were found to be suitable for biomedical applications.

Biocompatibility assessment of type-II collagen and its polypeptide for tissue engineering: effect of collagen's molecular weight and glycoprotein content on tumor necrosis factor (Fas/Apo-1) receptor activation in human acute T-lymphocyte leukemia cell line

Low molecular weight (LMW) (57, 40 and 25 kDa) collagens were separated and Fas cell surface death receptor activation were investigated based on its MW and glycoprotein content. The biochemical properties such as amino acid profile, FTIR and glycoprotein content of LMW collagens were slightly different and confirmed that limited digestion by proteolytic enzyme disrupted the triple helical structure of collagen. Cell adhesion pattern between collagens and immune cells were dose dependent manner. Better active cellular suppression of fish collagen than mammalian collagen was confirmed by increased secretion of immunological indexes (FAS/APO-1, cytokines (IL-2, IL-6) and caspases (casp-3, casp-8)) in T cell. Apoptotic regulatory gene expression (FAS, FAS-L, CASp-3, CASP-8, IL-2 and IL-6) of 6T-CEM cells was further confirmed the biocompatibility of fish collagens. Fluorescence microstructure revealed destruction of immune cells morphology and nuclei due to apoptosis by fish collagens. Consequently, this study disclosed that the apoptosis induction in T cells was based on the MW of collagens and also attachment of glycoprotein with tumor necrosis factor receptor (TNFRSF6) in cell membrane, which further activates DD, FAD, DED and caspase cascade pathway. Accordingly, this study created new platform for the better use of fish collagen in biomedical industries for the treatment of autoimmune diseases.

Biomedical and Pharmaceutical Application of Fish Collagen and Gelatin: A Review

In last decade, more research has been conducted in order to find the better way for utilizing the wastes product generated from food processing industries. The increasing demand of industrial by-products is one of the main reasons for the conversion of these wastes into valuable products. Among the different valuable products from the waste, the extraction of collagen and gelatin could be a better way of utilizing the wastes, due to their effective applications in biomedical and pharmaceutical industries. The most abundant source of collagen and gelatin are land-based animals, such as cow and pig. However, the extraction of collagen and gelatin from non-mammalian sources such as fish has been high influences in current society due to some religious and disease transmission issues. Many studies have dealt with the extraction and functional properties of collagen and gelatin from fish wastes. The present work is a compilation of information on biomedical and pharmaceutical application of collagen and gelatin from fish processing wastes.

Novel metabolites from Trichoderma atroviride against human prostate cancer cells and their inhibitory effect on Helicobacter pylori and Shigella toxin producing Escherichia coli

The present study aimed to purify and identify the metabolites from T. atroviride using high-performance liquid chromatography (HPLC) and 1H and 13C nuclear magnetic resonance spectrometer (NMR) followed by analyzing their toxicological, antibacterial and anticancer properties. This work identified two metabolites - TM1 and TM2. TM1 was in two forms: (i) 1, 3-dione-5, 5-dimethylcyclohexane; and, (ii) 2-enone-3hydroxy -5,5-dimethylcylohex, while TM2 was 4H-1,3-dioxin-4-one-2,3,6-trimethyl. These metabolites did not exhibit any irritant or allergic reaction as revealed by HET- CAM test. TM2 significantly inhibited the growth of H. pylori and Shigella toxin producing Escherichia coli (STEC) as evident by in vitro and microscopic observations of bacterial cell death. TM2 also induced the cell death and cytotoxicity, as revealed by cell viability test and western blot analysis. According to microscopic, flow cytometer and western blot analysis, TM2 treated cells displayed higher ROS, cell death, and apoptosis-related protein expression than TM1 and control. This study concluded that TM2 derived from T. atroviride was a potential therapeutic agent for anti-prostate cancer and antibiotic agent against MDR- H. pylori and STEC and it is also recommended to carry out further in vivo animal model experiments with improved stability of the metabolites for future pharmaceutical trails.

Fungal enzyme-mediated synthesis of chitosan nanoparticles and its biocompatibility, antioxidant and bactericidal properties

This paper reports the synthesis of chitosan nanoparticles (T-CSNPs) using the fungal enzyme of Trichoderma harzianum and its biocompatibility, antioxidant and bactericidal properties. The T-CSNPs synthesis was confirmed by absorbance at 280 nm using UV-Vis spectrophotometer. T-CSNPs were of spherical shape, as evident by field emission transmission electron microscopic (FETEM) analysis, and the average size of T-CSNPs was 90.8 nm, as calculated using particle size analyzer (PSA). The functional groups showed modifications of chitosan in T-CSNPs as evident by fourier-transform infrared spectroscopic (FTIR) analysis. T-CSNPs were found soluble at the wide range of pH, showing 100% solubility at pH 1-3 and 72% at pH 10. The T-CSNPs exhibited antioxidant property in a dose-dependent manner with pronounced activity at 100 mg·mL-1. The T-CSNPs also showed bactericidal activity against Staphylococcus aureus and Salmonella enterica Typhimurium by causing detrimental effects on bacterial cells. The T-CSNPs (50 μg·mL-1) did not display any cytotoxic effect on murine fibroblast NIH-3T3 cells, as evident by cell viability and acridine orange/ethidium bromide staining assays, which confirmed biocompatibility of the nanoparticles. This work suggested further investigations on the utilization of the mycosynthesized nanochitosan in biomedical applications.

Zinc-chitosan nanoparticles induced apoptosis in human acute T-lymphocyte leukemia through activation of tumor necrosis factor receptor CD95 and apoptosis-related genes

Chitosan (CS), a novel biomaterial is widely used as a drug nano-carrier for cancer treatments. Towards this aim, anticancer and antibacterial activities of CS-nanoparticles-linked zinc (Zn-CSNPs) were evaluated. The particle size of CSNPs was lowered (113.09 nm) compared to Zn-CSNPs (160.7 nm). Both nanoparticles (CSNPs and Zn-CSNPs) were spherical in shape, polydispersive and homogenous. Fourier transforms infrared spectrophotometer (FTIR) and energy dispersive X-ray spectroscopy (EDX) analysis confirmed the different molecular arrangement of NPs and the presence of Zn in Zn-CSNPs and CS in both NPs, respectively. Zn-CSNPs had higher inhibitory activity against tested pathogens with a minimal inhibitory concentration (MIC) of 9.25-13.5 μg·mL-1 and showed the complete inhibition of Staphylococcus aureus and Escherichia coli. Zn-CSNPs have triggered the apoptosis through activation of first apoptosis signal receptor/cluster of differentiation 95 (Fas/CD95), and apoptotic-regulatory genes and caused 65-70% of cellular damage in human acute T-lymphocyte leukemia (6T-CEM) cells. Overall, internalizing properties of Zn from CSNPs is a promising therapeutic approach to treat Zn-deficiency related diseases particularly human acute leukemia (HAL).