Balaraman Madhan

Molecular mechanics and dynamics studies on the interaction of gallic acid with collagen-like peptides

Abstract Molecular modelling approaches have been used to understand the interaction of collagen-like peptides with gallic acid, which mimic vegetable tanning processes involved in protein stabilization. Several interaction sites have been identified and the binding energies of the complexes have been calculated. The calculated binding energies for various geometries are in the range 6–13 kcal/mol. It is found that some complexes exhibit hydrogen bonding, and electrostatic interaction plays a dominant role in the stabilization of the peptide by gallic acid. The π-OH type of interaction is also observed in the peptide stabilization. Molecular dynamics (MD) simulation for 600 ps revealed the possibility of hydrogen bonding between the collagen-like peptide and gallic acid.

UV Damage of Collagen: Insights from Model Collagen Peptides

Fibrils of Type I collagen in the skin are exposed to ultraviolet (UV) light and there have been claims that collagen photo-degradation leads to wrinkles and may contribute to skin cancers. To understand the effects of UV radiation on collagen, Type I collagen solutions were exposed to the UV-C wavelength of 254 nm for defined lengths of time at 4°C. Circular dichroism (CD) experiments show that irradiation of collagen leads to high loss of triple helical content with a new lower thermal stability peak and SDS-gel electrophoresis indicates breakdown of collagen chains. To better define the effects of UV radiation on the collagen triple-helix, the studies were extended to peptides which model the collagen sequence and conformation. CD studies showed irradiation for days led to lower magnitudes of the triple-helix maximum at 225 nm and lower thermal stabilities for two peptides containing multiple Gly-Pro-Hyp triplets. In contrast, the highest radiation exposure led to little change in the T(m) values of (Gly-Pro-Pro)(10) and (Ala-Hyp-Gly)(10) , although (Gly-Pro-Pro)(10) did show a significant decrease in triple helix intensity. Mass spectroscopy indicated preferential cleavage sites within the peptides, and identification of some of the most susceptible sites of cleavage. The effect of radiation on these well defined peptides gives insight into the sequence and conformational specificity of photo-degradation of collagen.

Sol–Gel Assisted Fabrication of Collagen Hydrolysate Composite Scaffold: A Novel Therapeutic Alternative to the Traditional Collagen Scaffold

Collagen is one of the most widely used biomaterial for various biomedical applications. In this Research Article, we present a novel approach of using collagen hydrolysate, smaller fragments of collagen, as an alternative to traditionally used collagen scaffold. Collagen hydrolysate composite scaffold (CHCS) was fabricated with sol-gel transition procedure using tetraethoxysilane as the silica precursor. CHCS exhibits porous morphology with pore sizes varying between 380 and 780 μm. Incorporation of silica conferred CHCS with controlled biodegradation and better water uptake capacity. Notably, 3T3 fibroblast proliferation was seen to be significantly better under CHCS treatment when compared to treatment with collagen scaffold. Additionally, CHCS showed excellent antimicrobial activity against the wound pathogens Staphylococcus aureus, Bacillus subtilis, and Escherichia coli due to the inherited antimicrobial activity of collagen hydrolysate. In vivo wound healing experiments with full thickness excision wounds in rat model demonstrated that wounds treated with CHCS showed accelerated healing when compared to wounds treated with collagen scaffold. These findings indicate that the CHCS scaffold from collagen fragments would be an effective and affordable alternative to the traditionally used collagen structural biomaterials.

Capsaicin inhibits collagen fibril formation and increases the stability of collagen fibers

AbstractCapsaicin is a versatile plant product which has nbeen ascribed several health benefits and anti-inflammatory and analgesic properties. We have investigated the effect of capsaicin on the molecular stability, self-assembly, and fibril stability of type-I collagen. It was found that capsaicin suppresses collagen fibril formation, increases the stability of collagen fibers in tendons, and has no effect on the molecular stability of collagen. Turbidity assay data show that capsaicin does not promote disassembly of collagen fibrils. However, capsaicin moderately protects collagen fibrils from enzymatic degradation. Computational studies revealed the functions of the aromatic group and amide region of capsaicin in the collagen–capsaicin interaction. The results may have significant implications for capsaicin-based therapeutics that target excess collagen accumulation-linked pathology, for example thrombosis, fibrosis, and sclerosis.

2,2,2-Trifluoroethanol disrupts the triple helical structure and self-association of type I collagen.

Collagen, a fibrous structural protein, is a major component of skin, tendon, bone, and other connective tissues. Collagen is one of the dominant biomaterials used for tissue engineering and drug delivery applications. 2,2,2-Trifluoroethanol (TFE) has been used as a co-solvent in the preparation of collagen based biomaterials, which are used for tissue engineering applications. However, the basic knowledge about the structural behavior of collagen in TFE is necessary for an adequate application of collagen as a carrier system. In this work, the effect of TFE on the structure and self-association of collagen has been studied in detail using different spectroscopic methods such as circular dichroism (CD), Fourier transform infrared (FTIR), and UV-Vis absorption. The results obtained from CD and FTIR suggest that collagen transform its structure from triple helix to predominantly unordered conformation with increasing concentration of TFE. Thermal melting studies reveal that the stability of collagen triple helix decreases even at low concentration of TFE. Turbidity measurements indicate that TFE, at higher concentrations, inhibits the collagen fibril formation which arises due to the self-association of collagen molecules. TFE has conventionally been known to promote the ordered structures in proteins and peptides. Destabilization of collagen triple helix by TFE is first of its kind information on the effect of TFE to disrupt the native conformation of proteins.

Molecular Level Insights on Collagen–Polyphenols Interaction Using Spin–Relaxation and Saturation Transfer Difference NMR

Interaction of small molecules with collagen has far reaching consequences in biological and industrial processes. The interaction between collagen and selected polyphenols, viz., gallic acid (GA), pyrogallol (PG), catechin (CA), and epigallocatechin gallate (EGCG), has been investigated by various solution NMR measurements, viz., (1)H and (13)C chemical shifts (δH and δC), (1)H nonselective spin-lattice relaxation times (T1NS) and selective spin-lattice relaxation times (T1SEL), as well as spin-spin relaxation times (T2). Furthermore, we have employed saturation transfer difference (STD) NMR method to monitor the site of GA, CA, PG, and EGCG which are in close proximity to collagen. It is found that -COOH group of GA provides an important contribution for the interaction of GA with collagen, as evidenced from (13)C analysis, while PG, which is devoid of -COOH group in comparison to GA, does not show any significant interaction with collagen. STD NMR data indicates that the resonances of A-ring (H2, H5 and H6) and C-ring (H6 and H8) protons of CA, and A-ring (H2 and H6), C-ring (H6 and H8), and D-ring (H2″and H6″) protons of EGCG persist in the spectra, demonstrating that these protons are in spatial proximity to collagen, which is further validated by independent proton spin-relaxation measurement and analysis. The selective (1)H T1 measurements of polyphenols in the presence of protein at various concentrations have enabled us to determine their binding affinities with collagen. EGCG exhibits high binding affinity with collagen followed by CA, GA, and PG. Further, NMR results propose that presence of gallic acid moiety in a small molecule increases its affinity with collagen. Our experimental findings provide molecular insights on the binding of collagen and plant polyphenols.

Effect of aqueous ethanol on the triple helical structure of collagen

AbstractnCollagen, the most abundant protein in mammals, is widely used for making biomaterials. Recently, organic solvents have been used to fabricate collagen-based biomaterials for biological applications. It is therefore necessary to understand the behavior of collagen in the presence of organic solvents at low (≤50xa0%, v/v) and high (≥90xa0%, v/v) concentrations. This study was conducted to examine how collagen reacts when exposed to low and high concentrations of ethanol, one of the solvents used to make collagen-based biomaterials. Solubility testing indicated that collagen remains in solution at low concentrations (≤50xa0%, v/v) of ethanol but precipitates (gel-like) thereafter, irrespective of the method of addition of ethanol (single shot or gradual addition); this behavior is different from that observed recently with acetonitrile. Collagen retains its triple helix in the presence of ethanol but becomes thermodynamically unstable, with substantially reduced melting temperature, with increasing concentration of ethanol. It was also found that the CD ellipticity at 222xa0nm, characteristic of the triple-helical structure, does not correlate with the thermal stability of collagen. Time-dependent experiments reveal that the collagen triple helix is kinetically stable in the presence of 0–40xa0% (v/v) ethanol at low temperature (5xa0°C) but highly unstable in the presence of ethanol at elevated temperature (~34xa0°C). These results indicate that when ethanol is used to process collagen-based biomaterials, such factors as temperature and duration should be done taking into account, to prevent extensive damage to the triple-helical structure of collagenn.

κ-Carrageenan: An effective drug carrier to deliver curcumin in cancer cells and to induce apoptosis

The current study is to develop a natural drug carrier with seaweed derived polymers namely κ-Carrageenan (κ-Car) for drug delivery applications. κ-Car is a natural polysaccharide which derived from edible red seaweeds, they are easily available, non-toxic, cost effective, biodegradable and biocompatible nature. Curcumin (Cur) is a yellow-orange polyphenol existing in turmeric, which is predominantly used as spice and food coloring agent. The ultimate use of polymeric composites, especially those composed of natural polymers, has become a very interesting approach in recent drug delivery applications, due to their non-toxicity and biological origin. In this study the primary approach which depends on the loading of Curcumin into κ-Carrageenan was accomplished, and which (κ-Car-Cur) an active drug carrier was developed for drug delivery against selected lung cancer cells (A549). Thus, the κ-Car-Cur was synthesized by solvent evaporation method followed by freeze drying, and it was further characterized. From this study, it has been reported that the high encapsulation efficiency, good stability, and successful release of Cur from the carrier (κ-Car) was achieved. The drug release was more active at acidic pH 5.0 with the cumulative release of 78%, which is the favorable condition present in tumor microenvironments. The in vitro cellular applications studies of κ-Car-Cur demonstrated that, κ-Car-Cur composites induced higher cytotoxicity against selected cancer cells than free Cur and effectively involved to trigger cellular apoptosis in A549 cancer cells. Further, it was also possessed that inhibition of cell growth and changes in metabolic activity of cancer cells are the unique characteristic features of cellular apoptosis, through reactive oxygen species (ROS) generation. It also observed that there was a decrease in mitochondrial membrane potential (ΔψmΔψm) which leads to a cellular apoptosis during treatment with κ-Car-Cur. Hence, the study outcomes may provide the potential outline for the use of κ-Car-Cur as a promising tool to deliver drugs at intracellular level.

Density functional theory calculations on dipeptide–gallic acid interaction

Abstract In the present investigation, an attempt has been made to study the interaction of dipeptides with gallic acid, using Becke3 parameter Lee Yang Parr (B3LYP) method employing 3-21G*, 6-31G* and 6-31+G* basis sets. The interaction energies of the dipeptide–gallic acid complexes are in the range of −5 to −18 kcal/mol depending on the mode of intermolecular complexation. Calculated molecular electrostatic potential (MESP) for the various intermolecular complexes revealed the electrostatic nature of the interaction. Qualitative estimations based on chemical hardness and chemical potential demonstrated fractional electron transfer from dipeptide to gallic acid.

Collagen-fucoidan blend film with the potential to induce fibroblast proliferation for regenerative applications

Collagen is a unique protein abundantly present in the connective tissues of mammals and widely used for biomaterial preparation. In this study, we synthesized and characterized collagen-fucoidan blend films for tissue regenerative properties. Thermogravimetric analysis (TGA) and Differential Scanning Calorimetry (DSC) were used for thermal analysis of the blend films, and the films exhibited higher thermal stability and denaturation temperature (Td) than those of native collagen due to intramolecular hydrogen bonding interaction between collagen and fucoidan, which was analyzed by FTIR spectroscopy. Morphological evaluation of these films using Scanning Electron Microscopy (SEM) showed smaller pore size than the control. Moreover, fucoidan protects collagen against enzymatic degradation and thereby increases the structural stability of collagen. Further, the in vitro studies of the synthesized films showed that they effectively facilitated the proliferation and migration of fibroblast cells without exhibiting toxicity. These study results suggested that the collagen-fucoidan blend films are a favorable substrate for growth of fibroblast cells, and may have great potential for tissue engineering applications.