Angiogenesis, the process of formation of new blood vessels from pre-existing one, occurs in many physiological and pathological conditions, most of which are underlined by hypoxia and resultant accumulation of lactate. Although lactate is known to induce angiogenesis, the mechanism of its action on endothelial cells (ECs) is not known. The present study was designed to examine the response of ECs to lactate. Morphological analysis revealed that human umbilical vein endothelial cells (HUVECs) in culture respond to lactate by switching over to angiogenic phenotype concomitant with upregulation of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR2) as determined by reverse transcription-PCR (RT-PCR). Apart from increase in the levels of VEGF protein as determined by ELISA, chorio allantoic membrane (CAM) assay using the cell extracts revealed that lactate also increased the angiogenic potency of VEGF. Isolated VEGF, when blotted and subsequently probed with anti-PAR antibody, revealed considerable reduction in poly-adenosyl ribosylation of VEGF associated with a significant decrease in the levels of NAD(+), in presence of lactate. Thus it appears that ECs respond to lactate by increasing the production of VEGF and modulating its angiogenic potency through poly-ADP ribosylation (PAR)-dependent mechanism and thereby switch over to angiogenic phenotype.
Bacterial colonization on medical devices is a major concern in the healthcare industry. In the present study, we report synthesis of environmental sustainable reduced graphene oxide (rGO) on the large scale through biosynthetic route and its potential application for antibacterial coating on medical devices. HRTEM image depicts formation of graphene nanosheet, while DLS and ζ potential studies reveal that in aqueous medium the average hydrodynamic size and surface charge of rGO are 4410 ± 116 nm and -25.2 ± 3.2 mV, respectively. The Raman, FTIR, and XPS data suggest in situ conjugation of protein with rGO. The as-synthesized rGO protein nanoframework exhibits dose-dependent antibacterial activity and potential of killing of 94% of Escherichia coli when treated with 80 μg/mL of rGO for 4 h. The hemolytic and cytotoxicity studies demonstrate that rGO protein nanoframework is highly biocompatible at the same concentration showing significant antimicrobial properties. The rGO coated on the glass surface obtained through covalent bonding exhibits potent antibacterial activity. Antibacterial mechanism further demonstrates that rGO-protein nanoframework in dispersed state (rGO solution) exerts bactericidal effect through physical disruption accompanied by ROS-mediated biochemical responses. The rGO subsequently entering into the cytoplasm through the damaged membrane causes metabolic imbalance in the cells. In sharp contrast, physical damage of the cell membrane is the dominant antibacterial mechanism of rGO in the immobilized state (rGO coated glass). The obtained results help indepth understanding of the antibacterial mechanism of the biosynthesized rGO and a novel way to develop nontoxic antibacterial coating on medical devices to prevent bacterial infection.
This paper elucidates the development of a curcumin cross-linked collagen aerogel system with controlled anti-proteolytic activity and pro-angiogenic efficacy. The results of this study showed that in situ cross-linking of curcumin with collagen leads to the development of aerogels with enhanced physical and mechanical properties. The integrity of collagen after cross-linking with curcumin was studied via FTIR spectroscopy. The results confirmed that the cross-linking with curcumin did not induce any structural changes in the collagen. The curcumin cross-linked collagen aerogels exhibited potent anti-proteolytic and anti-microbial activity. Scanning electron and atomic force microscopic analysis of curcumin cross-linked collagen aerogels showed a 3D microstructure that enhanced the adhesion and proliferation of cells. The highly organized geometry of collagen-curcumin aerogels enhanced the permeability and water-retaining ability required for the diffusion of nutrients that aid cellular growth. The pro-angiogenic properties of collagen-curcumin aerogels were ascribed to the cumulative effect of the nutraceutical and the collagen molecule, which augmented the restoration of damaged tissue. Further, these aerogels exhibited controlled anti-proteolytic activity, which makes them suitable 3D scaffolds for biomedical applications. This study provides scope for the development of biocompatible and bioresorbable collagen aerogel systems that use a nutraceutical as a cross-linker for biomedical applications.
The present study illustrates the progress of the wheat grass bioactive-reinforced collagen-based aerogel system as an instructive scaffold for collagen turnover and angiogenesis for wound healing applications. The reinforcement of wheat grass bioactives in collagen resulted in the design and development of aerogels with enhanced physicochemical and biomechanical properties due to the intermolecular interaction between the active growth factors of wheat grass and collagen fibril. Differential scanning calorimetry analysis revealed an enhanced denaturation temperature when compared to those of native collagen aerogels. Fourier transform infrared spectroscopy analysis confirmed that the reinforcement of bioactives in the wheat grass did not affect the structural integrity of the collagen molecule. Additionally, the reinforced biomaterial with a systematic absorptive morphology resulted in a three-dimensional (3D) sponge-like aerogel exhibiting a potent highly oriented 3D structural assembly that showed increased water retention ability and substance permeability that would enable the passage of nutrients and gaseous components for cellular growth. Furthermore, the cumulative effect of the growth factors in wheat grass and the collagen molecule augments the angiogenic ability and collagen production of the aerogel by restoration of the damaged tissue thereby making it a potential 3D wound dressing scaffold. The results were confirmed by in vivo wound healing assays. This study shows the possibility for progress of a biocompatible, biodegradable, and nonadhesive nutraceutical-reinforced collagen aerogel as an instructive scaffold with good antimicrobial properties for collagen turnover and angiogenic response for wound healing applications.
Wound dressing material based on nano-biotechnological intervention by caging plumbagin on silver nanoparticle (PCSN) as a multi-site cross-linking agent of collagen scaffolds with potent anti-microbial and wound healing activity.
The surface of a living yeast cell (Saccharomyces cerevisiae strain W303-1A) has been labeled with silver (Ag) nanoparticles that can form nanoaggregates which have been shown to have surface-enhanced Raman scattering (SERS) activity. The cell wall of a single living yeast cell has been imaged by use of a Raman microspectroscope. The SERS spectra measured from different Ag nanoaggregates were found to be different. This can be explained on the basis of detailed spectral interpretation. The SERS spectral response originates from mannoproteins which cover the outermost regions of the yeast cell wall. Analysis of SERS spectra from the cell wall and the extracted mannoproteins from the yeast has been performed for the clarification of variation in SERS spectra.
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