RESUMO
The diverse application of collagen has created a need to discover renewable and economical sources with prevailing/improved physico-chemical properties. To address this scenario, the present study has extracted collagen from Human Amniotic Membrane (AM) and Umbilical cord, which are treated as medical waste and compared its physico-chemical properties. Collagen was extracted by pepsin solubilization using various salt concentrations (1 M, 2 M and 4 M). Umbilical Cord Collagen (UC) yield was 10% higher than Amniotic Membrane Collagen (AC). UC reported 58% higher sulphated glycosaminoglycan content than AC. Electrophoretic pattern of AC and UC in both disulphide bond reducing and non-reducing conditions showed bands corresponding to collagen type I, III, IV, V and XV. Collagen morphology was examined using SEM and the amino acid content was quantified by HPLC and LC-MS/MS. Triple helicity was confirmed by CD and FTIR spectra. Thermal transition temperature of AC and UC was found equivalent to animal collagen. Self-assembly, fibril morphology and spatial alignment was studied using AFM and DLS. Biocompatibility was analyzed using 3T3 fibroblast cells. In conclusion, UC with higher yield, presented with better physico-chemical, structural and biological properties than AC could serve as an efficient alternative to the existing animal collagen for diverse applications.
Assuntos
Proliferação de Células/efeitos dos fármacos , Colágeno/química , Glicosaminoglicanos/química , Células-Tronco Mesenquimais/efeitos dos fármacos , Âmnio/química , Cromatografia Líquida , Colágeno/classificação , Colágeno/isolamento & purificação , Colágeno/farmacologia , Fibroblastos/efeitos dos fármacos , Glicosaminoglicanos/isolamento & purificação , Humanos , Pepsina A/farmacologia , Espectrometria de Massas em Tandem , Cordão Umbilical/químicaRESUMO
This study reported the synthesis of Vicenin-2 gold nanoparticles (VN-AuNPs) and evaluated their effect on the glucose utilization efficiency of 3T3-L1 adipocytes. The VN-AuNPs were characterized by microscopic, DLS and spectral analysis. The bio-reducing efficiency of Vicenin-2 (VN) was computed and confirmed by HPLC analysis. The stability of VN-AuNPs in various physiological media was explored. The cytotoxicity and glucose uptake assays were performed in 3T3-L1 adipocytes. The docking of VN with PTP1B and AMPK was also performed. The color change and UV absorption at 537 nm preliminarily confirmed the VN reduced gold nanoparticles. The VN-AuNPs appeared as spherical particles (57 nm) and face centered cubic crystals under TEM and XRD analysis, respectively. Its zeta potential was found to be -6.53 mV. The FT-IR spectra of VN and its AuNPs confirmed its stability. The computed reducing potential of VN was similar to the extent of VN utilized during the synthesis of VN-AuNPs. The VN-AuNPs showed a remarkable stability in different physiological media. At 100 µM concentration, VN-AuNPs displayed 78.21% cell viability. A concentration dependent increase in glucose uptake was noted in 3T3-L1 adipocytes when incubated with VN-AuNPs. The docking data revealed a strong interaction of VN with the binding pockets of PTP1B and AMPK. This demonstrates that the fabricated VN-AuNPs might enhance the intracellular VN availability mediated cellular glucose utilization and this would serve as a novel nanodrug for the management of diabetes.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Apigenina/farmacologia , Glucose/metabolismo , Glucosídeos/farmacologia , Ouro/farmacologia , Nanopartículas Metálicas/química , Proteína Tirosina Fosfatase não Receptora Tipo 1/metabolismo , Adipócitos , Animais , Apigenina/química , Apigenina/toxicidade , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Biologia Computacional , Glucosídeos/química , Glucosídeos/toxicidade , Ouro/química , Ouro/toxicidade , Nanopartículas Metálicas/toxicidade , Camundongos , Simulação de Acoplamento MolecularRESUMO
Enzymatic browning by polyphenoloxidase (PPO) affects food quality and taste in fruits and vegetables. Thus, the study was designed to reduce browning in apple juice by coumarin. The ethanolic extract of cinnamon was prepared and its coumarin content was quantitated by HPLC, using authentic coumarin (AC) as standard. The effect of cinnamon extract (CE) and AC on enzymatic browning, its time dependent effects, and the specific activity of PPO and peroxidase (POD) were studied in apple juice. The docking of coumarin with PPO and POD was also performed to elucidate its antibrowning mechanism. The CE (73%) and AC (82%) showed better reduction in browning, maintained its antibrowning effect at all time points, and significantly (p < 0.05) reduced the specific activity of PPO and POD when compared with controls. Coumarin showed strong interaction with binding pockets of PPO and POD, suggesting its potential use as inhibitor to enzyme mediated browning in apple juice.
