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www.nature.com/scientificreports/ Figure1. (1.1)UV–visabsorptionspectrumof(A)silvernitrate(1mM)andsilvernanoparticles(B)synthesized using extract of fresh Aloe vera leaf. (1.2) HR-TEM micrograph (A) and EDX spectrum (B) of green synthesized silver nanoparticles. Figure 2. (1.1) DLS histogram showing particle size distribution of synthesized silver nanoparticles. (1.2) X-ray powder diffraction pattern of green synthesized silver nanoparticles. controlled cell holder attached to a Neslab’s RTE water bath with a temperature accuracy of ±0.1 °C. Solutions of control and glycated samples with or without AgNPs (20 μM) were placed in a 1 mm pathlength cuvette and profiles were recorded in the wavelength range of 200–250 nm. A scan speed of 100 nm/min and response time of 1 second was chosen to record the CD spectra. Two sets of each sample were studied under identical conditions to confirm reproducibility of the results36. Fourier transformation infrared (FTIR) spectroscopy. FTIR spectroscopy was used to investigate the changes in structure in MG-modified protein with various concentrations of AgNPs. Buffer subtracted transmis- sion spectra were recorded in the wave number range of 600–4000 cm−1 using a Perkin Elmer spectrum 100 FTIR spectrometer (PerkinElmer, Inc., USA). Results and Discussion Characterization of AgNPs. Prior to investigation of the antiglycating effect of AgNPs, characterization of synthesized AgNPs was performed as previously described, with slight modifications31. Briefly, Aloe Vera leaf extract was used in this study for one point green synthesis of AgNPs at room temperature. Upon adding the leaf extract to the silver nitrate solution, a faint yellow color was observed after 24 h of reaction (avoid light). The appearance of yellow color, which is due to the excitation of surface plasmon vibrations, represents spec- troscopic signature of AgNPs formation37. Characterization of biosynthesized AgNPs was accomplished using UV-Vis spectroscopy, XRD, DLS and HR-TEM (Figs 1 and 2). In the UV–Visible spectrum a strong broad peak was observed at 455 nm, and widening of the peak indicated that the particles were polydispersed (Fig. 1–1.1). AgNPs are known to exhibit a UV–Visible absorption maximum in the range of 400–500 nm because of surface plasmon resonance38. HR-TEM analysis revealed that the AgNPs were primarily spherical in shape. Their size ranges from in between 5–85 nm (Fig. 1–1.2), and the size distribution histogram of DLS indicated that the aver- age size was 30.5 nm (Fig. 2–1.1). The XRD pattern indicated that the nanoparticles had a face-centered cubic structure (JC-PDS No: 03-0921). All peaks of the XRD pattern could be indexed according to the Ag (Fig. 2–1.2). It is well known that blood proteins including albumin are glycated in diabetic patients by a variety of reduc- ing sugars, leading to the formation of AGEs and affecting the structure and function of proteins39. Following the addition of reducing sugars, various metabolites react with amino groups of biomolecules. Methylglyoxal (2-oxopropanal) is a reactive physiological metabolite40 that is mainly responsible for forming inter- and Scientific RepoRts | 6:20414 | DOI: 10.1038/srep20414 4PDF Image | Green synthesis of silver nanoparticles inhibitory effects on AGEs
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