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www.nature.com/scientificreports OPEN Received: 23 June 2015 Accepted: 04 January 2016 Published: 02 February 2016 Green synthesis of silver nanoparticles and characterization of their inhibitory effects on AGEs formation using biophysical techniques Jalaluddin M. Ashraf1,*, Mohammad Azam Ansari2,3,*, Haris M. Khan2, Mohammad A. Alzohairy3,4 & Inho Choi1 Advanced glycation end-products (AGEs) resulting from non-enzymatic glycation are one of the major factors implicated in secondary complications of diabetes. Scientists are focusing on discovering new compounds that may be used as potential AGEs inhibitors without affecting the normal structure and function of biomolecules. A number of natural and synthetic compounds have been proposed as AGE inhibitors. In this study, we investigated the inhibitory effects of AgNPs (silver nanoparticles) in AGEs formation. AgNPs (~30.5 nm) synthesized from Aloe Vera leaf extract were characterized using UV- Vis spectroscopy, energy-dispersive X-ray spectroscopy (EDX), high resolution-transmission electron microscopy, X-ray diffraction and dynamic light scattering (DLS) techniques. The inhibitory effects of AgNPs on AGEs formation were evaluated by investigating the degree of reactivity of free amino groups (lysine and arginine residues), protein-bound carbonyl and carboxymethyl lysine (CML) content, and the effects on protein structure using various physicochemical techniques. The results showed that AgNPs significantly inhibit AGEs formation in a concentration dependent manner and that AgNPs have a positive effect on protein structure. These findings strongly suggest that AgNPs may play a therapeutic role in diabetes-related complications. The Maillard reaction is a nonenzymatic reaction of reducing sugars with amino groups of biological macromol- ecules. This process, which is also known as glycation, involves post-translation protein modification and may be responsible for a variety of diseases. The reaction is initiated by the reversible formation of a Schiff base between a reducing sugar and the amino group of a protein, DNA and lipoproteins1–3. The relatively unstable Schiff base undergoes rearrangement to form a more stable Amadori product, which in turn undergoes a series of reac- tions to form advanced glycation end products (AGEs)4,5. The accumulation of these AGEs in long-lived tissue is thought to be involved in diabetic complications and aging6. The Maillard reaction is found to be instigated by several sugar and non-sugar metabolites. Methylglyoxal (MG) is one of the most reactive metabolites that are involved in the formation of AGEs. It is generated during several enzymatic and nonenzymatic processes like glycolytic pathway, autoxidation of sugars and during all stages of the Maillard reaction7,8. Very high MG concentration has been detected in the lens, blood and kidney of diabetic patients9. For instance, 5–6 and 2–3 fold increases of MG was noted in Type I and II diabetic patients, respectively, as compared to their normal counterparts7,9. Considering its high reactivity with proteins and pres- ence of significant amounts of MG in the plasma (0.1 mM), MG may play as one of the major glycating agents in 1School of Biotechnology, Yeungnam University, Gyeongsan, Republic of Korea. 2Nanotechnology and Antimicrobial Drug Resistance Research Laboratory, Department of Microbiology, Jawaharlal Nehru Medical College & Hospital, Aligarh Muslim University, Aligarh-202002, U.P., India. 3Department of Medical Laboratories, College of Applied Medical Science, Buraydah Colleges, Buraydah 51452, Saudi Arabia. 4Department of Medical Laboratories, College of Medical Science, Qassim University, Saudi Arabia. *These authors contributed equally to this work. Correspondence and requests for materials should be addressed to J.M.A. (email: jmashraf@gmail.com) or M.A.A. (email: azammicro@gmail.com) or I.C. (email: inhochoi@ynu.ac.kr) Scientific RepoRts | 6:20414 | DOI: 10.1038/srep20414 1PDF Image | Green synthesis of silver nanoparticles inhibitory effects on AGEs
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