Confocal Raman Spectroscopic Analysis of the Changes in Type I Collagen Resulting from Amide I Glycation
Journal Title: Biomedical Journal of Scientific & Technical Research (BJSTR) - Year 2017, Vol 1, Issue 3
Abstract
The process of glycation is a non - enzymatic reaction between sugar and free amino group of proteins, leading to skin aging. In this sense, the present study aimed to analyze the changes resulting from glycation in the dermis by Confocal Raman spectroscopy. The analysis was performed with 30 patients with skin types I and II, divided into 3 groups: 10 healthy young women aged 20-33 years, 10 healthy elderly women and 10 elderly women with diabetes type I and II in the age group of 65-80 years. Raman spectra were collected from 70-130 microns in depth and the data were analyzed. It was observed that the amide I band was centered at 1664 cm-1 for the young participants and healthy elderly which may be due to the presence of water and change in the direction of the shorter wavelength was observed which is centered around 1662 cm-1. Considerable changes were observed for the elderly participants with diabetes and the peak was centered around 1658 cm-1. Thus, it is inferred that hyperglycemia in elderly subjects with diabetes mellitus accelerate the formation of advanced glycation end products, which increase the type I collagen degradation in human skin.The effects of the glycation reaction in the skin`s proteins at cellular level accelerate the aging process and results in wrinkling and stiffness. Without the action of an enzyme, protein reacts with reducing sugar and [1-4] give rise to Advanced Glycation Endproducts (AGEs). These AGEs cause degradation of type I collagen and apoptosis of fibroblasts, resulting in alterations of the amide I bands which correspond to the secondary structure of this protein [5-12]. The Confocal Raman spectroscopy is a non invasive technique which uses a laser of 785 nm as an excitation source, with the ability to measure the biochemical composition of the skin, without causing damage and does not need the preparation of material for analysis [13-17]. By the objective lens of the inverted microscope, the laser is guided to the skin and collects the inelastic scattering of light in real time and at different depth profile [17-22] making it a suitable tool to probe human skin. Therefore, the main goal of this study is to characterize and compare the chronological effect of skin aging due to the process of glycation in amide I of type I collagen in the dermis of healthy young women (HYW), health elderly women (HEW) and diabetic elderly women (DEW).In Vivo Raman Data acquisition and analysis: Confocal Raman setup for in vivo human skin experiments were performed by a confocal Raman system from Rivers Diagnostics® (Model 3510 Skin Composition Analyzer) coupled to Laser light of 785 nm and is focused into the skin by a microscope objective. The power of the laser at the skin was set as 27 mW. The laser light was focused on the forearm skin with an objective of the microscope (40x) located under the quartz window. The Raman signal was collected by a CCD camera and recorded by a computer coupled to the system. The data were obtained in the fingerprint region (400- 1800 cm-1) from 70-130 microns below the skin surface with a spatial resolution of 3 microns, integration time of 60 seconds and 2 accumulations. Pre processing was done in the order normalization of the amide I band, smoothing and baseline correction using LabSpec 5 software.
Authors and Affiliations
Liliane Pereira, A Téllez S Claudio, Fávero Priscila, Airton A Martin
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