The Fabrication of a Low Cost Glycated Collagen Substrate

Introduction: Hyperglycemia, or diabetes, occurs when a person's body doesn't produce adequate insulin, or is refractory to insulin's effect of stimulating the intake of glucose into the body's cells. This persistent elevation of blood sugar results in glycation of long-lived proteins, especially Collagen typeI. As Collagen typeI is a major component of the extracellular matrix of tissues, this glycation of Collagen should be involved in the pathophysiology of diabetes such as impaired wound healing, peripheral neuropathy, and the reported increased severity of periodontal disease in diabetes patients. A first step in developing successful treatments for diabetic sequelae is the generation of a low cost disease-state-mimicking substrate to conduct experiments on. Gelatin (denatured Collagen) may be considered as a low cost substitute to native Collagen. Objective: to develop a low cost glycated Gelatin/Collagen substrate upon which experimentation can be performed to investigate its effects on cell types known to be affected in the Diabetic state and also as a wound healing medicament. Method: Bovine type A gelatin was incubated with glucose, glucose-6-phosphate, and glyoxylic acid with or without a reducing agent for 24-48 hours at 370C, then cast into thin films. Results: Fourier-transform infrared spectrometer (FTIR) analysis at the Amide I and Amide II wavelength positions indicated glycation of the gelatin had occurred to different degrees in the experimental groups compared to control. Spectrofluorescent analysis determined the percent glycation obtained. Conclusion: The glycation of bovine type A gelatin was successful. Gelatin is a low cost alternative to Collagen type I, and this new glycalted Gelatin/Collagen is now available for use in in vitro and in vivo assays to compare with glycalted native Collagen substrates. This study was supported by funds from the Faculty of Dentistry and 3Petroleum and Petrochemical College Chulalongkorn University Keywords: Collagen, Diabetes, Gelatin, Glycation, Wound Healing