Analysis and standardization of a new methodology for extraction, quantification and distinction of sulfated glycosaminoglycans of knee joint cartilage in rats

• Grant number: 12/08290-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2012
• End date: June 30, 2013
• Field of knowledge: Health Sciences - Medicine - Pathological Anatomy and Clinical Pathology
• Principal Investigator: Flavio Faloppa
• Grantee: Bruno Goncalves Pereira Paschoa
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

This study aims the standardization and analysis of a new methodology of extraction, distinction and quantitation of glycosaminoglycans of the articular cartilage. Analysis of glycosaminoglycans (GAG) components of articular cartilage is an essential process to understand the diseases and the pharmacology related to this tissue as well as understand the changes generated by excessive mechanical stress, load privation, imobilization and even physiological loads. The methods used for evaluating the GAGs of the articular cartilage are the system of Mankin or HHGS-Histological-Histochemical Grading System and the OARSI (Ostheoarthritis Research Society International) Classification. Both classifications have been traditionally applied, but use a subjective color intensity graduation determined by the examiner and also do not distinguish or quantify the GAGs individually. Therefore it is of great value the development of a new methodology involving extraction, distinction and quantification of these elements in the articular cartilage. For the present study it will be used rats divided into two groups, the control group (CG) and training group (TG), with a training period of 36 days. At the end of this period, both groups will be sacrificed, it will be realized the extraction of cartilage from five sections, subsequent electrophoretic analysis with improved enzymatic degradation by ²-elimination, and, by agarose gel electrophoresis, it will be possible to identify the GAGs, separating them in accordance with the buffer used and its subsequent quantification by densitometry. (AU)