Investigation of cell preservation using Tissue Microarray (TMA) on the histology of human exhumed skeletons

• Grant number: 13/17901-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: December 01, 2013
• End date: November 30, 2014
• Field of knowledge: Health Sciences - Medicine - Pathological Anatomy and Clinical Pathology
• Principal Investigator: Edna Sadayo Miazato Iwamura
• Grantee: Rafael Barrios de Mello
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

The Arrangement Tissue Matrix (Tissue Microarray or TMA) may be considered a simple but very effective technique since it is based on miniaturization and ordered grouping of tissue samples for large-scale surveys. This technique, described by Kononen et al (1998) and widely used, allows the investigation of fungi, proteins, DNA, and RNA by tests involving slide staining with Hematoxylin & Eosin (H&E), Periodic Acid Schiff (PAS), and Silver Methenamine, Immunohistochemistry, Fluorescence In Situ Hybridization (FISH) and In Situ Hybridization (CISH). Generally, conventional techniques of investigation in paraffin or frozen tissue are costly and require a relatively long time to be applied to hundreds of samples. Accordingly, the TMA has been used successfully in the simultaneous investigation of hundreds of tissue fractions in a single histologic section. Although the TMA was initially applied in researches involving cancer, other areas can also be benefited from this technology, such as forensic investigation. Once there are increasingly frequent situations in which genetic analysis in exhumed bodies is necessary, such as in criminal cases in which corpses are hidden, missing, or even in political or civil cases in which examinations of complex genetic kinship are required, the TMA can be an important tool in order to provide agility and skill in the processes of identification. The advantages obtained with this technique are: 1) Obtain of a large number of samples simultaneously; 2) Standardization (qualification) of reactions, and therefore ease of interpretation of results for purposes of comparison; 3) Economy of reagents and time for the completion of the protocols; 4) Use of the blocks in projects involving different visualization techniques, such as H&E, PAS, Silver Methenamine and Immunohistochemistry. The application of these methodologies in exhumed human bones could allow the classification of samples for molecular genetic Assay for human identification. However, the use of these techniques in forensic practice is rarely described in the literature. There are examples of CD31 (PECAM-1) and CD34 antibodies as endothelial markers commonly used in the study of neoplasia, but they could help out in forensic science indicating possible preservation of other cell types (beyond the osteocytes) in the bone tissue and skeletonized, however, are not used for this purpose. Therefore, it is necessary to test the applicability of the TMA technique and subsequent histopathologic methodologies (H&E, PAS, Silver Methenamine and Immunohistochemistry) to speed up processes, reduce costs and qualify the samples coming from human exhumed skeletons to use them in subsequent molecular techniques for genotyping profiles with the purpose of identification. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MELLO, RAFAEL BARRIOS; REGIS SILVA, MARIA REGINA; SEIXAS ALVES, MARIA TERESA; EVISON, MARTIN PAUL; GUIMARAES, MARCO AURELIO; FRANCISCO, RAFAELLA ARRABACA; ASTOLPHI, RAFAEL DIAS; MIAZATO IWAMURA, EDNA SADAYO. Tissue Microarray Analysis Applied to Bone Diagenesis. SCIENTIFIC REPORTS, v. 7, JAN 4 2017. (12/24918-4, 12/12344-3, 13/17901-0)