Fast and accurate protocol for histology and immunohistochemistry reactions in temporomandibular joint of rats

Autor(es)

Basting, Rosanna Tarkany ^[1] ; Napimoga, Marcelo Henrique ^[1] ; de Lima, Juliana Moreira ^[1] ; de Freitas, Nadir Severina ^[2] ; Clemente-Napimoga, Juliana Trindade ^[1]

Afiliação do(s) autor(es)

^[1] Sao Leopoldo Mandic Inst & Res Ctr, Fac Sao Leopoldo Mandic, Lab Neuroimmune Interface Pain Res, Rua Jose Rocha Junqueira 13, BR-13045755 Campinas, SP - Brazil

^[2] Sao Leopoldo Mandic Inst & Res Ctr, Fac Sao Leopoldo Mandic, Lab Pathol, Rua Jose Rocha Junqueira 13, BR-13045755 Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: ARCHIVES OF ORAL BIOLOGY; v. 126, JUN 2021.
• Citações Web of Science: 0

Resumo

Objective: Propose a standard, fast and accurate protocol for the processing of the temporomandibular joint (TMJ) of adults' rats for histology and immunohistochemistry reactions. Design: Wistar male rats were perfused with paraformaldehyde (4 %). The heads were fixed in formaldehyde 10 % solution for 48 h. After that, the heads were sectioned in a sagittal plane and fixed for plus 48 h. Decalcification was performed using 20 % formic acid for 96 h and delimitation of TMJ area was done. Detailed methodology to a standard extraction and processing of TMJ to histological sections is described. Different buffers, equipment, temperature and time were tested to optimize immunostaining. Morphological preservation and antigenicity were evaluated by hematoxylin and eosin staining and immunohistochemistry reaction. Results: The current findings demonstrated that TMJ fixed in 10 % formaldehyde and decalcified in 20 % formic acid optimized decalcification processing time with preservation of cell morphology. Antigen retrieval with citrate buffer in pressure cooker (2 min at 100 degrees C and 5 min at room temperature) demonstrated the best protocol to preservation of the structures of TMJ. Conclusions: This work demonstrates in detail a methodology of a fast and accurate TMJ processing for histology and immunohistochemistry reactions that guarantee tissue integrity and quality of staining. (AU)

• Processo FAPESP: 17/22334-9 - Uso de sistemas de liberação de fármacos para o desenvolvimento e aplicabilidade de agentes anti-inflamatórios com potencial efeito imunomodulador e neuroprotetor
• Beneficiário: Marcelo Henrique Napimoga
• Modalidade de apoio: Auxílio à Pesquisa - Temático