Proposal for experimental in vitro model to assess morphological alterations in erythrocytes exposed to 5.25% NaOCl

• Published in: Revista odontológica Mexicana 2016-10, Vol.20 (4), p.e241-e245
• Main Authors: Arroyo Cervantes, Roberto, Cuin Macedo, Sergio Iván, Calderón Rojas, Benigno Miguel, Rodríguez Zaragoza, Diana Ened, Ruiz Reyes, Héctor
• Format: Article
• Language: English
• Subjects: eritrocitos; erythrocytes; Experimental model; hipoclorito de sodio; Modelo experimental; sodium hypochlorite
• ISSN: 1870-199X
• DOI: 10.1016/j.rodmex.2016.11.014

Sodium hypochlorite (NaOCl) is the chemical agent most frequently used as irrigation solution during endodontic therapy. When extruded to periapical tissue, it is highly toxic. In endodontics, hemolysis caused by NaCOl has been proven using different models, nevertheless, there is little or no evidence of morphological alterations in the cellular membrane of erythrocytes. To propose an experimental model which might allow to assess morphological alterations suffered by erythrocytes when they are exposed to NaOCl used in the dental practice by means of high resolution scanning electron microscopy (SEM). In the present study, 20mL of peripheral blood were obtained and deposited in tubes with EDTA (ethylenediaminetetraacetic acid) anticoagulant. Rinses were conducted with a phosphate buffer solution (Evan's solution). Several dilutions of the erythrocyte sample were prepared (1:1, 1:2, 1:4, 1:8 and 1:16); 100μL of each of these dilutions was obtained to be then confronted with 100μL of dental use 5.25% NaOCl (Viarzoni-T, Medental®); 0.5μL of these samples were taken to then be deposited in a sample holder made of Zn-Cu alloy which was subjected to a process of Cu ion metallization bath, following the old Spluttering method. Microphotographs were obtained with SEM. Erythrocytes with alteration type anisocytosis and poikilocytosis (stomatocytes, elliptocytes and discocytes) were observed. Some structural characteristics of NaOCl crystals were equally observed. This experimental model allowed assessment of morphological changes experienced by erythrocytes when exposed to 5.25% NaOCl. El hipoclorito de sodio (NaOCl) es el agente químico más utilizado como solución irrigadora durante la terapia endodóntica. Es altamente tóxico cuando se extruye a tejido periapicales. En endodoncia la hemolisis causada por el NaOCl ha quedado demostrada utilizando diferentes modelos, sin embargo poca o ninguna evidencia se tiene de las alteraciones morfológicas en la membrana celular de los eritrocitos. Proponer un modelo experimental que permita evaluar las alteraciones morfológicas que sufren los eritrocitos cuando son expuestos a NaOCl utilizado en la práctica odontológica mediante microscopia electrónica de barrido de alta resolución (MEB). Se obtuvieron 20mL de sangre periférica y se depositaron en tubos con anticoagulante EDTA (ácido etilendiaminotetraacético). Se realizaron lavados con solución amortiguadora de fosfatos (solución Evan's). Se prepararon diferentes diluciones