Annexin v - affinity assay - apoptosis detection system in granular cell ameloblastoma

Abstract

Background: The aim of the study is to detect apoptosis in granular cell ameloblastoma by annexin v affinity assay, a novel technique.

Materials & methods: Excitional biopsy of two patients with granular cell ameloblastoma were fixed in buffered formalin and later embedded in paraffin wax. Blocks were sliced into 3m thick sections for routine histological and subsequent immunohistochemical examinations. For electron microscopic examination tissues were fixed in 2.5% glutaraldehyde.electron microscopic examination was carried out to find the nature of granularity in granular cell ameloblastoma which was finally confirmed by annexin v technique.

Results: Annexin v is a sensitive marker to detect early apoptosis. Fluorescence in granular cell clusters showed that apoptotic cell death is higher in granular cells. Both early and late events of apoptosis were identified in annexin v staining and electron microscopic study respectively.

Conclusion: Our study confirms that increased apoptotic cell death and subsequent phagocytosis is responsible for granular appearance of cells in granular cell ameloblastoma compared with that of conventional ameloblastoma. How to cite this article: Balaji N, Devy AS, Sumathi MK, Vidyalakshmi S, Kumar GS, D'Silva S. Annexin V - Affinity Assay - Apoptosis Detection System in Granular Cell Ameloblastoma. J Int Oral Health 2013; 5(6):25-30 .

Keywords: Affinity assay; annexinv; apoptosis; phagocytosis.

Figure 1: Odontogenic tumuor in connective tissues stroma.

Figure 2: Tall columnar cells.

Figure 3: EM examination, granular cell showed lysosomal aggregates.

Figure 4: Lysosomal vacules showed phagocytosed material.

Figure 5: Granular cell clusters, apoptotic cell fragments with condensed chromatin is identified.

Figure 6: Phosphotidyl serine an aminophospholipid present in inner phase of the plasma membrane is exposed to the outer leaflet during apoptosis. This acts as a tag for its specific recognition by macrophages and for phagocytosis of the dying cell.

Figure 7: Annexin V is a calcium dependent protein that binds with phospholipid and this FITC tagged annexin V floresed as it stained positive for apoptotic bodies.

Figure 8: Annexin V is a calcium dependent protein that binds with phospholipid and this FITC tagged annexin V floresed as it stained positive for apoptotic bodies.