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A Study to Determine the Effects of Aceclofenac Sodium on Angiogenesis by using Chorioallantoic Membrane (CAM) Assay

Anam Nazeer*, Lubna Shakir, Zaka ur Rehman, Owais Omer, Komal Najam, Mohsan Sohail, Nasira Saeed, Tayyaba Nazir, Shawana Aslam, Arshia Batool Khanum

The chick Chorioallantoic Membrane (CAM) is a possibly three-dimensional representation that can be used for in vivo as well as in situ studies. Its relatively easily available, relatively less expensive and consistent in quality render it an appropriate biological model for the use in experiments requiring live tissues. The aim of current research was to determine the angiogenic/antiangiogenic effect of Aceclofenac Sodium (AcS) and the effective dose of AcS. Total 30 fertilized chicken eggs (5 days old) were obtained from a Big bird Pvt. Ltd (local hatchery). Five groups were formed. They were incubated at 37°C with humidity of 55-60%. An opening about 2 cm of diameter was created under aseptic conditions by removing the shell and inner shell membrane. On 6thday of fertilization, Group A was given 0.1 ml Phosphate Buffer Saline (PBS) and served as control, group B, C, D and E were given 0.64 mg/0.1 ml, 0.32 mg/0.1 ml, 0.16 mg/0.1 ml, and 0.08 mg/0.1 ml of AcS respectively. Eggs were then again sealed with paraffin film under sterile environment and were kept in incubator for next 24 hours. After incubation period, the CAM was exposed and pictures were taken. The growth of blood vessels, diameter, branching pattern, surface roughness and other characteristics were evaluated by using Adobe Photoshop and Scan Probing Image Processing (SPIP). AsS showed dose dependent dual effect. At low concentration (0.08 mg/ml) it showed anti-angiogenic effect whereas at higher concentration (0.64 mg/ml) it produced angiogenic results. This indicates that AsS can be beneficial for development of strategies for prevention and therapy for several types of angiogenesis dependent diseases.