Angiogenesis is a physiological process which involves the formation of new blood vessels from vessels which are already present. It is a common and most important process in formation and development of vessels, so used in healing of wound and granulation of tissues. To maintain natural balance between formulation and inhibitory factors, body controls angiogenesis. When this balance is disturbed, the body results in either too much growth or extensive inhibition of expansion of blood vessels.
This study was designed to evaluate the effects of Punica granatum (pomegranate) fruit extract on angiogenesis using chick chorioallantoic membrane assay (CAM). The phytochemicals in pomegranate include polyphenoliccatechins, gallocatechins and anthocyanins like prodelphinidins, delphinidin, cyanidin and pelargonidin. CAM assay model is very helpful to proceed in research due to its easy and accurate observation of embryonic development and the process of angiogenesis.
The proposed hypothesis was, is there any Effect of Punica granatum (Pomegranate) fruit extract on angiogenesis? To check the authenticity of this hypothesis pomegranate (Punica granatum) was collected from the locality of Lahore. Aqueous extract was obtained by maceration of dried powder. Forty fresh fertilized eggs were collected from local hatchery and divided into four groups, having 10 eggs in each group. These eggs were incubated and were windowed at day 5 of incubation. On the same day a specified portion of albumin was removed with the help of syringe. On day 6 various dilutions of fruit extract was prepared and applied on developing CAM on day 6 after incubation while the controlled group was treated with distilled CHAPTER-6 65
water only. After 24 hours, the pattern of angiogenesis in the developing CAM was evaluated by taking images with the help of digital camera and SPIP software program.
3D surface roughness parameters showed the clear decline in values as compared to control. The parameters are; Sa (surface area), Sq (root mean square), Ssk (surface deviation), , Sy (lowest valley), Sz ( average absolute height), Smin (minimum height), Smax (maximum Height), Smean (mean height), Sdr (developed surface area ratio), Svk (Reduce valley depth), Sci (ratio of void volume of the unit sampling at core zone over root mean square deviation), Sk (core roughness depth), Stdi (texture index). All of these parameters were assessed thoroughly for the quantification of angiogenesis. The length and diameter of primary, secondary and tertiary blood vessels as well as the area of CAM were measured with the help of calibration and measurement command.
Changes in angiogenesis were observed with different percentage dilutions of fruit extracts and were statistically analyzed by using the analysis of variance where “p-value” of control and treated groups were calculated. With this application, significant differences in the results were observed at p<0.05 which collectively pointed toward antiangiogeic effect of the pomegranate fruit extract.