The development of new blood vessels from pre-existing capillaries is termed as angiogenesis Griffioen and Molema, 2000. This term angiogenesis (AG) was used in 1935 to define the development of new blood vessels in the placenta Hertig, 1935. All cells and tissues survival are solely dependent on AG. Therefore, AG is extremely meaningful to different physiological functions like wound healing, menstruation, development of embryo and normal tissue growth Griffioen and Molema, 2000. Physiologic AG involves many different pro- and anti-angiogenic stimuli including various environmental and growth factors Griffioen and Molema, 2000. The AG switch is also observed as a distinct stage of cancer development, that occurs at any phase of tumor development and its micro-environment Bergers and Benjamin, 2003. The use of anti-AG drugs for the management of AG established conditions have presented a lot of therapeutic potential. Numerous anti-AG medications are presently under development, involved in clinical trials or are being used in the treatment of AG complaints, mostly cancer Abdollahi and Folkman, 2010. CAM model is used in the experiments in this study in order to test anti-angiogenic agents alone and in combination Tufan and Satiroglu-Tufan, 2005. This model delivers a usual, in vivo setting of angiogenic blood vessels with all the multifaceted swarm communications on which angiogenic combinations can be verified Tufan and Satiroglu-Tufan, 2005.

Naproxen sodium is a non-steroidal anti-inflammatory drug (NSAID). It functions by decreasing hormones that provoke agony and soreness in the body. The aim of current research was to determine the angiogenic/anti-angiogenic effect of NapS and to determine the effective dose of NapS for angiogenesis.

The CAM model is widely used in angiogenic investigation and is principally appropriate for the study of diseases categorized by proliferative retinal vasculature, such as age-related macular degeneration Tufan and Satiroglu-Tufan, 2005. In the current effort, this model was used in order to determine the angiogenic and/or anti-angiogenic effect of Naproxen sodium (NapS) and determine the effective dose of NapS for angiogenesis.

The effects of drug visualized in Figure 1 show that NapS is angiogenic at higher concentration and anti-angiogenic at low concentrations. Similar studies were conducted by Blacher et al. (2005) Blacher et al., 2011. They used new advances in imaging investigation to quantify CAM angiogenic response, covering all vascular components, from the large supplying and feeding vessels down to the capillary plexus. Their morphometric investigation highlighted that a precise quantification of the CAM vasculature desires to be executed at several scales Blacher et al., 2011.

Quantitative analysis was performed including imaging studies, roughness analysis and diameter of blood vessels. The NapS was applied on CAM and images were captured at uniform time intervals. The images captured were then analyzed with the software. These were transformed into three dimensional views as shown in Figure 2 . These were prepared to know the better understanding of blood vessels networks.

There are no worldwide gauges for roughness investigation in light of surface pictures yet, however the actualized calculations are in assertion suggestions from driving specialists and a large portion of the definitions are regular augmentations of the ISO norms for surface profiles. With respect to all other expository outcomes the roughness parameters are composed to records, which can be transported in by worksheet suites and utilized for factual purposes. We also performed this parameter to check the roughness of images. These factors illuminate the variances in surface roughness between control and treated CAMs. The Abbott curve, a graphical picture of roughness, was also dignified to appraise even slight changes in the stature of blood vessels on the surface of CAMs. Abbott curve heights for control and treated CAMs were observed as shown in figure 3. This shows that roughness was found high in group A and B while in group C, D and E, roughness was comparatively low. Similarly, same roughness parameter was observed by Hussain et al. (2011) Hussain, 2011. In their study, the average values of roughness in the control group were high in relation to treated CAMs. It portrayed that surface roughness analysis, demonstrating neo-vascularization of treated CAMs was expressively (P<0.05) less than that of control CAMs Hussain, 2011.

The captured images were assessed with the software to determine the vessel diameters. Different segments in a selected area for measurement were identified. The respective diameters of each vessel portion located in distinct segments were identified using the software. Using this procedure, the diameter of a specific vessel was determined automatically at each time point for the respective drug concentration. The dispersal of the vessel diameter of the CAM was observed to be usual. From the investigations, it was obvious that the vessel diameters of the CAM did not modify meaningfully over time. It was also observed that in Group A and B diameter increased which shows that NapS increases diameter in higher concentrations. However, there is no significant statistical difference between both groups. On the other hand, in low concentrations, NapS worked as an anti-angiogenic agent and diameter of blood vessels decreased with a significant difference among control and treated groups (P=0.000) as shown in figure 4. Another study was conducted by Salas (2015) Salas, 2015 to assess the phytochemical components and properties of herbal plant extracts, such as Gynura nepalensis, Pandanus odoratissimus L. and Carmona retusa masam, as potential angiogenesis inhibitors using the CAM assay. They found similar reduction in diameter with Carmona retusa masam.

The current research was also executed to investigate that blood vessels are fashioned by the capillaries selection in the plexus. To recognize the developmental phenomena of blood vessel branching pattern, a series of radiographs was successfully recorded, and a computer simulation was carried out by the process of in vivo vascularization. The simulation established that a positive feedback system contributed in the development of a branching pattern. As the embryo grew, it was witnessed that in Groups A and B, there was a fork branching pattern while in group C, D and E, there was polygonal and/or tree capillary networks as shown in Figure 5 . An area where progress was rapidly received much blood flow and produced finer networks of capillaries.

It is concluded that the presentation of Naproxen sodium on CAM on 6th day of incubation showed angiogenic effects in high concentration and anti-angiogenic effect in low concentration. Results indicated noticeable modifications in CAM design, weakening of blood vessels, reduction in surface roughness parameters, and decrease in Abbott curve. The significant amounts of Naproxen sodium may reveal anti-angiogenic activity in the similar fashion as those seen in vitro and describe its clinical efficacy.

AG: Angiogenesis

CAM: Chorioallantoic membrane

NapS: Naproxen sodium

SPSS: Statistical package for social sciences

Hafiz Muhammad Shoaib Zahid conducted the research, Tanveer Ahmed Khan performed data analysis, Muhammad Ijaz-ul-Haq drafted the manuscript, Humayun Riaz reviewed the manuscript, Syed Atif Raza supervised the study, and Zia Mohy-ud-din Khan performed graphical interpretation.