INVESTIGATION OF THE EFFECT OF CAFFEIC-ACID-PHENETHYL ESTER TO ISCHEMIA/REPERFUSION DAMAGE IN TRAM PEDICULAR FLAP MODEL
Project no. 99 · Status: Kapanmış · 2017
Summary
Recent execution of Plastic and Reconstructive Surgery practice includes frequent use of free and pedicled flaps for reconstruction of tissue defects. Frequent use of this technique also brings many complications such as ischemia-reperfusion injury. In this study, altering effect of CAPE on flap loss in rat TRAM free and pedicled flap model was studied. The purpose of the study is to investigate if preoperative CAPE administration to animals with I/R risk can prevent abnormal changes including suspension of internal defence mechanisms and also if it enhances the vascularisation of the flap to enhance the tissue tolerance to I/R injury. In the study, 42 randomised Sprague-Dawley rats were grouped to 3 groups, including 14 animals at each, named as Pedicled/Free Flap, Pedicled- or Free- Flap-Ischemia and Pedicled- or Free-Flap—Ischemia-CAPE. Each group was subdivided to two groups by their flap procedure. Pedicled bilateral TRAM flaps were designed on the first seven rat of each group, and free bilateral TRAM flaps were designed on the rest of the group. 2x2 cm of skin and 4 cm long of rectus muscle was included in each flap. In the first group, flaps were returned to the original places without clamping or injection. The second group, ischemia was done with microclamps. Before the ischemic process, 2 cc i.p. injection of Saline was made 30 minutes before the application of the clamps. Ischemic term was lasted exactly for 2 hours. On the 3rd group, with the same ischemia protocol, 2 cc of CAPE solution was injected instead of Saline only. After the 2 hours, clamps were released. All of the flaps were harvested on the 7th postoperative day and flaps on the right were brought to biochemistry and the contralateral side, to the histology departments. (A total of 84 flaps.) With macroscopical examination, necrosis areas were calculated. Photos of the left flaps were evaluated for measurements were done. The calculations showed statistically significant difference of healthy flap areas between the groups, indicating less injury in the third group than the second group. (p<0.01) Left flaps of the rats were evaluated with histological examination for marks of injury with H&E ve Mallory Azan stains. Verhofstad scoring was done with necrosis, edema, PMNL infiltration, collagen density, vasculatisation and healthy glandula sebacea rates. Besides, VEGF, iNOS, BAX and Bcl-2 levels were showed with immunohistochemical staining. The results showed significant differences between the groups, showing less injury on the third group than the second group. (p<0.01) Biochemical data from the right-sided TRAM flaps were evaluated for SOD, MDA and CAT levels with ELISA. Results showed that the second group which has the highest MDA level, has the highest rate of ischemia-reperfusion injury. On the other hand, third group showed less MDA levels and higher SOD and CAT levels, indicating less injury than the second group. (p<0.01) In light of these information, preoperative CAPE administration prevents postreperfusion pathological processes that damages the tissues with significant difference and can be used for pharmacological pre-conditioning.