Natural extracts like C. asiatica, T. arjuna natural extracts were procured from Chemiloids, India. Collagen was obtained from Shevoroy’s Ltd India. 2,2 1 azo bisisobutyronitrile (AIBN) were purchased FRAX597 from Merck (India). All other chemicals used in this research

activity were of analytical grade. Collagen was soaked in 0.05 M glacial acetic acid at 25 mg/ml concentration for 24 h at 4 °C. The obtained viscous solution was homogenized for 5 min, deaerated for 15 min by using sonicator and squeezed through a muslin cloth to get rid of undissolved solid traces if any (Note: for cross-linking 0.8 ml of 25% v/v glutaraldehyde solutions were added to the formulation at this stage).7 Various solutions with different concentrations of C. asiatica and T. arjuna ( Table 1) were prepared by dissolving them in 3 ml of alcohol. Each of the prepared solutions

was mixed with 40 ml of the above cross-linked collagen Crizotinib solution separately. The obtained mixture was casted in petri plate (64 cm2) having polyethylene membrane base and placed in incubator at 37 °C until dried. The scaffold thus obtained was sterilized under UV-radiation for a period of 18 h. The thickness of the plain collagen, cross-linked collagen and different natural extracts (C. asiatica and T. arjuna) of varying concentration impregnated collagen based films was measured by using a screw gauge (LINKER-20 × 1/100 mm). The mean of 3 observations was calculated. Folding Endurance was measured manually for the prepared films. For this a strip of film (2 × 2 cm2) was cut evenly and repeatedly folded at the same place until it broke. The number of times the film could

be folded at the same place without breakage gave the exact value of folding endurance. The mean of 3 observations was calculated. The equilibrium swelling ratio (Es) was measured by the conventional gravimetric method. The dry weight of different scaffolds was measured before immersing in 0.05 M phosphate buffer saline (PBS) pH 7.4 at a temperature of 37 °C and excess surface phosphate buffer saline was blotted out with absorbent paper. The wet weight (Ws) of the film was determined after being incubated for Electron transport chain 24 h. The equilibrium swelling ratio of the films was defined as the ratio of weight increase (Ws − Wd) with respect to the initial weight (Wd) of dry samples. Each value was averaged from three parallel measurements. Es was calculated using the following equation: Es=Ws−WdWdwhere Ws and Wd denote the weights of swollen and dry sample, respectively. The Micro Shrinkage Temperature Studies were carried out for the collagen, cross-linked collagen and various natural extracts of different concentration impregnated collagen based films. For this study, the collagen films were stage fitted to an optical microscope.