Three Dimensional Polymeric Matrix and Terminalia chebula Synergitic Effect on Human Lung Cells

The proliferative rate of lung epithelial cells is very important for repair of alveolar surface injury which can be caused due to oxidant stress in local tissues leading to different pathological conditions. However, for testing of any antioxidant component it is critical to fabricate a three-dimensional culture that mimics the local tissue enviornment thereby any results achieved in the in-vitro could easily be translated in the biological system. In step towards this, we have combined 3-D polymeric matrix made by 2-hydroxyethylmethacrylate-gelatin-alginate along with natural herbal extract of Terminalia chebula and tested for its effect on lung epithelial cells. Scanning electron microscopy was performed to check the pore size and distribution within the matrix. Average pore size was found to be in range of 75 mu m with homogenous distributed interconnected pores. Dynamic mechanical analysis was performed to check of the mechanical stability of the scaffold and storage modulus was around 3 MPa +/- 0.01 for the scaffold is saturated with phosphate buffer saline. Terminalia chebula extracts was prepared by soxhlet extraction method in which acetone was used as an extraction solvent. Different concentration of this extract tested with DPPH assay and extract with highest antioxidant characteristics was loaded onto matrix for 24 hours before seeding human lung epithelial cells. Total metabolic activity of cells was checked using MU assay and live/dead viability staining for 4 weeks. Cellular attachment and proliferation was analyzed using SEM shows higher cell attachment and ECM production in presence of chebula extract in comparison to 2D and 3D system without extracts. The combination of herbal extracts with three-dimensional matrix has synergetic effect that aided in higher ECM production and cellular proliferation. This combination could be used as step in finding potential biomaterials based lung tissue regeneration therapy.