THE EFFECT OF ACTIVATED M1 MACROPHAGES ON FIBROTIC HUMAN LUNG MICROTISSUE ARRAY
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Pulmonary fibrosis is a disease which is caused due to formation of excess connective tissue which leads to stiffening of the lungs. This disease can be benign or lethal in some cases. In our study, we have taken primary human lung fibroblasts and seen how they react when treated with M1 macrophages. Human lung fibroblasts (HLF) are the best model for studying pulmonary function. This interaction is studied in the microdevice which is made up of Poly-Di-Methyl-Siloxane (PDMS) and produced with the help of photolithography. There are arrays of microtissue formed in the microdevice which show biomechanical changes in the morphology of cells. When M1 macrophages are added to HLF cells, the HLF cells contract to a certain extent. In the results, the increase in force of contraction is clearly shown over a period of 3 days. This contraction is because of the release of TGF-β1 by M1 macrophages. By immunostaining done on these microtissues we can see the α-Smooth Muscle Actin and the nuclei in the microtissue. The amount of TGF-β1 is measured later in the study with the help of ELISA Assay. The second part of the thesis deals with drug testing, where HLF cells are treated with anti-fibrosis drug, Pirfenidone. Pirfenidone is taken at two different concentrations which shows how the drug negates the effect of M1 macrophages on HLF cells and also help control fibrosis.