Avaliação in vitro de alterações biomecânicas e bioquímicas induzidas por fármacos em células utilizando microscopia de força atômica e espectroscopia Raman

Abstract

In this work, atomic force microscopy and Raman spectroscopy were used to evaluate druginduced biomechanical and biochemical changes in biological cells. First, an atomic force microscope was used to investigate the effects of treatment with Semaphorin 3A on the biomechanical properties of thymocytes, through the analysis of morphology, membrane roughness and cell elasticity. The results indicated that treatment with Semaphorin 3A directly affected these properties, inducing the formation of filopodia, increasing stiffness and roughness of the cell membrane due to rearrangement of the cytoskeleton. In the second study, based on the changes observed in the Raman spectra, the drug-induced anti-inflammatory response in A549 cells was evaluated through a comparative analysis of the actions of dexamethasone and p-coumaric acid. The results of this study indicated that both drugs were efficient in the inflammatory process. However, a joint analysis of the Raman technique with multivariate statistical methods revealed different action mechanisms of the drugs, indicating p-coumaric acid as a natural therapeutic compound for the treatment of inflammatory lung diseases. Additionally, the Raman technique was also used to investigate mechanisms of cell death induced by flavonoids in macrophages infected with Leishmania amazonensis promastigotes. The biochemical changes observed in Raman spectra were associated with markers of cell death. Such alterations evidenced the major molecular mechanisms induced by flavonoids in cell behavior.