Dupuytren’s disease investigation under 2D and 3D environment: Atomic Force Microscopy approach

Abstract

The general aim of this thesis was to study the differences between three primary fibroblasts from the Dupuytren’s disease in different environments using mostly AFM supported by other techniques like immunofluorescence. The three primary cell types were obtained from different tissue samples from the palm of the same patient who suffered from Dupuytren’s disease. The different natural of the samples was identified, showing healthy fibroblasts phenotype for the cells subtracted from dermal regions of the palm, ambiguous behavior of the cells from the scar tissue and myofibroblast phenotype for the cells from the nodules of the palmar fascia. Knowing the differences between the cells, different strategists were applied to learn more about the disease and to disentangle new scientific problems. Cell mechanics have been used for many years to obtain information about cell characteristics linked to mechanical properties. Lekka et al. were the first to use it as a diagnostic tool to distinguish between healthy and cancerous cells, discovering the softer nature of cancerous cells on hard supports. Cell mechanics has become a useful methodology to study cell behavior at single cell level, such as diseased cells response under pharmacological treatments. As mentioned before, cell mechanics enables the design of a multitude of experiments to assess single cell response in different conditions, like cell’s behavior in matrices of different mechanics as well as different biochemical composition, cells under pharmacological treatment, salt, pH and Tª variations, etc. However, in natural conditions cells are surrounded by neighboring cells and ECM. Therefore, cells mechanics in 2D environment differ from those in 3D environment. In 3D, cells are subjected to space limitations (constraints) and cell-cell and cell-ECM interactions. Neighboring cells communicate and interact to each other developing characteristic cell behavior in tissue, which will be different at single cell level due to the lack of cells cross-communication. Thus, research experiments turning into tissue-like conditions can help to decipher hidden mechanisms in diseases. In this line, this thesis evolves from single cell level experiments to more tissue-like conditions in an attempt to disentangle unknown characteristics of Dupuytren’s disease.