High-resolution topographic mapping of cells adhered to hydrogel matrices

• Grant number: 25/11693-4
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2025
• End date: July 31, 2026
• Field of knowledge: Biological Sciences - Biophysics - Molecular Biophysics
• Principal Investigator: Emerson Rodrigo da Silva
• Grantee: Kayky Novais Batista
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated research grant: 22/03056-6 - Ultrastructural characterization of bioactive nanoscaffolds, AP.PNGP.PI

Abstract

The development of scaffolds capable of modulating and stimulating cell growth is considered a frontier issue with major impacts on future medicine. Among other things, these matrices are aimed at stimulating tissue growth and producing, for example, artificial skin and prosthetic corneas. Naturally, an in-depth knowledge of how cells are distributed on these matrices and the membrane structures (e.g. microtubules) responsible for attachment to the substrate is an essential condition for the full development of the field. One of the most powerful ways of determining this information in great detail is atomic force microscopy, which allows three-dimensional information to be collected on a nanometric scale. However, obtaining adequate data requires appropriate preparation protocols, as it is not trivial to carry out tests on cells under hydrated conditions, often requiring them to be fixed. In this project, we intend to develop a fixation protocol for observing cells grown on peptide hydrogel matrices. Our aim will be to identify the best conditions and materials for obtaining viable samples, capable of providing quality and reproducible results. The work will consist of growing cells (HeLa, NIH3T3 and bacteria) on specimens covered with hydrogels. The samples will then be fixed using traditional microscopy methods (e.g. PFA, glutaraldehyde) and analyzed via AFM. The conditions for both sample preparation and equipment configuration will have to be optimized in order to obtain quality data.