Synchrotron FTIR Microspectroscopy Imaging for Evaluating Cell Distribution on Electrospun Fibrous Scaffolds

In tissue engineering and biomaterials research, reliable evaluation of cell adhesion and spatial distribution on fibrous scaffolds remains challenging. Synchrotron radiation-based Fourier transform infrared (SR-FTIR) microspectroscopy imaging provides a rapid, nondestructive, and label-free approach for mapping biochemical components through their characteristic functional groups. In this study, SR-FTIR imaging was employed as an analytical framework to evaluate NIH3T3 fibroblast adhesion and distribution on electrospun polylactic acid (PLA)/microcellulose fibrous scaffolds. Chemical mapping enabled direct visualization of cell localization on the fibers after 24 h of culture, while hierarchical cluster analysis (HCA) revealed variations in cell distribution associated with microcellulose content. A qualitative distribution index derived from normalized cluster-based metrics across replicated mapped areas showed a decrease from 100.61 to 95.58, 10.74, and 42.64 for microcellulose contents of 0, 1, 5, and 10 wt %, respectively, indicating the most homogeneous cell distribution at 5 wt %. These results demonstrate that SR-FTIR microspectroscopy imaging is an effective tool for assessing cell-material interactions on electrospun fibrous scaffolds and provides a practical methodology for biomaterials evaluation in tissue engineering applications.