Microstructural Evaluation of Eggshell Membrane Using Scanning Electron Microscopy Combined with Digital Image Analysis for Biomedical and Pharmaceutical Applications

Abstract

In the context of pharmaceutical innovation driven by digitalization and sustainable development, the valorization of agricultural by-products for biomedical material development has attracted increasing attention. Eggshell membrane (ESM) is a collagen  ̶  and carbohydrate-rich natural material, making it suitable for wound dressing and soft tissue regeneration applications. This study evaluates the effects of an acetic-citric acid mixture on the microstructure of ESM through scanning electron microscopy (SEM) observation and digital image analysis. ESM samples were treated with acid mixtures at different ratios and analyzed using Fiji/ImageJ software. The results showed that porosity increased from (36.09 ± 3.02) % to (42.76 ± 3.91) %, while the equivalent pore diameter increased from (22.00 ± 14.41) px to (27.12 ± 18.09) px. Whereas pore density decreased from (280 ± 17 to 216 ± 12) pores per field of view. These structural changes reflect the reorganization of the ESM fibrous network under the influence of the acetic-citric acid treatment, thereby affecting the material’s gas exchange and moisture retention properties. The digital image-based approach established a valuable quantitative dataset, contributing to the optimization of biomedical materials in line with innovation and sustainable development trends in the pharmaceutical industry in the digital era.