Kunza Naveed Asdaq, Hashmat Gul, Tayyaba Bari, Asima Asghar, Muhammad Kaleem, Fakhera Ikram
Plant-derived decellularized scaffolds have emerged as sustainable and structurally versatile biomaterials for tissue engineering; however, their potential for supporting early osteogenic responses remains insufficiently explored. In this study, decellularized scaffolds derived from Apple (Malus pumila), Carrot (Daucus carota), Capsicum (Capsicum annuum), and Zucchini (Cucurbita pepo) were characterized using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and Fourier transform infrared spectroscopy (FTIR), and their cellular responses were evaluated using the Alamar Blue assay and reverse transcription-polymerase chain reaction (RT-PCR). Alamar Blue data were analyzed using two-way ANOVA followed by Tukey's post hoc test (n = 3), while gene expression was evaluated using the comparative Ct (2^-ΔΔCt) method in a singlicate run (n = 1). Characterization confirmed preservation of interconnected porous architectures across all scaffold groups, with zucchini-derived scaffolds exhibiting the most uniform pore distribution. All scaffolds supported MC3T3-E1 cellular metabolic activity, which increased from Day 1 to Day 7. Expression of the osteogenic-associated markers RUNX-2, COL1A1, and ALP was detected at Day 7, indicating an early osteogenic response. However, the single-time-point gene-expression analysis does not establish progression of osteogenic differentiation or mineralization. Overall, the findings demonstrate that decellularized plant-derived scaffolds provide a cytocompatible three-dimensional platform that warrants further investigation for bone and craniofacial tissue engineering applications.