Synergistic Multi-Peptide Interfaces Enhance Early Osteogenic Differentiation of Mesenchymal Stem Cells

The extracellular matrix (ECM) orchestrates stem cell fate through a sophisticated interplay of biochemical and biophysical cues. While prior biomaterial strategies have typically employed one or two bioactive peptides, such approaches rarely replicate the multifaceted signaling environment of native ECM. Here, we present a biomaterial surface cofunctionalized with three distinct peptides─BMP2, RGD, and P15─through a novel spin-coating silanization strategy, providing an advanced level of ECM biomimicry. Surface functionalization was confirmed via polarization modulation-infrared reflection-absorption spectroscopy (PM-IRRAS) and fluorescence microscopy. Human mesenchymal stem cells (hMSCs) cultured on these multipeptide surfaces were systematically evaluated by RT-qPCR and immunocytochemistry to assess the expression of osteogenic markers at the gene and protein levels. Our results demonstrate that the concurrent presentation of BMP2, RGD, and P15 significantly accelerates early osteogenic gene expression and enhances the sustained differentiation of hMSCs compared to single- or dual-peptide modifications. These findings highlight the importance of multifactorial signaling for directing stem cell fate and establish multifunctionalized surfaces as promising platforms for improving biomaterial performance in regenerative medicine.