Effect of Osteoblast‐Extracellular Matrix‐Functionalized Nanostructured Titanium Surface on Osteoblastic Cell Behavior

Nanostructured titanium (Ti) facilitates osteoblast differentiation by modulating various signaling pathways and may enhance osseointegration, potentially through its influence on the formation and remodeling of the extracellular matrix (ECM). This study aimed to investigate the impact of osteoblast-ECM-functionalized nanostructured Ti surface on osteoblastic cell behavior. Initially, it was demonstrated that nanotopography upregulated the expression of genes associated with ECM formation and remodeling in MC3T3-E1 osteoblastic cells compared to a polished Ti surface. Then, MC3T3-E1 cells were cultured on nanotopography, and the cultures were decellularized to generate ECM-functionalized nanostructured Ti surface. Successful decellularization was verified by 4',6-diamidino-2-phenylindole staining for DNA, DNA quantification, fibronectin immunodetection and scanning electron microscopy. To investigate the effect of ECM functionalization on osteoblasts, calvaria-derived osteoblasts were cultured on both non-functionalized and osteoblast-ECM-functionalized nanostructured Ti surfaces. ECM functionalization did not affect cell morphology but reduced proliferation and the gene expression of osteoblastic markers while increasing alkaline phosphatase activity and ECM mineralization. Additionally, RAW 264.7 osteoclastic cells cultured on ECM-functionalized surface exhibited higher tartrate-resistant acid phosphatase activity. Functionalization of nanostructured Ti surfaces created a favorable microenvironment for bone cell activity and bone remodeling, contributing to the development of smart osseointegrated Ti implants.