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Objective To test the surface properties and in vitro effects of a new sequential release system on MC3T3‐E1 cells for improved osseointegration. Background BMP6‐loaded anodized titanium coated with PDGF containing silk fibroin (SF) may improve osseointegration. Methods Titanium surfaces were electrochemically anodized, and SF layer was covered via electrospinning. Five experimental groups (unanodized Ti (Ti), anodized Ti (AnTi), anodized + BMP6‐loaded Ti (AnTi‐BMP6), anodized + BMP6 loaded + silk fibroin‐coated Ti (AnTi‐BMP6‐SF), and anodized + BMP6‐loaded + silk fibroin with PDGF‐coated Ti (AnTi‐BMP6‐PDGF‐SF)) were tested. After SEM characterization, contact angle analysis, and FTIR analysis, the amount of released PDGF and BMP6 was detected using ELISA. Cell proliferation (XTT), mineralization, and gene expression (RUNX2 and ALPL) were also evaluated. Results After successful anodization and loading of PDGF and BMP6, contact angle measurements showed hydrophobicity for TiO2 and hydrophilicity for protein‐adsorbed surfaces. In FTIR, protein‐containing surfaces exhibited amide‐I, amide‐II, and amide‐III bands at 1600 cm‐1‐1700 cm‐1, 1520 cm‐1‐1540 cm‐1, and 1220 cm‐1‐1300 cm‐1 spectrum levels with a significant peak in BMP6‐ and/or SF‐loaded groups at 1100 cm‐1. PDGF release and BMP6 release were delayed, and relatively slower release was detected in SF‐coated surfaces. Higher MC3T3‐E1 proliferation and mineralization and lower gene expression of RUNX2 and ALPL were detected in AnTi‐BMP6‐PDGF‐SF toward day 28. Conclusion The new system revealed a high potential for an improved early osseointegration period by means of a better factor release curve and contribution to the osteoblastic cell proliferation, mineralization, and associated gene expression.

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Cell sheet engineering is an emerging field based on the acquisition of cells together with their extracellular matrix (ECM) and is used not only in vitro but also in regeneration studies of various tissues in the clinic. Within this scope, wide variety of cell types have been investigated in terms of sheet formation and underlying mechanism. MC3T3-E1 is a mouse pre-osteoblast cell line that has greatly attracted researchers' attention for bone tissue engineering (BTE) applications thanks to its high proliferation and differentiation properties. The potential of MC3T3-E1 cells on sheet formation and the effects of culture conditions have not been investigated in detail. This study aims to examine the effects of growth and osteogenic medium on cell sheet formation of MC3T3-E1. As a result of this study; intact, ECM-rich, transferable cell sheets at the beginning of the mineralization phase of the differentiation process were obtained by using osteogenic medium. Hereafter, 3D tissue model can be constructed by stacking MC3T3 cell sheets in vitro. This 3D model can conveniently be used for the development of novel biomaterials and in vitro drug screening applications to reduce the need for animal experiments.

Novel stable diphenylalaninamide peptide based nanocarriers were designed by layer-by-layer polyelectrolyte deposition to load siRNA for gene silencing.

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