Platelet-Rich Plasma Pretreatment on Grit-Blasted Titanium Alloy for Enhanced Osteogenic Differentiation of Human Adipose-Derived Stem Cells

ABSTRACT

BACKGROUND: Adequate bone formation around titanium alloy implants is integral to successful implantation surgery. Stem cell-coated implants may accelerate peri-implant bone formation. This study investigates the effect of platelet-rich plasma (PRP) pretreatment on a titanium-alloy surface in terms of proliferation and osteogenic differentiation of human adipose-derived stem cells (hADSCs). METHODS: Allogenic leukocyte-depleted PRP was obtained from blood supernatants. The hADSCs were isolated from thigh subcutaneous fat tissue. Grit-blasted titanium plugs were used in two different groups. In one group, 200 µL of PRP was added to the grit-blasted titanium plugs. The hADSCs were seeded in two groups grit-blasted titanium plugs with or without PRP. The number of hADSCs was measured after 4 hours, 3 days, and 7 days of culture using Cell Counting Kit-8. Osteogenesis of hADSCs was measured by using an alkaline phosphatase activity assay on days 7 and 14, and a calcium assay on days 14 and 21. Osteogenic gene expression was measured by using reverse transcription polymerase chain reaction analysis of alkaline phosphatase, osteocalcin, and type I collagen mRNA. The microscopic morphology of grit-blasted titanium plugs with or without PRP was examined with a field-emission scanning electron microscope using a JSM-7401F apparatus on days 1 and 7. RESULTS: Proliferation and osteogenic differentiation of hADSCs were found to be significantly higher on the grit-blasted titanium alloy preprocessed with PRP than the same alloy without pretreatment. Furthermore, a structural fibrillar mesh developed compactly on the grit-blasted titanium alloy with the PRP pretreatment. CONCLUSIONS: Our results demonstrate that a hADSC-based approach can be used for tissue-engineered peri-implant bone formation and that PRP pretreatment on the grit-blasted titanium alloy can improve proliferation and osteogenic differentiation of hADSCs.