3D-Printed PLA Scaffold with Fibronectin Enhances In Vitro Osteogenesis.

Abstract

Tricalcium phosphate (TCP, Molecular formula: Ca3(PO4)2) is a hydrophilic bone graft biomaterial extensively used for guided bone regeneration (GBR). However, few studies have investigated 3D-printed polylactic acid (PLA) combined with the osteo-inductive molecule fibronectin (FN) for enhanced osteoblast performance in vitro, and specialized bone defect treatments. This study evaluated PLA properties and efficacy following glow discharge plasma (GDP) treatment and FN sputtering for fused deposition modeling (FDM) 3D printed PLA alloplastic bone grafts. 3D trabecular bone scaffolds (8 × 1 mm) were printed by the 3D printer (XYZ printing, Inc. 3D printer da Vinci Jr. 1.0 3-in-1). After printing PLA scaffolds, additional groups for FN grafting were continually prepared with GDP treatment. Material characterization and biocompatibility evaluations were investigated at 1, 3 and 5 days. SEM images showed the human bone mimicking patterns, and EDS illustrated the increased C and O after fibronectin grafting, XPS and FTIR results together confirmed the presence of FN within PLA material. Degradation increased after 150 days due to FN presence. 3D immunofluorescence at 24 h demonstrated better cell spreading, and MTT assay results showed the highest proliferation with PLA and FN (p < 0.001). Cells cultured on the materials exhibited similar alkaline phosphatase (ALP) production. Relative quantitative polymerase chain reaction (qPCR) at 1 and 5 days revealed a mixed osteoblast gene expression pattern. In vitro observations over a period of five days, it was clear that PLA/FN 3D-printed alloplastic bone graft was more favorable for osteogenesis than PLA alone, thereby demonstrating great potential for applications in customized bone regeneration.