Layer-by-layer assembly of bioactive nanofilms for in situ gene transfection of a LAMA3 mutant on hydrogenated TiO₂ nanotubes: in vitro evaluation of epithelial adhesion and hemidesmosome formation
摘要
Reliable peri-implant epithelial sealing is paramount for the prevention of peri-implantitis. Remarkably, both hemidesmosomes (HDs) and the internal basal lamina, which notably includes laminin332—essential for transmucosal epithelial sealing—are significantly diminished compared to their natural tooth counterparts. In situ gene therapy has emerged as a promising approach for the spatial regulation of HD formation.
MethodsIn this study, we aim to develop a bioactive coating with an exceptional capacity for the in situ transfection of a recombinant adenovirus carrying a mutant of the laminin α3 chain gene (Ad-mLAMA3), thereby improving transmucosal epithelial sealing. Dopamine (DA), graphene oxide (GO), and type Ⅳ collagen (COL-Ⅳ) multilayers—(DA/GO/COL-Ⅳ)10, designated DGC—were assembled layer by layer on hydrogenated TiO2 nanotubes. The resulting construct was termed H‑DGC. We characterized H-DGC and evaluated its effect on HD formation in vitro following integration with Ad-mLAMA3.
ResultsThe DGC nanofilms were successfully established on hydrogenated TiO2 nanotubes, enhancing in situ transfection efficiency. The integration of H-DGC with Ad-mLAMA3 promoted epithelial cell adhesion and mature HD formation by upregulating the expression of laminin α3 and integrin β4.
ConclusionsOur findings demonstrate that H-DGC can be engineered as a bio-instructive and in situ interface, significantly enhancing epithelial cell adhesion and mature HD formation, and offering a strategy to prevent peri-implantitis.