Chitosan-miRNA functionalized microporous titanium oxide surfaces via a layer-by-layer approach with a sustained release profile for enhanced osteogenic activity

Abstract

Background The biofunctionalization of titanium implants for high osteogenic ability is a promising approach for the development of advanced implants to promote osseointegration, especially in compromised bone conditions. In this study, polyelectrolyte multilayers (PEMs) were fabricated using the layer-by-layer approach with a chitosan-miRNA (CS-miRNA) complex and sodium hyaluronate (HA) as the positively and negatively charged polyelectrolytes on microarc-oxidized (MAO) Ti surfaces via silane-glutaraldehyde coupling. Methods Dynamic contact angle and scanning electron microscopy measurements were conducted to monitor the layer accumulation. RiboGreen was used to quantify the miRNA loading and release profile in phosphate-buffered saline. The in vitro transfection efficiency and the cytotoxicity were investigated after seeding mesenchymal stem cells (MSCs) on the CS-antimiR-138/HA PEM-functionalized microporous Ti surface. The in vitro osteogenic differentiation of the MSCs and the in vivo osseointegration were also evaluated. Results The surface wettability alternately changed during the formation of PEMs. The CS-miRNA nanoparticles were distributed evenly across the MAO surface. The miRNA loading increased with increasing bilayer number. More importantly, a sustained miRNA release was obtained over a timeframe of approximately 2 weeks. In vitro transfection revealed that the CS-antimiR-138 nanoparticles were taken up efficiently by the cells and caused significant knockdown of miR-138 without showing significant cytotoxicity. The CS-antimiR-138/HA PEM surface enhanced the osteogenic differentiation of MSCs in terms of enhanced alkaline phosphatase, collagen production and extracellular matrix mineralization. Substantially enhanced in vivo osseointegration was observed in the rat model. Conclusions The findings demonstrated that the novel CS-antimiR-138/HA PEM-functionalized microporous Ti implant exhibited sustained release of CS-antimiR-138, and notably enhanced the in vitro osteogenic differentiation of MSCs and in vivo osseointegration. This novel miRNA-functionalized Ti implant may be used in the clinical setting to allow for more effective and robust osseointegration.

3D printed porous titanium filled with mineralized UV-responsive chitosan hydrogel promote cell proliferation and osteogenesis in vitro

“…Titanium (Ti) and its alloys have been widely applied for biomedical application due to their superior physicochemical properties and biocompatibilities for last several decades [1,2]. The fabrication of porous Ti implants by 3D printing technology are catching the interests as the strategy for hard tissue replacement, which could effectively avoid stress shielding, control mechanical properties to targeted implantation area and favor bone ingrowth [3][4][5][6].…”

“…Among them, the phase transition of photo-responsive hydrogels allows fast gelation and injection in situ. The precisely spatiotemporal control of photoresponsive hydrogels could regulate encapsulated drug/protein release and activate cell behaviors [5,17,18]. Chitosan (CS), derived from chitin, is a natural polysaccharide with controllable biodegradability and favorable biocompatibility.…”

“…Chitosan (CS), derived from chitin, is a natural polysaccharide with controllable biodegradability and favorable biocompatibility. Methacrylic anhydride (MA) can interact with amidogen of CS through acylation reaction, resulting in the formation of photo-responsive CSMA under UV radiation [5,16,19].…”

Dual-Targeted Nanoplatform Regulating the Bone Immune Microenvironment Enhances Fracture Healing

“…Research has highlighted the promising use of nanomedicine in osteogenic differentiation, with recent years seeing a surge in the development of nanomedicines for bone repair, in an effort to overcome the weaknesses of currently available strategies. − A number of nano drug delivery systems (DDS) have been designed to effectively locally deliver various drugs into fracture sites, with improved drug concentration and decreased side-effects. − Moreover, targeted drug delivery systems have been designed to enhance the therapeutic effects via surface modifications of specific recognition ligands. − Despite extensive strides, the field is still in its infancy, exemplified by a scarcity of effective nanomedicines for bone repair.…”

Magnetically-Targeted Nanoparticle-Guided M2 Macrophage Polarization For Fracture Healing Promotion

“…Research has highlighted the promising use of nanomedicine in osteogenic differentiation, with recent years seeing a surge in the development of nanomedicines for bone repair, in an effort to overcome the weaknesses of currently available strategies [13][14][15]. A number of nano-drug delivery systems have been designed to effectively locally deliver various drugs into fracture sites, with improved drug concentration and decreased side-effects [16][17][18].…”