Silk Fibroin/Hydroxyapatite Coating Improved Osseointegration of Porous Titanium Implants under Diabetic Conditions via Activation of the PI3K/Akt Signaling Pathway

The application of three-dimensional printed porous titanium implants (TIs) is compromised in patients suffering from diabetes mellitus (DM), which disturbs the normal process of implant osseointegration, resulting in fixation failure. It was possibly because of reactive oxygen species (ROS) overproduction at the bone-implant interface. A silk fibroin-based hydroxyapatite (SF/HA) hybrid material emerged as a novel biological material for accelerating new bone formation. We proposed that the SF/HA hybrid coated titanium implant (SHT) could mitigate DM-mediated impaired osseointegration, which had never been reported previously. To test this assumption and further elucidate the mechanisms, primary rabbit osteoblasts were seeded on TIs or SHTs and cultured with normal serum, diabetic serum (DS), DS + N-acetyl-L-cysteine (NAC) (a potent ROS inhibitor), and DS + LY294002 (a specific PI3K/Akt inhibitor) for osteoblast behavior examinations. An animal study was performed on diabetic rabbits implanted with the two kinds of implants for osseointegration tests. DM-mediated ROS overproduction caused osteoblastic biological dysfunctions and apoptotic injury, associated with suppression of PI3K/Akt signaling in osteoblasts cultured on a TI substrate. Of note, the SHT substrate significantly suppressed ROS overproduction under diabetic conditions, improved osteoblast functional recovery including ameliorative osteoblast adhesion and morphology, improved cellular proliferation and differentiation, and abrogated apoptosis, which exhibited the same effect as NAC administration on the TI. The in vitro results were further corroborated in vivo by enhanced osteogenesis and osseointegration of SHTs in diabetic rabbits. Moreover, the aforesaid promotive effects afforded by the SF/HA coating were totally abolished with administration of LY294002 for blocking PI3K/Akt signaling. The above results collectively demonstrated that the SF/HA hybrid coating significantly ameliorated DM-mediated impaired osseointegration of the TI via reactivation of the ROS-mediated PI3K/Akt signaling pathway. The hybrid coating elicited a novel surface biofunctionalization strategy to attain favorable clinical performance of TI in diabetics.