Three commercially available artificial bone substitutes with different compositions, hydroxyapatite (HAp; Neobone^®), carbonate apatite (CO₃Ap; Cytrans^®), and β-tricalcium phosphate (β-TCP; Cerasorb^®), were compared with respect to their physical properties and tissue response to bone, using hybrid dogs. Both Neobone^® (HAp) and Cerasorb^® (β-TCP) were porous, whereas Cytrans^® (CO₃Ap) was dense. Crystallite size and specific surface area (SSA) of Neobone^® (HAp), Cytrans^® (CO₃Ap), and Cerasorb^® (β-TCP) were 75.4 ± 0.9 nm, 30.8 ± 0.8 nm, and 78.5 ± 7.5 nm, and 0.06 m²/g, 18.2 m²/g, and 1.0 m²/g, respectively. These values are consistent with the fact that both Neobone^® (HAp) and Cerasorb^® (β-TCP) are sintered ceramics, whereas Cytrans^® (CO₃Ap) is fabricated in aqueous solution. Dissolution in pH 5.3 solution mimicking Howship’s lacunae was fastest in CO₃Ap (Cytrans^®), whereas dissolution in pH 7.3 physiological solution was fastest in β-TCP (Cerasorb^®). These results indicated that CO₃Ap is stable under physiological conditions and is resorbed at Howship’s lacunae. Histological evaluation using hybrid dog mandible bone defect model revealed that new bone was formed from existing bone to the center of the bone defect when reconstructed with CO₃Ap (Cytrans^®) at week 4. The amount of bone increased at week 12, and resorption of the CO₃Ap (Cytrans^®) was confirmed. β-TCP (Cerasorb^®) showed limited bone formation at week 4. However, a larger amount of bone was observed at week 12. Among these three bone substitutes, CO₃Ap (Cytrans^®) demonstrated the highest level of new bone formation. These results indicate the possibility that bone substitutes with compositions similar to that of bone may have properties similar to those of bone.

This study was aimed to investigate the osseointegration ability of poly(ether ether ketone) (PEEK) implants with modified surface roughness and/or surface chemistry. The roughened surface was prepared by a sandblast method, and the phosphate groups on the substrates were modified by a two-step chemical reaction. The in vitro osteogenic activity of rat mesenchymal stem cells (MSCs) on the developed substrates was assessed by measuring cell proliferation, alkaline phosphatase activity, osteocalcin expression, and bone-like nodule formation. Surface roughening alone did not improve MSC responses. However, phosphorylation of smooth substrates increased cell responses, which were further elevated in combination with surface roughening. Moreover, in a rabbit tibia implantation model, this combined surface modification significantly enhanced the bone-to-implant contact ratio and corresponding bone-to-implant bonding strength at 4 and 8 weeks post-implantation, whereas modification of surface roughness or surface chemistry alone did not. This study demonstrates that combination of surface roughness and chemical modification on PEEK significantly promotes cell responses and osseointegration ability in a synergistic manner both in vitro and in vivo. Therefore, this is a simple and promising technique for improving the poor osseointegration ability of PEEK-based orthopedic/dental implants.

Biomedical applications of poly(ether ether ketone) (PEEK) are hindered by its inherent bioinertness and lack of osseointegration capability. In the present study, to enhance osteogenic activity and, hence, the osseointegration capability of PEEK, we proposed a strategy of combined phosphate and calcium surface-functionalization, in which ozone-gas treatment and wet chemistry were used for introduction of hydroxyl groups and modification of phosphate and/or calcium, respectively. Surface functionalization significantly elevated the surface hydrophilicity without changing the surface roughness or topography. The cell study demonstrated that immobilization of phosphate or calcium increased the osteogenesis of rat mesenchymal stem cells compared with bare PEEK, including cell proliferation, alkaline phosphatase activity, and bone-like nodule formation. Interestingly, further enhancement was observed for samples co-immobilized with phosphate and calcium. Furthermore, in the animal study, phosphate and calcium co-functionalized PEEK demonstrated significantly enhanced osseointegration, as revealed by a greater direct bone-to-implant contact ratio and bond strength between the bone and implant than unfunctionalized and phosphate-functionalized PEEK, which paves the way for the orthopedic and dental application of PEEK.

Fracture-related infections require both treatments for bacteria removal and bone reconstruction. The use of combined broad-spectrum antibacterial silver compounds and artificial bone with high osteogenic activity is considered to be an effective strategy for achieving these treatments in one surgery. However, silver compounds are toxic for living tissues even at low concentrations. Herein, we investigated the no-observed-effect level (NOEL) of silver phosphate (Ag₃PO₄) in a bone substitute composed of carbonate apatite (CO₃Ap), a bone mineral, using in vitro and in vivo experiments. In vitro experiments demonstrated that the CO₃Ap artificial bone containing ≥0.1 wt % Ag₃PO₄ exerted antibacterial effects against Staphylococcus epidermidis, while those containing ≤0.3 wt % Ag₃PO₄ did not affect cellular adhesion, proliferation, differentiation, and calcification of osteoblast-like MC3T3-E1 cells. In vivo experiments demonstrated that the CO₃Ap artificial bone containing ≤0.3 wt % Ag₃PO₄ replaced a new bone to the same levels as those without Ag₃PO₄ 4 weeks after implantation into the bone defect of the rabbit femur condyle. However, the CO₃Ap artificial bone containing 0.3 wt % Ag₃PO₄ caused an inflammatory reaction, whereas those containing ≤0.1 wt % Ag₃PO₄ did not. Thus, both bone regeneration and infection control without any adverse effects were achieved using the CO₃Ap artificial bone containing 0.1 wt % Ag₃PO₄, indicating that the NOEL of Ag₃PO₄ was 0.1 wt %. Our results provide an effective strategy for the treatments of fracture-related infections.