Novel Osteogenic Behaviors around Hydrophilic and Radical-Free 4-META/MMA-TBB: Implications of an Osseointegrating Bone Cement.

Poly(methyl methacrylate) (PMMA)-based bone cement, which is widely used to affix orthopedic metallic implants, is considered bio-tolerant but lacks osteoconductivity and is cytotoxic. Implant loosening and toxic complications are significant and recognized problems. Here we devised two strategies to improve PMMA-based bone cement: (1) adding 4-methacryloyloxylethyl trimellitate anhydride (4-META) to MMA monomer to render it hydrophilic; and (2) using tri-n-butyl borane (TBB) as a polymerization initiator instead of benzoyl peroxide (BPO) to reduce free radical production. Rat bone marrow-derived osteoblasts were cultured on PMMA-BPO, common bone cement ingredients, and 4-META/MMA-TBB, newly formulated ingredients. After 24 h of incubation, more cells survived on 4-META/MMA-TBB than on PMMA-BPO. The mineralized area was 20-times greater on 4-META/MMA-TBB than PMMA-BPO at the later culture stage and was accompanied by upregulated osteogenic gene expression. The strength of bone-to-cement integration in rat femurs was 4- and 7-times greater for 4-META/MMA-TBB than PMMA-BPO during early- and late-stage healing, respectively. MicroCT and histomorphometric analyses revealed contact osteogenesis exclusively around 4-META/MMA-TBB, with minimal soft tissue interposition. Hydrophilicity of 4-META/MMA-TBB was sustained for 24 h, particularly under wet conditions, whereas PMMA-BPO was hydrophobic immediately after mixing and was unaffected by time or condition. Electron spin resonance (ESR) spectroscopy revealed that the free radical production for 4-META/MMA-TBB was 1/10 to 1/20 that of PMMA-BPO within 24 h, and the substantial difference persisted for at least 10 days. The compromised ability of PMMA-BPO in recruiting cells was substantially alleviated by adding free radical-scavenging amino-acid N-acetyl cysteine (NAC) into the material, whereas adding NAC did not affect the ability of 4-META/MMA-TBB. These results suggest that 4-META/MMA-TBB shows significantly reduced cytotoxicity compared to PMMA-BPO and induces osteoconductivity due to uniquely created hydrophilic and radical-free interface. Further pre-clinical and clinical validations are warranted.

Tuning of Titanium Microfiber Scaffold with UV-Photofunctionalization for Enhanced Osteoblast Affinity and Function.

Titanium (Ti) is an osteoconductive material that is routinely used as a bulk implant to fix and restore bones and teeth. This study explored the effective use of Ti as a bone engineering scaffold. Challenges to overcome were: (1) difficult liquid/cell infiltration into Ti microfiber scaffolds due to the hydrophobic nature of Ti; and (2) difficult cell attachment on thin and curved Ti microfibers. A recent discovery of UV-photofunctionalization of Ti prompted us to examine its effect on Ti microfiber scaffolds. Scaffolds in disk form were made by weaving grade 4 pure Ti microfibers (125 µm diameter) and half of them were acid-etched to roughen the surface. Some of the scaffolds with original or acid-etched surfaces were further treated by UV light before cell culture. Ti microfiber scaffolds, regardless of the surface type, were hydrophobic and did not allow glycerol/water liquid to infiltrate, whereas, after UV treatment, the scaffolds became hydrophilic and immediately absorbed the liquid. Osteogenic cells from two different origins, derived from the femoral and mandibular bone marrow of rats, were cultured on the scaffolds. The number of cells attached to scaffolds during the early stage of culture within 24 h was 3-10 times greater when the scaffolds were treated with UV. The development of cytoplasmic projections and cytoskeletal, as well as the expression of focal adhesion protein, were exclusively observed on UV-treated scaffolds. Osteoblastic functional phenotypes, such as alkaline phosphatase activity and calcium mineralization, were 2-15 times greater on UV-treated scaffolds, with more pronounced enhancement on acid-etched scaffolds compared to that on the original scaffolds. These effects of UV treatment were associated with a significant reduction in atomic carbon on the Ti microfiber surfaces. In conclusion, UV treatment of Ti microfiber scaffolds tunes their physicochemical properties and effectively enhances the attachment and function of osteoblasts, proposing a new strategy for bone engineering.

Success rate and strength of osseointegration of immediately loaded UV-photofunctionalized implants in a rat model.

STATEMENT OF PROBLEM: Despite its clinical benefits, the immediate loading protocol might have a higher risk of implant failure than the regular protocol. Ultraviolet (UV) photofunctionalization is a novel surface enhancement technique for dental implants. However, the effect of photofunctionalization under loading conditions is unclear. PURPOSE: The purpose of this animal study was to evaluate the effect of photofunctionalization on the biomechanical quality and strength of osseointegration under loaded conditions in a rat model. MATERIAL AND METHODS: Untreated and photofunctionalized, acid-etched titanium implants were placed into rat femurs. The implants were immediately loaded with 0.46 N of constant lateral force. The implant positions were evaluated after 2 weeks of healing. The strength of osseointegration was evaluated by measuring the bone-implant interfacial breakdown point during biomechanical push-in testing. RESULTS: Photofunctionalization induced hydrophilic surfaces on the implants. Osseointegration was successful in 28.6% of untreated implants and 100% of photofunctionalized implants. The strength of osseointegration in successful implants was 2.4 times higher in photofunctionalized implants than in untreated implants. The degree of tilt of untreated implants toward the origin of force was twice that of photofunctionalized implants. CONCLUSIONS: Within the limit of an animal model, photofunctionalization significantly increased the success of osseointegration and prevented implant tilt. Even for the implants that underwent successful osseointegration, the strength of osseointegration was significantly higher for photofunctionalized implants than for untreated implants. Further experiments are warranted to determine the effectiveness of photofunctionalization on immediately loaded dental implants.

Effect of Photofunctionalization on Ti6Al4V Screw Stability Placed in Segmental Bone Defects in Rat Femurs.

PURPOSE: Ultraviolet-mediated photofunctionalization is a new technology to improve bone and titanium integration. We hypothesized that photofunctionalization would enhance the stability of titanium screws used for segmental bone defects. MATERIALS AND METHODS: Disks were prepared of a titanium alloy (Ti6Al4V) for an in vitro study to evaluate the attachment, proliferation, and differentiation of osteoblasts. Commercially available Ti6Al4V screws were used in vivo. Segmental bone defects were created in rat femurs as an immediate loading reconstruction model. The defects were reconstructed with commercially available titanium plates and Ti6Al4V screws, with or without photofunctionalization. The screw survival rates and mechanical stability were evaluated at 2 and 4 weeks, and the bone formation around the screws was analyzed. RESULTS: Osteoblasts showed greater attachment, proliferation, and differentiation on the photofunctionalized Ti6Al4V disks. Photofunctionalized screws had significantly greater survival rates and mechanical stability at 2 and 4 weeks. The bone formation around the photofunctionalized screws was significantly greater than that around the untreated screws at 4 weeks. CONCLUSIONS: The results of the present study have demonstrated the efficacy of photofunctionalization on enhancing the survival and stability of Ti6Al4V screws under a loaded condition in the reconstruction of segmental defects. This was associated with increased bioactivity and bone formation around the photofunctionalized Ti6Al4V material.