Jo-Young Suh

Effects of phosphoric acid treatment of titanium surfaces on surface properties, osteoblast response and removal of torque forces

This study investigated the surface characteristics and biocompatibility of phosphate ion (P)-incorporated titanium (Ti) surfaces hydrothermally treated with various concentrations of phosphoric acid (H(3)PO(4)). The surface characteristics were evaluated by scanning electron microscopy, thin-film X-ray diffractometry, X-ray photoelectron spectroscopy, optical profilometry, contact angle and surface energy measurement and inductively coupled plasma mass spectroscopy (ICP-MS). MC3T3-E1 cell attachment, spreading, proliferation and osteoblastic gene expression on different surfaces were evaluated. The degree of bony integration was biomechanically evaluated by removal torque testing after 4 weeks of healing in rabbit tibiae. The H(3)PO(4) treatment produced micro-rough Ti surfaces with crystalline P-incorporated Ti oxide layers. High concentration H(3)PO(4) treatment (1% and 2%) produced significantly higher hydrophilic surfaces compared with low H(3)PO(4) treatment (0.5%) and untreated surfaces (P<0.01). ICP-MS analysis showed P ions were released from P-incorporated surfaces. Significant increased cell attachment (P<0.05) and notably higher mRNA expressions of Runx2, alkaline phosphatase, osteopontin and osteocalcin were observed in cells grown on P-incorporated surfaces compared with cells on untreated machined surfaces. P-incorporated surfaces showed significantly higher removal torque forces compared with untreated machined implants (P<0.05). Ti surfaces treated with 2% H(3)PO(4) showed increasing tendencies in osteoblastic gene expression and removal torque forces compared with those treated with lower H(3)PO(4) concentrations or untreated surfaces. These results demonstrate that H(3)PO(4) treatment may improve the biocompatibility of Ti implants by enhancing osteoblast attachment, differentiation and biomechanical anchorage.

Gene expression pattern during osteogenic differentiation of human periodontal ligament cells in vitro

Purpose Periodontal ligament (PDL) cell differentiation into osteoblasts is important in bone formation. Bone formation is a complex biological process and involves several tightly regulated gene expression patterns of bone-related proteins. The expression patterns of bone related proteins are regulated in a temporal manner both in vivo and in vitro. The aim of this study was to observe the gene expression profile in PDL cell proliferation, differentiation, and mineralization in vitro. Methods PDL cells were grown until confluence, which were then designated as day 0, and nodule formation was induced by the addition of 50 µg/mL ascorbic acid, 10 mM β-glycerophosphate, and 100 nM dexamethasone to the medium. The dishes were stained with Alizarin Red S on days 1, 7, 14, and 21. Real-time polymerase chain reaction was performed for the detection of various genes on days 0, 1, 7, 14, and 21. Results On day 0 with a confluent monolayer, in the active proliferative stage, c-myc gene expression was observed at its maximal level. On day 7 with a multilayer, alkaline phosphatase, bone morphogenetic protein (BMP)-2, and BMP-4 gene expression had increased and this was followed by maximal expression of osteocalcin on day 14 with the initiation of nodule mineralization. In relationship to apoptosis, c-fos gene expression peaked on day 21 and was characterized by the post-mineralization stage. Here, various genes were regulated in a temporal manner during PDL fibroblast proliferation, extracellular matrix maturation, and mineralization. The gene expression pattern was similar. Conclusions We can speculate that the gene expression pattern occurs during PDL cell proliferation, differentiation, and mineralization. On the basis of these results, it might be possible to understand the various factors that influence PDL cell proliferation, extracellular matrix maturation, and mineralization with regard to gene expression patterns.

Healing of rabbit calvarial bone defects using biphasic calcium phosphate ceramics made of submicron-sized grains with a hierarchical pore structure

OBJECTIVES This study investigated the efficacy of new bone graft substitutes - biphasic calcium phosphates (BCP) made of submicron-sized grains with fully interconnected wide-range micron-scale pores in two different macrodesigns: donut shaped with a 300-400 microm central macropore (n-BCP-1) or rod-shaped (n-BCP-2)--in the healing of rabbit calvarial defects, and compared their bone-healing properties with those of various commercial bone substitutes, which included substitutes with similar BCP composition (MBCP and Osteon), anorganic bovine bone (Bio-Oss), and beta-TCP (Cerasorb). MATERIAL AND METHODS The surface morphology of the bone substitutes was investigated using scanning electron microscopy (SEM). Defects 8 mm in diameter were created in the calvaria of 30 adult male New Zealand White rabbits and were filled with six types of bone substitutes. The percentage of newly formed bone (NB%) was evaluated histomorphometrically 4 and 8 weeks after implantation. RESULTS SEM observation showed submicron-sized grains with fully interconnected micropore structures in the n-BCP-1 and n-BCP-2 groups; these groups also showed considerable new bone formation in inner micropores as well as on the outer surfaces. The n-BCP-1 group exhibited enhanced new bone formation and direct ingrowth of bone tissue with blood vessels into central pores. Histomorphometric analysis showed significantly greater NB% in the n-BCP-1 group when compared with the other groups at 4 and 8 weeks (P<0.05). CONCLUSION A new BCP ceramics made of submicron-sized grains with a hierarchical pore structure was an effective osteoconductive material for the treatment of osseous defects of rabbit calvaria.

Osseointegration of commercial microstructured titanium implants incorporating magnesium: a histomorphometric study in rabbit cancellous bone

OBJECTIVE Recent studies have suggested that magnesium (Mg) ions exert a beneficial effect on implant osseointegration. This study assessed the osseointegration of nanoporous titanium (Ti) surface incorporating the Mg produced by hydrothermal treatment in rabbit cancellous bone to determine whether this surface would further enhance bone healing of moderately rough-surfaced implants in cancellous bone, and compared the result with commercially available micro-arc oxidized Mg-incorporated implants. MATERIAL AND METHODS The Mg-incorporated Ti surfaces (RBM/Mg) were obtained by hydrothermal treatment using an alkaline Mg-containing solution on grit-blasted moderately rough (RBM) implants. Untreated RBM and recently introduced Mg-incorporated microporous Ti implants produced by micro-arc oxidation (M) were used controls in this study. The surface characteristics were evaluated by scanning electron microscopy, X-ray photoelectron spectroscopy and optical profilometry. Twenty-four threaded implants with a length of 10 mm (eight RBM implants, eight RBM/Mg implants and eight M implants) were placed in the femoral condyles of 12 New Zealand White rabbits. Histomorphometric analysis was performed 4 weeks after implantation. RESULTS Hydrothermally treated and untreated grit-blasted implants displayed almost identical surface morphologies and R(a) values at the micron-scale. The RBM/Mg implants exhibited morphological differences compared with the RBM implants at the nano-scale, which displayed nanoporous surface structures. The Mg-incorporated implants (RBM/Mg and M) exhibited more continuous bone apposition and a higher degree of bone-to-implant contact (BIC) than the untreated RBM implants in rabbit cancellous bone. The RBM/Mg implants displayed significantly greater BIC% than untreated RBM implants, both in terms of the all threads region and the total lateral length of implants (P<0.05), but no statistical differences were found between the RBM/Mg and M implants except BIC% values in total lateral length. CONCLUSION These results indicate that a nanoporous Mg-incorporated surface may be effective in enhancing the osseointegration of moderately rough grit-blasted implants by increasing the degree of bone-implant contact in areas of cancellous bone.

Increased new bone formation with a surface magnesium-incorporated deproteinized porcine bone substitute in rabbit calvarial defects†

This study investigated the effects of magnesium ion (Mg) incorporation into the surface of deproteinized porcine cancellous bone in the bone healing of rabbit calvarial defects with the expectation of utilizing the integrin-ligand binding enhancement effect of Mg, and compared its bone healing capacity with that of untreated porcine cancellous bone and deproteinized bovine bone (Bio-Oss). Hydrothermal treatment was performed to produce Mg-incorporated porcine bone using an alkaline Mg-containing solution. The surface morphology and chemical composition of the samples were investigated using scanning electron microscopy, energy-dispersive X-ray spectrometry, and X-ray photoelectron spectroscopy. Defects 7 mm in diameter were created in the calvaria of 14 adult male New Zealand White rabbits and were filled with (1) untreated porcine bone (PB), (2) Bio-Oss, and (3) Mg-containing porcine bone (MG). The percentage of newly formed bone (NB%) was evaluated histomorphometrically at 2 and 4 weeks after implantation. Hydrothermal treatment resulted in a Mg-containing surface in porcine bone covered with nanostructures ~100 nm in size. The MG group supported better new bone formation compared with the other groups. Osteoconductive new bone formation was observed in the central defect area in the MG group at an early healing time-point. Histomorphometric analysis revealed significantly greater NB% in the MG group when compared with the untreated PB and Bio-Oss groups at 4 weeks (p < 0.05). The Mg-incorporated porcine bone with surface nanostructures achieved rapid new bone formation in the osseous defects of rabbit calvaria compared with untreated xenografts of porcine and bovine origin.

Surface characteristics and primary bone marrow stromal cell response of a nanostructured strontium-containing oxide layer produced on a microrough titanium surface †

Strontium (Sr) has been successfully used for the treatment of osteoporotic bone, increasing new bone formation while reducing bone resorption by stimulating proliferation and differentiation of osteoblastic cells and inhibiting osteoclast function. In this study, Sr-incorporated Ti oxide layer was produced on clinically relevant osteoconductive implant surface, that is, a grit-blasted microrough Ti surface, by a simple hydrothermal treatment with the expectation of utilizing the osteoblast response enhancement effect of Sr for the future applications as a more osteoconductive surface of the permanent load-bearing endosseous implants, without altering the original microrough surface features of grit-blasted Ti at the micron-scale. This surface exhibits a hierarchical structure (i.e., a nanoscale surface architecture of the Sr-incorporated Ti oxide layer (SrTiO(3)) imposed on micron-scale rough Ti structure) and Sr ion release into physiological solution. In vitro experiments using primary mouse bone marrow stromal cells (BMSCs) revealed that the hydrothermally produced SrTiO(3) coating promotes both the early and late cell response of BMSCs grown on a microrough Ti surface, with notably enhanced attachment, spreading, focal adhesion, alkaline phosphatase activity, and expression of critical integrins and osteoblastic phenotype genes. These results indicate that a hydrothermally produced SrTiO(3) coating improves the osteoconductivity of the microrough Ti surface by enhancing both the early and late cell response of BMSCs.

IL-17 inhibits osteoblast differentiation and bone regeneration in rat

OBJECTIVE The interleukin-17 (IL-17) family is a group of pro-inflammatory cytokines that are produced by a subset of helper T cells. IL-17 family members are not only involved in the immune response of tissues but also play a role in bone metabolism. Although the role of IL-17 in osteoclast-mediated bone resorption has been extensively studied, its role during osteoblast-mediated bone formation has rarely been investigated. In this study, we examined the effect of IL-17 on osteogenesis in rats both in vitro and in vivo. DESIGN To evaluate osteogenesis in vitro, rat calvarial osteoblast precursor cells were cultured for 14 days in osteogenic medium with or without 50ng/mL IL-17. Osteogenic activity was evaluated by alkaline phosphatase and alizarin red staining. The mRNA expression of alkaline phosphatase, osteocalcin, and osterix was also measured by using real-time PCR. To test whether IL-17 affects bone formation in vivo, bone filling was examined by micro-computed tomography and histological observations at 8 weeks after critical-sized defects were made in rat calvaria. RESULTS The presence of IL-17 significantly reduced alkaline phosphatase and alizarin red staining and the expression of alkaline phosphatase, osteocalcin, and osterix in vitro. IL-17 also significantly inhibited the filling of calvarial defects in vivo. CONCLUSION IL-17 exerted a negative effect on osteogenesis in a rat model. In contrast to the previously reported beneficial effect on osteogenic differentiation of human mesenchymal stem cells, our results suggest a species or cell type-specific role for IL-17 in bone formation.

The relative effect of surface strontium chemistry and super-hydrophilicity on the early osseointegration of moderately rough titanium surface in the rabbit femur.

OBJECTIVE It is unclear whether surface bioactive chemistry or hydrophilicity plays a more dominant role in the osseointegration of micro-structured titanium implants having the same surface topography at the micrometer and submicrometer scales. To understand their comparative effect on enhancing the early osseointegration of micro-rough-surfaced implants, this study compared the bone healing-promoting effect of surface strontium (Sr) chemistry that has been shown in numerous studies to super-hydrophilicity in the early osseointegration of moderately rough-surfaced clinical oral implants (SLA(®) implant) in rabbit cancellous bone. MATERIAL AND METHODS Hydrothermal treatment was performed to incorporate Sr ions into the surface of clinical SLA implants (SLA/Sr implant). The surface characteristics were evaluated by using field emission-scanning electron microscopy, X-ray photoelectron spectroscopy and optical profilometry. Twenty screw implants (10 control and 10 experimental) were placed in the femoral condyles of 10 New Zealand White rabbits. The early osseointegration of the SLA/Sr implant was compared with a chemically modified super-hydrophilic SLA implant (SLActive(®) implant) by histomorphometric and resonance frequency analysis after 2 weeks of implantation. RESULTS The SLA/Sr and SLActive implants exhibited an identical surface topography and average R(a) values at the micron and submicron scales. The SLA/Sr implant displayed a high amount of surface Sr content (15.6 at.%). There was no significant difference in the implant stability quotient (ISQ) values between the two groups. However, histomorphometric analysis revealed a significantly higher bone-to-implant contact percentage in the SLA/Sr implants compared with the SLActive implants in rabbit cancellous bone (P < 0.01). CONCLUSION The results indicate that the surface Sr chemistry surpasses the effect of super-hydrophilicity in promoting the early bone apposition of moderately rough Ti surface in cancellous bone.

Bone-added osteotome technique versus lateral approach for sinus floor elevation: a comparative radiographic study.

Purpose:The aim of this study was to evaluate and compare the radiographic results of bone-added osteotome sinus floor elevation (BAOSFE) and lateral approach sinus floor elevation techniques. Methods:The 43 patients who had undergone implant procedure with either BAOSFE or lateral approach method on their maxillary molar edentulous area were included. Their dental records were confirmative and the radiographicchanges using orthopantomographs were consistently checked up during 2 years after the procedure (immediately after procedure and 6 months, 12 months, and 24 months after implant placement). Results:The radiographic evaluation after 2 years of implantation with sinus elevation showed the significant amount of bone formation (6.75 mm for BAOSFE and 11.36 mm for lateral approach method). Largest amount of grafted height loss occurred during the first 6 months (62.8% of total amount of bone loss), but the resorption was minimal (1.35 mm for BAOSFE and 1.36 mm for lateral approach method) for overall 24 months. Conclusion:Long-term stability of graft height was achieved using both BAOSFE and lateral approach sinus floor elevation. Overall, graft height decreased gradually during 2 years after procedures, but the changes were minimal.

Improved pre‐osteoblast response and mechanical compatibility of ultrafine‐grained Ti–13Nb–13Zr alloy

OBJECTIVE Metallic implantation materials having high yield strength, low elastic modulus, and non-cytotoxic alloying elements would be advantageous for the long-term stability of implants. This study assessed the surface and mechanical properties, and also in vitro osteoconductivity of ultrafine-grained (UFG) Ti-13Nb-13Zr alloy produced by dynamic globularization without any severe deformation for future biomedical applications as an endosseous implant material. MATERIAL AND METHODS The surface characteristics and mechanical properties were investigated by orientation image microscopy, contact angle measurements, optical profilometry, and uniaxial tension tests. Mouse calvaria-derived pre-osteoblastic cell (MC3T3-E1) attachment, spreading, viability, alkaline phosphatase (ALP) activity, and quantitative analysis of osteoblastic gene expression on UFG Ti-13Nb-13Zr alloy were compared with coarse-grained (CG) Ti-13Nb-13Zr and CG Ti-6Al-4V alloys. RESULTS Dynamic globularized Ti-13Nb-13Zr alloy has an ultrafine grain size (0.3 μm) and an excellent combination of yield strength and elastic modulus compared with CG alloys, which displayed significantly lower water contact angles compared with CG alloys (P<0.05). The UFG and CG Ti-13Nb-13Zr alloys displayed significantly increased cellular attachment compared with CG Ti-6Al-4V alloy (P<0.05). The UFG Ti-13Nb-13Zr supported better cell spreading and more numerous focal adhesions. ALP activity (P<0.05) and mRNA expressions of the osteoblast transcription factor genes (osterix, Runx2) and marker gene for osteoblast differentiation (osteocalcin) were markedly increased in cells grown on the UFG substrate compared with CG substrates at early incubation timepoints. CONCLUSION Enhanced pre-osteoblast response to UFG Ti-13Nb-13Zr substrate is attributable to the non-cytotoxic alloying elements and the submicron scale grain size contributes to the superior surface hydrophilicity and abundant grain boundaries favorable for cell behavior. These findings indicate that dynamic globularized UFG Ti-13Nb-13Zr alloy is promising for load-bearing endosseous implant material because of excellent mechanical and biological compatibilites.