Suk-Won Lee

Factors influencing osteoblast maturation on microgrooved titanium substrata

In this study, we demonstrate surfaces with various dimensions of microgrooves fabricated by photolithography and subsequent acid etching that enhance various characteristics of titanium. Microgrooves with truncated V-shape in cross-section from 15 to 90 microm widths enabled us to report their exclusive effects on altering the surface chemistry and on enhancing the surface hydrophilicity, serum protein adsorption and osteoblast maturation on titanium substrata in a microgroove dimension-dependent manner. Further, acid etching and measurement direction separately affected the surface hydrophilicity results. By multiple correlation and regression analyses, surface chemistry, surface hydrophilicity and serum protein adsorption were determined to be the significant influential factors on osteoblast maturation. Within the limitations of this study, we conclude that combined submicron- and microtopography with relevant micro-dimension and structure enhance various characteristics of titanium, including surface hydrophilicity, which act as the essential factors influencing the osteoblast maturation on microgrooved titanium substrata.

Catechol-functionalized adhesive polymer nanoparticles for controlled local release of bone morphogenetic protein-2 from titanium surface.

We report on a novel surface functionalization approach to equip the titanium (Ti) surfaces with osteogenic properties. A key feature of the approach is the treatment of the Ti surfaces with Ti-adhesive nanoparticles that can stably load and controllably release bone morphogenetic protein-2 (BMP-2). Ti-adhesive nanoparticles were prepared by self-assembly of a catechol-functionalized poly(amino acid) diblock copolymer, catechol-poly(L-aspartic acid)-b-poly(L-phenylalanine) (Cat-PAsp-PPhe). The nanoparticles consist of Ti-adhesive peripheral catechol groups, anionic PAsp shells, and PPhe inner cores. Field-emission scanning electron microscopy (Fe-SEM) images showed that the Ti-adhesive nanoparticles could be uniformly immobilized on Ti surfaces. X-ray photoelectron spectroscopy (XPS) confirmed the successful anchoring of nanoparticles onto Ti surfaces. After surface immobilization of the nanoparticles, the static water contact angle of the Ti substrate decreased from 75.3° to 50.0° or 36.4°, depending on the surface nanoparticle. Fluorescence microscopic analysis showed that BMP-2 could be effectively incorporated onto the Ti surface with adhesive nanoparticles. BMP-2 was controllably released for up to 40 days. The Ti substrate functionalized with BMP-2-incorporated nanoparticles significantly promoted attachment, proliferation, spreading, and alkaline phosphatase (ALP) activity of human adipose-derived stem cell (hADSC). The catechol-functionalized adhesive nanoparticles may be applied to various medical devices to create surfaces for improved performance.

Compressive stress induced the up-regulation of M-CSF, RANKL, TNF-α expression and the down-regulation of OPG expression in PDL cells via the integrin-FAK pathway

OBJECTIVES This study was performed to elucidate the involvement of integrin-FAK (focal adhesion kinase) pathway in compressive stress-induced mRNA expression of macrophage colony stimulating factor (M-CSF), tumour necrosis factor (TNF)-α, receptor activator of nuclear factor κB (RANKL) and osteoprotegerin (OPG) and to further confirm the role of the integrin-FAK complex as a mechanoreceptor in PDL cells. DESIGN Periodontal ligament (PDL) cells were obtained from patients having healthy first premolars extracted for orthodontic purposes. Cultured PDL cells were divided into three groups: the control group in which compressive stress was administered; the negative control group in which mechanical stress was administered after transfection of negative control siRNA; and FAK knockdown group in which mechanical stress was administered after FAK siRNA treatment. Compressive stress (2g/cm(2)) was for various time durations (0.5, 2, 6, 24, 48h). Total RNA was collected after the experiment and real-time PCR analysis was performed to determine the mRNA expression levels of M-CSF, TNF-α, RANKL and OPG. Also the supernatant was analysed with ELISA to detect the corresponding cytokine concentrations. RESULTS The cells of the control group and the negative control group expressed higher mRNA levels of M-CSF, TNF-α, and RANKL but a lower mRNA level of OPG compared to those of baseline. FAK knockdown cells showed lower mRNA expression levels of M-CSF, TNF-α, and RANKL but a higher mRNA expression level of OPG than that in the control. The OPG mRNA expression level in FAK knockdown cells was even higher than that of baseline. ELISA results showed similar pattern of cytokine concentration changes. CONCLUSIONS Results of this study indicate that the integrin-FAK pathway regulates compressive stress-induced expression of M-CSF, TNF-α, RANKL and OPG and suggests that the integrin-FAK complex acts as a mechanoreceptor in PDL cells.

Differences in implant stability associated with various methods of preparation of the implant bed: An in vitro study

STATEMENT OF PROBLEM It is difficult to achieve the primary stability necessary for immediate loading in the posterior maxilla because of thin cortical bone, low density trabecular bone, and inadequate bone height due to the presence of the maxillary sinus. PURPOSE The purpose of this study was to examine the primary stability of dental implants placed by using different methods of preparation for in vitro monocortical and bicortical models of the posterior maxilla. MATERIAL AND METHODS Sixty screw-shaped implants (4.0 × 10 mm) were inserted into solid rigid polyurethane blocks. The implants were divided into 6 groups (n=10) to test 2 variables: 1) location (monocortical or bicortical block) and 2) preparation method (standard preparation, underpreparation, or the osteotome technique). The insertion and removal torques were measured and resonance frequency analysis (RFA) was performed to determine the primary stability of each implant. Insertion and removal torque data were analyzed by 2-way ANOVA, followed by the post hoc Tukey HSD multiple comparison test. RFA data were analyzed by 2-way and 1-way ANOVAs and the Tukey HSD multiple comparison test (α=.05). The Pearson correlation analysis was also performed to examine correlations among the values. RESULTS The preparation method had a significant effect on insertion torque, RFA value, and removal torque; however location had a significant effect only on the removal torque (P<.001). There was a significant interaction between location and preparation method for RFA values (P=.045) and a significant difference in standard preparation method according to the location (P=.039); however, there was no significant difference in underpreparation (P=1.00) and osteotome technique (P=1.00). Statistically significant correlations were found between insertion torque and RFA values (r=0.529, P< .001), insertion torque and removal torque values (r=0.517, P< .001), and removal torque and RFA values (r=0.481, P<.001). CONCLUSIONS Underpreparation and bicortical fixation significantly increased implant stability and the osteotome technique decreased implant stability in synthetic bone models that mimicked the posterior maxillary region. The primary stability values had statistically significant correlations to each other.

Surface microgrooves and acid etching on titanium substrata alter various cell behaviors of cultured human gingival fibroblasts.

OBJECTIVE This study aimed to investigate the influence of surface microgrooves and acid etching on titanium substrata on cell proliferation and gene expression of cultured human gingival fibroblasts. MATERIAL AND METHODS Ti substrata with various dimensions of microgrooves and further acid etching comprised four experimental groups (E15/3.5, E30/5, E60/10, and E90/15), whereas smooth and acid etched Ti discs were both used as control (NE0 and E0). The chemical composition of the control and experimental cp Ti substrata was analyzed by X-ray photoelectron spectroscopy (XPS). Cell proliferation, gene expression, and protein expression of cultured human gingival fibroblasts were analyzed between all groups using bromodeoxyuridine (BrdU) assay, reverse-transcriptase polymerase chain reaction (RT-PCR), and Western blotting, respectively. RESULTS The XPS results showed that the Ti substrata used in this study showed no significant differences in the expression of surface chemical composition. BrdU assay showed that cell proliferation was significantly increased in E60/10 compared with that in any other group. In RT-PCR, E60/10 was noted to increase the expression of various genes involved in cell-matrix adhesion and adhesion-dependent cell cycle progression. In Western blotting, increased expression of fibronectin and Rho A was noted in E60/10 compared with that in NE0 or E15/3.5. CONCLUSION This study indicates that surface microgrooves 60 mum in width and 10 mum in depth, and further acid etching on Ti substrata trigger the proliferation and alter the expression of both genes and proteins in cultured human gingival fibroblasts.

Influence of etched microgrooves of uniform dimension on in vitro responses of human gingival fibroblasts

OBJECTIVE The purpose of this study was to investigate the influence of titanium (Ti) substrata with etched surface microgrooves on in vitro responses of human gingival fibroblasts. MATERIAL AND METHODS Commercially pure Ti discs with surface microgrooves 15, 30, and 60 microm in width were fabricated using photolithography. Microgrooves 15 microm in widths were designed to be 3.5 microm in depth, whereas microgrooves 30 and 60 microm in width varied between 5 and 10 microm in depth. The entire surface of the microgrooved Ti substrata was further acid etched and used as the five experimental groups in this study (E15/3.5, E30/5, E30/10, E60/5, and E60/10), whereas the smooth and acid-etched Ti discs were both used as the control (NE0 and E0). Human gingival fibroblasts were cultured on all groups of substrata on successive timelines. Fibroblast adhesion and morphology was analyzed using confocal laser scanning microscopy (CLSM). Cell adhesion and proliferation were analyzed and compared using crystal violet staining and sulforhodamine B protein staining assays, respectively. RESULTS In CLSM, the cells in E30/10 and E60/10 were observed to be able to readily descend into and form focal adhesions inside the microgrooves, whereas the cells in E15/3.5 were mostly found on the ridge tops. Cell adhesion was significantly increased in E60/10 compared with that in NE0 or E0 at 4-h incubation. Cell proliferation was significantly increased in E60/10 and E15/3.5 compared with NE0 or E0 after 72 and 96 h of culture. CONCLUSION This study indicates that Ti substrata with etched microgrooves 60 microm in width and 10 microm in depth significantly enhance human gingival fibroblast adhesion and proliferation.

Effects of moderate intensity static magnetic fields on human bone marrow-derived mesenchymal stem cells.

This study aimed to explore effects of static magnetic fields (SMFs) of moderate intensity (3-50 mT) as biophysical stimulators of proliferation and osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells (MSCs). MSCs were exposed to SMFs of three intensities: 3, 15, and 50 mT. Proliferation was assessed by cell counting and bromodeoxyuridine incorporation, and differentiation by measuring alkaline phosphatase (ALP) activity, calcium content, mineralized nodule formation, and transcripts of osteogenic markers. Moderate intensity SMFs increased cell proliferation, ALP activity, calcium release, and mineralized nodule formation in a dose- and time-dependent manner, which peaked at 15 mT. In the same manner, they upregulated expression of osteogenic marker genes such as ALP, bone sialoprotein 2 (BSP2), collagen1a1 (COL1a1), osteocalcin (OCN), osteonectin (ON), osteopontin (OPN), osterix (OSX), and runt-related transcription factor 2 (RUNX2) with peak at 15 mT after 14 or 21 days of exposure. Results demonstrate that moderate intensity SMFs promote proliferation and osteoblastic differentiation of MSCs. This effect could help to improve MSC responses during osseointegration between a dental implant and surrounding bone.

The significance of differential expression of genes and proteins in human primary cells caused by microgrooved biomaterial substrata

We demonstrate that etched microgrooves, with truncated V-shape in cross-section and subsequent acid etching, on titanium substrata alter the expression of various genes and proteins in human primary cells. Etched microgrooves with 30 or 60 μm width and 10 μm depth promoted human gingival fibroblast proliferation and significantly enhanced the osteoblast differentiation of human bone marrow-derived mesenchymal stem cells and human periodontal ligament cells by inducing differential expression of various genes involved in cell adhesion, migration, proliferation, mitosis, cytoskeletal reorganization, translation initiation, vesicular trafficking, proton transportation, transforming growth factor-β signaling, mitogen-activated protein kinase signaling, simvastatins anabolic effect on bone, inhibitory guanine nucleotide binding protein (G protein)s action, sumoylation pathway, survival/apoptosis, mitochondrial distribution, type I collagen production, osteoblast differentiation, and bone remodeling that were verified by the differential display PCR and quantitative real-time PCR. The most influential genes on the enhancement of fibroblast proliferation or osteoblast differentiation were determined by multiple regression analysis, and the expression of relevant proteins was confirmed by western blotting and protein quantitation.

Texture direction of combined microgrooves and submicroscale topographies of titanium substrata influence adhesion, proliferation, and differentiation in human primary cells

OBJECTIVE This study aimed to identify the optimal micro- and submicroscale topographies of titanium (Ti) substrata that would most significantly influence adhesion, proliferation, and other activities of these cells. DESIGN Truncated V-shaped microgrooves in 60 μm-wide and 10 μm-deep cross-sections with 0°, 30°, 60°, or 90° angles between the microgrooves and ridge-top submicroscale texture were created on the Ti substrata (designated NE60/10-0°, NE60/10-30°, NE60/10-60° and NE60/10-90°, respectively). Ground titanium with submicroscale texture but with no microgrooves was used as the control substratum, NE0. Scanning electron microscopic observation and the assays determining the cell adhesion, cell proliferation and osteoblast differentiation were performed. RESULTS Cells more actively migrated into the microgrooves on NE60/10-30° than into the microgrooves on any other substrata tested, suggesting that the cells utilise the increased surface area of the substrata at the microscale level. NE60/10-0° and NE60/10-30° substrata generally enhanced adhesion, proliferation, alkaline phosphatase activity, and osteoblast differentiation of human primary cells when compared to other Ti substrata, and significant correlations were observed between these cellular activities. CONCLUSIONS Here, we show that the contact guidance of human primary cells grown on Ti substrata can be controlled more by specific submicroscale textures on ridge tops than by the dimensions of surface microgrooves only. Also, the degree of angles created between the submicroscale textures and microgrooves on Ti substrata significantly affect cell adhesion, proliferation and differentiation in human primary cells.

The Effects of a Static Magnetic Field on Bone Formation Around a Sandblasted, Large-Grit, Acid-Etched–Treated Titanium Implant

The purpose of this study was to compare the bone formation around commercial sandblasted, large-grit, acid-etched (SLA)–treated titanium implants with or without a neodymium magnet in a rabbit tibia through histomorphometric analysis. Commercial SLA-treated implants with or without neodymium magnets were placed in 10 rabbits. After incising the flat part of the rabbits tibia and installation of the specimens of titanium implants, the nonmagnet group was stitched without magnet insertion. On the other hand, the magnet group was inserted with neodymium magnet, fixed with pattern resin, and stitched. At 3 and 6 weeks after surgery, the animals were sacrificed, and the specimens were obtained. Undecalcified specimens were prepared for histomorphometric analysis of the bone-to-implant contact ratio (BIC) and bone volume (BV). The histomorphometric findings of the cortical bone showed that the mean BVs of the magnet group (3 weeks, 75.99%; 6 weeks, 82.94%) were higher than those of the nonmagnet group (3 week...