Cellular Response of Anodized Titanium Surface by Poly(Lactide-co-Glycolide)/Bone Morphogenic Protein-2

BACKGROUND: The purpose of this study is to examine physical characteristics of and initial biological properties to anodized titanium treated with poly(D,L-lactide-co-glycolide) (PLG) mixed with recombinant human bone morphogenic protein-2 (rhBMP-2). METHODS: Titanium specimens were prepared in groups of four as follows: group NC was anodized under 300 V as control; group PC was anodized then dropped and dried with solution 0.02 ml PLG; group D was anodized then dropped and dried with solution 0.02 ml PLG/rhBMP-2 (3.75 µg per disc); and group E was anodized then coated with 0.02 ml PLG/rhBMP-2 (3.75 µg per disc) by electrospray. Human osteoblastic-like sarcoma cells were cultured. Cell proliferation and alkaline phosphatase (ALP) activity test were carried out. Runx-2 gene was investigated by the reverse transcription-polymerase chain reaction. Immunofluorescence outcome of osteogenic proteins was observed. RESULTS: After 3 days, there were significantly higher proliferations compared rhBMP-2 loaded titanium discs with rhBMP-2 unloaded discs. The ALPase activity on rhBMP-2 loaded titanium discs was significantly higher than in rhBMP-2 unloaded discs. The expression level of Runx2 mRNA presented the highest on the PLG/rhBMP-2-coated surface. CONCLUSION: PLG polymers mixed with rhBMP-2 might improve proliferation, differentiation and osteogenic protein formation of cells on the anodized titanium.

Effect of immobilization of the recombinant human bone morphogenetic protein 2 (rhBMP-2) on anodized implants coated with heparin for improving alveolar ridge augmentation in beagle dogs: Radiographic observations / 대한치과보철학회지

PURPOSE: The aim of this study was to evaluate the effect of immobilization of the recombinant human bone morphogenetic protein 2 (rhBMP-2) on anodized titaum implants coated with heparin to enhance the vertical alveolar ridge augmentation in the supraalveolar peri-implant defect region. MATERIALS AND METHODS: 18 pure titanium implants (7.0 mm in length, 3.5 mm in diameter) were manufactured for this study. All implants were anodized and designed insertion reference line marked with laser at the apical 2.5 mm from the fixture platform. Implantation of 6 noncoated anodized implants (Control group), 6 anodized implants physically adsorbed with rhBMP-2 by dip and dry method (BMP group) and 6 anodized implants chemically immobilized 3,4-dihydroxyphenylalanine (DOPA)-heparin/rhBMP-2 (Hep-BMP group) was performed in the both mandibular of three male adult beagle dogs using split-mouth design. Radiologic examinations were performed immediately after implant placement and 4 and 8 weeks after implant placement. The amount of mesio-distal bone augmentation was evaluated by measuring the vertical distance from the platform to the marginal bone. Statistical analysis was performed using one-way analysis of variance (SPSS version 18.0) and multiple comparison analysis of The Kruskal-Wallis test and the Mann-Whitney U test. Statistical significance was established at the 5%significant level. RESULTS: At the 4 weeks vertical alveolar ridge augmentation of Control group, BMP group and Hep-BMP group is 0.09 +/- 0.22 mm, 1.02 +/- 0.72 mm, and 1.29 +/- 0.51 mm, At the 8 weeks 0.11 +/- 1.26 mm, 1.11 +/- 0.58 mm, 1.59 +/- 0.79 mm according to radiographic observations. The two experimental groups showed a significantly increasing in vertical bone height compared with the control group (P.05). CONCLUSION: The rhBMP-2 coated implants were enhanced the vertical bone growth in the supraalveolar peri-implant defect area. However, there is no significant difference between chemically and physically coating method.

Osteogenic Gene Expression on Anodizing Titanium Surface

Analysis of attachment, proliferation and differentiation response of human mesenchymal stem cell to various implant surfaces coated with rhBMP-2 / 대한치과보철학회지

PURPOSE: In this paper we tried to evaluate the most appropriate surface for rhBMP-2 coating among 4 rough titanium surfaces. MATERIALS AND METHODS: We used machined surface as a control group and anodized, RBM and SLA surfaces as test groups. We coated rhBMP-2 on the 4 surfaces and with uncoated surfaces for each case, we cultured human mesenchymal stem cells on all 8 surfaces. 24 hours after we measured the stem cell'attachment with SEM, and on 3rd, 7th, and 14th days, we checked the cell proliferation and differentiation by using MTT and ALP activity assay. And on the 7th day after the culture, we performed RT-PCR assay to determine whether the expression levels of Type I collagen, osteocalcin, osteopontin were changed. RESULTS: We observed with SEM that 4 rhBMP-2 coated surfaces exhibited wider and tighter cell attachment and more cell process spreading than uncoated surfaces. The anodized rhBMP-2 surface caused robustest effects. In MTT assay we could not find any meaningful difference. In ALP assay there was a significant increase (P<.05) in the ALP activity of anodized rhBMP-2 coated surface compared with that of the control (3rd and 14th days) and with that of the RBM rhBMP-2 coated surface (14th day). In RT-PCR assay there was increased expressions in the anodized rhBMP-2 coated surface for osteocalcin, and osteopontin. CONCLUSION: We found that the anodized rhBMP-2 coated surface were most prominent stem cell attachment and differentiation in compared to control and Machined rhBMP-2 coated, RBM rhBMP-2 coated surface.

Effect of RGD peptide coating of implant titanium surface on human mesenchymal stem cell response / 대한치과보철학회지

PURPOSE: The aim of this in vitro study was to estimate surface characteristic after peptide coating and investigate biological response of human mesenchymal stem cell to anodized titanium discs coated with RGD peptide by physical adhesion and chemical fixation. MATERIALS AND METHODS: Fluorescence isothiocyanate (FITC) modified RGD-peptide was coated on the anodized titanium discs (diameter 12 mm, height 3 mm) using two methods. One was physical adhesion method and the other was chemical fixation method. Physical adhesion was performed by dip and dry procedure, chemical fixation was performed by covalent bond via silanization. In this study, human mesenchymal stem cell was used for experiments. The experiments consisted of surface characteristic evaluation after peptide coating, analysis about cell adhesion, proliferation, differentiation, and mineralization. Obtained data are statistically treated using Kruskal-Wallis test and Bonferroni test was performed as post hoc test (P=.05). RESULTS: The evaluation of FE-SEM images revealed no diffenrence at micro-surfaces between each groups. Total coating dose was higher at physical adhesion experimental group than at chemical fixation experimental group. In cell adhesion and proliferation, RGD peptide coating did not show a statistical significance compared with control group (P>.05). In cell differentiation and mineralization, physical adhesion method displayed significantly increased levels compared with control group and chemical fixation method (P<.05). CONCLUSION: RGD peptide coating seems to enhance osseointegration by effects on the response of human mesenchymal stem cell. Especially physical adhesion method showed more effective than chemical fixation method on response of human mesenchymal stem cell.

Effect of Various Inodizing Characteristics on Bone Integration of Titanium Implant Surface Design

A study on the biological characteristics of modified titanium surface / 대한치주과학회지

PURPOSE: The purpose of this research is to study about initial adhesion, proliferation and activation of osteoblast to titanium surface treated with machined, hydroxyapatite coating, resorbable blast material blasting and anodizing method. MATERIAL AND METHODS: After treating the titanium surface of each block with machined, impurities were removed and sterilized. The number of cells attached from cultured osteoblast of respective experimental groups were measured at 1, 4, 7, and 14day and alkaline phosphatase, calcium, and inorganic phosphate concentration of cultured solution was measured. RESULT: Anodizing group showed the highest rate of cell attachment and proliferation activity. RBM treated group showed the highest increasing on their alkaline phosphatase activity, on the calcium apposition, on inorganic phosphate apposition of 1 and 4 days in cultured osteoblast to compare with other groups. CONCLUSION: On the basis of these findings, we conclude that surface modification of titanium was profoundly effected on the attachment, proliferation and activation of osteoblast in initial stage osseointegration.

Cellular responses on anodized titanium discs coated with 1 alpha,25-dihydroxyvitamin D3 incorporated Poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles / 대한치과보철학회지

STATEMENT OF PROBLEM: A biochemical approach for surface modification has offered an alternative for physicochemical and morphological methods to obtain desirable bone-implant interfaces. PURPOSE: The purpose of the present study was to investigate cell responses to poly (D,L-lactide-co-glycolide) (PLGA)/1alpha, 25-(OH)2D3 coating with reference to cellular proliferation and differentiation in vitro. MATERIAL AND METHODS: 96 titanium discs were fabricated and divided into four groups. Group 1 was anodized under 300 V as control. Group 2, 3 and 4 were anodized then coated with 3 ml PLGA/1alpha, 25-(OH)2D3 solutions. Amount of the solutions were 2 ul, 20 ul and 200ul respectively. The osteoblast-like Human Osteogenic Sarcoma (HOS) cells were seeded and cultured for 1, 3 and 7 days. MTSbased cell proliferation assay and ALPase activity test were carried out. RESULTS: PLGA nanoparticles were observed as fine, smooth and round and HOS cells attached to the anodized surfaces through strand-like and sheet-like filopodia. After 3 days of culture, the dendritic filopodia were exaggerated and sheet-like cytoplasmic projections covered the coated titanium surfaces. After 3 days of culture, all of the groups showed increased cellular proliferation and the lowest proliferation rate was measured on group 2. Higher amount of incorporated 1 alpha, 25-(OH)2D3 (Group 3 and 4) improved cellular proliferation but the differences were not significant statistically (P > .05). But they increased the rate of ALP activities than the control group at day 3 (P < .05). CONCLUSION: Biodegradable PLGA nanoparticles incorporated with vitamin D metabolite positively affected proliferation and differentiation of cells on the anodized titanium surface.

DNA microarray analysis of gene expression of MC3T3-E1 osteoblast cell cultured on anodized- or machined titanium surface / 대한치주과학회지

PURPOSE: The aim of this study was to evaluate adhesion and gene expression of the MC3T3-E1 cells cultured on machined titanium surface (MS) and anodized titanium surface (AS) using MTT test, Scanning electron micrograph and cDNA microarray. MATERIALS AND METHODS: The MTT test assay was used for examining the proliferation of MC3T3-E1 cells, osteoblast like cells from Rat calvaria, on MS and AS for 24 hours and 48 hours. Cell cultures were incubated for 24 hours to evaluate the influence of the substrate geometry on both surfaces using a Scanning Electron Micrograph (SEM). The cDNA microarray Agilent Rat 22K chip was used to monitor expressions of genes. RESULTS: After 24 hours of adhesion, the cell density on AS was higher than MS (p0.05). AS had the irregular, rough and porous surface texture. After 48 hours incubation of the MC3T3-E1 cells, connective tissue growth factor (CTGF) was up-regulated on AS than MS (more than 2 fold) and the insulin-like growth factor 1 receptor was down-regulated (more than 2 fold) on AS than MS. CONCLUSION: Microarray assay at 48 hours after culturing the cells on both surfaces revealed that osteoinductive molecules appeared more prominent on AS, whereas the adhesion molecules on the biomaterial were higher on MS than AS, which will affect the phenotype of the plated cells depending on the surface morphology.

Effects of different oxidized surfaces of implant on osseointegration; resonance frequency and histomorphometric analysis study in mini-pigs

Bone response of two different surface titanium subperiosteal implants: anodized surface, ibad ha coating surface / 대한치과보철학회지

STATEMENT OF PROBLEM: In case of poor bone quality or immediately loaded implant, various strategies have been developed focusing on the surface of materials to improve direct implant fixation to the bone. The microscopic properties of implant surfaces play a major role in the osseous healing of dental implant. PURPOSE OF STUDY: This study was undertaken to evaluate bone response of ion beam-assisted deposition(IBAD) of hydroxyapatite(HA) on the anodized surface of subperiosteal titanium implants. MATERIAL AND METHODS: Two half doughnut shape subperiosteal titanium implants were made. The control group was treated with Anodized surface treatment and the test group was treated with IBAD of HA on control surface. Then two implants inserted together into the subperiosteum of the skull of 30 rats and histological response around implant was observed under LM(light microscope) and TEM(transmission electron microscope) on 4th, 6th and 8th week. RESULTS: Many subperiosteal implants were fixed with fibrous connective tissue not with bony tissue because of weak primary stability. The control group observed poor bone response and there was no significant change at any observation time. However the test group showed advanced bone formation and showed direct bone to implant contact under LM on 8th week. The test group observed much rER in the cell of osteoblast but the control group showed little rER under TEM. CONCLUSIONS: The test group showed better bone formation than the control group at the condition of weak primary stability. With these results IBAD surface treatment method on Anodized surface, may be good effect at the condition of weak primary stability.

Evaluation of bone response by resonance frequency analysis of anodized implants / 대한치과보철학회지

STATEMENT OF PROBLEM: Resonance frequency analysis has been increasingly served as a noninvasive and objective method for clinical monitoring of implant stability. Many clinical studies must be required for standardized data using RFA. PURPOSE: This study was performed to evaluate RFA value changes in two anodized implant groups. MATERIAL AND METHOD: Among a total of 24 implants, twelve screw shaped implants as a test group (H2-R8.5) were manufactured, which had a pitch-height of 0.4 mm, an outer diameter of 4.3 mm, a length of 8.5 mm, and external hexa-headed, were turned from 5 mm rods of commercially pure titanium (ASTM Grade IV, Warantec Co., Seoul, Korea), and another twelve implants as a control group were Branemark Ti-Unite MK4 (diameter 4.0 mm, length 8.5 mm). Each group was installed in tibia of rabbit. Two implants were placed in each tibia (four implants per rabbit). Test two implants were inserted in right side and control two in left side. ISQ values were measured using Osstell(TM) (Integration Diagnostics Ltd. Sweden) during fixture installation, and 12 weeks later and evaluated the RFA changes. RESULTS: Mean and SD of baseline ISQ values of test group were 75.0 +/- 3.4 and 68.7 +/- 8.1 for control group. Mean and SD of ISQ values 12 weeks after implant insertion were 73.2 +/- 4.7 for test group and 72.6 +/- 3.9 for control group. There were no statistically significant differences between groups in ISQ values after 3months (P>0.05). From the data, RFA gains after 3 months were calculated, and there was statistically significant difference between groups (P<0.05). CONCLUSION: Although there were RFA changes between groups, implant stability after experimental period shows alike tendency and good bone responses.