Effect of various implant coatings on biological responses in MG63 using cDNA microarray

During the process of bone formation, titanium (Ti) surface is an important factor in the modulation of osteoblastic function. This study was conducted in order to determine the effects of different Ti surfaces on the biological responses of a human osteoblast-like cell line (MG63). MG63 cells were cultured on smooth (S), sandblasted large-grit and acid etching (SLA), hydroxyapatite (HA), hydroxyfluoride (HF), titanium nitrate (TIN), and diamond-like carbon (DLC) Ti. The morphology of these cells were assessed by SEM. The cDNAs prepared from the total RNAs of the MG63 were hybridized into a human cDNA microarray (1152 elements). The appearances of the surfaces observed by SEM were different on each of the six dental substrate types. The SLA and HA surfaces were determined to be rougher than the others. MG63 cells cultured on SLA and HA exhibited cell-matrix interactions. In the expression of genes involved in osseointegration, several genes, including bone morphogenetic protein, cadherin, integrin, and insulin-like growth factors, were upregulated on the different surfaces. Several genes, including fibroblast growth factor receptor 4, Bcl 2-related protein, and collagen, were downregulated on the different surfaces. The attachment and expression of key osteogenic regulatory genes were enhanced by the surface roughness of the dental materials used.     

Differentiation and cytokine synthesis of human alveolar osteoblasts compared to osteoblast-like cells (MG63) in response to titanium surfaces.

OBJECTIVES: The aim of this study was to investigate the influence of different implant surface topographies and chemistries on the expression of differentiation/proliferation markers on MG63 cells and primary human alveolar osteoblasts. METHODS: Hydrophobic acid-etched (A) and hydrophobic coarse-grit-blasted, acid-etched (SLA) surfaces and hydrophilic acid-etched (modA) and hydrophilic coarse-grit-blasted (modSLA) surfaces were produced. Thereby, modA and modSLA surfaces were rinsed under nitrogen protection and stored in a sealed glass tube containing isotonic NaCl solution at pH 4-6. Tissue culture plates without specimens served as controls. The behavior of MG63 cells and primary human alveolar osteoblasts (AOB) grown on all surfaces was compared through determination of alkaline phosphatase (ALP) activity, cell proliferation ((3)H-thymidin incorporation, MTT colorimetric assay) and expression of osteocalcin (OC), osteoprotegerin (OPG), transforming growth factor-beta1 (TGF-beta(1)) and vascular endothelial growth factor (VEGF), detected with commercial available test kits. RESULTS: Proliferation of MG63 and primary cells was highest on controls, followed by A surfaces, modA and SLA surfaces being almost on the same level and lowest on modSLA surfaces. modSLA surfaces exhibited highest ALP and OC production, followed by SLA, modA and A surfaces. Proliferation and OC production were comparable for MG63 cells and AOB. OPG, TGF-beta(1) and VEGF produced on primary cells showed a slightly different rank order on different surfaces compared to MG63 cells. modSLA still showed the highest production of OPG, TGF-beta(1) and VEGF, but was followed by modA, SLA and A. Statistical significance was checked by ANOVA (p<0.0035). SIGNIFICANCE: MG63 cells and primary human alveolar osteoblasts showed similar proliferation and differentiation characteristics on different titanium surfaces. Only modA surfaces showed enhanced expression of OPG, TGF-beta(1) and VEGF on MG63 cells compared to primary human alveolar osteoblasts. Overall, the lowest proliferation rates and the highest expressions of differentiation markers and growth factor productions were observed on modSLA.     

The angiogenic behaviors of human umbilical vein endothelial cells (HUVEC) in co-culture with osteoblast-like cells (MG-63) on different titanium surfaces

Objectives

Interaction between osteogenesis and angiogenesis plays an important role in implant osseointegration. In the present study we investigated the influence of titanium surface properties on the angiogenic behaviors of endothelial cells grown in direct contact co-culture with osteoblasts.

Methods

Human umbilical vein endothelial cells (HUVECs) and osteoblast-like cells (MG-63 cells) were grown in direct co-culture on the following titanium surfaces: acid-etched (A), hydrophilic A (modA), coarse-gritblasted and acid-etched (SLA) and hydrophilic SLA (SLActive). Cell proliferation was evaluated by cell counting combined with flow cytometry. The expression of von Willebrand Factor (vWF), thrombomodulin (TM), endothelial cell protein C receptor (EPCR), E-Selectin, as well as vascular endothelial growth factor (VEGF) receptors Flt-1 and KDR in HUVECs and VEGF in MG-63 were measured by qPCR. The dynamic behavior of endothelial cells was recorded by time-lapse microscopy.

Results

Proliferation of HUVECs was highest on A, followed by SLA, modA and SLActive surfaces. The expression of vWF, TM, EPCR, E-Selectin and Flt-1 in HUVECs was significantly higher on A than on all other surfaces. The expression of KDR in HUVECs grown on A surface was below detection limit. VEGF expression in MG-63 cells was significantly higher on SLActive vs SLA and modA vs A surfaces. Time-lapse microscopy revealed that HUVECs moved quickest and formed cell clusters earlier on A surface, followed by SLA, modA and SLActive surface.

Conclusions

In co-culture conditions, proliferation and expression of angiogenesis associated genes in HUVECs are promoted by smooth hydrophobic Ti surface, which is in contrast to previous mono-culture studies.     

Biological responses in osteoblast-like cell line according to thin layer hydroxyapatite coatings on anodized titanium

Several features of the implant surface, such as roughness, topography and composition play a relevant role in implant integration with bone. This study was conducted in order to determine the effects of various thin layer hydroxyapatite (HA) coatings on anodized Ti surfaces on the biological responses of a human osteoblast-like cell line (MG63). MG63 cells were cultured on 100 nm HA (100 nm HA coating on anodized surface), 500-700 nm HA (500-700 nm HA coating on anodized surface), 1 mum HA (1 mum HA coating on anodized surface) and anodize (non-HA coating on anodized surface) Ti. The morphology of these cells was assessed by scanning electron microscopy (SEM). The cDNAs prepared from the total RNAs of the MG63 were hybridized into a human cDNA microarray (1152 elements). The appearances of the surfaces observed by SEM were different on each of the four dental substrate types. MG63 cells cultured on 100 nm HA, 1 mum HA and anodize exhibited cell-matrix interactions. It was 500-700 nm HA surface showing cell-cell interaction. In the expression of genes involved in osseointegration, several genes, including bone morphogenetic protein 2, latent transforming growth factor beta binding protein 1, catenin (cadherin-associated protein), integrin, PDGFRB and GDF-1 growth differentiation factor 1 were up-regulated on the different surfaces. Several genes, including fibroblast growth factor receptor 3, fibroblast growth factor 12 and CD4 were down-regulated on the different surfaces. The attachment and expression of key osteogenic regulatory genes were enhanced by the surface morphology of the dental materials used.     

Effect of recombinant human bone morphogenetic protein-7 (rhBMP-7) on the viability, proliferation and differentiation of osteoblast-like cells cultured on a chemically modified titanium surface

Aim: The aim of the present study was to assess the influence of the chemical characteristics and roughness of titanium surfaces on the viability, proliferation and differentiation of osteoblast-like cells cultured in a medium supplemented with recombinant human bone morphogenetic protein-7 (rhBMP-7).
Material and methods: Osteo-1 cells were grown on titanium disks presenting with the following surfaces: (1) machined, (2) coarse grit-blasted and acid-attacked (SLA) and (3) chemically modified SLA (SLAmod) in the absence or presence of 20 ng/ml rhBMP-7 in culture medium. The viability and number of osteo-1 cells were evaluated after 24 h. Analyses of total protein content (TP) and alkaline phosphatase (AP) activity at 7, 14 and 21 days, collagen content at 7 and 21 days and mineralized matrix formation at 21 days were performed.
Results: Cell viability ( P =0.5516), cell number ( P =0.3485), collagen content ( P =0.1165) and mineralized matrix formation ( P =0.5319) were not affected by the different surface configurations or by the addition of rhBMP-7 to the medium. Osteo-1 cells cultured on SLA surfaces showed a significant increase in TP at 21 days. The ALPase/TP ratio ( P =0.00001) was affected by treatment and time.
Conclusion: The results suggest that the addition of rhBMP-7 to the culture medium did not exert any effect on the viability, proliferation or differentiation of osteoblast-like cells grown on the different surfaces tested. All titanium surfaces analyzed allowed the complete expression of the osteoblast phenotype such as matrix mineralization by osteo-1 cells.     

Behavior of two osteoblast-like cell lines cultured on machined or rough titanium surfaces

Background: Two osteosarcoma-derived cell lines have been extensively used to investigate the biological events occurring on titanium surfaces: MG63 and Saos-2. However, the behavior of the two lines on different titanium surfaces has never been compared.
Aim: The aim of the present study was to compare the behavior of MG63 and Saos-2 cells on two different titanium surfaces, machined and rough (sandblasting and acid-etched). We compared cell proliferation and morphology, alkaline phosphatase (ALP) activity and secretion of osteocalcin (OC).
Results: The most pronounced difference between the two cell lines was that ALP activity in the Saos-2 cells was 10-fold higher than in the MG63 cells. The proliferation rate of the MG63 cells was much higher than that of the Saos-2 cells at all the tested cell concentrations. MG-63 cells, but not Saos-2 cells, grown on rough surface titanium proliferated more rapidly than cells grown on machined surfaces. Morphological analysis revealed that Saos-2 cells and cells grown on the rougher surface, displayed a more mature phenotype. The level of OC secreted by the Saos-2 cells, but not the MG63 cells, were higher on the rough surface than on the machined surface.
Conclusions: This study shows that Saos-2 cells exhibit a more mature osteoblast phenotype, compared with that of MG63 cells, rendering them a good candidate for an in vitro model of osseointegration.     

Influence of gaseous ozone in peri-implantitis: bactericidal efficacy and cellular response. An in vitro study using titanium and zirconia

Dental implants are prone to bacterial colonization which may result in bone destruction and implant loss. Treatments of peri-implant disease aim to reduce bacterial adherence while leaving the implant surface intact for attachment of bone-regenerating host cells. The aims of this study were to investigate the antimicrobial efficacy of gaseous ozone on bacteria adhered to various titanium and zirconia surfaces and to evaluate adhesion of osteoblast-like MG-63 cells to ozone-treated surfaces. Saliva-coated titanium (SLA and polished) and zirconia (acid etched and polished) disks served as substrates for the adherence of Streptococcus sanguinis DSM20068 and Porphyromonas gingivalis ATCC33277. The test specimens were treated with gaseous ozone (140 ppm; 33 mL/s) for 6 and 24 s. Bacteria were resuspended using ultrasonication, serially diluted and cultured. MG-63 cell adhesion was analyzed with reference to cell attachment, morphology, spreading, and proliferation. Surface topography as well as cell morphology of the test specimens were inspected by SEM. The highest bacterial adherence was found on titanium SLA whereas the other surfaces revealed 50-75% less adherent bacteria. P. gingivalis was eliminated by ozone from all surfaces within 24 s to below the detection limit (≥99.94% reduction). S. sanguinis was more resistant and showed the highest reduction on zirconia substrates (>90% reduction). Ozone treatment did not affect the surface structures of the test specimens and did not influence osteoblastic cell adhesion and proliferation negatively. Titanium (polished) and zirconia (acid etched and polished) had a lower colonization potential and may be suitable material for implant abutments. Gaseous ozone showed selective efficacy to reduce adherent bacteria on titanium and zirconia without affecting adhesion and proliferation of osteoblastic cells. This in vitro study may provide a solid basis for clinical studies on gaseous ozone treatment of peri-implantitis and revealed an essential base for sufficient tissue regeneration.     

Sandblasted-acid-etched titanium surface influences in vitro the biological behavior of SaOS-2 human osteoblast-like cells

Osseointegrated dental implants have been successfully used over the past several years, allowing functional replacement of missing teeth. Surface properties of titanium dental implants influence bone cell response. Implant topography appears to modulate cell growth and differentiation of osteoblasts thus affecting the bone healing process. Optimal roughness and superficial morphology are still controversial and need to be clearly defined. In the present study we evaluated in vitro the biological behavior of SaOS-2 cells, a human osteoblast-like cell line, cultured on two different titanium surfaces, smooth and sandblasted-acid-etched, by investigating cell morphology, adhesion, proliferation, expression of some bone differentiation markers and extracellular matrix components. Results showed that the surface topography may influence in vitro the phenotypical expression of human osteoblast-like cells. In particular the tested sandblasted-acid-etched titanium surface induced a significantly increased Co I deposition and α2-β1 receptor expression as compared to the relatively smooth surface, promoting a probable tendency of SaOS-2 cells to shift toward a mature osteoblastic phenotype. It is therefore likely that specific surface properties of sandblasted-acid-etched titanium implants may modulate the biological behavior of osteoblasts during bone tissue healing.     

Biofilm and cell adhesion strength on dental implant surfaces via the laser spallation technique

ObjectiveThe aims of this study are to quantify the adhesion strength differential between an oral bacterial biofilm and an osteoblast-like cell monolayer to a dental implant-simulant surface and develop a metric that quantifies the biocompatible effect of implant surfaces on bacterial and cell adhesion.MethodsHigh-amplitude short-duration stress waves generated by laser pulse absorption are used to spall bacteria and cells from titanium substrates. By carefully controlling laser fluence and calibration of laser fluence with applied stress, the adhesion difference between Streptococcus mutans biofilms and MG 63 osteoblast-like cell monolayers on smooth and rough titanium substrates is obtained. The ratio of cell adhesion strength to biofilm adhesion strength (i.e., Adhesion Index) is determined as a nondimensionalized parameter for biocompatibility assessment.ResultsAdhesion strength of 143 MPa, with a 95% C.I. (114, 176), is measured for MG 63 cells on smooth titanium and 292 MPa, with a 95% C.I. (267, 306), on roughened titanium. Adhesion strength for S. mutans on smooth titanium is 320 MPa, with a 95% C.I. (304, 333), and remained relatively constant at 332 MPa, with a 95% C.I. (324, 343), on roughened titanium. The calculated Adhesion Index for smooth titanium is 0.451, with a 95% C.I. (0.267, 0.622), which increased to 0.876, with a 95% C.I. (0.780, 0.932), on roughened titanium.SignificanceThe laser spallation technique provides a platform to examine the tradeoffs of adhesion modulators on both biofilm and cell adhesion. This tradeoff is characterized by the Adhesion Index, which is proposed to aid biocompatibility screening and could help improve implantation outcomes. The Adhesion Index is implemented to determine surface factors that promote favorable adhesion of cells greater than biofilms. Here, an Adhesion Index ? 1 suggests favorable biocompatibility.     

Primäre Osteoblastenreaktionen auf SLA- und mikrostrukturierten Implantatoberflächen

The osseointegration process of dental implants depends on the tissue reaction at the tissue-implant interface. Osteoblasts are the main cells responsible for the regulation of osteoinduction. The manner and kinetics of the tissue reaction crucially depend on the interaction between osteoblasts and the morphology of the implant surface. The aim of this study was to investigate osteoblast behaviour on different implant surfaces (smooth, microgrooved, SLA) under standardized conditions. For this in vitro investigation we used primary bovine osteoblasts. Attachment kinetics, proliferation rate and synthesis of bone-associated proteins were used as parameters for cell reaction. The results demonstrate that both attachment and adhesion strength of the primary cell surface interaction was higher on the microgrooved surfaces than on SLA surfaces. The proliferation rate of cells and the synthesis of bone-specific proteins were higher on microgrooved surfaces in contrast to SLA surfaces. Ultrastructural analysis revealed phenotypic osteoblast-like cells on smooth and microgrooved surfaces, whereas cells on SLA surfaces showed a more fibroblastic appearance. This study demonstrates that the morphology of the implant surface determined the subsequent osteoblast reaction. An optimal cell reaction was found at surfaces which are smooth in the microenvironment of osteoblasts.     

阳极氧化处理后的新型钛合金体外诱导实验及对成骨细胞早期附着的影响

目的：研究新型医用钛合金Ti-24Nb-4Zr-7．9Sn（TNZS）经过阳极氧化（AD）技术处理后体外诱导活性及对人成骨样MG63细胞早期附着的影响。方法：采用阳极氧化处理TNZS合金，对其进行模拟体液（SBF）浸泡实验，分析其表面元素成分及相结构的变化；并将人成骨样MG63细胞接种于Ti6Al4V、TNZS、AD—TNZS表面，观察细胞的早期附着及形态学变化。结果：AD—TNZS在SBF中浸泡6d后，材料表面有羟基磷灰石形成。在MG63成骨样细胞接种60、120min时，人成骨样MG63细胞在AD—TNZS表面的附着率均明显高于Ti6Al4V和TNZS表面附着率（P〈0．05），并表现出良好的形态。结论：阳极氧化后的新型医用钛合金在模拟体液中可诱导羟基磷灰石形成；对成骨细胞的早期附着有一定的促进作用。     

Type I collagen grafting on titanium surfaces using low-temperature glow discharge

To improve the bioactivity of titanium surfaces, glow discharge was used to facilitate collagen grafting on titanium disks. Titanium test specimens were pre-treated by glow discharge fed with a mixture of argon and allylamine (AA) gases. Treated titanium disks were then grafted with type I collagen using glutaraldehyde (GA) as a crosslinking agent. The surfaces of collagen-grafted titanium disks were evaluated using scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) and X-ray photoelectron spectroscopy (XPS). MG-63 osteoblast-like cells were cultured on the grafted titanium surfaces to examine the effect of collagen grafting in terms of cell morphology. Our results demonstrated that collagen component elements could be detected on the titanium surfaces. Morphology of the cells on the surfaces of collagen-grafted titanium disks indicated differentiation. These findings showed that type I collagen could be successfully grafted onto titanium surfaces using glow discharge technology, with enhanced biofunctionality demonstrated on osteoblastic cells.     

Immunolocalization of fibronectin and vitronectin receptors in human gingival fibroblasts spreading on titanium surfaces

The adhesion and spreading of human gingival fibroblasls on glass and differently processed titanium surfaces was studied by immunolocalization of vinculin and the alpha and beta subunits of the fibronectin (α₅β₁) and vitronectin (α_vβ3) receptors. Vinculin-containing focal contacts were present both at 4 and 24 h of spreading in cells grown on glass or electropolished or etched titanium surfaces but not in cells spreading on sandblasted titanium surfaces. Immunostaining for the α5 and β1 subunits of the fibronectin receptor showed only a diffuse membrane fluorescence after 4 h of cell spreading irrespective of the growth surface. The α_v and β₃ subunits of the vitronectin receptor were at this stage detected in focal contacts in cells spreading on glass or electropolished or etched titanium surfaces. In cells spreading on sandblasted titanium surfaces, however, the vitronectin receptor had only a diffuse distribution. In cells that had been allowed to spread for 24 h on glass or electropolished or etched titanium surfaces the α₅ and β₁ integrin subunits were either diffusely distributed or showed a localization typical of extracellular matrix contacts. The α_v and β₃ integrin subunits were, as earlier, localized to typical focal contacts in cells grown on glass or electropolished or etched titanium surfaces. Cells attached to sandblasted titanium surfaces still expressed all the integrin subunits only diffusely. The results show that the surface texture of the substratum can affect the expression of integrin subunits in human gingival fibroblasts. As evidenced by the recruitment of integrin subunits to focal and extracellular matrix contacts, smooth or finely grooved titanium surfaces appear to be optimal in supporting the attachment of human gingival fibroblasts.     

人脱细胞真皮基质的结构及与MG63成骨样细胞生物相容性的研究

目的观察脱细胞真皮基质（ADM）的结构及MG63成骨样细胞在其上黏附与增殖的情况。方法ADM为实验组,膨体聚四氟乙烯（e- PTFE）膜为对照组,扫描电镜和光学显微镜下观察两组膜的结构。在两组膜上分别接种MG63成骨样细胞,并设空白对照组。采用细胞活力分析仪检测3组细胞增殖活力,酶联免疫法检测细胞内碱性磷酸酶（ALP）表达,扫描电镜观察细胞在膜上接种后第1天和第5天的黏附及增殖情况。结果ADM分为基底膜面和组织面,组织面为鳞片状的结构,基底膜面可见指突结构和毛囊孔。e- PTFE成行排列,由直径比较均一的长椭圆形的裂隙组成。与空白对照组相比,ADM和e- PTFE对MG63成骨样细胞增殖活力和ALP活性无显著影响。扫描电镜观察细胞在两种膜上生长良好,但在ADM膜上,MG63成骨样细胞伸展更充分。结论ADM适于作引导骨再生膜材料,对MG63成骨样细胞的生长无抑制作用,相比较e- PTFE,ADM具有更优良的结构及生物学性能。     

Proliferation, differentiation, and protein synthesis of human osteoblast‐like cells (MG63) cultured on previously used titanium surfaces

This study compared osteoblast proliferation, differentiation, and protein synthesis on new and used titanium (Ti) disks to test the hypothesis that cleaning and resterilization of previously used Ti disks does not alter cell response to a particular surface. Ti disks of varying roughness were prepared by one of five different treatment regimens. Standard tissue culture plastic was used as a control. Human osteoblast-like cells (MG63) were cultured on the Ti disks and cell proliferation, cell differentiation, RNA synthesis and matrix production (collagen and noncollagen protein; proteoglycans) measured. After their first use, the disks were cleaned, re-sterilized by autoclaving. and MG63 cells cultured on them as before. At confluence, the same parameters were measured and cell behaviour on new and used disks compared. When confluent cultures of cells on plastic were compared to those cultured on new Ti surfaces, cell number was reduced on the roughest surfaces and equivalent to plastic on the other surfaces. Cell number was further reduced when disks with the roughest surfaces were re-used; no differences in cell number could be discerned after cleaning and re-sterilization. Cell proliferation was inversely related to surface roughness and was less than seen on tissue culture plastic. Re-use of the Ti disks resulted in no change in cell proliferation rate. Alkaline phosphatase specific activity in isolated cells was lowest on the rougher surfaces; no differences between new and used disks were observed. Similarly, enzyme activity in the cell layer was decreased in cultures grown on rougher surfaces, with no effect of prior disk use being noted. RNA synthesis was decreased with respect to plastic in cultures on smoother surfaces and increased on rougher surfaces; prior disk use did not alter RNA synthesis. Collagen production by the cells was decreased on smoother surfaces, but was comparable to tissue culture plastic when grown on rougher surfaces. Non-collagen protein production was unaffected by culture surface and whether or not the disk had been previously used. Proteoglycan synthesis by cells was decreased on all surfaces studied and comparable on both new and used disks. The results of this study indicate that Ti implant surfaces are unaffected by cleaning and resterilization, although rougher surfaces may require more extensive cleaning than smoother ones. This suggests the possibility that implants, in the same patient, could be safely reused. In vivo studies in animals, however, need to be performed before clinical application can be considered.     

Osteoblast proliferation and differentiation on dentin slices are modulated by pretreatment of the surface with tetracycline or osteoclasts

BACKGROUND: Implant surface roughness and chemical composition, as well as other factors, affect the ability of osteogenic cells to form bone adjacent to an implant. The same principles may also apply to the tooth root and some reports have shown that surface modification of the root may lead to improved restoration of the periodontal apparatus. The most common of these surface modification techniques involves demineralization with citric acid or treatment with tetracycline to expose collagen fibrils. In addition, during normal bone remodeling, osteoclasts demineralize the extracellular matrix, leaving resorption pits and exposed collagen fibrils. In this study, the effect of different dentin surface-preparation techniques on osteoblasts were compared. METHODS: Slices of sperm whale dentin were mechanically polished and surfaces were treated with tetracycline-HCl (TCN) or were cultured with mouse bone marrow cells to create a surface with osteoclast (OC) resorption pits or left untreated. Profilometry, x-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM) were used to evaluate the 3 different dentin surfaces. MG63 osteoblast-like cells were cultured on the 3 different surfaces and the effect of dentin surface preparation technique on MG63 cell proliferation (cell number), differentiaton (alkaline phosphatase specific activity of isolated cells and cell layer lysates; osteocalcin production), and local factor production (transforming growth factor (TGF)-beta1 and prostaglandin E2 (PGE2) compared. RESULTS: Profilometry showed the polished and TCN surfaces were smooth with comparable Ra values, whereas the OC surfaces were slightly rougher due to resorption pits which covered 3.7% of the surface. XPS measurements showed that TCN treatment reduced the Ca and P content of the surface, indicating that it had dissolved the mineral. Osteoclast-resorption also reduced the Ca and P content, but to a lesser extent. MG63 cell proliferation on polished dentin and tissue culture polystyrene was equivalent. In contrast, cells grown on the TCN- and OC-treated surfaces exhibited increased proliferation. No effect of surface treatment on cell alkaline phosphatase activity was observed, but activity in the cell layer lysates was increased on the TCN- and OC-treated surfaces. Osteocalcin production was reduced on all dentin surfaces, but the greatest reduction was found on the TCN-treated surface. Production of both TGF-beta1 and PGE2 was increased on the treated surfaces. All effects were greatest in cultures grown on the TCN-treated dentin. CONCLUSIONS: These data indicate that demineralization of the dentin surface promotes proliferation of osteoblasts and early differentiation events like production of alkaline phosphatase and autocrine mediators such as PGE2 and TGF-beta1. However, later differentiation events like osteocalcin production are decreased. Osteoclast-mediated bone resorption elicits similar responses; less than 4% of the dentin surface resulted in approximately 75% of the response caused by TCN treatment. These observations suggest that greater attention should be paid to the effects of osteoclastic resorption in designing methods for enhancing bone and cementum formation adjacent to root surfaces.     

Corrosion resistance and biocompatibility of a new porous surface for titanium implants

Alterations of the commercially pure titanium (cpTi) surface may be undertaken to improve its biological properties. The aim of this study was to investigate the biocompatibility of cpTi when submitted to a new, porous titanium, surface treatment (porous Ti). Five types of surface treatments, namely sintered microspheres porous titanium (porous Ti), titanium plasma spray (TPS), hydroxyapatite (HA), sandblasted and acid etched (SBAE), and resorbable blast medium, sandblasted with hydroxyapatite (RBM) were made. In the experimental methods, the corrosion potentials were measured over time, and then a linear sweep voltammetric analysis measured the polarization resistances and corrosion currents. For biocompatibility evaluation, MG63 osteoblast-like cells were used. Cell morphology, cell proliferation, total protein content, and alkaline phosphatase (ALP) activity were evaluated after 2 h, and after 2, 4 and 7 d. Porous Ti and SBAE showed a better corrosion resistance, with a weak corrosion current and a high polarization resistance, than the other surfaces. Cell attachment, cell morphology, cell proliferation, and ALP synthesis were influenced by the surface treatments, with a significant increase observed of the activity of osteoblast cells on the porous coating (porous Ti). Based on these results, it is suggested that the porous Ti surface has a significantly better biocompatibility than the other surface treatments and an excellent electrochemical performance.     

微弧氧化AZ91D镁合金对成骨细胞早期粘附的影响

目的体外观察探讨新型口腔植入材料微弧氧化(Micro-arc oxidation,MAO)AZ91D镁合金对人成骨样MG63细胞早期粘附的影响。方法实验分为三组:微弧氧化AZ91D镁合金材料实验组A,纯钛材料对照组B和细胞直接生长在培养板对照组C,利用表面轮廓仪、接触角测量仪、扫描电镜(SEM)和能谱分析(EDS)分别测量研究试件表面粗糙度、表面能大小、形貌特点和元素成分;将MG63细胞培养于各组表面,分析比较成骨细胞的早期粘附率及形态学变化。对数据采用SPSS13.0统计软件进行单因素方差分析。结果A组表面为一层粗糙多孔、起伏不平的薄膜,主要元素有Mg、O、Si,粗糙度及表面能均比B组增加(P<0.05);0.5h、1h和2h三个时间点三组细胞粘附率差异有统计学意义(P<0.05),A组>B组>C组;培养4h时A、B两组材料表面成骨细胞形态良好。结论微弧氧化AZ91D镁合金具有良好的成骨细胞相容性,有利于细胞的早期粘附。     

Enhanced biological responses of a hydroxyapatite/TiO2 hybrid structure when surface electric charge is controlled using radiofrequency sputtering

We investigated whether surface roughness and control of surface electric charge of a hydroxyapatite (HA)/titanium oxide (TiO2) hybrid coating could enhance biological responses associated with bone formation. After acid etching, a titanium surface was modified with HA and TiO2 by the dual sputtering deposition technique using radiofrequency sputtering. These surfaces were analyzed for surface roughness and surface electric charge intensity. Rat bone marrow-derived osteoblast-like cells were cultured on HA/TiO2 hybrid surfaces with different electric charges. The attachment and spreading behavior of these cells were significantly increased on the hybrid surface (p<0.05). In vivo experiment, the strength of bone-titanium implant integration with a hybrid surface was 3 times that of a control (p<0.05). The dual sputtering deposition technique created a HA/TiO2 hybrid structure. Our results show that the surface electric charge on a titanium surface is an important factor for enhancing biological responses.     

Effects of fluoride-modified titanium surfaces on osteoblast proliferation and gene expression

PURPOSE: The objective of this study was to test the hypothesis that fluoride-modified titanium surfaces would enhance osteoblast differentiation. Osteoblast growth on a moderately rough etched fluoride-modified titanium surface (alteration in cellular differentiation) was compared to osteoblast growth on the same surface grit-blasted with titanium dioxide. The potential role of nanometer-level alterations on cell shape and subsequent differentiation was then compared. MATERIALS AND METHODS: Human embryonic palatal mesenchymal (HEPM) cultures were incubated on the respective surfaces for 1, 3, and 7 days, followed by analysis for cell proliferation, alkaline phosphatase (ALP) -specific activity, and mRNA steady-state expression for bone-related genes (ALP, type I collagen, osteocalcin, bone sialoprotein [BSP] II, Cbfa1, and osterix) by real-time polymerase chain reaction (PCR). RESULTS: The different surfaces did not alter the mRNA expression for ALP, type I collagen, osterix, osteocalcin, or BSP II. However, Cbfa1 expression on the fluoride-modified titanium surface was significantly higher (P < .001) at 1 week. The number of cells on this surface was 20% lower than the number of cells on the surface TiO2-blasted with 25-microm particles but not significantly different from the number of cells on the surface TiO2-blasted with 125-microm particles. Cells grown on all the titanium surfaces expressed similar levels of ALP activity. CONCLUSIONS: The results indicated that a fluoride-modified surface topography, in synergy with surface roughness, may have a greater influence on the level of expression of Cbfa1 (a key regulator for osteogenesis) than the unmodified titanium surfaces studied.