Surface properties and biocompatibility of acid-etched titanium

The purpose of this study was to evaluate the effects of acid-etched titanium on the biological responses of osteoblast-like MC3T3-E1 cells. Four types of treatments (polishing, sandblasting, concentrated H2SO4 etching, and concentrated H2SO4 etching with vacuum firing) were carried out on the surfaces of commercially pure titanium (cpTi) disks. MC3T3-E1 cells were then cultured on the treated cpTi surfaces. Through surface roughness measurement and SEM analysis, it was found that the acid-etched surfaces showed higher roughness values than the sandblasted ones. Scanning electron microscope analysis showed that the cells on the disks treated with acid-etching and acid-etching with vacuum firing spread as well as the sandblasted ones. There were no significant differences in cell proliferation and collagen production on cpTi among the four different surface treatments. Based on the results of this study, it was concluded that etching with concentrated sulfuric acid was a simple and effective way to roughen the surface of titanium without compromising its biocompatibility.     

Behavior of CAL72 osteoblast-like cells cultured on zirconia ceramics with different surface topographies

OBJECTIVES: Because of its inherent strength, biocompatibility, and tooth-like color, zirconia ceramics have the potential to become an alternative to titanium as dental implant material. This study aimed at investigating the osteoblastic response to yttrium-stabilized tetragonal zirconia polycrystal (Y-TZP) with different surface topographies. METHODS: CAL72 osteoblast-like cells were cultured on machined (TZP-m), airborne particle abraded (TZP-s), and airborne particle abraded and acid-etched Y-TZP (TZP-sa) surfaces. Polystyrene and airborne particle abraded with large grit and acid-etched (SLA) titanium served as a reference control. The surface topography was examined by scanning electron microscopy (SEM) and profilometry. At culture days 3, 6, and 12, cell proliferation, at day 12 cell morphology, and cell-covered surface area were determined. RESULTS: The surface roughness of Y-TZP was increased by airborne particle abrasion and additionally by acid etching. No statistically significant differences were found between average roughness (R(a)) and maximum peak-to-valley height (R(p-v)) values of airborne particle abraded and acid-etched Y-TZP and SLA titanium. Whereas the cell proliferation assay revealed statistically significant greater values at day 3 for surface-treated Y-TZP and polystyrene cultures as compared with machined Y-TZP, no differences between the Y-TZP groups, SLA titanium, and polystyrene were observed at culture days 6 and 12. CONCLUSIONS: Cell morphology and cell-covered surface area were not affected by the type of substrate. The results suggest that roughened Y-TZP is an appropriate substrate for the proliferation and spreading of osteoblastic cells.     

Direct assessment of profilometric roughness variability from typical implant surface types

PURPOSE: Protocols for quantifying the surface roughness of implants are varied and dependent upon the roughness parameter produced by the particular measurement device. The objective of this study was to examine the accuracy and precision of typical roughness characterization instruments used in the dental implant industry. MATERIALS AND METHODS: The average roughness (Ra) was measured using 2 common surface characterization instruments: an interferometer and a stylus profilometer. Titanium disks were prepared to represent 4 typical dental implant surfaces: machined, acid-etched, hydroxyapatite-coated, and titanium plasma-sprayed. Repeated measurements from multiple sites on each surface were undertaken to establish statistical inferences. Qualitative images of the surfaces were also acquired using a laser scanning confocal microscope. After surface measurements were conducted, the disks were diametrically cut and cross-sectional profiles were examined using a scanning electron microscope (SEM) as a comparative measure of surface topography. An analysis of variance was applied to isolate the effects of the measurement site, measurement sequence, surface treatment, and instrument type on Ra values. RESULTS: The results indicated that surface treatment (P = .0001) and instrument (P = .0001) strongly influenced Ra data. By design, measurement site (diametrical: P = .9859; area: P = .9824) and measurement sequence (P = .9990) did not influence roughness. In the assessment of individual instrument accuracy, the interferometer was the most accurate in predicting SEM-based roughness (P = .6688) compared with the stylus (P = .0839). As a measure of aggregate precision over all measurements, the most repeatable instrument was the stylus (coefficient of variation [CV] = 0.108), followed by the interferometer (CV = 0.125) and SEM (CV = 0.273). DISCUSSION: These results indicate dependencies in accuracy and precision related to the surface characterization technique. CONCLUSION: Instrument variability may obscure functional correlations between implant surface topography and osseointegration.     

The effect of titanium surface roughness on the adhesion of monocytes and their secretion of TNF-alpha and PGE2

Dental implant surfaces are important in determining the tissue/surface interaction. One of the first cells to adhere to the implant surface is the monocyte. This study examines the effect of surface roughness on monocyte adhesion and cytokine secretion. Monocyte adherence to titanium discs of 4 different degrees of surface roughness and plastic surfaces was assayed. Blood mononuclear cells were incubated for 1.5 h in 16 mm culture wells into which titanium discs had been placed. Non-adherent cells were washed off and the numbers of remaining adherent monocyte determined by DNA quantification. TNF-alpha and PGE2 secretion in media from overnight cultures of attached monocytes stimulated with lipopolysaccharide (LPS) was quantified using ELISA and RIA, respectively. Monocyte adherence to rough titanium surfaces was greater than to turned titanium surfaces, while the lowest adherence was to the plastic surface. No significant differences in adherence to 250, 75 or 25 microm blasted surfaces could be detected. The number of adherent monocytes increased with time, with maximum adhesion after 2 h of incubation. Incubation of monocytes adherent to titanium surfaces resulted in a decrease of less than 30% in their numbers over 7 days, whereas cells attached to plastic surfaces decreased to non-detectable numbers after 48 h. Porphyromonas gingivalis LPS stimulation upregulated TNF-alpha and PGE2 secretion into the media. The LPS-induced TNF-alpha and PGE2 secretion was independent of the titanium surface roughness, however the lowest amounts of TNF-alpha and PGE2 were secreted from cells attached to plastic surfaces. The results of this study indicate that the number of monocytes attached to blasted titanium surfaces is significantly greater than to machined titanium surfaces. PGE2 and TNF-alpha secretion is less influenced by titanium surface roughness.     

钛表面形貌和亲水性表面对成骨细胞增殖、分化的影响

目的:通过体外实验研究普通喷砂酸蚀纯钛表面和亲水性喷砂酸蚀纯钛表面对成骨细胞增殖、分化等生物学行为的影响。方法:纯钛片表面分别采用光滑处理(smooth pretreated Ti,PT)、大颗粒喷砂酸蚀表面处理(sand-blasted,large-grit,acid-etched,SLA)及亲水性化学活化大颗粒喷砂酸蚀表面处理(chemically-modified SLA,modSLA/SLActive),在表面接种MC3T3-E1成骨细胞,采用MTT、碱性磷酸酶半定量测试以及茜素红染色检测其对成骨细胞增殖、分化的影响,并采用实时荧光定量PCR检测成骨细胞在不同材料表面骨功能基因表达的差异。应用SAS 9.0软件包对数据进行统计学分析。结果:与光滑钛表面相比,普通喷砂酸蚀钛表面能通过促进ALP、钙基质的分泌和成骨功能基因(Runx2、OSX、OCN和OPN)的表达而显著抑制成骨细胞增殖并促进其分化。在表面粗糙度的基础上增加亲水性,可使这一效应更加明显。结论:表面粗糙度和亲水性是影响成骨细胞生物学行为的重要因素,粗糙钛表面能显著抑制成骨细胞增殖,促进其分化,亲水性的粗糙钛表面促进成骨细胞分化的作用更加显著。     

Harder and stiffer bone osseointegrated to roughened titanium

Mechanisms underlying the beneficial anchorage of roughened titanium implants have not been identified. We hypothesized that the implant surface roughness alters intrinsic biomechanical properties of bone integrated to titanium. Nano-indentation performed on two- and four-week post-implantation bone specimens of rats revealed that bone integrated to acid-etched titanium was approximately 3 times harder than that integrated to the machined titanium, both at the osseointegration interface and at the inner area of the peri-implant bone. The hardness of the acid-etched surface-associated bone was equivalent to that of untreated cortical bone at week 4, while the bone hardness around the machined surface was equivalent to that of the untreated trabecular bone. The elastic modulus of the integrated bone was 1.5 to 2.5 times greater around the acid-etched surface than around the machined surface. Analysis of the data suggests that the implant surface roughness affects the biomechanical quality of osseo-integrated bone, and that the bone integrated to the acid-etched surface is harder and stiffer than the bone integrated to the machined surface.     

Plaque formation on surface modified dental implants

Bacterial adhesion on titanium implant surfaces has a strong influence on healing and long-term outcome of dental implants. Parameters like surface roughness and chemical composition of the implant surface were found to have a significant impact on plaque formation. The purpose of this study was to evaluate the influence of two physical hard coatings on bacterial adhesion in comparison with control surfaces of equivalent roughness. Two members of the oral microflora, Streptococcus mutans and Streptococcus sanguis were used. Commercially pure titanium discs were modified using four different surface treatments: physical vapour deposition (PVD) with either titanium nitride (TiN) or zirconium nitride (ZrN), thermal oxidation and structuring with laser radiation. Polished titanium surfaces were used as controls. Surface topography was examined by SEM and estimation of surface roughness was done using a contact stylus profilometer. Contact angle measurements were carried out to calculate surface energy. Titanium discs were incubated in the respective bacterial cell suspension for one hour and single colonies formed by adhering bacteria were counted by fluorescence microscopy. Contact angle measurements showed no significant differences between the surface modifications. The surface roughness (Ra) of all surfaces examined was between 0.14 and 1.00 microm. A significant reduction of the number of adherent bacteria was observed on inherently stable titanium hard materials such as TiN and ZrN and thermically oxidated titanium surfaces compared to polished titanium. In conclusion, physical modification of titanium implant surfaces such as coating with TiN or ZrN may reduce bacterial adherence and hence improve clinical results.     

The role of surface roughness in promoting osteointegration

Scientific evidence that has been gathered in the past 20 years established that certain endosseous dental implants--primarily screw-type implants made of commercially pure titanium can be successfully utilized as anchorage for dental prostheses. In recent years, an effort has been made to simplify the surgical procedure, in order to modify clinical treatment modalities. One of the trends is to increasingly utilize microrough titanium implants. Roughened implant surfaces have a long history in implant dentistry, and the most prominent surface is titanium plasma-sprayed (TPS). In recent years new implant surfaces have emerged, so-called microrough titanium surfaces produced with reducing techniques such as grit-blasting with Al2O3 or TiO2 particles, sandblasting and acid-etching, or acid-etching alone. These different titanium surfaces have been tested in numerous in-vivo studies utilizing different animal models. Summarizing the results of these studies, it can be concluded that there is currently sufficient evidence that titanium implants with a microrough surfaces achieve a faster bone integration, a higher percentage of Bone implant Contact (BIC), and a higher resistance to shear documented with higher Removal Torque Values (RTV) when compared with titanium implants with a polished or machined surface. In order to understand the mechanism through which surface roughness modulates its effects mentioned above, recent studies used in-vitro experimental methods to study cell response to implant surface topography. These studies have shown that osteoblasts are sensitive to surface roughness, exhibiting decreased proliferation and a more differentiated phenotype on rougher surfaces. PGE2 production is enhanced on rough surfaces, as is the production of TGF beta 1, suggesting that surface roughness can mediate autocrine and paracrine regulation of osteogenesis. Moreover, surface roughness was found to modulate the effect of systemic hormones like 1,25-(OH)2D3 on osteoblasts. The clinical advantages of implants with rough surface were observed in recently conducted clinical trials. It was found, in humans, that roughened titanium implants need shorter healing period before loading, 6-8 (SLA and Osseotite respectively) weeks instead of 12 weeks. The clinical advantages of shorter healing periods are obvious. Moreover, it was found that certain roughened implants can be used in shorter sizes (6-8 mm) then accepted today. The utilization of shorter implants offers the avoidance of extensive surgical procedures such as nerve lateralization in the mandible or sinus grafting in the maxilla. However, sufficient long term documentation is still lacking, and the predictability of such modalities has yet to be examined in long term prospective clinical trials.     

A multi-center study comparing dual acid-etched and machined-surfaced implants in various bone qualities

BACKGROUND: A major reason for the success of modern dental implant systems has been the development of implant designs that enhance direct bone-implant interface. Surface roughness has been a factor in this success and different systems have utilized very different implant surface roughness. The major purpose of this study was to evaluate 2 similar implants with different surface roughness characteristics. METHODS: Two similarly designed, screw-type, commercially pure titanium implants, one dual acid-etched (DAE) and the other machined-surfaced (MS), were compared in this prospective, randomized-controlled, multi-center study, in which a total of 97 patients were enrolled at a private dental practice or a university dental clinic. Both implant types were placed in each patient using a 2-stage approach with a conventional 4- to 6-month healing period. Implants supported fixed prostheses, hybrid prostheses, and overdentures as dictated by the individual patient's need. All of the cases were followed using clinical and radiographic examinations. Criteria of success were the absence of peri-implant radiolucency, mobility, and persistent signs or symptoms of pain or infection. RESULTS: Of the 432 implants (247 dual acid-etched, 185 machined-surfaced), 36 implants (12 dual acid-etched and 24 machined-surfaced) have failed. The pre-loading integration success rate of the dual acid-etched implants (95.0%) was statistically higher (P < 0.01) than the success rate of the machined-surfaced implants (86.7%). At 36 months, the cumulative success rates (CSR) are 95.0% for the dual acid-etched implants and 86.7% for the machined-surfaced implants. CONCLUSIONS: The difference in success rates is most likely attributed to the acid-etched surface characteristics. The greatest performance difference is observed in the conditions of poor quality or soft bone where the 3-year post-loading CSR are 96.8% (dual acid-etched) and 84.8% (machined-surfaced).     

The mucosal attachment to titanium implants with different surface characteristics: an experimental study in dogs

BACKGROUND: Findings from in vitro studies have indicated that the orientation and proliferation of cells on titanium surfaces may be influenced by the topography of the surface on which they are grown. It may be argued, therefore, that differences may occur in the mucosal attachment to titanium implants with different surface roughness. AIM: The present experiment was performed to study the composition of the soft tissue barrier that formed to implants prepared with well-defined smooth or rough surfaces. MATERIAL AND METHODS: Five beagle dogs were used. Four implants made of c.p. titanium were placed in the right edentulous mandibular premolar region. After 3 months, two different types of abutments were connected: one experimental (OA) with a dual, thermal acid-etched surface ('Osseotite'), and one regular (RA) abutment with a 'turned' surface. At the end of a 6-month period during which proper plaque control had been maintained, biopsies including the implant and the surrounding soft and hard tissues were obtained, decalcified and processed for light and electron microscopy. A confocal He-Ne laser profilometer was used to study the surface topography of the abutments. RESULTS: The attachment between the peri-implant mucosa and titanium abutments with either a turned (RA; 'smooth') or acid-etched (OA; 'rough') surface was similar from both a quantitative and a qualitative aspect. The attachment comprised a barrier epithelium and a zone of connective tissue attachment of similar dimension at RA and OA. It was further observed that the 'inner' zone of the connective tissue attachment at both types of abutment was composed of about 30-33% fibroblasts and 63-66% collagen. CONCLUSION: It was demonstrated that the soft tissue attachment that formed to implants made of c.p. titanium was not influenced by the roughness of the titanium surface.     

Influence of an erbium, chromium-doped yttrium, scandium, gallium, and garnet (Er,Cr:YSGG) laser on the reestablishment of the biocompatibility of contaminated titanium implant surfaces

BACKGROUND: The aim of the present study was to evaluate the influence of an erbium, chromium-doped yttrium, scandium, gallium, and garnet (Er,Cr:YSGG laser [ERCL]) on 1) the surface structure and biocompatibility of titanium implants and 2) the removal of plaque biofilms and reestablishment of the biocompatibility of contaminated titanium surfaces. METHODS: Intraoral splints were used to collect an in vivo supragingival biofilm on sand-blasted and acid-etched titanium disks for 24 hours. ERCL was used at an energy output of 0.5, 1.0, 1.5, 2.0, and 2.5 W for the irradiation of 1) non-contaminated (20 and 25 Hz) and 2) plaque-contaminated (25 Hz) titanium disks. Unworn and untreated non-irradiated, sterile titanium disks served as untreated controls (UC). Specimens were incubated with SaOs-2 osteoblasts for 6 days. Treatment time, residual plaque biofilm (RPB) areas (%), mitochondrial cell activity (MA) (counts per second), and cell morphology/surface changes (scanning electron microscopy [SEM]) were assessed. RESULTS: 1) ERCL using either 0.5, 1.0, 1.5, 2.0, or 2.5 W at both 20 and 25 Hz resulted in comparable mean MA values as measured in the UC group. A monolayer of flattened SaOs-2 cells showing complete cytoplasmatic extensions and lamellopodia was observed in both ERCL and UC groups. 2) Mean RPB areas decreased significantly with increasing energy settings (53.8 +/- 2.2 at 0.5 W to 9.8 +/- 6.2 at 2.5 W). However, mean MA values were significantly higher in the UC group. CONCLUSION: Within the limits of the present study, it was concluded that even though ERCL exhibited a high efficiency to remove plaque biofilms in an energy-dependent manner, it failed to reestablish the biocompatibility of contaminated titanium surfaces.     

Adsorption of human plasma proteins to modified titanium surfaces

OBJECTIVES: The aim of this study was to examine the effect of modified titanium (Ti) surfaces on the initial events of plasma proteins adsorption. MATERIALS AND METHODS: 'Ti disks' with three types of surface modifications were compared: machined, acid-etched and acid-etched and blasted. Physical and chemical characterizations of the surfaces were performed via scanning electron microscopy (SEM), atomic force microscopy (AFM) used for analysis of surface topography, characterization of the titanium oxide (TiO2) layer was carried out by X-ray photoelectron spectroscopy (XPS) and characterization of surface energy by the determination of contact angles. Evaluation of plasma proteins' adsorption to the treated Ti surfaces was performed by mass spectrometry, confocal laser scanning microscopy and XPS. Quantitative proteins' assessment was carried out by enzyme-linked immunosorbent assay. RESULTS: SEM images revealed major differences in the topography of the examined surfaces. Acid-etched and blasted Ti surfaces were found to have higher roughness values and a thicker TiO2 layer as compared with acid-etched and machined surfaces. Moreover, acid-etched and blasted surfaces showed high surface area differentiation, pointing to a high increase in the three-dimensional (3D) surface area over the 2D surface area compared with the other surfaces. Adsorption of plasma proteins to the acid-etched and blasted Ti surfaces was both qualitatively and quantitatively more intense compared with the machined and acid-etched surfaces. This was shown for each examined protein, total proteins and by the removal degree of the protein coat. CONCLUSIONS: The preferential adsorption of plasma proteins to the acid-etched and blasted Ti surfaces may be explained by its topographical characteristics and by the increase of the 3D surface area of this modified surface.     

Production of transforming growth factor beta1 and prostaglandin E2 by osteoblast-like cells cultured on titanium surfaces blasted with TiO2 particles

The surface roughness of an implant to which osteoblasts attach may influence endogenous expression of growth factor and cytokines at the implant-tissue interface, modulating the healing process and affecting the quality of bone formation. The present study, using cells derived from human mandibular bone, investigated the effect of varying roughness of titanium surfaces on production of transforming growth factor beta1 (TGF-beta1) and prostaglandin E2 (PGE2). The titanium surfaces were turned (control) and then roughened by blasting with 63-90 micro m, 106-180 micro m or 180-300 micro m TiO2 particles. The cells were cultured onto the surfaces till confluence was achieved. Fresh media were added in the presence or absence of 1,25-dihydroxyvitamin D3[1,25-(OH)2D3], the stimulator of osteogenic differentiation, and aliquots of conditioned cell media were used for assay 24 h later. Cellular morphology was determined by scanning electron microscopy. Cellular production of TGF-beta1 and PGE2 on each surface was assessed by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA), respectively, using commercially available kits. All blasted surfaces showed significantly higher production of TGF-beta1 than the turned surfaces (P < 0.05). In response to stimulation by 1,25-(OH)2D3 cellular production of TGF-beta1, was also significantly greater (P < 0.05) on the blasted surfaces than on the turned one. The total amount of PGE2 in the conditioned media was higher than on the turned surfaces in the presence or absence of 1,25-(OH)2D3. There were no significant differences among the three blasted surfaces with respect to production of the local factors. However, we could not show a synergistic effect of surface roughness and vitamin D on the production of both TGF-beta1 and PGE2 using primary cell culture model.     

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.     

Surface analysis of machined versus sandblasted and acid-etched titanium implants

Initially, implant surface analyses were performed on 10 machined implants and on 10 sandblasted and acid-etched implants. Subsequently, sandblasted and acid-etched implant cytotoxicity (using L929 mouse fibroblasts), morphologic differences between cells (osteoblast-like cells MG63) adhering to the machined implant surfaces, and cell anchorage to sandblasted and acid-etched implant surfaces were evaluated. Results indicated that acid etching with 1% hydrofluoric acid/30% nitric acid after sandblasting eliminated residual alumina particles. The average roughness (Ra) of sandblasted and acid-etched surfaces was about 2.15 microns. Cytotoxicity tests showed that sandblasted and acid-etched implants had non-cytotoxic cellular effects and appeared to be biocompatible. Scanning electron microscopic examination showed that the surface roughness produced by sandblasting and acid etching could affect cell adhesion mechanisms. Osteoblast-like cells adhering to the machined implants presented a very flat configuration, while the same cells adhering to the sandblasted and acid-etched surfaces showed an irregular morphology and many pseudopodi. These morphologic irregularities could improve initial cell anchorage, providing better osseointegration for sandblasted and acid-etched implants.     

Osteoblast-like cells are sensitive to submicron-scale surface structure

OBJECTIVES: Studies showing that osteoblasts exhibit a more differentiated phenotype on rough titanium (Ti) surfaces and osteoclast-resorbed bone surfaces used materials characterized by average peak to valley distance (Ra). Other surface features impacting the cells include distance between peaks, curvature of the valleys, and relative distribution of flat and smooth regions. We used novel Ti surfaces prepared by electrochemical micromachining as models to examine specific contributions of individual design features to osteoblast response. Results show that micron-scale topography modulates cell number, cell morphology and prostaglandin E2 (PGE2). In the presence of the appropriate microtopography, submicron-scale rugosity modulates differentiation and transforming growth factor-beta1 (TGF-beta1) levels. In this study, we examined the role of different types of submicron-scale structures. MATERIAL AND METHODS: Thirty micrometer diameter craters on Ti disks were produced by photolithography resulting in an electropolished smooth surface, and arranged so that inside crater area vs. outside flat area was 6 (30/6). Submicron-scale structures were superposed by acid etching and porous anodization. Ra's were 700, 400, 60 nm on acid-etched, porous anodized and smooth 30/6 surfaces, respectively. RESULTS: MG63 osteoblast-like cells were sensitive to submicron-scale architecture. Cell morphology on anodized surfaces was similar to morphology on smooth surfaces, whereas on etched surfaces, cells had a more elongated differentiated shape. Cell number was greatest on smooth surfaces > anodized > etched. Osteocalcin and PGE2 were affected in a reverse manner. Active TGF-beta1 was greatest on etched 30/6 surfaces > anodized > smooth; latent TGF-beta1 was elevated on all rough surfaces. CONCLUSIONS: These results support our previous observations that submicron-scale structures modulate osteoblastic phenotype and show that the physical properties of the submicron-scale structures are important variables in determining osteoblast response to substrate topography.     

The effect of titanium surface roughening on protein absorption, cell attachment, and cell spreading

PURPOSE: The purpose of this study was to compare properties of roughened and polished titanium with respect to their ability to attach to cells and bind to protein as well as their cell spreading behavior. MATERIALS AND METHODS: Three different titanium surface treatments were compared for their ability to support cell attachment and spreading: sandblasted and acid-etched, resorbable blast media, and machine-polished titanium. The surface of the materials was characterized for surface roughness, surface energy, and surface chemistry. Osteoblast-like MG-63 cells were tested for in vitro attachment and spreading in the presence of serum proteins. Cell attachment was assessed by direct counting, dye binding, and microculture titanium assays. Cell spreading was determined by measuring area/cell in phalloidin-AlexaFluor 488 stained cells. Absorption of bovine serum albumin was determined by assay. RESULTS: Scanning electron micrography and x-ray diffractometry confirmed increased surface roughness of the roughened materials. All 3 materials had similar albumin binding kinetics. Three different methods confirmed that roughened surfaces enhance early cell attachment to titanium in the presence of serum. Cells spread better on smoother machined surfaces than on the roughened surfaces. CONCLUSION: Roughened titanium surfaces exhibited better early cell attachment than smooth surfaces in the presence of serum. The cells attached to roughened titanium were less spread than those attached to machined titanium. Although albumin binding was not different for roughened surfaces, it is possible that roughened surfaces preferentially bound to serum adhesive proteins to promote early cell attachment.