Dual Acid-etched Research Articles

Osseointegration of implant surfaces in metabolic syndrome and type-2 diabetes mellitus.

This in vivo study evaluated the bone healing response around endosteal implants with varying surface topography/chemistry in a preclinical, large transitional model induced with metabolic syndrome (MS) and type-2 diabetes mellitus (T2DM). Fifteen Göttingen minipigs were randomly distributed into two groups: (i) control (normal diet, n = 5) and (ii) O/MS (cafeteria diet for obesity induction, n = 10). Following obesity induction, five minipigs from the obese/metabolic syndrome(O/MS) group were further allocated, randomly, into the third experimental group: (iii) T2DM (cafeteria diet + streptozotocin). Implants with different surface topography/chemistry: (i) dual acid-etched (DAE) and (ii) nano-hydroxyapatite coating over the DAE surface (NANO), were placed into the right ilium of the subjects and allowed to heal for 4 weeks. Histomorphometric evaluation of bone-to-implant contact (%BIC) and bone area fraction occupancy (%BAFO) within implant threads were performed using histomicrographs. Implants with NANO surface presented significantly higher %BIC (~26%) and %BAFO (~35%) relative to implants with DAE surface (%BIC = ~14% and %BAFO = ~28%, p < .025). Data as a function of systemic condition presented significantly higher %BIC (~28%) and %BAFO (~42%) in the control group compared with the metabolically compromised groups (O/MS: %BIC = 14.35% and %BAFO = 26.24%, p < .021; T2DM: %BIC = 17.91% and %BAFO = 26.12%, p < .021) with no significant difference between O/MS and T2DM (p > .05). Statistical evaluation considering both factors demonstrated significantly higher %BIC and %BAFO for the NANO surfacerelative to DAE implant, independent of systemic condition (p < .05). The gain increase of %BIC and %BAFO for the NANO compared with DAE was more pronounced in O/MS and T2DM subjects. Osseointegration parameters were significantly reduced in metabolically compromised subjects compared with healthy subjects. Nanostructured hydroxyapatite-coated surfaces improved osseointegration relative to DAE, regardless of systemic condition.

Adipogenesis-Related Metabolic Condition Affects Shear-Stressed Endothelial Cells Activity Responding to Titanium

Obesity has increased around the world. Obese individuals need to be better assisted, with special attention given to dental and medical specialties. Among obesity-related complications, the osseointegration of dental implants has raised concerns. This mechanism depends on healthy angiogenesis surrounding the implanted devices. As an experimental analysis able to mimic this issue is currently lacking, we address this issue by proposing an in vitro high-adipogenesis model using differentiated adipocytes to further investigate their endocrine and synergic effect in endothelial cells responding to titanium. Firstly, adipocytes (3T3-L1 cell line) were differentiated under two experimental conditions: Ctrl (normal glucose concentration) and High-Glucose Medium (50 mM of glucose), which was validated using Oil Red O Staining and inflammatory markers gene expression by qPCR. Further, the adipocyte-conditioned medium was enriched by two types of titanium-related surfaces: Dual Acid-Etching (DAE) and Nano-Hydroxyapatite blasted surfaces (nHA) for up to 24 h. Finally, the endothelial cells (ECs) were exposed in those conditioned media under shear stress mimicking blood flow. Important genes related to angiogenesis were then evaluated by using RT-qPCR and Western blot. Firstly, the high-adipogenicity model using 3T3-L1 adipocytes was validated presenting an increase in the oxidative stress markers, concomitantly with an increase in intracellular fat droplets, pro-inflammatory-related gene expressions, and also the ECM remodeling, as well as modulating mitogen-activated protein kinases (MAPKs). Additionally, Src was evaluated by Western blot, and its modulation can be related to EC survival signaling. Our study provides an experimental model of high adipogenesis in vitro by establishing a pro-inflammatory environment and intracellular fat droplets. Additionally, the efficacy of this model to evaluate the EC response to titanium-enriched mediums under adipogenicity-related metabolic conditions was analyzed, revealing significant interference with EC performance. Altogether, these data gather valuable findings on understanding the reasons for the higher percentage of implant failures in obese individuals.

Nanohydroxyapatite-Blasted Bioactive Surface Drives Shear-Stressed Endothelial Cell Growth and Angiogenesis

Nanosized crystalline hydroxyapatite coating (HAnano®) accelerates the osteointegration of dental implants which is hypothesized to drive angiogenesis. In order to test this hypothesis, we have subjected shear-stressed human umbilical vein endothelial cells (HUVECs) to a HAnano®-enriched medium, as well as to surface presenting dual acid etching (DAE) as a control. To note, the titanium implants were coated with 10 nm in diameter HA particles using the Promimic HAnano method. Our data reveals that HAnano® modulates higher expression of genes related with endothelial cell performance and viability, such as VEGF, eNOS, and AKT, and further angiogenesis in vitro by promoting endothelial cell migration. Additionally, the data shows a significant extracellular matrix (ECM) remodeling, and this finding seems developing a dual role in promoting the expression of VEGF and control endothelial cell growth during angiogenesis. Altogether, these data prompted us to further validate this phenomenon by exploring genes related with the control of cell cycle and in fact our data shows that HAnano® promotes higher expression of CDK4 gene, while p21 and p15 genes (suppressor genes) were significantly lower. In conjunction, our data shows for the first time that HAnano®-coated surfaces drive angiogenesis by stimulating a proliferative and migration phenotype of endothelial cells, and this finding opens novel comprehension about osseointegration mechanism considering nanosized hydroxyapatite coating dental implants.

The Sigmoidal Nature of Bone Anchorage.

The purpose of this study was to show the full evolution of bone anchorage caused by the growth of secondary stability and to determine which empirical model would provide the best quantitative description of this growth. The retention and anchorage of machined (M), grit-blasted and dual acid etched (BAE), and BAE implants with discrete crystals of calcium phosphate (+DCD) were evaluated with both ex vivo and in vivo methods. Ex vivo evaluation of implant retention was tested by measuring the force required to pull implants out of blood-filled osteotomies formed in bovine bone for up to 1 hour. In vivo measurements of bone anchorage were evaluated by reverse torque testing of implants placed in the proximal metaphysis of rat tibiae up to 28 days after initial placement. Four models were fit to the reverse torque results, and fits were evaluated by Bayesian and Akaike information criteria (BIC and AIC) and analysis of variance (ANOVA). AIC and BIC were 655.53 and 684.78, 472.53 and 512.74, 477.40 and 513.96, and 470.60 and 507.16 for the monomolecular, Richards, Gompertz, and logistic curves, respectively. Comparison of the Richards and logistic curves by analysis of variance (ANOVA) resulted in a P value of .78. A comparison of the three implant types using the logistic curve found that M implants had an earlier inflection point compared with BAE implants (P = .038), and the BAE+DCD implants had the greatest peak anchorage and were significantly greater than both M (P < .0001) and BAE implants (P = .005). Bone anchorage was found to follow sigmoidal growth, which was best described by the logistic function. Further comparison of the fit values for the logistic curve shows that both overall anchorage and timing of bone anchorage are influenced by implant surface topography.

Peri-Implant Tissue Behaviour Next to Different Titanium Surfaces: 16-Year Post-Trial Follow-Up

The present post-trial follow-up investigated the influence of titanium implants with different surface treatments on clinical behavior of soft and hard peri-implant tissues. Each of the 18 included patients received at least two adjacent implants: one control implant with a dual acid-etched (DAE) surface in their apical portion and a machined coronal part, and one test implant with a DAE surface up to its coronal portion. Peri-implant bone level change (BLC), probing depth (PD), bleeding on probing (BOP) and plaque index (PI) were recorded. A total of 42 implants was inserted. The mean follow-up period was 9.3 years (range: 5–16 years) and there were six dropouts. No implant failed. Moderate crestal bone remodeling occurred during the first year after implant insertion, with lower bone loss next to test implants compared to control ones (0.80 vs. 1.39 mm; p = 0.002). This difference was also detected at the 5- (p = 0.011), 6- (p = 0.008) and 7-year follow-up appointment (p = 0.027). No statistically significant differences were found in bone resorption between implants rehabilitated with ceramic vs. composite resin veneering material. No statistically significant differences were detected between test and control implants for BOP, PI, and PD at any time point. The results of the present study suggest that DAE surfaces reduce peri-implant bone loss in the initial phase of healing compared to machined surfaces, while they do not significantly affect soft peri-implant tissue and bone maintenance in the long-term. In conclusion, the minimally rough surfaces favour peri-implant bone maintenance and their effect is greater in the first year post implant insertion.

Microstructured titanium functionalized by naringin inserted multilayers for promoting osteogenesis and inhibiting osteoclastogenesis

Osteoporosis is the most common cause of fractures in middle-aged and elderly people. Fracture repair can be difficult due to the decreased bone volume in osteoporosis patients and implants are often required. In this study, a slow-release system for microstructured titanium (Micro-Ti) was designed to promote osteogenesis and inhibit osteoclastogenesis. Firstly, Micro-Ti was prepared on titanium surfaces by dual acid etching. Micro-Ti was covered with naringin (NA), chitosan (CHI) and gelatin (GEL) multilayers through layer by layer technique, which is denoted as LBL (NA) coated-Ti. Osteoblasts (ME3T3-E1) and macrophages (RAW 264.7) were cultured on untreated and treated titanium surfaces in vitro. Osteoblasts grown on LBL (NA) coated-Ti showed higher alkaline phosphatase (ALP) and mineralization, consistent with qRT-PCR analysis of osteoblast genes including runt-related transcription factor 2 (Runx2), ALP, collagen I (Col I), osteocalcin (OCN), osteopontin (OPN), and osteoprotegerin (OPG). In contrast, acid tartarate-resistant phosphatase activity and the expression of osteoclastic differentiation related genes comprising of cathepsin K (CTSK), nuclear factor of activated T cells (NFAT), tartrate resistant acid phosphatase (TRAP) and V-ATPase (VATP) in osteoclasts were significantly reduced on LBL (NA) coated-Ti surfaces compared with other groups. These results indicate that microstructured titanium functionalized by naringin inserted multilayers enhanced the differentiation of osteoblasts and inhibited osteoclast formation. The proposed approach in this research provides a novel way to modify titanium-based implants for fracture repair in osteoporosis patients.

Metabolic effects of CoCr-enriched medium on shear-stressed endothelial cell and osteoblasts: A possible mechanism involving a hypoxic condition on bone healing

Cobalt-chromium (CoCr)-based alloys have emerged as an interesting biomaterial within biomedical field, mainly considering their biocompatibility, resistance to corrosion and absence of magnetism; however, its effect on cell metabolism is barely known and this prompted us better evaluating whether CoCr-enriched medium affects the metabolism of both osteoblast and endothelial cells, and also if there is a coupling between them. This is also considered here the already-known effect of Cobalt (Co) as a hypoxic element. Firstly, discs of CoCr [subjecting (W) or not (Wo) to dual acid-etched (DAE)] were incubated into FBS-free cell culture medium up to 24 h (37 °C). This CoCr-enriched medium was further used to treat shear-stressed endothelial cells cultures up to 72 h. Thereafter, the conditioned medium containing metabolites of shear-stressed endothelial cells in response to CoCr-enriched medium was further used to subject osteoblast's cultures, when the samples were properly harvested to allow the analysis of the molecular issues. Our data shows that CoCr-enriched medium contains 1.5 ng–2.0 ng/mL of Co, which was captured by endothelial cells and osteoblasts in about 30% in amount and it seems modulate their metabolic pathways: shear-stressed endothelial cells expressed higher profile of HIF1α, VEGF and nNOS genes, while their global profile of protein carbonylation was lower than the control cultures, suggesting lower oxidative stress commitment. Additionally, osteoblasts responding to metabolites of CoCr-challenged endothelial cells show dynamic expression of marker genes in osteogenic differentiation, with alkaline phosphatase (ALP), osteocalcin, and bone sialoprotein (BSP) genes being significantly increased. Additionally, tensional shear-stress forces decrease the stimulus for ColA1gene expression in osteoblasts responding to endothelial cells metabolites, as well as modifying the extracellular matrix remodeling related genes. Analyzing the activities of matrix metalloproteinases (MMPs), the data shows that shear-stressed endothelial cells metabolites increase the activities of both MMP9 and MMP2 in osteoblasts. Altogether, our data shows for the first time that shear-stressed endothelial metabolites responding to CoCr discs contribute to osteogenic phenotype in vitro, and this predicts an active crosstalk between angiogenesis and osteogenesis during osseointegration of CoCr alloy and bone healing, maybe guided by the Co-induced hypoxic condition.

A super-hydrophilic surface enhanced by the hierarchical reticular porous structure on a low-modulus Ti–24Nb–4Zr–8Sn alloy

ABSTRACT In this study, we fabricated a super-hydrophilic (θ < 5°) hierarchical reticular porous structure on the surface of Ti–24Nb–4Zr–8Sn (Ti2448) by combining sandblasting, dual acid-etching, and alkali thermal treatment. The super-hydrophilic hierarchical structure on the surface contained fine well-distributed nanopores with random microscale structures, leading to extremely high surface hydrophilicity. The sand blasting and dual acid-etching processes (SLA) and alkaline treatment (AT) resulted in the formation of microscale structures and nanopores, respectively. The formation mechanism of the hierarchical reticular porous structures was also examined. The average size of the nanopores on the surface of the SLA + AT Ti2448 sample was 86.3 nm, and the hierarchical structure contained several biocompatible components, including TiO2, Ti2ZrO, ZrTiO4, Nb2O5, Ti2O3, and ZrO2. Because of their super-hydrophilic and chemically biocompatible characteristics, hierarchical Ti2448 structural candidates are expected to be more suitable as implants for successful osseointegration.

PI3K/AKT signaling drives titanium-induced angiogenic stimulus

Although osseointegration and clinical success of titanium (Ti)-implanted materials depend on neovascularization in the reactional peri-implant tissue, very little has been achieved considering the Ti-molecules release on the behavior of endothelial cells. To address this issue, we challenged endothelial cells (HUVECs) with Ti-enriched medium obtained from two types of commercial titanium surfaces [presenting or not dual-acid etching (DAE)] up to 72 h to allow molecular machinery analysis. Our data show that the Ti-enriched medium provokes significant stimulus of angiogenesis-related machinery in endothelial cells by upexpressing VEGFR1, VEGFR2, VEGF, eNOS, and iNOS genes, while the PI3K/Akt signaling pathway was also significantly enhanced. As PI3K/AKT signaling was related to angiogenesis in response to vascular endothelial growth factor (VEGF), we addressed the importance of PI3K/Akt upon Ti-enriched medium responses by concomitantly treating the cells with wortmannin, a well-known PI3K inhibitor. Wortmannin suppressed the angiogenic factors, because VEGF, VEGFR1, and eNOS genes were downregulated in those cells, highlighting the importance of PI3K/AKT signaling on driving angiogenic phenotype and angiogenesis performance within the peri-implant tissue reaction. In conjunction, these data reinforce that titanium-implantable devices modify the metabolism of surrounding cells, such as endothelial cells, probably coupling osteogenesis and angiogenesis processes in peri-implant tissue and then contributing to successfully osseointegration of biomedical titanium-based devices.

VEGF/VEGF-R/RUNX2 Upregulation in Human Periodontal Ligament Stem Cells Seeded on Dual Acid Etched Titanium Disk.

In restorative dentistry, the main implants characteristic is the ability to promote the osseointegration process as the result of interaction between angiogenesis and osteogenesis events. On the other hand, implants cytocompatibility remains a necessary feature for the success of surgery. The purpose of the current study was to investigate the interaction between human periodontal stem cells and two different types of titanium surfaces, to verify their cytocompatibility and cell adhesion ability, and to detect osteogenic and angiogenic markers, trough cell viability assay (MTT), Confocal Laser Scanning Microscopy (CLSM), scanning electron microscopy (SEM), and gene expression (RT-PCR). The titanium surfaces, machined (CTRL) and dual acid etched (TEST), tested in culture with human periodontal ligament stem cells (hPDLSCs), were previously treated in two different ways, in order to evaluate the effects of CTRL and TEST and define the best implant surface. Furthermore, the average surface roughness (Ra) of both titanium surfaces, CTRL and TEST, has been assessed through atomic force microscopy (AFM). The vascular endothelial growth factor (VEGF) and Runt-related transcription factor 2 (RUNX2) expressions have been analyzed by RT-PCR, WB analysis, and confocal laser scanning microscopy. Data evidenced that the different morphology and topography of the TEST disk increased cell growth, cell adhesion, improved osteogenic and angiogenic events, as well osseointegration process. For this reason, the TEST surface was more biocompatible than the CTRL disk surface.

Implant surface alters compartmental-specific contributions to fixation strength in rats.

Mechanical fixation of the implant to host bone is an important contributor to orthopedic implant survivorship. The relative importance of bone-implant contact, trabecular bone architecture, and cortical bone geometry to implant fixation strength has never been directly tested, especially in the settings of differential implant surface properties. Thus, using a rat model where titanium rods were placed into the intramedullary canal of the distal femur, we determined the relative contribution of bone-implant contact and peri-implant bone architecture to the fixation strength in implants with different surface roughness: highly polished and smooth (as-received) and dual acid-etched (DAE) implants. Using a training set that maximized variance in implant fixation strength, we initially examined correlation between implant fixation strength and outcome parameters from microcomputed tomography and found that osseointegration volume per total volume (OV/TV), trabecular bone volume per total volume (BV/TV), and cortical thickness (Ct.Th) were the single best compartment-specific predictors of fixation strength. We defined separate regression models to predict implant fixation strength for as-received and DAE implants. When the training set models were applied to independent validation sets, we found strong correlations between predicted and experimentally measured implant fixation strength, with r2 = .843 in as received and r2 = .825 in DAE implants. Interestingly, for as-received implants, OV/TV explained more of the total variance in implant fixation strength than the other variables, whereas in DAE implants, Ct.Th had the most explanatory power, suggesting that surface topography of implants affects which bone compartment is most important in providing implant fixation strength.

A "best fit" approach for synergistic surface parameters to guide the design of candidate implant surfaces.

Human bone resorption surfaces can provide a template for endosseous implant surface design. We characterized the topography of such sites using four synergistic parameters (fractal dimension, lacunarity, porosity, and surface roughness) and compared the generated values with those obtained from two groups of candidate titanium implant surfaces. For the first group (n = 5/group): grit-blasted acid etched (BAE), BAE with either discrete calcium phosphate crystal deposition or nanotube formation, machined titanium with nanotubes, or a nanofiber surface; each measured synergistic parameter was statistically compared with that of the resorbed bone surface and scored for inclusion in a "best fit" analysis. The analysis informed changes that could be made to a candidate implant surface to render it a closer "best fit" to that of the resorbed bone surface. In a second group of either titanium or titanium alloy implants their micro-topography, created by dual acid etching, was the same for each material substrate; but their nanotopographic complexity was changed by varying the degree of calcium phosphate crystalline deposits. These implants were also used in vivo where bone anchorage was tested using a tensile disruption test; and the "best fit" of synergistic parameters coincided with the best biological outcome for both titanium and titanium alloy implants. In conclusion, the four chosen synergistic parameters can be used to guide the sub-micron surface design of candidate implants, and our "best fit" approach is capable of identifying the surfaces with the best biological outcomes. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2165-2177, 2019.

Synergistic Effects of Implant Macrogeometry and Surface Physicochemical Modifications on Osseointegration: An In Vivo Experimental Study in Sheep.

This study evaluates the influence of two surface physicochemical modifications on osseointegration parameters of a healing chamber implant design. We examine dental implants with internal and external trapezoidal threads that have the following surface modifications: dual acid etching (DAE) and nano-hydroxyapatite (HA) coating over DAE surface (Nano). We installed implants in the right ilium of sheep and conducted histologic/metric analyses after 3 and 12 wk in vivo. We quantified the percentage of bone-to-implant contact (%BIC) and bone area fraction occupancy (%BAFO) within implant threads. Histologic micrographs indicate early bone formation within the healing chambers of implants with Nano surface relative to DAE surface. Histomorphometric analysis demonstrates there to be no significant differences in %BIC between 3 and 12 wk (p = 0.298). Compared to DAE, Nano shows more bone formation in contact with implant, regardless of time (p < 0.025). We observe > %BAFO at 12 wk relative to 3 wk, which differs significantly for Nano (p < 0.038). Implant surface treatment affects the amounts of bone formation within healing chambers, with Nano significantly outperforming DAE at 12 wk (p < 0.025). Nano presents a synergistic effect with implant design, improving osseointegration parameters.

Titanium-enriched medium drives low profile of ECM remodeling as a pre-requisite to pre-osteoblast viability and proliferative phenotype

Titanium is widely used for biomedical applications, but little information is being delivered regarding the cellular/molecular mechanisms explaining their efficacy, mainly considering the effects of the Ti-released trace elements on pre-osteoblasts. We addressed this issue by investigating decisive intracellular signal transduction able to modulate cytoskeleton rearrangement, proliferative phenotype and extracellular matrix (ECM) remodeling. We considered titanium grades IV and V, submitted or not to dual acid-etching (w/DAE or wo/DAE, respectively). Our results showed there is no cytotoxicity, preserving AKT involvement. Additionally, Ti-enriched medium promoted a diminution of the downstream signaling upon integrin activation (phosphorylating Rac1 and cofilin), guaranteeing a dynamic cytoskeleton rearrangement. Moreover, the low profile of ECM remodeling obtained in response to trace molecules released by Ti-based devices seems contributing to the osteoblast performance in mediating extracellular support to cell anchorage. This hypothesis was validated by the up-expression of ß1-integrin, src and Focal adhesion kinase (fak) genes, mainly in response to titanium grade V. Proliferative phenotype showed an unbalance between cyclin-dependent kinases (CDKs) and p15INK4b/p21Cip1. In conjunction, we showed for the first time that trace elements from Ti-based biomedical devices provoke important modulation of the osteoblast biology, driving cell anchoring, viability, and proliferative phenotype. Certainly, these biological outcomes compromise implant osseointegration.

Investigation of osteogenic responses of Fe-incorporated micro/nano-hierarchical structures on titanium surfaces.

Effective and fast osseointegration is important for the survival of titanium-based orthopaedic implants. Previous studies confirmed that topographic features combined with inorganic ions showed positive effects on the biological functions of osteoblastic cells. In this study, we report an approach for fabricating Fe-incorporated micro-nano hierarchical structures on titanium substrates, which was realized by dual acid etching and subsequent hydrothermal treatment. The surface morphology, surface chemistry and wettability of the titanium substrates were characterized using scanning electron microscopy, X-ray photoelectron spectroscopy and contact angle measurements, respectively. The Fe-incorporated micro-nano hierarchical titanium substrates were probed to be biocompatible and positively improved protein adsorption, cell proliferation and cell differentiation of osteoblasts in vitro. Furthermore, the Fe-incorporated titanium substrates significantly enhanced the expressions of osteogenic genes (such as Runx2, Col I, OPN, and OCN), which were attributed to the synergistic effects of micro-nano structures and Fe ions. More importantly, the Fe-incorporated titanium implants with micro-nano hierarchical structures promoted new bone formation in vivo. This study provides an alternative for the development of orthopaedic implants with improved osseointegration for potential clinical applications.

Nano hydroxyapatite-blasted titanium surface creates a biointerface able to govern Src-dependent osteoblast metabolism as prerequisite to ECM remodeling

Over the last several years, we have focused on the importance of intracellular signaling pathways in dynamically governing the biointerface between pre-osteoblast and surface of biomaterial. Thus, this study investigates the molecular hallmarks involved in the pre-osteoblast relationship with different topography considering Machined (Mc), Dual Acid-Etching (DAE), and nano hydroxyapatite-blasted (nHA) groups. There was substantial differences in topography of titanium surface, considering Atomic Force Microscopy and water contact angle (Mc = 81.41 ± 0.01; DAE = 97.18 ± 0.01; nHA = 40.95 ± 0.02). Later, to investigate their topography differences on biological responses, pre-osteoblast was seeded on the different surfaces and biological samples were collected after 24 h (to consider adhesion signaling) and 10 days (to consider differentiation signaling). Preliminary results evidenced significant differences in morphological changes of pre-osteoblasts mainly resulting from the interaction with the DAE and nHA, distinguishing cellular adaptation. These results pushed us to analyze activation of specific genes by exploring qPCR technology. In sequence, we showed that Src performs crucial roles during cell adhesion and later differentiation of the pre-osteoblast in relationship with titanium-based biomaterials, as our results confirmed strong feedback of the Src activity on the integrin-based pathway, because integrin-ß1 (∼5-fold changes), FAK (∼12-fold changes), and Src (∼3.5-fold changes) were significantly up-expressed when Src was chemically inhibited by PP1 (5 μM). Moreover, ECM-related genes were rigorously reprogrammed in response to the different surfaces, resulting on Matrix Metalloproteinase (MMP) activities concomitant to a significant decrease of MMP inhibitors. In parallel, we showed PP1-based Src inhibition promotes a significant increase of MMP activity. Taking all our results into account, we showed for the first time nano hydroxyapatite-blasted titanium surface creates a biointerface able to govern Src-dependent osteoblast metabolism as pre-requisite to ECM remodeling

Nano hydroxyapatite-blasted titanium surface affects pre-osteoblast morphology by modulating critical intracellular pathways.

Although, intracellular signaling pathways are proposed to predict the quality of cell-surface relationship, this study addressed pre-osteoblast behavior in response to nano hydroxyapatite (HA)-blasted titanium (Ti) surface by exploring critical intracellular pathways and pre-osteoblast morphological change. Physicochemical properties were evaluated by atomic force microscopy (AFM) and wettability considering water contact angle of three differently texturized Ti surfaces: Machined (Mac), Dual acid-etching (DAE), and nano hydroxyapatite-blasted (nHA). The results revealed critical differences in surface topography, impacting the water contact angle and later the osteoblast performance. In order to evaluate the effect of those topographical characteristics on biological responses, we have seeded pre-osteoblast cells on the Ti discs for up to 4 h and subjected the cultures to biological analysis. First, we have observed pre-osteoblasts morphological changes resulting from the interaction with the Ti texturized surfaces whereas the cells cultured on nHA presented a more advanced spreading process when compared with the cells cultured on the other surfaces. These results argued us for analyzing the molecular machinery and thus, we have shown that nHA promoted a lower Bax/Bcl2 ratio, suggesting an interesting anti-apoptotic effect, maybe explained by the fact that HA is a natural element present in bone composition. Thereafter, we investigated the potential effect of those surfaces on promoting pre-osteoblast adhesion and survival signaling by performing crystal violet and immunoblotting approaches, respectively. Our results showed that nHA promoted a higher pre-osteoblast adhesion supported by up-modulating FAK and Src activations, both signaling transducers involved during eukaryotic cell adhesion. Also, we have shown Ras-Erk stimulation by the all evaluated surfaces. Finally, we showed that all Ti-texturing surfaces were able to promote osteoblast differentiation up to 10 days, when alkaline phosphatase (ALP) activity and osteogenic transcription factors were up-modulated. Altogether, our results showed for the first time that nano hydroxyapatite-blasted titanium surface promotes crucial intracellular signaling network responsible for cell adapting on the Ti-surface.Biotechnol. Bioeng. 2017;114: 1888-1898. © 2017 Wiley Periodicals, Inc.

Early Bone Response to Dual Acid-Etched and Machined Dental Implants Placed in the Posterior Maxilla

To compare the early bone response to implants with dual acid-etched (DAE) and machined (MA) surface, when placed in the posterior human maxilla. Fourteen patients received 2 implants in the posterior maxilla: 1 DAE and 1 MA. After 2 months, the implants were retrieved for histologic/histomorphometric evaluation. The bone-to-implant contact (BIC%), bone density in the threaded area (BDTA%), and the bone density (BD%) were calculated. The Wilcoxon matched-pairs signed rank test was used to evaluate differences (BIC%, BDTA%, and BD%) between the surfaces. In the MA implants, a mean (±SD) BIC%, BDTA%, and BD% of 21.76 (±12.79), 28.58 (±16.91), and 21.54 (±11.67), respectively, was reported. In the DAE implants, a mean (±SD) BIC%, BDTA%, and BD% of 37.49 (±29.51), 30.59 (±21.78), and 31.60 (±18.06), respectively, was reported. Although the mean BIC% of DAE implants value was almost double than that of MA implants, no significant differences were found between the 2 groups with regard to BIC% (P = 0.198) and with regard to BDTA% (P = 0.778) and BD% (P = 0.124). The DAE surface increased the periimplant endosseous healing properties in the native bone of the posterior maxilla.

THE EFFECT OF LOW INTENSITY ULTRASOUND THERAPY ON OSSEOINTEGRATION OF TITANIUM DENTAL IMPLANT PLACED IN THE ESTHETIC ZONE

Background: Endosseous dental implants may be considered as the most popular treatment option for partially and fully edentulous patients. It is generally accepted that implant success is primarily dependent upon or achieved by osseointegration, a direct contact between the implant surfaces and living bone and gum. For decrease the time of osseointegration there is a different modalities used as altering the surface and/or shape of the implant has been frequently researched and it has been shown that rough implant surfaces allow a higher percentage of bone-to-implant contact compared to implants with smooth surfaces. Also dual acid etching of titanium, engineering of dental pulp cells on various implant surfaces and biomimetic implant coatings containing bone morphogenetic protein-2. Other methods studied to enhance endogenous bone healing around biomaterials are different forms of biophysical stimulations such as pulsed electromagnetic fields and low intensity pulsed ultrasounds (LIPU). Ultrasound is a form of energy that is transmitted through biological tissues as high-frequency acoustic waves, which is widely used in medicine as a diagnostic, therapeutic and operative tool. The aim of the present study was to evaluate The effect of low-intensity pulsed ultrasound on the osseointegration of titanium dental implants .Patient and methods: Ten implants were placed in ten patients each one receives one implant. The patients were 6 males and 4 females ranged in ages from 25 to 40 years. All the implants were placed in maxilla. Observations were made postoperatively at time of implant placement then after one month, 3 months, 6 months and one year follow up periods for crestal bone loss and stability, Data analysis was performed in several steps. Initially, descriptive statistics for each case results. One way ANOVA followed by pair-wise Tukey post-hoc tests were performed to detect significance between preparation cases. Statistical analysis was performed using Graph-Pad Prism-4 statistics software for Windows. P values ≤0.05 are considered to be statistically significant in all tests.Conclusion: From the a aforementioned review it became clear that Low-intensity pulsed ultrasound could be accelerate the osteointegration of dental implant .

Microarray expression in peri-implant tissue next to different titanium implant surfaces predicts clinical outcomes: a split-mouth study.

This split-mouth study evaluated miRNA expression of tissues around implants with different surface treatments. Each patient of the sample (five men and five women) received two implants (one control and one test) into an edentulous quadrant to support fixed partial dentures. The control implants (Osseotite) had a dual acid-etched (DAE) surface in the apical portion and a machined coronal part, test implants (Full Osseotite, FOSS) were completely DAE. Machined healing abutments were placed on control implants and DAE abutments on test ones. All implants were assigned codes for blinding. Standardized periapical radiographs were taken at baseline, 2 and 6months, and 1year after surgery. Plaque index (PI), bleeding on probing (BOP), and probing depth (PD) were recorded at 3 and 6weeks, and 2, 3, 6, and 12months post-implant placement. After 3 months, a mini-invasive sample of soft tissue was collected from seven patients (four women and three men) for miRNA microarray analysis. Control implants showed greater bone resorption (BR) and lower PI: this was not statistically significant. No statistically significant differences in BOP and PD appeared. miRNA modulated by implant surfaces as well as by other clinical conditions has been identified. miRNA microarray analysis revealed that: (i) implant sites with low PI and absence of BOP had a miRNA expression profile similar to those with plaque and absence of BOP; sites with high PI and high BOP had a different profile. (ii) Implant sites with BOP presented similar profiles independently from implant surface. (iii) Implant sites with high PI and normal BR differed from others for miRNA expression profile. (iv) Implant sites with normal BR despite high BOP differed from others. This profile resembled that of FOSS implants. (v) Implant surface affected BR; groups having similar BR clusterized differently according to the implant type. DAE surfaces induced lower BR and more plaque accumulation: This did not affect the health of soft tissues. miRNA analysis indicated that soft tissue inflammation is more related to gene expression profile than to plaque or to implant surface. Specific miRNA profile can protect implant sites from bleeding and BR irrespective of plaque accumulation.