Effect of a gel containing green tea extract and hyaluronic acid on palate pain scores and wound healing after free gingival graft: a quasi-randomized controlled clinical trial.

OBJECTIVES: The aim of this study was to evaluate the effect of a gel containing green tea extract and hyaluronic acid (HA) on pain scores and wound healing in donor sites after free gingival graft (FGG). MATERIALS AND METHODS: Forty-two patients requiring FGG were included in three groups: (1) control group (n = 14), no material was placed in the donor area; only the clot was kept in position by sutures; (2) placebo group (n = 14), vehicle gel applied 3 times a day for 7 days; and (3) test group (n = 14), gel containing green tea extract and HA applied 3 times a day for 7 days. The wound size by clinical measurement (WS-CM) and photographic image (WS-PI), complete wound epithelialization (CWE), and palatal mucosa color were evaluated after 3 days and 1, 2, and 4 weeks postoperatively. The visual analog scale (VAS) for pain and analgesic consumption were used to assess participant's perception in the same postoperative periods. RESULTS: A similar progressive reduction in the wound size, associated with an improvement in the color pattern, was observed in all groups (p > 0.05). No significant differences were found for CWE and pain assessment between the examined groups (p > 0.05). CONCLUSION: The gel containing green tea extract and HA application in palatal wounds after FGG removal does not provide clinical healing benefits using this investigated protocol. CLINICAL RELEVANCE: This is the first clinical study evaluating the effect of gel containing green tea extract and HA on the palate postoperative pain control and wound healing after FGG. TRIAL REGISTRATION: http://clinicaltrials.gov : NCT05270161.

Rehabilitation of Atrophic Maxilla With Immediate Loading of Extrasinus Zygomatic Implant.

ABSTRACT: The aim of this case series was to evaluate the long-term success rate of immediate occlusal loading of extrasinus zygomatic dental implants after a 3-year follow-up. The sample consisted of 31 patients (mean age of 64 years) with atrophic maxillae rehabilitated with 1 to 4 extrasinus zygomatic implants, placed unilaterally or bilaterally. All the patients received complete implant-supported dental prostheses with immediate loading by associating zygomatic implants with conventional implants. None of the procedures were associated with bone grafts. During the 3-year period of follow-up in the present study, all the patients attended clinical sessions and underwent radiographic exams every 6 months. In total 55 zygomatic and 69 conventional implants were placed, where 1 zygomatic and 2 conventional implants were lost, representing success rates of 98.18% and 97.20%, respectively. None of the studied patients had signs of sinusitis or changes in the maxillary sinuses. All the patients showed occlusal contact on natural antagonist teeth or implant-supported dental prostheses. Therefore, it was concluded that the use of exteriorized zygomatic implants with immediate loading represented a feasible option with high success rates for the treatment of atrophic maxilla.

Effect of osseodensification on the increase in ridge thickness and the prevention of buccal peri-implant defects: an in vitro randomized split mouth pilot study.

BACKGROUND: Implant installation with conventional drilling can create buccal bone defects in areas of limited ridge thickness. Implant installation with osseodensification may aid in preventing buccal bone defects in these situations. This in vitro pilot study evaluated the impact of osseodensification on the increase in alveolar ridge thickness and the prevention of buccal peri-implant defects. METHODS: Ten fresh pig mandibles with limited bone thickness were selected for use in an experimental randomized split mouth pilot study. Two site-preparation protocols were used: conventional drilling with cutting burs (CTL, n = 10) and osseodensification with Densah® burs (OD, n = 10). After implant bed preparation, 20 implants (4.5 × 10 mm) were placed in the prepared sites and the insertion torque was recorded. Clinical and photographic analysis evaluated ridge thickness and the extent (height, width, and area) of bone defects in the buccal and lingual bone walls following implant placement. Three-dimensional measurements were performed using STL files to analyze the increase in buccal ridge thickness following site preparation and implant placement. The height of the buccal bone defect was considered as the primary outcome of this study. Defect width, area, implant insertion torque, and linear buccal ridge increase after implant site preparation and installation were also assessed. Non-parametric evaluations were carried out with the Mann-Whitney test to verify intergroup differences. RESULTS: There was no statistically significant difference between groups in the baseline ridge thickness. OD presented a significantly higher insertion torque, associated with reduced buccal and lingual bone defect width, in comparison to CTL. CONCLUSIONS: The increase in buccal ridge thickness after site preparation and implant placement was significantly higher in OD compared to CTL. Osseodensification increased the ridge thickness through expansion and reduced buccal bone defects after implant installation.

The Impact of Surgical Guide Fixation and Implant Location on Accuracy of Static Computer-Assisted Implant Surgery.

PURPOSE: To evaluate the accuracy of static computer-assisted implant surgery (sCAIS) for tooth-supported free-end dental implantation with the aid/and without the aid of fixation pins to secure the surgical template through comparison between planned, 3D printed guide position and placement implant position. MATERIALS AND METHODS: Thirty-two duplicated maxillary resin models were used in the present in vitro study. Digital planning was performed and fabrication of a surgical template that allowed implant placement on the distal extension edentulous site of the model (maxillary left side). A first optical scan was performed after fitting the surgical template on the model to assess the deviation at the surgical guide level. After placing implants in the model using the surgical guide, scan bodies were attached to the implants, and a second scan was performed to record the position of placed implants. The digital representations were later superimposed to the pre-operative scan and measurements of implant deviations were performed. Global (coronal and apical), horizontal (coronal and apical), depth and angular deviations were recorded between planned implant position, guide position, and placement implant position. Three-way ANOVA was used to compare implant location (#13, 14, and 15), fixation pin (with or without pin), and guide comparison (planned, guided, and placement). RESULTS: Final implant placement based on the digital plan and based on the 3D printed guide were very similar except for depth deviation. Use of fixation pin had a statistically significant effect on the depth and angular deviation. Overall, without fixation pins and based on guide versus placement, mean global coronal (0.88 ± 0.36 mm), horizontal coronal (0.55 ± 0.32 mm), and apical (1.44 ± 0.75 mm), and angular deviations (4.28 ± 2.01°) were similar to deviations with fixation pins: mean global coronal (0.88 ± 0.36 mm); horizontal coronal (0.67 ± 0.22 mm) and apical (1.60 ± 0.69 mm); and angular deviations (4.53 ± 2.04°). Horizontal apical without pins (1.63 ± 0.69 mm) and with fixation pins (1.72 ± 0.70 mm) was statistically significant (p = 0.044). Depth deviation without pins (-0.5 ± 0.5 mm) and with fixation pins (-0.16 ± 0.62 mm) was also statistically significant (p = 0.005). Further analysis demonstrated that the final sleeve position on the 3D printed guide was on average 0.5 mm more coronal than the digital plan. CONCLUSIONS: The use of surgical guides with or without fixation pins can provide clinically acceptable outcomes in terms of accuracy in implant position. There was a statistically significant difference in the accuracy of implant position when utilizing fixation pins only for horizontal apical and depth deviation. Additionally, a statistically significant difference between the planned and the 3D printed surgical guide when considering the sleeve position was detected.

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.

Monosodium urate crystal interleukin-1ß release is dependent on Toll-like receptor 4 and transient receptor potential V1 activation.

OBJECTIVE: The present study aimed to elucidate the mechanisms involved in MSU-induced IL-1ß release in a rodent animal model of acute gout arthritis. METHODS: Painful (mechanical and thermal hypersensitivity, ongoing pain and arthritis score) and inflammatory (oedema, plasma extravasation, cell infiltration and IL-1ß release) parameters were assessed several hours after intra-articular injection of MSU (100 µg/articulation) in wild-type or knockout mice for Toll-like receptor 4 (TLR4), inducible nitric oxide synthase (iNOS), transient receptor potential (TRP) V1 and the IL-1 receptor (IL-1R). Also, wild-type animals were treated with clodronate, lipopolysaccharide from Rhodobacter sphaeroides (LPS-RS) (TLR4 antagonist), spleen tyrosine kinase (SYK) inhibitor (iSYK), aminoguanidine (AMG, an iNOS inhibitor) or SB366791 (TRPV1 antagonist). Nitrite/nitrate and IL-1ß levels were measured on the synovial fluid of wild-type mice, 2 h after intra-articular MSU injections, or medium from macrophages stimulated for MSU (1000 µg) for 2 h. RESULTS: Intra-articular MSU injection caused robust nociception and severe inflammation from 2 up to 6 h after injection, which were prevented by the pre-treatment with clodronate, LPS-RS, iSYK, AMG and SB366791, or the genetic ablation of TLR4, iNOS, TRPV1 or IL-1R. MSU also increased nitrite/nitrate and IL-1ß levels in the synovial fluid, which was prevented by clodronate, LPS-RS, iSYK and AMG, but not by SB366791. Similarly, MSU-stimulated peritoneal macrophages released nitric oxide, which was prevented by LPS-RS, iSYK and AMG, but not by SB366791, and released IL-1ß, which was prevented by LPS-RS, iSYK, AMG and SB366791. CONCLUSION: Our data indicate that MSU may activate TLR4, SYK, iNOS and TRPV1 to induce the release of IL-1ß by macrophages, triggering nociception and inflammation during acute gout attack.

Effects of bovine serum albumin (BSA) on the excited-state properties of meso-tetrakis(sulfonatophenyl) porphyrin (TPPS4).

To infer changes in the photophysical properties of porphyrins due to complexation with albumin, a combination of Z-scan and conventional spectroscopic techniques was employed. We measured the characteristics of excited states of meso-tetrakis(sulfonatophenyl) porphyrin bound to bovine serum albumin and observed that the binding reduces the intersystem crossing quantum yield and increases the internal conversion one. A reverse saturable absorption process was observed in the nanosecond timescale. These results are important for prediction of the efficiency of this complex in medical and optical applications, because associating porphyrins to proteins enables better accumulation in tumors and improves its stability in optical devices, but at the same time, decreases its triplet quantum yield.

Photodynamic inactivation of Bovine herpesvirus type 1 (BoHV-1) by porphyrins.

In this work, the photodynamic efficiency of anionic meso-tetrakis sulfonophenyl (TPPS4), cationic meso-tetrakis methylpyridiniumyl (TMPyP) and their zinc complexes (ZnTPPS4 and ZnTMPyP) in the inactivation of Bovine herpesvirus type 1 (BoHV-1) was evaluated. At a non-cytotoxic concentration, all porphyrins showed significant antiviral activity after irradiation using a halogen lamp. The efficiency of the cationic porphyrins was higher than that of the anionic ones. Porphyrin complexation with zinc increases its lipophilicity and the number of absorbed photons, dramatically reducing the time for complete virus inactivation. The high superposition of the compound optical absorption and light source emission spectra played a key role in the virus inactivation efficiency. The results demonstrated the high effectivity of the photodynamic inactivation of BoHV-1. This method can be used as an auxiliary in the treatment of disorders attributed to BoHV-1 infection, and the porphyrins are promising photosensitizers for this application.

Zirconia stimulates ECM-remodeling as a prerequisite to pre-osteoblast adhesion/proliferation by possible interference with cellular anchorage.

The biological response to zirconia (ZrO2) is not completely understood, which prompted us to address its effect on pre-osteoblastic cells in both direct and indirect manner. Our results showed that zirconia triggers important intracellular signaling mainly by governing survival signals which leads to cell adhesion and proliferation by modulating signaling cascade responsible for dynamic cytoskeleton rearrangement, as observed by fluorescence microscopy. The phosphorylations of Focal Adhesion Kinase (FAK) and Rac1 decreased in response to ZrO2 enriched medium. This corroborates the result of the crystal violet assay, which indicated a significant decrease of pre-osteoblast adhesion in responding to ZrO2 enriched medium. However, we credit this decrease on pre-osteoblast adhesion to the need to govern intracellular repertory of intracellular pathways involved with cell cycle progression, because we found a significant up-phosphorylation of Mitogen-Activated Protein Kinase (MAPK)-p38 and Cyclin-dependent kinase 2 (CDK2), while p15 (a cell cycle suppressor) decreased. Importantly, Protein phosphatase 2 A (PP2A) activity decreased, guaranteeing the significant up-phosphorylation of MAPK -p38 in response to ZrO2 enriched medium. Complementarily, there was a regulation of Matrix Metalloproteinases (MMPs) in response to Zirconia and this remodeling could affect cell phenotype by interfering on cell anchorage. Altogether, our results show a repertory of signaling molecules, which suggests that ECM remodel as a pre-requisite to pre-osteoblast phenotype by affecting their anchoring in responding to zirconia.

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.

Use of Spectroscopic Techniques for Evaluating the Coupling of Porphyrins on Biocompatible Nanoparticles. A Potential System for Photodynamics, Theranostics, and Nanodrug Delivery Applications.

Modern medicine has been searching for new and more efficient strategies for diagnostics and therapeutics applications. Considering this, porphyrin molecules have received great interest for applications in photodiagnostics and phototherapies, even as magnetic nanoparticles for drug-delivery systems and magnetic-hyperthermia therapy. Aiming to obtain a multifunctional system, which combines diagnostics with therapeutic functions on the same platform, the present study employed UV/vis absorption and fluorescence spectroscopies to evaluate the interaction between meso-tetrakis(p-sulfonatofenyl)porphyrin (TPPS) and maghemite nanoparticles (γ-Fe2O3). These spectroscopic techniques allowed us to describe the dynamics of coupling porphyrins on nanoparticles and estimate the number of 21 porphyrins per nanoparticle. Also, the binding parameters, such as the association constants (Ka = 8.89 × 105 M-1) and bimolecular quenching rate constant (kq = 2.54 × 1014 M-1 s-1) were obtained. These results suggest a static quenching process where the electrostatic attraction plays an essential role. The work shows that spectroscopic techniques are powerful tools to evaluate the coupling of organic molecules and nanoparticles. Besides, the system studied provides a relevant background for potential applications in bionanotechnology and nanomedicine, such as (1) nanodrug delivery system, (2) photodiagnostics/theranostics, and/or (3) a combined action of photodynamic and hyperthermia therapies, working in a synergetic way.

A Technical Note on a Modified Subcrestal Buccal Flap Approach to All-on-Four: A Case Report.

This case report describes a dual full-arch rehabilitation focusing on a modified buccal incision for installation of four implants for full-arch rehabilitation of an edentulous maxilla. A modified buccal incision was performed in the subcrestal buccal region to promote direct access to the periosteum without incising the muscles in the region. For the installation of anterior implants, an 8.5 mm implant was locked in the cortical bone of the alveolar ridge and in the cortical bone of the floor of the pyriform cavity. The drilling point of the posterior implants was defined using the anterior implants as a visual reference, and the entry point could be visually estimated from the topography of the palatal surface of the maxilla. After bone leveling, the drilling enlargement sequence was carried out using drills that allowed the installation of long implants (18 mm). Straight mini-abutments were installed in the anterior implants and angled at 30º in the posterior implants. The flap was then perforated in the exact region where the mini-abutments were located. The buccal incision line was sutured with continuous 5-0 nylon suture. On the following day, aesthetic tests were carried out with teeth mounting. The patient presented minimal edema, and the lip motricity and smile width were completely preserved. The prosthesis was delivered five days after surgery. The suture was removed, and the prosthesis was installed while maintaining compression on the gingival tissue. The patient reported no pain during the prosthesis installation. The modified buccal flap enables implant placement for full-arch rehabilitation of an edentulous maxilla.

Survival analysis of dental implants placed in horizontally severely resorbed maxillae after reconstruction with xenogeneic graft: a case series.

PURPOSE: To evaluate the survival rates of dental implants with a hybrid macrostructure and the surface biomimetically coated with nanohydroxyapatite, placed in horizontally atrophic maxillae previously submitted to the guided bone regeneration (GBR) procedure, associated with the use of a deproteinized bovine bone graft (DBB). METHODS: Twenty-five patients who received 196 implants were involved in this study. First, these patients were submitted to GBR procedures and maxillary sinus lift, where DBB was used as the grafting material. The dental implants were placed after a minimum period of 6 months of the grafting procedures. The patients were followed up every six months and clinical/radiographic examinations were performed to assess the implants, using the following indicators as a reference: (1) Absence of mobility; (2) Absence of pain. Data about the age, surgery time, smoking status, implant size, and time between the grafting procedure and implant placement were correlated with implant failures. RESULTS: Twelve implants failed, generating a survival rate of 94.23%. None of the variables analysed correlated with the implant failures. CONCLUSION: Implants with a hybrid macrostructure and surface biomimetically coated with nanohydroxyapatite present good survival rates in horizontally atrophic maxillae grafted with DBB.

Mechanosignaling-related angiocrine factors drive osteoblastic phenotype in response to zirconia.

BACKGROUND: The growing use of zirconia as a ceramic material in dentistry is attributed to its biocompatibility, mechanical properties, esthetic appearance, and reduced bacterial adhesion. These favorable properties make ceramic materials a viable alternative to commonly used titanium alloys. Mimicking the physiological properties of blood flow, particularly the mechanosignaling in endothelial cells (ECs), is crucial for enhancing our understanding of their role in the response to zirconia exposure. METHODS: In this study, EC cultures were subjected to shear stress while being exposed to zirconia for up to 3 days. The conditioned medium obtained from these cultures was then used to expose osteoblasts for a duration of 7 days. To investigate the effects of zirconia on osteoblasts, we examined the expression of genes associated with osteoblast differentiation, including Runx2, Osterix, bone sialoprotein, and osteocalcin genes. Additionally, we assessed the impact of mechanosignaling-related angiocrine factors on extracellular matrix (ECM) remodeling by measuring the activities of matrix metalloproteinases 2 and 9 (MMP2 and MMP9) during the acquisition of the osteogenic phenotype, which precedes mineralization. RESULTS: Our data revealed that mechanosignaling-related angiocrine factors play a crucial role in promoting an osteoblastic phenotype in response to zirconia exposure. Specifically, exposed osteoblasts exhibited significantly higher expression levels of genes associated with osteoblast differentiation, such as Runx2, Osterix, bone sialoprotein, and osteocalcin genes. Furthermore, the activities of MMP2 and MMP9, which are involved in ECM remodeling, were modulated by mechanosignaling-related angiocrine factors. This modulation is likely an initial event preceding the mineralization phase. CONCLUSION: Based on our findings, we propose that mechanosignaling drives the release of angiocrine factors capable of modulating the osteogenic phenotype at the biointerface with zirconia. This process creates a microenvironment that promotes wound healing and osseointegration. Moreover, these results highlight the importance of considering the mechanosignaling of endothelial cells in the modulation of bone healing and osseointegration in the context of blood vessel effects. Our data provide new insights and open avenues for further investigation into the influence of mechanosignaling on bone healing and the osseointegration of dental devices.

Green tea and hyaluronic acid gel enhance fibroblast activation and improves the gingival healing post-third molar extraction.

This study evaluates the effects of a green tea (Camellia sinensis) and hyaluronic acid gel on fibroblast activity and alveolar bone repair following third molar extractions. By examining the gene expression related to cell survival, proliferation, and angiogenesis, the study bridges in vitro findings with clinical outcomes in a split-mouth randomized trial. Human fibroblasts were exposed to the treatment gel, analysing gene expression through RT-qPCR. Twenty participants undergoing bilateral third molar extractions received the test gel on one side and a placebo on the other. Assessments included patient-reported outcomes, professional evaluations, and radiographic analyses at multiple postoperative intervals. The test gel significantly enhanced AKT, CDKs, and VEGF gene expressions, indicating a positive effect on angiogenesis and cell proliferation. Clinically, it resulted in reduced exudate, swelling, and secondary interventions, with radiographs showing improved alveolar bone density after 90 days. The green tea and hyaluronic acid gel significantly improves soft tissue and bone healing post-extraction, offering a promising adjunctive therapy for enhancing postoperative recovery. This gel represents a novel adjuvant treatment option for facilitating improved healing outcomes after third molar extractions, highlighting its potential utility in clinical dental practice.

Nanohydroxyapatite-Coated Titanium Surface Increases Vascular Endothelial Cells Distinct Signaling Responding to High Glucose Concentration.

AIM: The success of dental implants depends on osseointegration can be compromised by well-known related adverse biological processes, such as infection and diabetes. Previously, nanohydroxyapatite-coated titanium surfaces (nHA_DAE) have been shown to contain properties that promote osteogenesis by enhancing osteoblast differentiation. In addition, it was hypothesized to drive angiogenesis in high-glucose microenvironments, mimicking diabetes mellitus (DM). On the other hand, the null hypothesis would be confirmed if no effect was observed in endothelial cells (ECs). MATERIALS AND METHODS: Titanium discs presenting the differential surfaces were previously incubated in an FBS-free cell culture medium for up to 24 h, which was, thereafter, supplemented with 30.5 mM of glucose to expose human umbilical vein endothelial cells (HUVECs, ECs) for 72 h. They were then harvested, and the sample was processed to provide molecular activity of specific genes related to EC survival and activity by using qPCR, and the conditioned medium by ECs was used to evaluate the activity of matrix metalloproteinases (MMPs). RESULTS: Our data guaranteed better performance of this nanotechnology-involved titanium surface to this end once the adhesion and survival characteristics were ameliorated by promoting a higher involvement of ß1-Integrin (~1.5-fold changes), Focal Adhesion Kinases (FAK; ~1.5-fold changes) and SRC (~2-fold changes) genes. This signaling pathway culminated with the cofilin involvement (~1.5-fold changes), which guaranteed cytoskeleton rearrangement. Furthermore, nHA_DAE triggered signaling that was able to drive the proliferation of endothelial cells once the cyclin-dependent kinase gene was higher in response to it, while the P15 gene was significantly down-regulated with an impact on the statement of angiogenesis. CONCLUSIONS: Altogether, our data show that a nanohydroxyapatite-coated titanium surface ameliorates the EC performance in a high-glucose model in vitro, suggesting its potential application in DM patients.

In vivo osseointegration evaluation of implants coated with nanostructured hydroxyapatite in low density bone.

OBJECTIVE: This in vivo study, aimed to biomechanically, histomorphometrically and histologically evaluate an implant surface coated with nanostructured hydroxyapatite using the wet chemical process (biomimetic deposition of calcium phosphate coating) when compared to a dual acid-etching surface. MATERIAL AND METHODS: Ten sheep (2-4 years old) received 20 implants, 10 with nanostructured hydroxyapatite coating (HAnano), and 10 with dual acid-etching surface (DAA). The surfaces were characterized with scanning electron microscopy and energy dispersive spectroscopy; insertion torque values and resonance frequency analysis were measured to evaluate the primary stability of the implants. Bone-implant contact (BIC) and bone area fraction occupancy (BAFo) were evaluated 14 and 28 days after implant installation. RESULTS: The HAnano and DAA groups showed no significant difference in insertion torque and resonance frequency analysis. The BIC and BAFo values increased significantly (p<0.05) over the experimental periods in both groups. This event was also observed in BIC value of HAnano group. The HAnano surface showed superior results compared to DAA after 28 days (BAFo, p = 0.007; BIC, p = 0.01). CONCLUSION: The results suggest that the HAnano surface favors bone formation when compared to the DAA surface after 28 days in low-density bone in sheep.

The Action of Angiocrine Molecules Sourced from Mechanotransduction-Related Endothelial Cell Partially Explain the Successful of Titanium in Osseointegration.

Since Branemark's findings, titanium-based alloys have been widely used in implantology. However, their success in dental implants is not known when considering the heterogenicity of housing cells surrounding the peri-implant microenvironment. Additionally, they are expected to recapitulate the physiological coupling between endothelial cells and osteoblasts during appositional bone growth during osseointegration. To investigate whether this crosstalk was happening in this context, we considered the mechanotransduction-related endothelial cell signaling underlying laminar shear stress (up to 3 days), and this angiocrine factor-enriched medium was harvested further to use exposing pre-osteoblasts (pOb) for up to 7 days in vitro. Two titanium surfaces were considered, as follows: double acid etching treatment (w_DAE) and machined surfaces (wo_DAE). These surfaces were used to conditionate the cell culture medium as recommended by ISO10993-5:2016, and this titanium-enriched medium was later used to expose ECs. First, our data showed that there is a difference between the surfaces in releasing Ti molecules to the medium, providing very dynamic surfaces, where the w_DAE was around 25% higher (4 ng/mL) in comparison to the wo_DAE (3 ng/mL). Importantly, the ECs took up some of this titanium content for up to 3 days in culture. However, when this conditioned medium was used to expose pOb for up to 7 days, considering the angiocrine factors released from ECs, the concentration of Ti was lesser than previously reported, reaching around 1 ng/mL and 2 ng/mL, respectively. Thereafter, pOb exposed to this angiocrine factor-enriched medium presented a significant difference when considering the mechanosignaling subjected to the ECs. Shear-stressed ECs showed adequate crosstalk with osteoblasts, stimulating the higher expression of the Runx2 gene and driving higher expressions of Alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin. Mechanotransduction-related endothelial cell signaling as a source of angiocrine molecules also stimulated the higher expression of the Col3A1 gene in osteoblasts, which suggests it is a relevant protagonist during trabecular bone growth. In fact, we investigated ECM remodeling by first evaluating the expression of genes related to it, and our data showed a higher expression of matrix metalloproteinase (MMP) 2 and MMP9 in response to mechanosignaling-based angiocrine molecules, independent of considering w_DAE or the wo_DAE, and this profile reflected on the MMP2 and MMP9 activities evaluated via gelatin-based zymography. Complimentarily, the ECM remodeling seemed to be a very regulated mechanism in mature osteoblasts during the mineralization process once both TIMP metallopeptidase inhibitor 1 and 2 (TIMP1 and TIMP2, respectively) genes were significantly higher in response to mechanotransduction-related endothelial cell signaling as a source of angiocrine molecules. Altogether, our data show the relevance of mechanosignaling in favoring ECs' release of bioactive factors peri-implant, which is responsible for creating an osteogenic microenvironment able to drive osteoblast differentiation and modulate ECM remodeling. Taking this into account, it seems that mechanotransduction-based angiocrine molecules explain the successful use of titanium during osseointegration.

Evaluation of Implant Surface Modification with Nanohydroxyapatite Associated with the Use of L-PRF: In Vivo Study in Rats.

Leukocyte-platelet-rich fibrin (L-PRF) contains growth factors that stimulate bone regeneration. This study evaluated the bone repair in a tibia rat model around two implant surfaces in combination or not with L-PRF by assessing microtomographic and histomorphometric parameters. A total of 48 female rats were used in the study, in which 24 received implants with two types of surface treatments (dual acid etched-DAE or nanohydroxyapatite-nanoHA), and the other 24 received the same mini implants with L-PRF, which was collected by cardiac puncture, centrifugated, and inserted in the bone bed. The animals were euthanized 7 and 30 days after implant placement, and the retrieved samples were prepared for microtomographic and histomorphometric (bone-to-implant contact-BIC; and Bone Area Fraction Occupancy-BAFO) analyses. The adhesion of the nanoHA surface onto the implant surface was investigated by insertion and removal in simulated bone medium (Sawbones). The adhesion evaluation revealed that the loss of nanoHA after this procedure (as measured with SEM) from the implant surface was less than 1%. Overall, the nanoHA surface presented more bone in contact and in proximity to the implant, a higher bone surface/tissue volume fraction, a higher number of bone trabeculae, as well as trabecular separation relative to the DAE surface. Such results were more evident when the nanoHA surface was combined with L-PRF and after 30 days in vivo. The nanoHA surface presented higher BAFO when compared to DAE, with or without association with L-PRF. Therefore, implants with a nanoHA surface potentially benefit from the association to L-PRF.