Saliva exposure reduces gingival keratinocyte growth on TiO2-coated titanium.

Bioactive, nanoporous TiO2-coating has been shown to enhance cell attachment on titanium implant surface. The aim of this study was to evaluate, whether the saliva proteins affect the epithelial cell adhesion on TiO2-coated and non-coated titanium. Grade V titanium discs were polished. Half of the discs were provided with TiO2-coating produced in sol with polycondensation method. Half of the TiO2-coated and non-coated discs were treated with pasteurized saliva for 30 min. After saliva treatment, the total protein amounts on surfaces were measured. Next, the hydrophilicity of discs were measured with water contact angle measurements. Further, the gingival keratinocyte adhesion strength was measured after 2 and 6 h of cultivation using serial trypsinization. In addition, cell growth and proliferation were measured after 1, 3, and 7 days of cell culture. Finally, cell morphology, spreading and adhesion protein signals were detected with high resolution confocal microscopy. As a result, in sol coated TiO2-surface had significantly higher hydrophilicity when compared to non-coated titanium, meanwhile both non-coated and TiO2-coated surfaces with saliva treatment had a significant increase in hydrophilicity. Importantly, the amounts of adhered saliva proteins were equal between TiO2-coated and non-coated surfaces. Adhesion strength against enzymatic detachment was weakest on non-coated titanium after saliva exposure. Cell proliferation and cell spreading were highest on TiO2-coated titanium, but saliva exposure significantly decreased cell proliferation and spreading on TiO2-coated surface. To conclude, even though saliva exposure makes titanium surfaces more hydrophilic, it seems to neutralize the bioactive TiO2-coating and decrease cell attachment to TiO2-coated surface.

Effect of TiO2 Abutment Coatings on Peri-Implant Soft Tissue Behavior: A Systematic Review of In Vivo Studies.

Establishing a proper soft tissue adhesion around the implant abutment is essential to prevent microbial invasion, inhibit epithelial downgrowth, and obtain an optimal healing process. This systematic review aims to evaluate the real potential of TiO2 coating on the behavior of peri-implant soft tissue health and maintenance. A specific aim was to evaluate clinically and histologically the effect of TiO2 abutment coating on epithelial and connective tissue attachment. Electronic database searches were conducted from 1990 to 2023 in MEDLINE/PubMed and the Web of Science databases. In total, 15 out of 485 publications were included. Eight studies involved humans, and seven were animal studies. Exposure time ranges from 2 days to 5 years. The peri-implant soft tissue evaluations included clinical assessment (plaque index (PI), peri-implant probing pocket depth (PPD), and bleeding on probing (BoP)), histological as well as histomorphometric analysis. The Office of Health Assessment and Translation (OHAT) Risk of Bias Rating Tool for Human and Animal Studies was used to evaluate the overall quality of the studies included in the review. The results showed some variation but remained within acceptable limits. Within the limitations of this systematic review, the present findings suggest that TiO2 coatings seem to influence soft tissue healing. TiO2-coated abutments with a roughness value between 0.2 and 0.5 µm enhance soft tissue health. Sol-gel-derived TiO2 coatings induced better soft tissue attachment than noncoated machined abutment surfaces. The anodized titanium abutments demonstrate comparable clinical and histological outcomes to conventional machined abutments. However, there was variation among the included studies concerning TiO2 coating characteristics and the measured outcomes used to evaluate the soft tissue response, and therefore, quantitative analysis was not feasible. Long-term in vivo studies with standardized soft tissue analysis and coating surface parameters are necessary before a definitive conclusion can be drawn. OSF Registration No.: 10.17605/OSF.IO/E5RQV.

Osteoblast Attachment on Bioactive Glass Air Particle Abrasion-Induced Calcium Phosphate Coating.

Air particle abrasion (APA) using bioactive glass (BG) effectively decontaminates titanium (Ti) surface biofilms and the retained glass particles on the abraded surfaces impart potent antibacterial properties against various clinically significant pathogens. The objective of this study was to investigate the effect of BG APA and simulated body fluid (SBF) immersion of sandblasted and acid-etched (SA) Ti surfaces on osteoblast cell viability. Another goal was to study the antibacterial effect against Streptococcus mutans. Square-shaped 10 mm diameter Ti substrates (n = 136) were SA by grit blasting with aluminum oxide particles, then acid-etching in an HCl-H2SO4 mixture. The SA substrates (n = 68) were used as non-coated controls (NC-SA). The test group (n = 68) was further subjected to APA using experimental zinc-containing BG (Zn4) and then mineralized in SBF for 14 d (Zn4-CaP). Surface roughness, contact angle, and surface free energy (SFE) were calculated on test and control surfaces. In addition, the topography and chemistry of substrate surfaces were also characterized. Osteoblastic cell viability and focal adhesion were also evaluated and compared to glass slides as an additional control. The antibacterial effect of Zn4-CaP was also assessed against S. mutans. After immersion in SBF, a mineralized zinc-containing Ca-P coating was formed on the SA substrates. The Zn4-CaP coating resulted in a significantly lower Ra surface roughness value (2.565 µm; p < 0.001), higher wettability (13.35°; p < 0.001), and higher total SFE (71.13; p < 0.001) compared to 3.695 µm, 77.19° and 40.43 for the NC-SA, respectively. APA using Zn4 can produce a zinc-containing calcium phosphate coating that demonstrates osteoblast cell viability and focal adhesion comparable to that on NC-SA or glass slides. Nevertheless, the coating had no antibacterial effect against S. mutans.

Bioactive Glasses in Periodontal Regeneration: A Systematic Review.

Bioactive glasses (BAGs) are surface-active ceramic materials that can be used in bone regeneration due to their known osteoconductive and osteoinductive properties. This systematic review aimed to study the clinical and radiographic outcomes of using BAGs in periodontal regeneration. The selected studies were collected from PubMed and Web of Science databases, and included clinical studies investigating the use of BAGs on periodontal bone defect augmentation between January 2000 and February 2022. The identified studies were screened using Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. A total of 115 full-length peer-reviewed articles were identified. After excluding duplicate articles between the databases and applying the inclusion and exclusion criteria, 14 studies were selected. The Cochrane risk of bias tool for randomized trials was used to assess the selected studies. Five studies compared using BAGs with open flap debridement (OFD) without grafting materials. Two of the selected studies were performed to compare the use of BAGs with protein-rich fibrin, one of which also included an additional OFD group. Also, one study evaluated BAG with biphasic calcium phosphate and used a third OFD group. The remaining six studies compared BAG filler with hydroxyapatite, demineralized freeze-dried bone allograft, autogenous cortical bone graft, calcium sulfate ß-hemihydrate, enamel matrix derivatives, and guided tissue regeneration. This systematic review showed that using BAG to treat periodontal bone defects has beneficial effects on periodontal tissue regeneration. OSF Registration No.: 10.17605/OSF.IO/Y8UCR.

Influence of Surface Characteristics of TiO2 Coatings on the Response of Gingival Cells: A Systematic Review of In Vitro Studies.

The soft tissue-implant interface requires the formation of epithelium and connective tissue seal to hinder microbial infiltration and prevent epithelial down growth. Nanoporous titanium dioxide (TiO2) surface coatings have shown good potential for promoting soft tissue attachment to implant surfaces. However, the impact of their surface properties on the biological response of gingival cells needs further investigation. This systematic review aimed to investigate the cellular behavior of gingival cells on TiO2-implant abutment coatings based on in vitro studies. The review was performed to answer the question: "How does the surface characteristic of TiO2 coatings influence the gingival cell response in in vitro studies?". A search in MEDLINE/PubMed and the web of science databases from 1990 to 2022 was performed using keywords. A quality assessment of the studies selected was performed using the SciRAP method. A total of 11 publications were selected from the 289 studies that fulfilled the inclusion criteria. The mean reporting and methodologic quality SciRAP scores were 82.7 ± 6.4/100 and 87 ± 4.2/100, respectively. Within the limitations of this in vitro systematic review, it can be concluded that the TiO2 coatings with smooth nano-structured surface topography and good wettability improve gingival cell response compared to non-coated surfaces.

Bioactive glass air-abrasion promotes healing around contaminated implant surfaces surrounded by circumferential bone defects: An experimental study in the rat.

OBJECTIVES: The present study aimed to evaluate the healing of experimentally induced bone defects around contaminated dental implants after air-abrasion using 45S5 or zinc oxide (ZnO)-containing bioactive glasses (BAGs). MATERIALS AND METHODS: One maxillary first molar was extracted from each Sprague-Dawley rat (n = 30). After 4-week healing, a titanium implant was placed in the extraction site with a circumferential bone defect. The rats were randomized into five different groups: (1) implants with Fusobacterium nucleatum and Porphyromonas gingivalis dual-species biofilm (IB); (2) implants with biofilm subjected to inert glass air-abrasion (inert); (3) sterile implants (S); (4) implants with biofilm subjected to 45S5 BAG air-abrasion (45S5); and (5) implants with biofilm subjected to ZnO-containing BAG air-abrasion (Zn4). After 8-week healing, maxillae were dissected, and histomorphometric analyses were performed. RESULTS: The first bone-to-implant contact was significantly shorter for the inert (1.58 ± 1.16 mm; p = 0.016), S (0.28 ± 0.13 mm; p < 0.001), 45S5 (0.41 ± 0.28 mm; p < 0.001), and Zn4 (0.26 ± 0.16 mm; p < 0.001) groups compared to IB group. Also, significantly more bone-to-implant contact was seen for S (72.35% ± 8.32%; p < 0.001), 45S5 (57.91% ± 24.10%; p = 0.002), and Zn4 (70.49% ± 12.74%; p < 0.001) groups than the IB group. The bone volume with the threads demonstrated significantly higher value for S (69.32% ± 9.15%; p < 0.001), 45S5 (58.93% ± 23.53%; p = 0.001), and Zn4 (68.65% ± 12.41%; p < 0.001) groups compared to the IB group. The bone volume within the defects was significantly higher for S (68.79% ± 11.77%; p < 0.001), 45S5 (62.51% ± 20.51%; p = 0.002), and Zn4 (73.81% ± 15.07%; p < 0.001) groups compared to the IB group. CONCLUSIONS: This study suggests that air-abrasion of contaminated moderately rough implant surfaces with either 45S5 or ZnO-containing BAGs enhances osseointegration and bone defect regeneration.

Organotypic in vitro block culture model to investigate tissue-implant interface. An experimental study on pig mandible.

In vitro studies of implant-tissue attachment are primarily based on two-dimensional cell culture models, which fail to replicate the three-dimensional native human oral mucosal tissue completely. Thus, the present study aimed to describe a novel tissue culture model using pig mandibular block including alveolar bone and gingival soft tissues to evaluate the tissue attachment to titanium implant provided with hydrothermally induced TiO2 coating. Tissue attachment on TiO2 coated and non-coated implants were compared. Ti-6Al-4V alloy posts were used to function as implants that were inserted in five pig mandibles. Implants were delivered with two different surface treatments, non-coated (NC) titanium and hydrothermal induced TiO2 coated surfaces (HT). The tissue-implant specimens were cultured at an air/liquid interface for 7 and 14 days. The tissue-implant interface was analyzed by histological and immunohistochemical stainings. The microscopic evaluation suggests that pig tissue explants established soft and hard tissue attachment to both implant surfaces. The epithelial cells appeared to attach to the coated implant. The epithelium adjacent to the implant abutment starts to change its phenotype during the early days of the healing process. New bone formation was seen within small pieces of bone in close contact with the coated implant. In conclusion, this in vitro model maintains the viability of pig tissue and allows histologically and immunohistochemically evaluate the tissue-implant interface. HT-induced TiO2 coating seems to have a favorable tissue response. Moreover, this organotypic tissue culture model is applicable for further studies with quantitative parameters to evaluate adhesion molecules present at the implant-tissue interface.

Effect of ultraviolet light treatment on surface hydrophilicity and human gingival fibroblast response on nanostructured titanium surfaces.

This study was designed to investigate the effect of nanostructured TiO2 coatings on human gingival fibroblast and to explore the influence of ultraviolet (UV) light on surface wettability and cellular response. Ti-6Al-4V titanium alloy discs (n = 96) were divided into three groups: a sol-gel-derived MetAlive™ (MA) coating; hydrothermal (HT) coating; and a non-coated (NC) group. Forty-eight titanium substrates were further treated with UV light for 15 min. The water contact angles of the substrates were measured using the sessile drop method. Human gingival fibroblasts were used to evaluate the cell adhesion strength and cell proliferation on experimental surfaces. The strength of cell adhesion against enzymatic detachment was studied after 6 hr of adhesion using gentle trypsinization for 15 min at room temperature. A fluorescence microscope was used for cell imaging (Zeiss-stereo-lumar-v12), and images were analyzed for cell counting, and the percentage of detached cells were calculated. The proliferation of cultured cells up to 10 days was determined according to the cell activity using Alamar Blue™assay. The HT group had the lowest contact angle value (31.1°) followed by MetAlive™ (35.3°), whereas the NC group had the highest contact angle (50.3°). After UV light treatment, all surfaces become considerably more hydrophilic. There was a significant difference in the amount of adherent cells between sol-gel and HT groups when compared with the NC group (p < .05) with detachment percentages of 35.8%, 36.4%, and 70.7%, respectively. All substrate types showed an increase in cell proliferation rate until 10 days. It can be concluded that nanostructured titanium oxide implant surfaces, obtained by sol-gel and HT coating methods, enhance the surface wettability and improve human gingival fibroblast function in terms of adhesion and proliferation rate when compared with non-coated surfaces. UV light treatment clearly enhances the wettability of all titanium surfaces.

Ultraviolet photofunctionalization of nanostructured titanium surfaces enhances thrombogenicity and platelet response.

The purpose of this study was to evaluate blood and platelet response to nanostructured TiO2 coatings and to investigate the effect of Ultraviolet (UV) light treatment on blood clotting ability, platelet activation and protein adhesion. Ti-6Al-4V titanium alloy plates (n = 138) were divided into three groups; a sol-gel derived MetAliveTM coating (MA); hydrothermal coating (HT); and a non-coated group (NC). Sixty nine titanium substrates were further treated with UV light for 1 h. The thrombogenicity of the titanium substrates was assessed using fresh human blood with a whole blood kinetic clotting time method. The platelet adhesion test was conducted to evaluate the morphology and adhesion behavior of the platelets on the titanium substrates. Human diluted plasma and bovine fibronectin were used to evaluate protein adsorption. Total clotting time for the UV treated HT, MA and NC titanium substrates was almost 40 min compared to 60 min for non-UV substrates, the total clotting time for the UV treated groups were significantly lower than that of the non UV NC group (p < 0.05). UV light treatment had significantly enhanced coagulation rates. The HT and MA substrates presented more platelet aggregation, spreading and pseudopod formation in comparison with the NC substrates. UV treatment did not affect the platelet activation and protein adsorption. This in vitro study concluded that nanostructured titanium dioxide implant surfaces obtained by sol-gel and hydrothermal coating methods increased coagulation rates and enhanced platelet response when compared with non-coated surfaces. UV light treatment clearly improved thrombogenicity of all examined Ti-6Al-4V surfaces.

Early Biofilm Formation on UV Light Activated Nanoporous TiO2 Surfaces In Vivo.

PURPOSE: To explore early S. mutans biofilm formation on hydrothermally induced nanoporous TiO2 surfaces in vivo and to examine the effect of UV light activation on the biofilm development. MATERIALS AND METHODS: Ti-6Al-4V titanium alloy discs (n = 40) were divided into four groups with different surface treatments: noncoated titanium alloy (NC); UV treated noncoated titanium alloy (UVNC); hydrothermally induced TiO2 coating (HT); and UV treated titanium alloy with hydrothermally induced TiO2 coating (UVHT). In vivo plaque formation was studied in 10 healthy, nonsmoking adult volunteers. Titanium discs were randomly distributed among the maxillary first and second molars. UV treatment was administered for 60 min immediately before attaching the discs in subjects' molars. Plaque samples were collected 24h after the attachment of the specimens. Mutans streptococci (MS), non-mutans streptococci, and total facultative bacteria were cultured, and colonies were counted. RESULTS: The plaque samples of NC (NC + UVNC) surfaces showed over 2 times more often S. mutans when compared to TiO2 surfaces (HT + UVHT), with the number of colonized surfaces equal to 7 and 3, respectively. CONCLUSION: This in vivo study suggested that HT TiO2 surfaces, which we earlier showed to improve blood coagulation and encourage human gingival fibroblast attachment in vitro, do not enhance salivary microbial (mostly mutans streptococci) adhesion and initial biofilm formation when compared with noncoated titanium alloy. UV light treatment provided Ti-6Al-4V surfaces with antibacterial properties and showed a trend towards less biofilm formation when compared with non-UV treated titanium surfaces.