A Novel High-Energy Vacuum Ultraviolet Light Photofunctionalization Approach for Decomposing Organic Molecules around Titanium.

Titanium undergoes biological aging, represented by increased hydrophobicity and surface accumulation of organic molecules over time, which compromises the osseointegration of dental and orthopedic implants. Here, we evaluated the efficacy of a novel UV light source, 172 nm wavelength vacuum UV (VUV), in decomposing organic molecules around titanium. Methylene blue solution used as a model organic molecule placed in a quartz ampoule with and without titanium specimens was treated with four different UV light sources: (i) ultraviolet C (UVC), (ii) high-energy UVC (HUVC), (iii) proprietary UV (PUV), and (iv) VUV. After one minute of treatment, VUV decomposed over 90% of methylene blue, while there was 3-, 3-, and 8-fold more methylene blue after the HUVC, PUV, and UVC treatments, respectively. In dose-dependency experiments, maximal methylene blue decomposition occurred after one minute of VUV treatment and after 20-30 min of UVC treatment. Rapid and effective VUV-mediated organic decomposition was not influenced by the surface topography of titanium or its alloy and even occurred in the absence of titanium, indicating only a minimal photocatalytic contribution of titanium dioxide to organic decomposition. VUV-mediated but not other light source-mediated methylene blue decomposition was proportional to its concentration. Plastic tubes significantly reduced methylene blue decomposition for all light sources. These results suggest that VUV, in synergy with quartz ampoules, mediates rapid and effective organic decomposition compared with other UV sources. This proof-of-concept study paves the way for rapid and effective VUV-powered photofunctionalization of titanium to overcome biological aging.

Influence of Surface Contaminants and Hydrocarbon Pellicle on the Results of Wettability Measurements of Titanium.

Hydrophilicity/hydrophobicity-or wettability-is a key surface characterization metric for titanium used in dental and orthopedic implants. However, the effects of hydrophilicity/hydrophobicity on biological capability remain uncertain, and the relationships between surface wettability and other surface parameters, such as topography and chemistry, are poorly understood. The objective of this study was to identify determinants of surface wettability of titanium and establish the reliability and validity of the assessment. Wettability was evaluated as the contact angle of ddH2O. The age of titanium specimens significantly affected the contact angle, with acid-etched, microrough titanium surfaces becoming superhydrophilic immediately after surface processing, hydrophobic after 7 days, and hydrorepellent after 90 days. Similar age-related loss of hydrophilicity was also confirmed on sandblasted supra-micron rough surfaces so, regardless of surface topography, titanium surfaces eventually become hydrophobic or hydrorepellent with time. On age-standardized titanium, surface roughness increased the contact angle and hydrophobicity. UV treatment of titanium regenerated the superhydrophilicity regardless of age or surface roughness, with rougher surfaces becoming more superhydrophilic than machined surfaces after UV treatment. Conditioning titanium surfaces by autoclaving increased the hydrophobicity of already-hydrophobic surfaces, whereas conditioning with 70% alcohol and hydrating with water or saline attenuated pre-existing hydrophobicity. Conversely, when titanium surfaces were superhydrophilic like UV-treated ones, autoclaving and alcohol cleaning turned the surfaces hydrorepellent and hydrophobic, respectively. UV treatment recovered hydrophilicity without exception. In conclusion, surface roughness accentuates existing wettability and can either increase or decrease the contact angle. Titanium must be age-standardized when evaluating surface wettability. Surface conditioning techniques significantly but unpredictably affect existing wettability. These implied that titanium wettability is significantly influenced by the hydrocarbon pellicle and other contaminants inevitably accumulated. UV treatment may be an effective strategy to standardize wettability by making all titanium surfaces superhydrophilic, thereby allowing the characterization of individual surface topography and chemistry parameters in future studies.

Disparity in the Influence of Implant Provisional Materials on Human Gingival Fibroblasts with Different Phases of Cell Settlement: An In Vitro Study.

The development of healthy peri-implant soft tissues is critical to achieving the esthetic and biological success of implant restorations throughout all stages of healing and tissue maturation, starting with provisionalization. The purpose of this study was to investigate the effects of eight different implant provisional materials on human gingival fibroblasts at various stages of cell settlement by examining initial cell attachment, growth, and function. Eight different specimens-bis-acrylic 1 and 2, flowable and bulk-fill composites, self-curing acrylic 1 and 2, milled acrylic, and titanium (Ti) alloy as a control-were fabricated in rectangular plates (n = 3). The condition of human gingival fibroblasts was divided into two groups: those in direct contact with test materials (contact experiment) and those in close proximity to test materials (proximity experiment). The proximity experiment was further divided into three phases: pre-settlement, early settlement, and late settlement. A cell culture insert containing each test plate was placed into a well where the cells were pre-cultured. The number of attached cells, cell proliferation, resistance to detachment, and collagen production were evaluated. In the contact experiment, bis-acrylics and composites showed detrimental effects on cells. The number of cells attached to milled acrylic and self-curing acrylic was relatively high, being approximately 70% and 20-30%, respectively, of that on Ti alloy. There was a significant difference between self-curing acrylic 1 and 2, even with the same curing modality. The cell retention ability also varied considerably among the materials. Although the detrimental effects were mitigated in the proximity experiment compared to the contact experiment, adverse effects on cell growth and collagen production remained significant during all phases of cell settlement for bis-acrylics and flowable composite. Specifically, the early settlement phase was not sufficient to significantly mitigate the material cytotoxicity. The flowable composite was consistently more cytotoxic than the bulk-fill composite. The harmful effects of the provisional materials on gingival fibroblasts vary considerably depending on the curing modality and compositions. Pre-settlement of cells mitigated the harmful effects, implying the susceptibility to material toxicity varies depending on the progress of wound healing and tissue condition. However, cell pre-settlement was not sufficient to fully restore the fibroblastic function to the normal level. Particularly, the adverse effects of bis-acrylics and flowable composite remained significant. Milled and self-curing acrylic exhibited excellent and acceptable biocompatibility, respectively, compared to other materials.

Porphyromonas gingivalis impairs glucose uptake in skeletal muscle associated with altering gut microbiota.

Skeletal muscles have a high metabolic capacity, which play key roles in glucose metabolism. Although periodontal disease increases the risk of metabolic syndrome, the relationship between periodontal bacterial infection and skeletal muscle metabolic dysfunction is unclear. We found that anti-Porphyromonas gingivalis (Pg) antibody titers positively correlated with intramuscular adipose tissue content (IMAC), fasting blood glucose, and HOMA-IR in metabolic syndrome patients. In C57BL/6J mice fed a high-fat diet, recipients of oral Pg (HFPg) had impaired glucose tolerance, insulin resistance, and higher IMAC compared to recipients of saline (HFco). The soleus muscle in HFPg mice exhibited fat infiltration and lower glucose uptake with higher Tnfa expression and lower insulin signaling than in HFco mice. Gene set enrichment analysis showed that TNFα signaling via NFκB gene set was enriched in the soleus muscle of HFPg mice. Moreover, TNF-α also decreased glucose uptake in C2C12 myoblast cells in vitro. Based on 16S rRNA sequencing, Pg administration altered the gut microbiome, particularly by decreasing the abundance of genus Turicibacter. Microbial network of the gut microbiome was dramatically changed by Pg administration. Our findings suggest that infection with Pg is a risk factor for metabolic syndrome and skeletal muscle metabolic dysfunction via gut microbiome alteration.

High-frequency pulsed diode laser irradiation inhibits bone resorption in mice with ligature-induced periodontitis.

AIM: The purpose of this study was to elucidate the suppressive effect of high-frequency pulsed diode laser irradiation on bone resorption and its biological effects on gene expression and microbiome composition on the gingival tissue in ligature-induced periodontitis in mice. MATERIALS AND METHODS: Ligating ligature around the teeth and/or laser irradiation was performed on the gingival tissue in mice as follows: Co (no ligature and no laser irradiation), Li (ligation without laser irradiation), La (no ligature but with laser irradiation), and LiLa (ligation with laser irradiation). Bone resorption was evaluated using micro-computed tomography. RNA-seq analysis was performed on gingival tissues of all four groups at 3 days after ligation. The differences in microbial composition between Li and LiLa were evaluated based on the number of 16S rRNA gene sequences. RESULTS: Bone resorption caused by ligation was significantly suppressed by laser irradiation. RNA-seq in Co and La gingival tissue revealed many differentially expressed genes, suggesting diode laser irradiation altered gene expression. Gene set enrichment analysis revealed mTORC1 signalling and E2F target gene sets were enriched in gingival tissues both in La and LiLa compared with that in Co and Li, respectively. The amount of extracted DNA from ligatures was reduced by laser irradiation, and bacterial network structure was altered between the Li and LiLa. CONCLUSIONS: High-frequency pulsed diode laser irradiation showed biological effects and suppressed bone resorption in ligature-induced periodontitis.

N-Acetyl Cysteine-Mediated Improvements in Dental Restorative Material Biocompatibility.

The fibroblast-rich gingival tissue is usually in contact with or adjacent to cytotoxic polymer-based dental restoration materials. The objective of this study was to determine whether the antioxidant amino acid, N-acetyl cysteine (NAC), reduces the toxicity of dental restorative materials. Human oral fibroblasts were cultured with bis-acrylic, flowable composite, bulk-fill composite, self-curing acrylic, and titanium alloy test specimens. Cellular behavior and function were analyzed on and around the materials. Impregnation of the bulk-fill composite and self-curing acrylic with NAC reduced their toxicity, improving the attachment, growth, and function of human oral fibroblasts on and around the materials. These mitigating effects were NAC dose dependent. However, NAC impregnation of the bis-acrylic and flowable composite was ineffective, with no cells attaching to nor around the materials. Although supplementing the culture medium with NAC also effectively improved fibroblast behaviors, direct impregnation of materials with NAC was more effective than supplementing the cultures. NAC-mediated improvements in fibroblast behavior were associated with reduced production of reactive oxygen species and oxidized glutathione together with increased glutathione reserves, indicating that NAC effectively directly scavenged ROS from materials and reinforced the cellular antioxidant defense system. These results establish a proof of concept of NAC-mediated improvements in biocompatibility in the selected dental restorative materials.

Patient-specific establishment of bacterial composition within the peri-implant microbiota during the earliest weeks after implant uncovering.

BACKGROUND AND OBJECTIVE: Dysbiosis, a loss of balance in the microbiota, is a potential factor of peri-implantitis. However, compositional change of the peri-implant microbiota soon after implant uncovering is still unknown. In this study, bacterial composition in the peri-implant sulcus was examined to understand the establishment of bacterial composition within the peri-implant microbiota during the earliest weeks after implant uncovering. METHODS: Microbiota samples were collected at weeks 1, 2, 4, and 6 after stage-two surgery. Bacterial DNA was isolated from the samples, and a 16S rRNA gene library was constructed. Sequence reads were obtained using a high-throughput sequencing platform and were taxonomically assigned at the phylum and genus levels. RESULTS: Alpha diversity indices, which did not include taxonomic information, were at similar levels throughout the four time points. At 1 and 2 weeks, the bacterial composition was similar among patients with the predominance of Firmicutes and Proteobacteria. However, the composition was diverse at 4 and 6 weeks and significantly dissimilar to the composition at 1 week. CONCLUSIONS: At 1 week, the peri-implant microbiota was already formed with alpha diversity as high as that at the later time points. However, the bacterial composition was not highly dissimilar among patients at 1 week. The composition changed over the passage of several weeks and was specific for each patient.

Application of Ligature-Induced Periodontitis in Mice to Explore the Molecular Mechanism of Periodontal Disease.

Periodontitis is an inflammatory disease characterized by the destruction of the periodontium. In the last decade, a new murine model of periodontitis has been widely used to simulate alveolar bone resorption and periodontal soft tissue destruction by ligation. Typically, 3-0 to 9-0 silks are selected for ligation around the molars in mice, and significant bone loss and inflammatory infiltration are observed within a week. The ligature-maintained period can vary according to specific aims. We reviewed the findings on the interaction of systemic diseases with periodontitis, periodontal tissue destruction, the immunological and bacteriological responses, and new treatments. In these studies, the activation of osteoclasts, upregulation of pro-inflammatory factors, and excessive immune response have been considered as major factors in periodontal disruption. Multiple genes identified in periodontal tissues partly reflect the complexity of the pathogenesis of periodontitis. The effects of novel treatment methods on periodontitis have also been evaluated in a ligature-induced periodontitis model in mice. This model cannot completely represent all aspects of periodontitis in humans but is considered an effective method for the exploration of its mechanisms. Through this review, we aimed to provide evidence and enlightenment for future studies planning to use this model.

Achieving Complete Human Gingival Fibroblast Collagen Coverage on Implant Abutments through Vacuum Ultraviolet (VUV) Photofunctionalization.

PURPOSE: The formation of a biological seal between implant abutments and the surrounding soft tissue is a preventive strategy against peri-implantitis. The aim of this study is to test the hypothesis that surfaces of prosthetic implant abutments treated with vacuum ultraviolet (VUV) light enhance the growth and function of human gingival fibroblasts. MATERIALS AND METHODS: Implant abutments were treated with 172 nm VUV light for one minute. Untreated abutments were subjected as controls. Their surface properties were characterized using SEM, contact angle measurements, and chemical composition analysis. Human gingival fibroblasts were cultured on both untreated and VUV-treated abutments to evaluate cell attachment, proliferation, distribution, and collagen production. Cell detachment assays were also performed under various mechanical and chemical stimuli. RESULTS: After VUV treatment, implant abutments demonstrated a notable transition from hydrophobic to hydrophilic wettability. Surface element analysis revealed a considerable reduction in surface carbon and increases in oxygen and titanium elements on the VUV-treated surfaces. On day 1 of culture, 3.9 times more fibroblasts attached on VUV-treated abutments than on untreated control abutments. Fibroblastic proliferation increased 1.9-3.1 times on VUV-treated abutments, along with a significant improvement in the distribution of populating cells. Collagen production on VUV-treated abutments increased by 1.5-1.7 times. While untreated abutment surfaces showed voids and limited spread of collagen deposition, dense and full coverage of collagen was observed on VUV-treated abutments, with a great contrast in the challenging axial surface zone. Cell retention against mechanical and chemical detaching stimuli was increased 11.3 and 4.3 times, respectively, by VUV treatment. CONCLUSION: Treatment of implant abutments with VUV light for one minute resulted in a reduction of surface carbon and a transformation of the surface from hydrophobic to hydrophilic. This led to enhanced attachment, proliferation, and retention of human gingival fibroblasts, along with nearly complete collagen coverage on implant abutments. These in vitro results indicate the promising potential of utilizing VUV photofunctionalized implant abutments to enhance soft tissue reaction and sealing mechanisms.

Optimizing implant osseointegration, soft tissue responses, and bacterial inhibition: A comprehensive narrative review on the multifaceted approach of the UV photofunctionalization of titanium.

Titanium implants have revolutionized restorative and reconstructive therapy, yet achieving optimal osseointegration and ensuring long-term implant success remain persistent challenges. In this review, we explore a cutting-edge approach to enhancing implant properties: ultraviolet (UV) photofunctionalization. By harnessing UV energy, photofunctionalization rejuvenates aging implants, leveraging and often surpassing the intrinsic potential of titanium materials. The primary aim of this narrative review is to offer an updated perspective on the advancements made in the field, providing a comprehensive overview of recent findings and exploring the relationship between UV-induced physicochemical alterations and cellular responses. There is now compelling evidence of significant transformations in titanium surface chemistry induced by photofunctionalization, transitioning from hydrocarbon-rich to carbon pellicle-free surfaces, generating superhydrophilic surfaces, and modulating the electrostatic properties. These changes are closely associated with improved cellular attachment, spreading, proliferation, differentiation, and, ultimately, osseointegration. Additionally, we discuss clinical studies demonstrating the efficacy of UV photofunctionalization in accelerating and enhancing the osseointegration of dental implants. Furthermore, we delve into recent advancements, including the development of one-minute vacuum UV (VUV) photofunctionalization, which addresses the limitations of conventional UV methods as well as the newly discovered functions of photofunctionalization in modulating soft tissue and bacterial interfaces. By elucidating the intricate relationship between surface science and biology, this body of research lays the groundwork for innovative strategies aimed at enhancing the clinical performance of titanium implants, marking a new era in implantology.

Human Gingival Fibroblast Growth and Function in Response to Laser-Induced Meso-and Microscale Hybrid Topography on Dental Implant Healing Abutments.

PURPOSE: Laser-created titanium surface topographies enhance soft tissue attachment and implant stability. However, knowledge about the underlying mechanisms governing the tissue-level reaction is lacking. The objective of this study was to examine the behavior and function of human gingival fibroblasts growing on healing abutments with or without laser-textured topography. MATERIALS AND METHODS: Human primary gingival connective tissue fibroblasts were cultured on healing abutments with machined or laser-textured (Laser-Lok, BioHorizons) surfaces. Cellular and molecular responses were evaluated by cell density assay (WST-1), fluorescence microscopy, qRT-PCR, and detachment test. RESULTS: The machined surface showed mono-directional traces and scratches from milling, whereas the laser-textured surface showed a distinct morphology consisting of mono-directional meso-scale channels (15 µm pitch) and woven, oblique micro-ridges formed within the channel. There were no differences in initial fibroblast attachment, subsequent fibroblast proliferation, nor collagen production between the machined and laser-textured surfaces. Fibroblasts growing on laser-textured surface spread mono-directionally along the meso-channels, while cells growing on machined surfaces spread randomly. Fibroblasts on laser-textured surfaces were 1.8-times more resistant to detachment than those on machined surfaces. An adhesive glycoprotein (fibronectin) and trans-membrane adhesion linker gene (integrin beta-1) were upregulated on laser-textured surfaces. CONCLUSIONS: The increased fibroblast retention, uniform growth, increased transcription of cell adhesion proteins compellingly explain the enhanced tissue-level response to laser-created, hybrid textured titanium surfaces. These results provide a cellular and molecular rationale for the tissue reaction to this unique surface and support its extended use from implant fixtures and healing abutments to diverse prosthetic components where enhanced soft tissue responses would be desirable.

Novel Flowchart Guiding the Non-Surgical and Surgical Management of Peri-Implant Complications: A Narrative Review.

Peri-implant diseases, such as peri-implant mucositis and peri-implantitis, are induced by dysbiotic microbiota resulting in the inflammatory destruction of peri-implant tissue. Nonetheless, there has yet to be an established protocol for the treatment of these diseases in a predictable manner, although many clinicians and researchers have proposed various treatment modalities for their management. With the increase in the number of reports evaluating the efficacy of various treatment modalities and new materials, the use of multiple decontamination methods to clean infected implant surfaces is recommended; moreover, the use of hard tissue laser and/or air abrasion techniques may prove advantageous in the future. Limited evidence supports additional effects on clinical improvement in antimicrobial administration for treating peri-implantitis. Implantoplasty may be justified for decontaminating the implant surfaces in the supracrestal area. Surgical treatment is employed for advanced peri-implantitis, and appropriate surgical methods, such as resection therapy or combination therapy, should be selected based on bone defect configuration. This review presents recent clinical advances in debridement methods for contaminated implant surfaces and regenerative materials for treating peri-implant bone defects. It also proposes a new flowchart to guide the treatment decisions for peri-implant disease.

Genome-wide transcriptional responses of osteoblasts to different titanium surface topographies.

This is the first genome-wide transcriptional profiling study using RNA-sequencing to investigate osteoblast responses to different titanium surface topographies, specifically between machined, smooth and acid-etched, microrough surfaces. Rat femoral osteoblasts were cultured on machine-smooth and acid-etched microrough titanium disks. The culture system was validated through a series of assays confirming reduced osteoblast attachment, slower proliferation, and faster differentiation on microrough surfaces. RNA-sequencing analysis of osteoblasts at an early stage of culture revealed that gene expression was highly correlated (r = 0.975) between the two topographies, but 1.38 % genes were upregulated and 0.37 % were downregulated on microrough surfaces. Upregulated transcripts were enriched for immune system, plasma membrane, response to external stimulus, and positive regulation to stimulus processes. Structural mapping confirmed microrough surface-promoted gene sharing and networking in signaling pathways and immune system/responses. Target-specific pathway analysis revealed that Rho family G-protein signaling pathways and actin genes, responsible for the formation of stress fibers, cytoplasmic projections, and focal adhesion, were upregulated on microrough surfaces without upregulation of core genes triggered by cell-to-cell interactions. Furthermore, disulfide-linked or -targeted extracellular matrix (ECM) or membranous glycoproteins such as laminin, fibronectin, CD36, and thrombospondin were highly expressed on microrough surfaces. Finally, proliferating cell nuclear antigen (PCNA) and cyclin D1, whose co-expression reduces cell proliferation, were upregulated on microrough surfaces. Thus, osteoblasts on microrough surfaces were characterized by upregulation of genes related to a wide range of functions associated with the immune system, stress/stimulus responses, proliferation control, skeletal and cytoplasmic signaling, ECM-integrin receptor interactions, and ECM-membranous glycoprotein interactions, furthering our knowledge of the surface-dependent expression of osteoblastic biomarkers on titanium.

Vacuum Ultraviolet (VUV) Light Photofunctionalization to Induce Human Oral Fibroblast Transmigration on Zirconia.

Soft tissue adhesion and sealing around dental and maxillofacial implants, related prosthetic components, and crowns are a clinical imperative to prevent adverse outcomes of periodontitis and periimplantitis. Zirconia is often used to fabricate implant components and crowns. Here, we hypothesized that UV treatment of zirconia would induce unique behaviors in fibroblasts that favor the establishment of a soft tissue seal. Human oral fibroblasts were cultured on zirconia specimens to confluency before placing a second zirconia specimen (either untreated or treated with one minute of 172 nm vacuum UV (VUV) light) next to the first specimen separated by a gap of 150 µm. After seven days of culture, fibroblasts only transmigrated onto VUV-treated zirconia, forming a 2.36 mm volume zone and 5.30 mm leading edge. Cells migrating on VUV-treated zirconia were enlarged, with robust formation of multidirectional cytoplastic projections, even on day seven. Fibroblasts were also cultured on horizontally placed and 45° and 60° tilted zirconia specimens, with the latter configurations compromising initial attachment and proliferation. However, VUV treatment of zirconia mitigated the negative impact of tilting, with higher tilt angles increasing the difference in cellular behavior between control and VUV-treated specimens. Fibroblast size, perimeter, and diameter on day seven were greater than on day one exclusively on VUV-treated zirconia. VUV treatment reduced surface elemental carbon and induced superhydrophilicity, confirming the removal of the hydrocarbon pellicle. Similar effects of VUV treatment were observed on glazed zirconia specimens with silica surfaces. One-minute VUV photofunctionalization of zirconia and silica therefore promotes human oral fibroblast attachment and proliferation, especially under challenging culture conditions, and induces specimen-to-specimen transmigration and sustainable photofunctionalization for at least seven days.

Impact of nano-scale trabecula size on osteoblastic behavior and function in a meso-nano hybrid rough biomimetic zirconia model.

PURPOSE: A novel implant model consisting of meso-scale cactus-inspired spikes and nano-scale bone-inspired trabeculae was recently developed to optimize meso-scale roughness on zirconia. In this model, the meso-spike dimension had a significant impact on osteoblast function. To explore how different nano-textures impact this model, here we examined the effect of different nano-trabecula sizes on osteoblast function while maintaining the same meso-spike conformation. METHODS: Zirconia disks with meso-nano hybrid surfaces were created by laser etching. The meso-spikes were fixed to 40 µm high, whereas the nano-texture was etched as large and small trabeculae of average Feret diameter 237.0 and 134.1 nm, respectively. A polished surface was also prepared. Rat bone marrow-derived and human mesenchymal stromal cell-induced osteoblasts were cultured on these disks. RESULTS: Hybrid rough surfaces, regardless of nano-trabecula dimension, robustly promoted the osteoblastic differentiation of both rat and human osteoblasts compared to those on polished surfaces. Hybrid surfaces with small nano-trabeculae further enhanced osteoblastic differentiation compared with large nano-trabeculae. However, the difference in osteoblastic differentiation between small and large nano-trabeculae was much smaller than the difference between the polished and hybrid rough surfaces. The nano-trabecula size did not influence osteoblast attachment and proliferation, or protein adsorption. Both hybrid surfaces were hydro-repellent. The atomic percentage of surface carbon was lower on the hybrid surface with small nano-trabeculae. CONCLUSIONS: Small nano-trabeculae promoted osteoblastic differentiation more than large nano-trabeculae when combined with meso-scale spikes. However, the biological impact of different nano-trabeculae was relatively small compared with that of different dimensions of meso-spikes.

Peri-implantitis management by resective surgery combined with implantoplasty and Er:YAG laser irradiation, accompanied by free gingival graft: a case report.

The optimal method for decontamination of implant surfaces for peri-implantitis treatment remains controversial. In recent years, erbium-doped yttrium aluminum garnet (Er:YAG) laser irradiation and implantoplasty (IP) (i.e. mechanical modification of the implant) have been reported to be effective in decontaminating implant surfaces during the surgical treatment. Also, a lack of adequate keratinized mucosa (KM) around the implant is known to be associated with more plaque accumulation, tissue inflammation, attachment loss, and mucosal recession, increasing the risk of peri-implantitis. Therefore, free gingival graft (FGG) has been recommended for gaining adequate KM around the implant. However, the necessity of acquiring KM for the treatment of peri-implantitis using FGG remains unclear. In this report, we applied the apically positioned flap (APF) as resective surgery for peri-implantitis treatment in conjunction with IP and Er:YAG laser irradiation to polish/clean the implant surface. Furthermore, FGG was conducted simultaneously to create additional KM, which increased the tissue stability and contributed to the positive results. The two patients were 64 and 63 years old with a history of periodontitis. The removal of granulation tissue and debridement of contaminated implant surfaces were performed with Er:YAG laser irradiation post flap elevation and then modified smooth surfaces mechanically using IP. Er:YAG laser irradiation was also utilized to remove the titanium particles. In addition, we performed FGG to increase the width of KM as a vestibuloplasty. Peri-implant tissue inflammation and progressive bone resorption were not observed, and both patients maintained good oral hygiene conditions until the 1-year follow-up appointment. Bacterial analysis via high-throughput sequencing revealed proportional decreases in bacteria associated with periodontitis (Porphyromonas, Treponema, and Fusobacterium). To the best of our knowledge, this study is the first to describe peri-implantitis management and bacterial change before and after procedures by resective surgery combined with IP and Er:YAG laser irradiation for peri-implantitis treatment, accompanied by FGG for increasing KM around the implants.

Conditional Mitigation of Dental-Composite Material-Induced Cytotoxicity by Increasing the Cure Time.

Light-cured composite resins are widely used in dental restorations to fill cavities and fabricate temporary crowns. After curing, the residual monomer is a known to be cytotoxic, but increasing the curing time should improve biocompatibility. However, a biologically optimized cure time has not been determined through systematic experimentation. The objective of this study was to examine the behavior and function of human gingival fibroblasts cultured with flowable and bulk-fill composites cured for different periods of time, while considering the physical location of the cells with regard to the materials. Biological effects were separately evaluated for cells in direct contact with, and in close proximity to, the two composite materials. Curing time varied from the recommended 20 s to 40, 60, and 80 s. Pre-cured, milled-acrylic resin was used as a control. No cell survived and attached to or around the flowable composite, regardless of curing time. Some cells survived and attached close to (but not on) the bulk-fill composite, with survival increasing with a longer curing time, albeit to <20% of the numbers growing on milled acrylic even after 80 s of curing. A few cells (<5% of milled acrylic) survived and attached around the flowable composite after removal of the surface layer, but attachment was not cure-time dependent. Removing the surface layer increased cell survival and attachment around the bulk-fill composite after a 20-s cure, but survival was reduced after an 80-s cure. Dental-composite materials are lethal to contacting fibroblasts, regardless of curing time. However, longer curing times mitigated material cytotoxicity exclusively for bulk-fill composites when the cells were not in direct contact. Removing the surface layer slightly improved biocompatibility for cells in proximity to the materials, but not in proportion to cure time. In conclusion, mitigating the cytotoxicity of composite materials by increasing cure time is conditional on the physical location of cells, the type of material, and the finish of the surface layer. This study provides valuable information for clinical decision making and novel insights into the polymerization behavior of composite materials.

Blue Light-Emitting Diode Irradiation Without a Photosensitizer Alters Oral Microbiome Composition of Ligature-Induced Periodontitis in Mice.

Objective: This study investigated the suppressive effects of blue light-emitting diode (LED) irradiation on bone resorption and changes in the oral microbiome of mice with ligature-induced periodontitis. Background: Wavelength of blue light has antimicrobial effects; however, whether blue LED irradiation alone inhibits the progression of periodontitis remains unclear. Methods: Nine-week-old male mice ligated ligature around the right maxillary second molar was divided into ligation alone (Li) and ligation with blue LED irradiation (LiBL) groups. The LiBL group underwent blue LED (wavelength, 455 nm) irradiation four times in a week at 150 mW/cm2 without a photosensitizer on the gingival tissue around the ligated tooth at a distance of 5 mm for 5 min. The total energy density per day was 45 J/cm2. Bone resorption was evaluated using micro-computed tomography at 8 days. Differences in the oral microbiome composition of the collected ligatures between the Li and LiBL groups were analyzed using next-generation sequencing based on the 16S rRNA gene from the ligatures. Results: Blue LED irradiation did not suppress bone resorption caused by ligature-induced periodontitis. However, in the LiBL group, the α-diversity, number of observed features, and Chao1 were significantly decreased. The relative abundances in phylum Myxococcota and Bacteroidota were underrepresented, and the genera Staphylococcus, Lactococcus, and Lactobacillus were significantly overrepresented by blue LED exposure. Metagenomic function prediction indicated an increase in the downregulated pathways related to microbial energy metabolism after irradiation. The co-occurrence network was altered to a simpler structure in the LiBL group, and the number of core genera decreased. Conclusions: Blue LED irradiation altered the composition and network of the oral microbiome of ligature-induced periodontitis in mice.

Novel and Efficient Method for Diagnosing Infective Endocarditis Using 16S Ribosomal RNA Gene Amplicon Sequence.

We present 2 cases that both developed infective endocarditis and underwent mitral valve replacement. In addition to positive blood culture and echocardiographic findings, such as vegetation or mitral valve perforation, the 16S ribosomal RNA gene amplicon sequence approach used was helpful for disease diagnosis. (Level of Difficulty: Intermediate.).

Decomposing Organic Molecules on Titanium with Vacuum Ultraviolet Light for Effective and Rapid Photofunctionalization.

Ultraviolet (UV) photofunctionalization counteracts the biological aging of titanium to increase the bioactivity and osseointegration of titanium implants. However, UV photofunctionalization currently requires long treatment times of between 12 min and 48 h, precluding routine clinical use. Here, we tested the ability of a novel, xenon excimer lamp emitting 172 nm vacuum UV (VUV) to decompose organic molecules coated on titanium as a surrogate of photofunctionalization. Methylene blue as a model organic molecule was coated on grade 4 commercially pure titanium and treated with four UV light sources: (i) ultraviolet C (UVC), (ii) high-energy UVC (HUVC), (iii) proprietary UV (PUV), and (iv) VUV. After one minute of treatment, VUV decomposed 57% of methylene blue compared with 2%, 36%, and 42% for UVC, HUVC, and PUV, respectively. UV dose-dependency testing revealed maximal methylene blue decomposition with VUV within one minute. Equivalent decomposition was observed on grade 5 titanium alloy specimens, and placing titanium specimens in quartz ampoules did not compromise efficacy. Methylene blue was decomposed even on polymethyl methacrylate acrylic specimens at 20-25% lower efficiency than on titanium specimens, indicating a relatively small contribution of titanium dioxide-mediated photocatalytic decomposition to the total decomposition. Load-testing revealed that VUV maintained high efficacy of methylene blue decomposition regardless of the coating density, whereas other UV light sources showed low efficacy with thin coatings and plateauing efficacy with thicker coatings. This study provides foundational data on rapid and efficient VUV-mediated organic decomposition on titanium. In synergy with quartz ampoules used as containers, VUV has the potential to overcome current technical challenges hampering the clinical application of UV photofunctionalization.