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The junctional epithelium (JE) has been recognized as a defensive organ rich in polymorphonuclear leukocytes (PMNs). However, the migration of PMNs through the JE has not been clearly documented. For mucosal defense, PMNs migrate outwards over the epithelium to defend the intestinal or respiratory tract on the epithelial surface. With this background, the present study investigated whether there is any structural evidence showing the transepithelial migration of PMNs through the JE in gingival mucosa.Three-dimensional modeling of gingiva surrounding mouse molars at varying ages was performed by array tomography. Images of the serial sections for array tomography at the 800 nm thickness were obtained using back scattered electron (BSE) detector equipped in the field-emission scanning electron microscopy (FESEM). Expressions of neutrophil marker or CD47 were immunohistochemically examined on the frozen sections.Array tomography using FESEM and 3-dimensional modeling clearly showed that a system of epithelial channels developed between keratinocytes and generally ran along the long axis of the JE. Most PMNs were found inside the channels, rather than being scattered throughout the JE. The channels could be traced from the base of the JE to the bottom of the gingival sulcus, although some channels were fragmented and interrupted with short intercellular gaps.These findings suggest that the JE may be an organ for transepithelial migration of PMNs to the bottom of the gingival sulcus through epithelial channels, as occurs in the epithelial lining of other organs such as the intestinal or respiratory tract.

Common periodontal disease treatment procedures often fail to restore the structural integrity of the junctional epithelium (JE), the epithelial attachment of the gum to the tooth, leaving the tooth-gum interface prone to bacterial colonization. To address this issue, we introduced a novel bio-inspired protein complex comprised of a proline-rich enamel protein, SCPPPQ1, and laminin 332 (LAM332) to enhance the JE attachment. Using quartz crystal microbalance with dissipation monitoring (QCM-D), we showed that SCPPPQ1 and LAM332 interacted and assembled into a protein complex with high-affinity adsorption of 5.9e

Teeth, long-lasting percutaneous organs, feature soft tissue attachment through adhesive structures, hemidesmosomes, in the junctional epithelium basement membrane adjacent to teeth. This soft tissue attachment prevents bacterial infection of the tooth despite the rich - and harsh - microbial composition of the oral cavity. Conversely, millions of percutaneous devices (catheters, dental, and orthopedic implants) fail from infection yearly. Standard of care antibiotic usage fuels antimicrobial resistance and is frequently ineffective. Infection prevention strategies, like for dental implants, have failed in generating durable soft tissue adhesion - like that seen with the tooth - to prevent bacterial colonization at the tissue-device interface. Here, inspired by the impervious natural attachment of the junctional epithelium to teeth, we synthesized four cell adhesion peptide (CAPs) nanocoatings, derived from basement membranes, to promote percutaneous device soft tissue attachment. The two leading nanocoatings upregulated integrin-mediated hemidesmosomes, selectively increased keratinocyte proliferation compared to fibroblasts, which cannot form hemidesmosomes, and expression of junctional epithelium adhesive markers. CAP nanocoatings displayed marked durability under simulated clinical conditions and the top performer CAP nanocoating was validated in a percutaneous implant murine model. Basement membrane CAP nanocoatings, inspired by the tooth and junctional epithelium, may provide an alternative anti-infective strategy for percutaneous devices to mitigate the worldwide threat of antimicrobial resistance. STATEMENT OF SIGNIFICANCE: Prevention and management of medical device infection is a significant healthcare challenge. Overzealous antibiotic use has motivated alternative material innovations to prevent infection. Here, we report implant cell adhesion peptide nanocoatings that mimic a long-lasting, natural "medical device," the tooth, through formation of cell adhesive structures called hemidesmosomes. Such nanocoatings sidestep the use of antimicrobial or antibiotic elements to form a soft-tissue seal around implants. The top performing nanocoatings prompted expression of hemidesmosomes and defensive factors to mimic the tooth and was validated in an animal model. Application of cell adhesion peptide nanocoatings may provide an alternative to preventing, rather that necessarily treating, medical device infection across a range of device indications, like dental implants.

Epstein‐Barr virus (EBV)‐associated gastric cancer belongs to 1 of the 4 subtypes of gastric cancer and accounts for 10% of total gastric cancers. However, most cases of gastric cancer have a history of Helicobacter pylori infection. Therefore, we investigated the possibility that H. pylori infection promotes the development of EBV‐associated gastric cancer. H. pylori was exposed to principal EBV receptor, CD21, negative gastric epithelial cells, and then infected with EBV recombinant expressing enhanced green fluorescent protein. Changes in EBV infectivity due to prior H. pylori exposure were analyzed using flow cytometry. The treatment of gastric epithelial cells with H. pylori increased the efficiency of EBV infection. An increase was also observed when CagA‐deficient, VacA‐deficient, and FlaA‐deficient H. pylori strains were used, but not when cag pathogenicity island‐deficient H. pylori was used. The treatment of epithelial cells with H. pylori induced the expression of accessory EBV receptors, EphA2 and NMHC‐IIA, and increased the efficiency of EBV infection depending on their expression levels. When gastric epithelial cells were treated with EPHA2 or NMHC‐IIA siRNA, EBV infection via H. pylori attachment was decreased. The adhesion of H. pylori induced the expression of accessory EBV receptors in gastric epithelial cells and increased the efficiency of EBV infection., We investigated the possibility that H. pylori infection promotes the development of EBV‐associated gastric cancer (EBVaGC). H. pylori adheres to gastric epithelial cells and induces low‐affinity EBV receptor expression in epithelial cells. A continuous increase in the number of EBV‐infected cells is expected during life‐long gastric inflammation caused by H. pylori. Therefore, H. pylori infection is thought to increase the incidence of EBVaGC by increasing the efficiency of EBV infection.

OBJECTIVE: To evaluate the type of wound healing following modified crown lengthening surgery in dog model to provide a biological basis for its clinical application. METHODS: Flap surgery, traditional crown lengthening procedure and modified crown lengthening procedure were performed on the right maxillary central incisor, the left maxillary central incisor and the left maxillary first lateral incisor respectively of five male beagle dogs. The right maxillary first lateral incisors with no surgical intervention were used as controls. Thirty-six weeks after the experimental procedure, tissue blocks were harvested and prepared for histological examination and analysis. RESULTS: Histometric examination of buccolingual sections stained with hematoxylin-eosin demonstrated that the type of wound healing in the flap surgery group was re-attachment, similar to the control group. For the traditional crown lengthening surgery group, all of the five beagle dogs had lamellar cementum defects on root surface, the wound healing of four beagle dogs was new attachment accompanied by new cementum formation at cementum defect areas and the suprac-restal connective tissue was functionally oriented perpendicular to the new cementum. The wound healing of the other beagle dog was long junctional epithelial attachment, in which the junctional epithelium extended to the apical terminus of the cementum defect. In the modified crown lengthening surgery group, four beagle dogs had cementum defects on root surface (two lamellar cementum defects and two shallow platform-like cementum defects), the wound healing of three beagle dogs was new attachment, however, the supracrestal connective tissue was parallel to the root surface. The type of wound healing of another one beagle dog was long junctional epithelial attachment. Wound healing of one beagle dog in this group could not be characterized due to incomplete dissection. CONCLUSION: Wound healing in the modified crown lengthening surgery group was similar to the traditional crown lengthening surgery group, and two types of wound healing were observed: new attachment and long junctional epithelium attachment. Neither type of root treatment procedure (root planing or root reshaping) nor root surface defect pattern (the lamellar cementum defect or shallow platform-like cementum defect) influenced the observed type of wound healing.

Subgingival dental restorations and periodontal health have been studied for many years; however, there is a low histological evidence on the behavior of new materials in the supracrestal tissue attachment. The aim of this study is to analyze the periodontal response when a tricalcium silicate material (TSM) or composite margin restorations is placed to 0.5 mm and 1.5 mm from the bone crest with a histomorphometric analysis in dogs. Nine mongrel dogs were used in this study: four dogs (8 canine teeth) for TSM group, 4 dogs (8 canine teeth) for composite group, and 1 dog (2 canine teeth) with cavities without restorations. Cavity preparation of 2×2×1 mm was created on the buccal aspect of the canines at 0.5 and 1.5 mm of the crestal bone. Cavities were restored with composite and TSM or were left unrestored as control. After 12 weeks of healing, the dogs were euthanized and blocks containing the tooth and soft tissues were processed. In all the specimens, the junction epithelium was stablished apical to the tooth preparations. A shorter distance to the bone (0.5 cavity) implies greater apical periodontal migration regardless of the material used. In the TSM groups, the connective tissue height and the distance between bone level and apical margin preparation were greater than those in the composite groups, while the epithelium height was less. However, there were no statistically significant differences comparing TSM and composite groups at either 0.5 mm or 1.5 mm (p > 0.05). Histologic analysis did not show periodontal reattachment to TSM or composite. In both cases, bone crest migrates apically. For that reason, it is recommended to perform composite restorations at the subgingival level whenever the distance to the bone crest is at least 2 mm. Both composite and TSM do not achieve reinsertion of the connective tissue in the biological width.

Objective In this study, we investigated the potential and mechanism of odontogenic ameloblast-associated protein (ODAM) in the promoting junctional epithelium-related gene expression in an ameloblast-like cell line ALC. Background ODAM is expressed in ameloblasts and JE and acts as a component of the inner basal lamina (IBL) and intercellular matrix of JE. ODAM KO mice showed destruction of the integrity of the JE, which detaches from teeth. ODAM was confirmed to regulate the cytoskeleton through the ODAM-ARHGEF5-RhoA signaling pathway of the JE. Whether ODAM contributes to the regulation of ameloblast differentiation in JE remains unclear. After the formation of enamel, the ameloblast undergoes a series of morphological changes. Whether ODAM will affect the biological behavior of ameloblasts making them have the characteristics of JE is unclear. Methods A murine ameloblast-like cell line, ALC, was used to investigate the effects of ODAM on the JE-like changes of ALC cells in an epithelium-induced environment by generating ODAM overexpression and ODAM knockdown cells through a lentivirus transduction approach. The biomarkers of junctional epithelium CK19, SLPI, and ODAM and the potential regulatory gene WNT1 were investigated by real-time PCR, western blot, immunocytochemistry, immunostaining, luciferase reporter, and rescue assays. Results ODAM, CK19, and SLPI were significantly upregulated after epithelial induction. Overexpression of ODAM in ALC cells markedly increased CK19 and SLPI expression, while knockdown of ODAM in ALC cells clearly decreased CK19 and SLPI expression. A reporter luciferase assay showed that ODAM activated the WNT signaling pathway, especially through WNT1. Exogenous overexpression of ODAM upregulated WNT1 expression, while knockdown of ODAM reversed this effect. The WNT1 inhibition assay further confirmed the above results and showed that the WNT1 pathway was positively correlated with biomarkers of junctional epithelium CK19 and SLPI expression. Rescue studies showed that knocking down WNT1 in the ODAM-overexpressing ALC cells decreased the expression of CK19 and SLPI. Immunocytochemistry showed that ODAM colocalized with CK19, SLPI, and WNT1 in the cells. Conclusion In conclusion, the research work showed that ODAM promotes junctional epithelium-related gene expression in ALC via the ODAM-WNT1 axis, which may provide new insight into the function of ODAM and JE formation.

Aim To identify the molecular mechanisms mediating the persistent defensive functions of the self-renewing junctional epithelium (JE). Materials and methods Two strains of Wnt reporter mice, Axin2CreErt2 /+ ;R26RmTmG /+ and Axin2LacZ /+ , were employed, along with three clinically relevant experimental scenarios where the function of the JE is disrupted: after tooth extraction, after a partial gingivectomy, and after a complete circumferential gingivectomy. Results Using transgenic Wnt reporter strains of mice, we established the JE is a Wnt-responsive epithelium beginning at the time of its formation and that it maintains this status into adulthood. After tooth extraction, progeny of the initial Wnt-responsive JE population directly contributed to healing and ultimately adopted an oral epithelium (OE) phenotype. In the traditional partial gingivectomy model, the JE completely regenerated and did so via progeny of the original Wnt-responsive population. However, following circumferential gingivectomy, the OE was incapable of re-establishing a functional JE. Conclusions A Wnt-responsive niche at the interface between tooth and oral epithelia is required for a functional JE.

Objective Mechanical plaque removal may alter the surface morphology of the gingival penetration part of the implant. We applied an air-powered abrasive system (AP), titanium curette (TC), stainless curette (SC), ultrasound scaler (US), and titanium brush (TB) which are commonly used to remove plaque, to titanium or zirconia and the changes in surface morphology and the epithelial attach against substrata. Materials and methods (a) The morphological changes of titanium and zirconia after mechanical cleaning were assessed by scanning electron microscopy and a roughness analyser. (b) Oral epithelial cells of rats were inoculated on the surface of the materials after mechanical cleaning, and the adherence of epithelial cells was observed. (c) The maxillary first molars were extracted from the rats and replaced by experimental titanium or zirconia implants. The length of the immunoreactive laminin-332 band was observed at the implant-peri-implant epithelium interface. Results (a) The surface roughness increased in experimental groups except the AP group. (b) Among the experimental groups, the AP group showed the highest number of attached cells. (c) The length of the immunoreactive laminin-332 band was longer in the control group than those in all five experimental groups. Among the experimental groups, the AP group showed the longest band. Conclusion All mechanical cleaning methods increased the surface roughness of the materials except AP. AP did not cause distinct implant surface alterations. Surface alteration caused by mechanical cleaning may evoke inferior for epithelial attachment and reduce resistance against foreign infiltration.

Extracellular matrix (ECM) proteins are highly abundant in the human body and can be found in various tissues, most prominently in connective tissue and basement membrane. For invasive bacterial pathogens, these structures function as physical barriers that block access to underlying tissues. The ability to bind and degrade these barriers is important for the establishment of infections and migration to other body sites. In the oral cavity, the ECM and the basement membrane (BM) are important components of the Junctional epithelium (JE) that closes the gap between the teeth surface and the mucosa. In periodontitis, the JE is breached by invading pathogenic bacteria, particularly strict anaerobic species. In periodontitis, invading microorganisms induce an unregulated and destructive host response through polymicrobial synergism and dysbiosis that attracts immune cells and contributes to the destruction of connective tissue and bone in the periodontal pocket. Colonization of the periodontal pocket is the first step to establish this infection, and binding to ECM is a major advantage in this site. Several species of strict anaerobic bacteria are implicated in acute and chronic periodontitis, and although binding to ECM proteins was studied in these species, few adhesins were identified so far, and the mechanisms involved in adhesion are largely unidentified. This review summarizes the data available on the interaction of strict anaerobic bacteria and components of the ECM.

Objective The aim of this study was to examine the potential of periodontal ligament (PDL) cells sheet and arginine-glycyl-aspartic acid (RGD)-modified chitosan scaffold for periodontal tissue regeneration in horizontal periodontal defect model. Materials and Methods PDL cell cytotoxicity was tested with 3–[4,5- dimethylthiazol-2yl]–2,5-diphenyl-2H-tetrazolium bromide assay. Cell migration toward the chitosan-based materials was analyzed with trans-well migration assay. Horizontal periodontal defect model was created in four maxillary and mandibular lateral incisors of Macaque nemestrina. Following periodontal therapy, the sites were transplanted with various regenerative materials: (1) chitosan, (2) RGD-modified chitosan, (3) PDL cell sheet with chitosan, (4) PDL cell sheet with RGD-modified chitosan. The periodontal tissue regeneration was evaluated clinically and radiographically. Gingival crevicular fluids were collected each week to evaluate cementum protein-1 (CEMP-1) expression with enzyme-linked immunosorbent assay, while the biopsies were retrieved after 4 weeks for histological and microcomputed tomography evaluation. Statistical Analysis Data was statistically analyzed using GraphPad Prism 6 for MacOS X. Normality was tested using the Shapiro–Wilk normality test. The Kruskal–Wallis test was used to compare the groups. Significance was accepted when p < 0.05. Results Clinical examination revealed more epithelial attachment was formed in the group with PDL cell sheet with RGD-modified chitosan. Similarly, digital subtraction radiography analysis showed higher gray scale, an indication of higher alveolar bone density surrounded the transplanted area, as well as higher CEMP-1 protein expression in this group. The incorporation of RGD peptide to chitosan scaffold in the group with or without PDL cells sheet reduced the distance of cement–enamel junction to the alveolar bone crest; hence, more periodontal tissue formed. Conclusions Horizontal periodontal defect model could be successfully created in M. nemestrina model. Combination of PDL cell sheet and RGD-modified chitosan resulted in the higher potential for periodontal tissue regeneration. The results of this study highlight the PDL cell sheet and RGD-modified chitosan as a promising approach for future clinical use in periodontal regeneration.

Background Gingival tissue attachment is known to be important for long-term prognosis of implants. This in vitro study evaluated the gingival attachment to zirconia implants and zirconia implants modified with sol-gel derived TiO2 coatings. Methods Zirconia endodontic posts (n = 23) were used to function as implants that were inserted into the center of full-thickness porcine gingival explants (n = 31). The tissue/implant specimens were then individually placed at an air/liquid interface on a stainless-steel grid in cell culture wells containing a nutrient solution. The tissue cultures were incubated at 37°C in a 5% CO2 environment and at days 7 and 14, the specimens were harvested and analyzed by dynamic mechanical analysis (DMA) measurements under dynamic loading conditions mimicking natural mastication. Specimens were also analyzed by immunohistochemical staining identifying the laminin (Ln) γ2 chain specific for Ln-332, which is known to be a crucial molecule for the proper attachment of epithelium to tooth/implant surface. Results Tissue attachment to TiO2 -coated zirconia demonstrated higher dynamic modulus of elasticity and higher creep modulus, meaning that the attachment is stronger and more resistant to damage during function over time. Laminin γ2 was identified in the attachment of epithelium to TiO2 -coated zirconia. Conclusions Both DMA and histological analysis support each other, so the gingival tissue is more strongly attached to sol-gel derived TiO2 -coated zirconia than uncoated zirconia. Immunohistochemical staining showed that TiO2 coating may enhance the synthesis and deposition of Ln-332 in the epithelial attachment to the implant surface.

Follicular dendritic cell-secreted protein (FDC-SP) is secreted protein expressed in follicular dendritic cells, periodontal ligament and junctional epithelium (JE). Its expression could be controlled during inflammatory process of gingiva; however, responsible mechanism for gingival overgrowth and involvement of FDC-SP in clinical condition is still unclear. We hypothesized that JE-specific genes are associated with the initiation of drug-induced gingival enlargement (DIGE) called gingival overgrowth, and investigated the changes of JE-specific gene's expression and their localization in overgrown gingiva from the patients. Immunohistochemical analysis revealed that the FDC-SP localization was spread in overgrown gingival tissues. FDC-SP mRNA levels in GE1 and Ca9-22 cells were increased by time-dependent nifedipine treatments, similar to other JE-specific genes, such as Amelotin (Amtn) and Lamininβ3 subunit (Lamβ3), whereas type 4 collagen (Col4) mRNA levels were decreased. Immunocytochemical analysis showed that FDC-SP, AMTN, and Lamβ3 protein levels were increased in GE1 and Ca9-22 cells. Transient transfection analyses were performed using luciferase constructs including various lengths of human FDC-SP gene promoter, nifedipine increased luciferase activities of -345 and -948FDC-SP constructs. These results raise the possibility that the nifedipine-induced FDC-SP may be related to the mechanism responsible for gingival overgrowth does not occur at edentulous jaw ridges.

The peri-implant soft tissue seal consists of a connective tissue cuff and a junctional epithelium that is different from the arrangement of periodontium around a natural tooth. However, the peri-implant soft tissue complex lacks Sharpey's fibers, thus offering less resistance to clinical probing and biofilm penetration compared to the natural dentition. Therefore, the proper restorative emergence profile design is essential to facilitate favorable esthetic outcomes and maintain peri-implant health. The aim of this article is to review the currently available evidence related to the design of subgingival (critical and subcritical) and supragingival contours of the implant restorative emergence profile (IREP) as well as provide a flowchart for decision-making in clinical practice. Theoretically, the subgingival contours of the crown/abutment complex should mimic the morphology of the root and the cervical third of the anatomic crown as much and as often as possible. However, this is highly dependent upon the three-dimensional spatial position of the implant relative to the hard and soft tissue complex, in addition to the location of the definitive restoration. Frequently, a convex critical contour is required on the facial aspect of a palatally or incisally positioned implant to support an adequate gingival-margin architecture. Conversely, if the implant is placed too far facially, then a flat or concave contour is recommended. In instances where soft tissue support is not needed, the subcritical area may be undercontoured to increase the thickness, height, and stability of the soft tissue cuff.

Sodium fluoride (NaF) is widely used in clinical dentistry. However, the administration of high or low concentrations of NaF has various functions in different tissues. Understanding the mechanisms of the different effects of NaF will help to optimize its use in clinical applications. Studies of NaF and epithelial cells, osteoblasts, osteoclasts, and periodontal cells have suggested the significant roles of fluoride treatment. In this review, we summarize recent studies on the biphasic functions of NaF that are related to both soft and hard periodontal tissues, multiple diseases, and clinical dentistry.

Background and Objectives: Orthodontic tooth movement is associated with inflammatory responses. The aim of this study was to identify gingival microcirculation using laser Doppler flowmetry in patients with orthodontic treatment. Materials and Methods: A longitudinal pilot study was performed. The participants were selected using a non-probability consecutive sampling. Of the twenty-five subjects, a total of six (four women and two men) complied with the criteria. Before and during the treatment, the oral hygiene index, gingival index, probing depth, level of epithelial attachment, and gingival microcirculation were evaluated with laser Doppler flowmetry (integrated parameters: 1. integrated primary basal flow (IPBF), 2. integrated total secondary real flow (ITSRF), and 3. difference between integration (DBI)) in all of the participants). Results: (a) An increase in gingival blood flow was identified at all time intervals with different arches during orthodontic treatment. (b) The IPBF and ITSRF (with treatment) identified after 20 min (treatment initial stage) were compared with the different time intervals, and we observed an increase in gingival perfusion at the 24th, 48th, and 72nd hours in some arches. (c) In the DBI, we found statistically significant differences (p &lt, 0.005) in the Nitinol group of 0.016 inches among all the time intervals (24 h, 48 h, and 72 h) within the 30-day interval, observing a flow increase three times greater than the basal flow after 30 days. Conclusions: Healthcare professionals must identify the inflammatory processes in treatment to observe and discontinue use of harmful methods in clinical practice.

OBJECTIVE: Calprotectin, the heterdimer of S100A8 and S100A9, is the major cytoplasmic protein of neutrophils, which is also expressed or induced in gingival epithelial cells, activated mononuclear macrophages and vascular endothelial cells. Calprotectin is intimately associated with the initiation and progression of periodontitis, but the in vivo expression patterns of calprotectin in healthy and inflamed periodontal tissue are not fully understood. To observe the expression, distribution and cellular localization of calprotectin in the samples of healthy periodontal tissues and experimental periodontitis tissues of Beagles and to explore their relationship with periodontal inflammation and possible effect. METHODS: Experimental periodontitis model was established by ligation around the mandibular second molar of the Beagle dogs, while the contralateral teeth were healthy controls. Induction duration was 12 weeks, before the dogs were executed. Tissue specimens were demineralized and serial sections were made conventionally. The in vivo expression of calprotectin in the healthy and inflamed periodontal tissues were examined by immunohistochemistry. The in vitro expression of calprotectin in human primary gingival fibroblasts (GFs) and periodontal ligament (PDL) cells were detected by immunocytochemistry. RESULTS: Immunohistochemistry analysis indicated that calprotectin was expressed in gingival epithelial cells and infiltrated neutrophils in the healthy periodontium within the gingival epithelium, S100A8/A9 was most strongly expressed in the junctional epithelium, followed by surface epithelium, and least expressed in the sulcular epithelium. The S100A8/A9 expression levels were sharply defined at the junction between the junctional epithelium and the sulcular epithelium. In periodontal inflammatory lesions, the expression level of calprotectin in sulcular epithelium and junctional epithelium was up-regulated than that in the healthy gingival epithelium. Calprotectin was inducibly expressed in fibroblast-like cells in gingival connective tissue and periodontal ligament tissue, microvascular endothelial cells (ECs) and bone marrow fibroblasts under inflammatory conditions. Additionally, the expression of calprotectin in primary human GFs and PDL cells was confirmed by immunnocytochemistry staining. CONCLUSION: Constitutively expressed in neutrophils and gingival epithelial cells, and calprotectin might maintain the homeostasis and integrity of periodontium. Inflammation-induced expression of calprotectin in GFs, PDL cells, microvascular ECs and bone marrow fibroblasts might process anti-microbial function and promote leukocytes transmigration to defend the host against the microorganisms.

At maturation stage of enamel development, a specialized basal lamina (sBL) was built between ameloblasts and enamel. After the teeth eruption, the ameloblasts transform into the inner cell layer of junctional epithelium. The inner cell layer forms the internal basal lamina of junctional epithelium. However, the composition of the sBL and internal basal lamina was not clarified. The objective of our study was to make a description of the localization of amelotin (AMTN), laminin γ2 (LAMC2) and Odontogenesis-associated phosphoprotein (ODAPH) on the sBL and internal basal lamina. In immunohistochemical study, AMTN, LAMC2 and ODAPH were detected on the sBL at maturation stage. AMTN was also detected in ameloblasts at maturation stage. The expression of AMTN decreased from early-to-late maturation stage. In contrast, the expression of LAMC2 and ODAPH was stable. Immunofluorescence double-staining showed the localization of AMTN was close to enamel surface. However, the localization of ODAPH was close to ameloblasts. LAMC2 and ODAPH were observed on internal basal lamina of junctional epithelium. In contrast, no expression of AMTN was detected on internal basal lamina of junctional epithelium. Our results suggested that ODAPH might participate in enamel maturation and periodontal health, which might provide a better understanding of enamel defects and periodontal disease in clinic.

Odontogenic ameloblast-associated protein (ODAM) is produced by maturation stage ameloblasts and junctional epithelium (JE). The function of ODAM is thought to be involved in the attachment of teeth and JE. To elucidate transcriptional regulation of human ODAM gene in inflamed gingiva, we have analyzed the effects of TNF-α on the expression of ODAM gene in Ca9-22 and Sa3 gingival epithelial cells. Total RNAs were extracted from Ca9-22 and Sa3 cells after stimulation by TNF-α (10 ng/ml). ODAM mRNA and protein levels were analyzed by qPCR and Western blotting. Luciferase (LUC) analyses were performed using LUC constructs inserted in various lengths of ODAM gene promoter. Gel shift and chromatin immunoprecipitation (ChIP) assays were carried out. TNF-α increased ODAM mRNA and protein levels at 3 to 24 h. TNF-α induced LUC activities of the ODAM gene promoter constructs, and the activities were inhibited by protein kinase A, tyrosine kinase, MEK1/2, PI3-kinase and NF-κB inhibitors. Gel shift and ChIP assays revealed that TNF-α increased CCAAT/enhancer-binding protein (C/EBP) β and Yin Yang1 (YY1) binding to three kinds of C/EBPs and YY1 elements. These results demonstrate that TNF-α stimulates ODAM gene transcription via C/EBPs and YY1 elements in the human ODAM gene promoter.

The junctional epithelium (JE) is an epithelial component that attaches directly to the tooth surface and performs the unique function of protecting against bacterial infections; its destruction causes inflammation of the periodontal tissue and loss of alveolar bone. A recent study that used the single-color lineage tracing method reported that JE is maintained by its stem cells. However, the process by which individual stem cells form the entire JE around a whole tooth remains unclear. Using a 4-color lineage tracing method, we performed a detailed examination of the dynamics of individual stem cells that constitute the entire JE. The multicolor lineage tracing method showed that single-color areas, which were derived from each cell color, replaced all the constituent JE cells 168 d after the administration of tamoxifen. The horizontal section of the first molar showed that the single-color areas in the JE expanded widely. We detected putative stem cells at the external basal layer farthest from the enamel. In this study, JE cells that were supplied from different stem cells were visualized as individual monochromatic regions, and the JE around the first molar was maintained by several JE-specific stem cells. These findings indicated that the JE consisted of several cell populations that were supplied from their multiple stem cells and could help to explore the mechanisms involved in periodontal tissue homeostasis.

Background and Objective As the interface between the oral cavity and the teeth, the junctional epithelial barrier is critical for gingival defense. The junctional epithelium is subject to mechanical stresses from biting force or external insults such as bacterial attacks, but little is known about the effects of mechanical stimuli on epithelial functions. Transient receptor potential vanilloid 4 (TRPV4) functions as a mechanosensitive nonselective cation channel. In the present study, based on marked expression of TRPV4 in the mouse junctional epithelium, we aimed to clarify the putative links between TRPV4 and junctional complexes in the junctional epithelium. Methods and Results Histological observations revealed that the junctional epithelium in TRPV4‐deficient (TRPV4−/−) mice had wider intercellular spaces than that in wild‐type (TRPV4+/+) mice. Exogenous tracer penetration in the junctional epithelium was greater in TRPV4−/− mice than in TRPV4+/+ mice, and immunoreactivity for adherens junction proteins was suppressed in TRPV4−/− mice compared with TRPV4+/+ mice. Analysis of a mouse periodontitis model showed greater bone volume loss in TRPV4−/− mice compared with TRPV4+/+ mice, indicating that an epithelial barrier deficiency in TRPV4−/− mice may be associated with periodontal complications. Conclusion The present findings identify a crucial role for TRPV4 in the formation of adherens junctions in the junctional epithelium, which could regulate its permeability. TRPV4 may be a candidate pharmacological target to combat periodontal diseases.

Background Oral gingival tissue, especially the junctional epithelium (JE), is constantly exposed to sub-gingival plaque. A key component of gingival health is the regulation of the number of neutrophils that migrate into the gingival crevice to counteract its harmful effects. This report investigates the contribution of innate defense receptors, Toll-like receptor (TLR)2, TLR4, and both (TLR2/4) to the maintenance of neutrophil homeostasis in the JE. Methods Bacterial composition was analyzed from whole oral swabs collected from 12- to 14-week-old TLR2, TLR4, TLR2/4 double knock-out (KO) mice using a MiSeq platform targeting the V3-V4 region of the 16S ribosomal RNA gene. Mandibles were histologically examined for quantification of neutrophils in the JE and bone loss. Lastly, total bacterial load was quantitated using quantitative real-time PCR. Results Compared with wild-type, all TLR KO mice displayed significantly increased recruitment of neutrophils (P = 0.0079) into the JE. In addition, TLR4 and TLR2/4 KO mice demonstrated a significant increase in the number of bacteria (P = 0.0022 and P = 0.0152, respectively). Lastly, comparative compositional analyses of the oral microbiome revealed that each KO strain harbored unique microbial communities that are distinct from each other but maintained similar levels of alveolar bone. Conclusions Neutrophil migration into healthy mouse JE does not require TLR2 or TLR4. However, a significant increase in the number of neutrophils as well as a significant change in the oral microbial composition in both TLR2 and TLR4 KO mice demonstrate that these TLRs contribute to the homeostatic relationship between bacteria and the host in healthy mice periodontal tissue.

The wound healing process in the soft tissues adjacent to percutaneous implants induces “epithelial downgrowth”, and subsequently, a sinus tract around the device. This provides an optimal environment for bacterial colonization and proliferation. In an attempt to arrest downgrowth and achieve epithelial attachment to a device surface, we have sought to mimic the most common and successful percutaneous organ, the tooth. Since teeth are composed of partially and fully fluoridated forms of hydroxyapatite (HA), it was hypothesized that the surface properties of fluoridated apatites, fluorohydroxyapatite (FHA) and fluorapatite (FA), would improve epithelial cellular adhesion and differentiation when compared to HA and titanium (Ti) surfaces. In this study, the apatites (HA, FHA, and FA) were synthesized and characterized. Following a high-temperature sintering treatment of these apatites, keratinocyte and fibroblast adhesion and differentiation properties were analyzed in vitro, revealing a statistically significant increase in keratinocyte adhesion and terminal differentiation on FA surfaces sintered at 1050–1150 °C as compared to Ti or HA. Moreover, fibroblasts displayed enhanced adhesion on FHA surfaces. This data suggests that percutaneous devices coated with, or fabricated from, fluoridated apatites may induce improved epithelial cellular adhesion and differentiation, potentially limiting deeply penetrating epithelial downgrowth and resultant bacterial ingress.

The junctional epithelium (JE) is a specialized portion of the gingiva that seals off the tooth‐supporting tissues from the oral environment. This relationship is achieved via a unique adhesive extracellular matrix that is, in fact, a specialized basal lamina (sBL). Three unique proteins – amelotin (AMTN), odontogenic ameloblast‐associated (ODAM), and secretory calcium‐binding phosphoprotein proline‐glutamine rich 1 (SCPPPQ1) – together with laminin‐332 structure the supramolecular organization of this sBL and determine its adhesive capacity. Despite the constant challenge of the JE by the oral microbiome, little is known of the susceptibility of the sBL to bacterial degradation. Assays with trypsin‐like proteases, as well as incubation with Porphyromonas gingivalis, Prevotella intermedia, and Treponema denticola, revealed that all constituents, except SCPPPQ1, were rapidly degraded. Porphyromonas gingivalis was also shown to alter the supramolecular network of reconstituted and native sBLs. These results provide evidence that proteolytic enzymes and selected gram‐negative periodontopathogenic bacteria can attack this adhesive extracellular matrix, intimating that its degradation could contribute to progression of periodontal diseases.

Aim To evaluate the similarities and differences in barrier function of a peri-implant epithelium (PIE) versus a native junctional epithelium (JE). Materials and methods A mouse model was used wherein titanium implants were placed sub-occlusally in healed extraction sites. The PIE was examined at multiple timepoints after implant placement, to capture and understand the temporal nature of its assembly and homeostatic status. Mitotic activity, hemidesmosomal attachment apparatus, and inflammatory responses in the PIE were compared against a JE. Additionally, we evaluated whether the PIE developed a Wnt-responsive stem cell niche like a JE. Results The PIE developed from oral epithelium (OE) that had, by the time of implant placement, lost all characteristics of a JE. Compared with a JE, an established PIE had more proliferating cells, exhibited lower expression of attachment proteins, and had significantly more inflammatory cells in the underlying connective tissue. Wnt-responsive cells in the OE contributed to an initial PIE, but Wnt-responsive cells and their descendants were lost as the PIE matured. Conclusions Although histologically similar, the PIE lacked a Wnt-responsive stem cell niche and exhibited characteristics of a chronically inflamed tissue. Both features contributed to suboptimal barrier functions of the PIE compared with a native JE.

Periodontitis is an oral chronic inflammatory disease that is initiated by periodontal microbial communities and requires disruption of the homeostatic responses. The prevalence of periodontal disease increases with age; more than 70% of adults 65 years and older have periodontal disease. A pathogenic microbial community is required for initiating periodontal disease. Dysbiotic immune-inflammatory response and bone remodeling are characteristics of periodontitis. The transcription factor forkhead box protein O1 (FOXO1) is a key regulator of a number of cellular processes, including cell survival and differentiation, immune status, reactive oxygen species (ROS) scavenging, and apoptosis. Although accumulating evidence indicates that FOXO1 activity can be induced by periodontal pathogens, the roles of FOXO1 in periodontal homeostasis and disease have not been well documented. The present review summarizes how the FOXO1 signaling axis can regulate periodontal bacteria-epithelial interactions, immune-inflammatory response, bone remodeling, and wound healing.

There is a need to modify the definition of attached gingiva (AG) as it applies to healthy and diseased teeth and implants. There are two parts to this new definition: Part A is when the biologic width is supracrestal (epithelial attachment and gingival fibers) and is attached to a healthy tooth or tissue-level implant, and the zone of AG is measured from the base of the sulcus to the mucogingival junction (MGJ); Part B is when the biologic width is subcrestal-as with infrabony defects on periodontally involved teeth, periodontally involved tissue-level implants, and bone-level implants placed at or below the bone crest-and the zone of AG is measured from the bone crest (not the base of the sulcus) to the MGJ. Further, what the AG is actually attached to around teeth and different types of implants, and the clinical significance of these differences, are thoroughly discussed.

Increases in adhesive and invasive commensal bacteria, such as Escherichia coli, and subsequent disruption of the epithelial barrier is implicated in the pathogenesis of inflammatory bowel disease (IBD). However, the protective systems against such barrier disruption are not fully understood. Here, we show that secretion of luminal glycoprotein 2 (GP2) from pancreatic acinar cells is induced in a TNF–dependent manner in mice with chemically induced colitis. Fecal GP2 concentration is also increased in Crohn’s diease patients. Furthermore, pancreas-specific GP2-deficient colitis mice have more severe intestinal inflammation and a larger mucosal E. coli population than do intact mice, indicating that digestive-tract GP2 binds commensal E. coli, preventing epithelial attachment and penetration. Thus, the pancreas–intestinal barrier axis and pancreatic GP2 are important as a first line of defense against adhesive and invasive commensal bacteria during intestinal inflammation., Glycoprotein-2 (GP-2) can protect the intestinal epithelial barrier from bacteria and is associated with protection against Crohn’s disease. Here, the authors show pancreatic GP-2 is the source of the intestine’s luminal GP-2 that binds bacteria and prevents them from attaching to the epithelium, also limiting pathology in a DSS colitis mouse model.

Summary The colonic microbiota exhibits cross-sectional heterogeneity, but the mechanisms that govern its spatial organization remain incompletely understood. Here we used Citrobacter rodentium, a pathogen that colonizes the colonic surface, to identify microbial traits that license growth and survival in this spatial niche. Previous work showed that during colonic crypt hyperplasia, type III secretion system (T3SS)-mediated intimate epithelial attachment provides C. rodentium with oxygen for aerobic respiration. However, we find that prior to the development of colonic crypt hyperplasia, T3SS-mediated intimate attachment is not required for aerobic respiration but for hydrogen peroxide (H2O2) respiration using cytochrome c peroxidase (Ccp). The epithelial NADPH oxidase NOX1 is the primary source of luminal H2O2 early after C. rodentium infection and is required for Ccp-dependent growth. Our results suggest that NOX1-derived H2O2 is a resource that governs bacterial growth and survival in close proximity to the mucosal surface during gut homeostasis.

The most fundamental function of an epithelial tissue is to act as a barrier, regulating interactions between the external environment and the body. This barrier function typically requires a contiguous cell layer but since teeth penetrate the oral epithelium, a modified barrier has evolved, called the junctional epithelium (JE). In health, the JE attaches to the tooth, sealing the inside of the body against oral micro-organisms. Breakdown of the JE barrier results in periodontal ligament (PDL) disintegration, alveolar bone resorption, and ultimately tooth loss. Using lineage tracing and DNA pulse-chase analyses, we identified an anatomical location in the JE that supported both fast- and slow-cycling Wnt-responsive stem cells that contributed to self-renewal of the tissue. Stem cells produced daughter cells with an extraordinarily high rate of turnover that maintained JE integrity for 1.4 y in mice. Blocking cell proliferation via a chemotherapeutic agent 5-fluorouracil (5-Fu) eliminated fast-cycling stem cells, which caused JE degeneration, PDL destruction, and bone resorption. Upon removal of 5-Fu, slow-cycling stem cells regenerated both the structure and barrier function of the JE. Taken together, our studies identified a stem cell population in the JE and have potential clinical implications for prevention and treatment of periodontitis.

Purpose The concept of biological width has been proposed and widely used in oral implantation. This review aimed to summarize the biological width around implant in detail. Study selection An electronic search of the literature prior to March 2019 was performed to identify all articles related to biological width in periimplant soft tissue. The search was conducted in the MEDLINE (National Library of Medicine) database accessed through PubMed with no date restriction. The following main keywords were used: "implant", "biological width", "soft tissue", "junctional epithelium", "peri-implant epithelium", "connective tissue", "gingiva", "mucosa" (connecting multiple keywords with AND, OR). Results The identified researches focused on several aspects related to biological width in oral implantation, namely the concept, formation, remodeling, dimension, structure and function. Conclusions Based on of the reviewed literature, the concept, formation, remodeling, structure, dimension, and functional significances of periimplant biological width are explored in this narrative review. The formation of biological width around implant is a complex process after several weeks of healing. The biological width around implant is a 3-4mm distance from the top of the peri-implant mucosa to the first bone-to-implant contact or the stabilized top of the adjacent bone, consisting of sulcular epithelium, junctional epithelium and fibrous connective tissue between the epithelium and the first bone-to-implant contact or the stabilized top of the adjacent bone. The biological width forms a biological barrier against the bacteria, influences the remodeling of soft and hard tissue around implant and has implications for clinical aspects of dental implantation.

Objective The aim of this investigation was to study the effects of Runt-related transcription factor 2 (Runx2) on the junctional epithelium and alveolar bone. Methods The attachment level of the junctional epithelium and the resorption of alveolar bone were analyzed by histology and scanning electron microscopy. The expression of amelotin was determined by immunohistochemistry, Western blot, and real-time PCR. The ultrastructure of the dentogingival interface was observed by transmission electron microscopy. Results The cKO mice demonstrated remarkable attachment loss, epithelial hyperplasia, and alveolar bone loss. The relative protein and mRNA expression of amelotin was increased in the junctional epithelium of the cKO mice. The attachment apparatus of the cKO mice showed ultrastructural deficiency. Conclusions Loss of Runx2 led to the junctional epithelium and alveolar bone defects in mice. Runx2 may play a crucial role in maintaining the integrity of the dentogingival junction and the normal structure of alveolar bone.

Purpose Anterior stromal puncture is an inexpensive technique for treating recurrent corneal erosions but is often ineffective and cannot be used in the optical axis because of scarring. These studies tested a novel microfabricated imprinting instrument to assess its potential efficacy for the treatment of corneal epithelial disease in the optical axis. Methods The device is made using glass rods, bundled and drawn through multiple iterative cycles, and then fused under high heat to generate a solid rod comprised of many parallel, aligned, cladded fibers. The rods are sliced into discs and then etched to yield designable spikes based on the borosilicate composition of the glass. Results Imprinting the cornea yields a regular pattern of imprints. Histologic studies showed both nonpenetrating stable deformations of Bowman layer, with formation of stable epithelial attachments, and full thickness penetration, with superficial ingrowth of the basal epithelium. Conclusions Microimprinted corneal tissue shows focal subepithelial scarring without evidence of optically evident anterior stromal scarring, and may be an effective way of treating recurrent corneal erosions in the optical axis, which is not currently possible using standard anterior stromal puncture methods.