Microcomputed-Tomographic Analysis of the Root Canal Morphology of the Mandibular First Molars in a Malaysian Subpopulation Using Two Classification Systems / Análisis por Tomografía Microcomputada de la Morfología del Conducto Radicular de los Primeros Molares Mandibulares en una Subpoblación de Malasia Utilizando dos Sistemas de Clasificación

SUMMARY: This work investigated the morphology of the root canal system of the mandibular first molar in a Malaysian subpopulation. Using micro-computed tomography with an isotropic resolution of 22 µm, 140 mandibular first molars were scanned. MIMICS software was used for segmentation, 3-D reconstruction and analysis of the acquired images. The canal configuration was described using Vertucci [supported by the supplementary configurations proposed by Sert & Bayirli (2004)] and Ahmed et al. (2027), coding systems. The chi-square test was used to assess the association between qualitative variables. By non-considering intercanal communications, Vertucci types IV (17.1%) and I (76.4%) were the most frequently reported configurations in the mesial and distal roots, respectively. Of the reported configurations, 24.3% and 4.3% were non-classifiable by Vertucci system in the mesial and distal roots, respectively. Up to 63.6% and 9.3% of the reported configurations were non- classifiable, and type I was the most frequent when considering intercanal communications (7.1% and 76.4% in the mesial and distal roots, respectively). According to Ahmed et al., system, almost half of the sample had more than four digits (47.9%), followed by the 3-digits category (20.71%). In both systems, a significant association was found between the canal configuration and the root type (p<0.001). The mandibular first molar of this Malaysian subpopulation demonstrated a wide range of root canal morphology. When compared to the Vertucci system, the system developed by Ahmed et al., successfully classified all molars configurations despite their level of complexity. The complex canal anatomy of mandibular first molars in this subpopulation warrants special attention during root canal treatment procedures.

En este trabajo se investigó la morfología del sistema de conductos radiculares del primer molar mandibular en una subpoblación de Malasia. Utilizando tomografía microcomputada con una resolución isotrópica de 22 µm, se escanearon 140 primeros molares mandibulares. Se utilizó el software MIMICS para segmentar (enmascarar), reconstruir en 3D, visualizar y analizar las imágenes adquiridas. La configuración del canal se describió utilizando Vertucci respaldado por las configuraciones complementarias propuestas por Sert & Bayirli (2004)] y Ahmed et al. (2017, 2020), sistemas de codificación. Se utilizó la prueba de chi-cuadrado para evaluar la asociación entre variables cualitativas. Sin considerar las comunicaciones intercanales, los tipos Vertucci IV (17,1%) y I (76,4%) fueron las configuraciones reportadas con mayor frecuencia en las raíces mesiales y distales, respectivamente. De las configuraciones reportadas, el 24,3 % y el 4,3 % fueron no clasificables por el sistema de Vertucci en las raíces mesial y distal, respectivamente. Hasta el 63,6 % y el 9,3 % de las configuraciones reportadas fueron no clasificables, siendo la tipo I la más frecuente al considerar las comunicaciones intercanales (7,1 % y 76,4 % en las raíces mesiales y distales, respectivamente). Según Ahmed et al. (2017, 2020) en el sistema, casi la mitad de la muestra tenía más de cuatro dígitos (47,9 %), seguido por la categoría de 3 dígitos (20,71 %). En ambos sistemas se encontró una asociación significativa entre la configuración del canal y el tipo de raíz (p<0,001). El primer molar mandibular de esta subpoblación de Malasia demostró una amplia gama morfológica del conducto radicular. En comparación con el sistema Vertucci, el sistema desarrollado por Ahmed et al. (2017, 2020) clasificaron con éxito todas las configuraciones de los molares a pesar de su nivel de complejidad. La compleja anatomía del canal de los primeros molares mandibulares en esta subpoblación merece una atención especial durante los procedimientos de tratamiento de conducto.

Contour Analysis of Three-Dimensional Peri-Implant Mucosal Model as an Endpoint Analysis of Photofunctionalization Effects on Implant Abutment Materials.

INTRODUCTION: The objective of this study was to examine the effect of photofunctionalization on the soft-tissue contour formed at the interface of various abutment materials using end-point analyses obtained from the three-dimensional oral mucosal model (3D-OMMs). METHODS: Commercially pure titanium (CPTi), alumina-toughened zirconia (ATZ), and yttria-stabilized zirconia (YSZ) made into discs shapes were classified into two groups: UV-treated (PTx) and non-treated (NTx). The materials in PTx groups were exposed to UV light for 12 min. Human gingival fibroblasts and TR146 epithelial cell lines co-cultured on the acellular dermal membrane were used to construct the 3D-OMM. After 4 days of culture, the discs were inserted into the holes prepared within the membrane of 3D-OMMs. The contour formed by the tissue was evaluated after 14 days of culture. RESULTS: The UV treatment of abutment materials resulted in the formation of more non-pocket-tissue types among the PTx group (p = 0.002). Of all materials tested, soft tissue contour around YSZ showed higher scores for the non-pocket type in both non- and UV-treated groups. CONCLUSIONS: The non-pocket type of tissue attachment was frequently found in all surfaces modified by photofunctionalization, particularly zirconia. The 3D-OMM can be used to evaluate the biological endpoints of implant surface modifications.

Root and canal anatomy of mandibular first molars using micro-computed tomography: a systematic review.

BACKGROUND: A thorough understanding of root and canal anatomy is crucial for successful root canal treatment outcomes. This systematic review aims to explore the published micro-CT studies investigated the anatomy of root and canal system in permanent mandibular first molars. METHOD: An electronic search was performed on Web of science, PubMed, and Scopus. Micro-CT journal studies investigated the root and canal anatomy of permanent double-rooted mandibular first molars were included. Data on study characteristics, objectives of interest, specifications of the studies, and micro-CT specifications were extracted. Risk of bias assessment (ROB) of the included studies was performed using Anatomical Quality Assessment (AQUA) tool. The extracted data were presented in tables and figures to present and synthesise the results. A meta-analysis was performed for the studies related to the prevalence of Vertucci's canal configurations, middle mesial canal (MMC) configurations, and Fan's isthmus types. RESULTS: Amongst 1358 identified studies, thirty met the inclusion criteria. In terms of the objectives, the selected studies showed high anatomical variability in mandibular first molars. Twenty-two (73%), 25 (83%), and 12 (40%) of the studies reported the population/ethnicity, micro-CT specifications, and ethical approval, respectively. 28 (93%) studies did not disclose the method of sample size estimation. In only 6 (20%) of the studies, the authors had calibrated the assessment approaches. Mostly, a potential ROB was reported in domain 1 (objective(s) and subject characteristics) and domain 3 (methodology characterization). Whilst, low risk was reported in domains 2 (study design), 4 (descriptive anatomy), and 5 (reporting of results). The overall ROB was reported to be ''moderate'' in the vast majority of the studies (27/30). Meta-analysis results showed high levels of heterogeneity among the studies related to MMCs (I2 = 86%) and Fan's isthmus (I2 = 87%). As for the root canal configuration, pooled prevalence showed that Vertucci type IV and type I were the most prevalent in mesial and distal root canals, respectively. CONCLUSION: Based on moderate risk of bias level of evidence, micro-CT studies have shown wide range of qualitative and quantitative data presentations of the roots and canals in mandibular first molars. Protocol and registration. The protocol of this systematic review was prospectively registered in the Open Science Framework database ( https://osf.io ) on 2022-06-20 with the registration number 10.17605/OSF.IO/EZP7K.

Expression of Interleukin-1ß and Histological Changes of the Three-Dimensional Oral Mucosal Model in Response to Yttria-Stabilized Nanozirconia.

Over the years, advancement in ceramic-based dental restorative materials has led to the development of monolithic zirconia with increased translucency. The monolithic zirconia fabricated from nano-sized zirconia powders is shown to be superior in physical properties and more translucent for anterior dental restorations. Most in vitro studies on monolithic zirconia have focused mainly on the effect of surface treatment or the wear of the material, while the nanotoxicity of this material is yet to be explored. Hence, this research aimed to assess the biocompatibility of yttria-stabilized nanozirconia (3-YZP) on the three-dimensional oral mucosal models (3D-OMM). The 3D-OMMs were constructed using human gingival fibroblast (HGF) and immortalized human oral keratinocyte cell line (OKF6/TERT-2), co-cultured on an acellular dermal matrix. On day 12, the tissue models were exposed to 3-YZP (test) and inCoris TZI (IC) (reference material). The growth media were collected at 24 and 48 h of exposure to materials and assessed for IL-1ß released. The 3D-OMMs were fixed with 10% formalin for the histopathological assessments. The concentration of the IL-1ß was not statistically different between the two materials for 24 and 48 h of exposure (p = 0.892). Histologically, stratification of epithelial cells was formed without evidence of cytotoxic damage and the epithelial thickness measured was the same for all model tissues. The excellent biocompatibility of nanozirconia, as evidenced by the multiple endpoint analyses of the 3D-OMM, may indicate the potential of its clinical application as a restorative material.

An In-Vitro Analysis of Peri-Implant Mucosal Seal Following Photofunctionalization of Zirconia Abutment Materials.

The presence of epithelial and connective tissue attachment at the peri-implant-soft tissue region has been demonstrated to provide a biological barrier of the alveolar bone from the oral environment. This barrier can be improved via surface modification of implant abutment materials. The effect of photofunctionalization on creating a bioactive surface for the enhancement of the epithelial and connective tissue attachment of zirconia implant abutment's peri-implant mucosal interface using organotypic model has not been investigated. Therefore, this study aimed to evaluate the soft tissue seal around peri-implant mucosa and to understand the effect of photofunctionalization on the abutment materials. Three types of abutment materials were used in this study; yttria-stabilized zirconia (YSZ), alumina-toughened zirconia, and grade 2 commercially pure titanium (CPTi) which were divided into nontreated (N-Tx) and photofunctionalized group (UV-Tx). The three-dimensional peri-implant mucosal model was constructed using primary human gingival keratinocytes and fibroblasts co-cultured on the acellular dermal membrane. The biological seal was determined through the concentration of tritiated water permeating the material-soft tissue interface. The biological seal formed by the soft tissue in the N-Tx group was significantly reduced compared to the UV-treated group (p < 0.001), with YSZ exhibiting the lowest permeability among all materials. Photofunctionalization of implant abutment materials improved the biological seal of the surrounding soft tissue peri-implant interface.

The clinical significance of the retromolar canal and foramen in dentistry.

The mandibular canal is nowadays acknowledged as a major trunk with multiple smaller branches running roughly parallel to it. Most of these accessory canals contain branches of the inferior alveolar neurovascular bundle that supplies the dentition, jawbone, and soft tissue around the gingiva and lower lip. This article reviews the prevalence, classification and morphometric measurements of the retromolar canal and its aperture. A retromolar canal is a bifid variation of the mandibular canal that divides from above this main canal, and travels anterosuperiorly within the bone to exit via a single foramen or multiple foramina into the retromolar fossa. This foramen, termed the retromolar foramen, allows accessory branches of the inferior alveolar neurovascular bundles to supply tissues at the retromolar trigone. Clinically, it is of the utmost importance to determine the exact location of the mandibular canal and to identify its retromolar accessory branches when surgery in the posterior mandible is to be performed.

Deregulation of lysophosphatidic acid metabolism in oral cancer promotes cell migration via the up-regulation of COX-2.

Oral squamous cell carcinoma (OSCC) is the sixth most common cancer worldwide and accounts for 300,000 new cases yearly. The five-year survival rate is approximately 50% and the major challenges to improving patient prognosis include late presentation, treatment resistance, second primary tumours and the lack of targeted therapies. Therefore, there is a compelling need to develop novel therapeutic strategies. In this study, we have examined the effect of lysophosphatidic acid (LPA) on OSCC cell migration, invasion and response to radiation, and investigated the contribution of cyclooxygenase-2 (COX-2) in mediating the tumour promoting effects of LPA. Using the TCGA data set, we show that the expression of the lipid phosphate phosphatases (LPP), LPP1 and LPP3, was significantly down-regulated in OSCC tissues. There was no significant difference in the expression of the ENPP2 gene, which encodes for the enzyme autotaxin (ATX) that produces LPA, between OSCCs and control tissues but ENPP2 levels were elevated in a subgroup of OSCCs. To explore the phenotypic effects of LPA, we treated OSCC cell lines with LPA and showed that the lipid enhanced migration and invasion as well as suppressed the response of the cells to irradiation. We also show that LPA increased COX-2 mRNA and protein levels in OSCC cell lines and inhibition of COX-2 activity with the COX-2 inhibitor, NS398, attenuated LPA-induced OSCC cell migration. Collectively, our data show for the first time that COX-2 mediates some of the pro-tumorigenic effects of LPA in OSCC and identifies the ATX-LPP-LPA-COX-2 pathway as a potential therapeutic target for this disease.

Profiling lysophosphatidic acid levels in plasma from head and neck cancer patients.

Head and neck squamous cell carcinoma (HNSCC) represents a significant world health problem, with approximately 600,000 new cases being diagnosed annually. The prognosis for patients with HNSCC is poor and, therefore, the identification of biomarkers for screening, diagnosis and prognostication would be clinically beneficial. A limited number of studies have used lipidomics to profile lipid species in the plasma of cancer patients. However, the profile and levels of lysophosphatidic acid (LPA) species have not been examined in HNSCC. In this study, a targeted lipidomics approach using liquid chromatography triple quadrupole mass spectrometry (LCMS/MS) was used to analyse the concentration of LPA (16:0 LPA, 18:0 LPA, 18:1 LPA, 18:2 LPA and 20:4 LPA) in the plasma of patients with oral squamous cell carcinoma (OSCC) and nasopharyngeal carcinoma (NPC), together with healthy controls. The levels of three LPA species (18:1 LPA, 18:2 LPA and 20:4 LPA) were significantly lower in the plasma of OSCC patients, whilst the concentrations of all five LPA species tested were significantly lower in plasma from NPC patients. Furthermore, the order of abundance of LPA species in plasma was different between the control and cancer groups, with 16:0 LPA, 18:0 LPA levels being more abundant in OSCC and NPC patients. Medium to strong correlations were observed using all pairs of LPA species and a clear separation of the normal and tumour groups was observed using PCA analysis. In summary, the results of this study showed that the levels of several LPA species in the plasma of patients with OSCC and NPC were lower than those from healthy individuals. Understanding these variations may provide novel insights into the role of LPA in these cancers.

The clinical anatomy of accessory mandibular canal in dentistry.

The mandibular canal is a conduit that allows the inferior alveolar neurovascular bundle to transverse the mandible to supply the dentition, jawbone and soft tissue around the gingiva and the lower lip. It is not a single canal but an anatomical structure with multiple branches and variations. The branches are termed accessory, bifid or trifid canals depending on their number and configuration. A bifid mandibular canal is an anatomical variation reported more commonly than the trifid variant. Because of these variations, it is of the utmost importance to determine the exact location of the mandibular canal and to identify any branches arising from it prior to performing surgery in the mandible. This article reviews the prevalence, classification and morphometric measurements of these accessory mandibular canals, emphasizing their clinical significance.

Angiogenic effect of platelet-rich concentrates on dental pulp stem cells in inflamed microenvironment.

OBJECTIVE: In this study, we aimed to determine the suitable concentrations of human platelet lysate (HPL) and platelet-rich plasma (PRP) for maintaining the in vitro proliferative and angiogenic potential of inflamed dental pulp stem cells. MATERIALS AND METHODS: Lipopolysaccharide (LPS)-induced inflamed dental pulp-derived stem cells (iDPSCs) were treated with different concentrations of HPL and PRP (10% and 20%) followed by determination of viability using Alamar Blue assay. Expression of angiogenesis-, adhesion-, and inflammation-regulating genes was also analyzed using RT-qPCR array. Furthermore, expression of growth factors at protein level in the cell culture microenvironment was measured using multiplex assay. RESULTS: Viability of iDPSCs was significantly (p < 0.05) higher in 20% HPL-supplemented media compared to iDPSCs. Expression of 10 out of 12 selected angiogenic genes, four out of seven adhesion molecules, and seven out of nine cytokine-producing genes were significantly (p < 0.05) higher in cells maintained in 20% HPL-supplemented media compared to that in FBS-supplemented media. Furthermore, expression of all the selected growth factors was significantly higher (p < 0.05) in the supernatants from 20% HPL media at 12 and 24 h post-incubation. CONCLUSION: This study suggests that 20% HPL could be optimum to stimulate angiogenesis-related factors in iDPSCs while maintaining their viability. CLINICAL RELEVANCE: This data may suggest the potential use of 20% HPL for expanding DPSCs scheduled for clinical trials for regenerative therapies including dental pulp regeneration.

Immune responses of human dental pulp stem cells in lipopolysaccharide-induced microenvironment.

This study aimed to investigate the effect of inflammatory stimuli on dental pulp stem cells (DPSCs) by assessing their proliferation and expression of genes as well as proteins in lipopolysaccharide (LPS)-induced microenvironment (iDPSCs). DPSCs were first characterized for their mesenchymal properties prior to challenging them with a series of LPS concentrations from 12 to 72 h. Following to this, their proliferation and inflammatory based genes as well as protein expression were assessed. iDPSCs had demonstrated significant expression of mesenchymal markers. Upon exposure to LPS, the viability dropped distinctly with increasing concentration, as compared to control (P < 0.05). The expression of pro-inflammatory genes such as interleukin 6, interleukin 8 were augmented with exposure to LPS (P < 0.05). Similarly, cytokines like tumour necrosis factor (TNF) α and interleukin 1α had increased in dose dependant manner upon LPS exposure (P < 0.05). Our results suggest that LPS concentration between 1 and 2 µg/mL demonstrated inflammation induction in DPSCs that may simulate inflamed microenvironment of dental pulp in clinical scenario. Thus, optimizing iDPSCs secretome profile could be a promising approach to test various regenerative protocols in inflamed microenvironment.

Modified porous scaffolds of silk fibroin with mimicked microenvironment based on decellularized pulp/fibronectin for designed performance biomaterials in maxillofacial bone defect.

Maxillofacial bone defect is a critical problem for many patients. In severe cases, the patients need an operation using a biomaterial replacement. Therefore, to design performance biomaterials is a challenge for materials scientists and maxillofacial surgeons. In this research, porous silk fibroin scaffolds with mimicked microenvironment based on decellularized pulp and fibronectin were created as for bone regeneration. Silk fibroin scaffolds were fabricated by freeze-drying before modification with three different components: decellularized pulp, fibronectin, and decellularized pulp/fibronectin. The morphologies of the modified scaffolds were observed by scanning electron microscopy. Existence of the modifying components in the scaffolds was proved by the increase in weights and from the pore size measurements of the scaffolds. The modified scaffolds were seeded with MG-63 osteoblasts and cultured. Testing of the biofunctionalities included cell viability, cell proliferation, calcium content, alkaline phosphatase activity (ALP), mineralization and histological analysis. The results demonstrated that the modifying components organized themselves into aggregations of a globular structure. They were arranged themselves into clusters of aggregations with a fibril structure in the porous walls of the scaffolds. The results showed that modified scaffolds with a mimicked microenvironment of decellularized pulp/fibronectin were suitable for cell viability since the cells could attach and spread into most of the pores of the scaffold. Furthermore, the scaffolds could induce calcium synthesis, mineralization, and ALP activity. The results indicated that modified silk fibroin scaffolds with a mimicked microenvironment of decellularized pulp/fibronectin hold promise for use in tissue engineering in maxillofacial bone defects. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1624-1636, 2017.

An injectable poly(caprolactone trifumarate-gelatin microparticles) (PCLTF-GMPs) scaffold for irregular bone defects: Physical and mechanical characteristics.

Recently, a modified form of a three-dimension (3D) porous poly(caprolactone-trifumarate) (PCLTF) scaffold has been produced using a fabrication technique that involves gelatin microparticles porogen leaching. This poly(caprolactone trifumarate-gelatin microparticles) (PCLTF-GMPs) scaffold has been shown to be biocompatible, more flowable clinically, and has a shorter degradation time as compared to its existing predecessors. In this report, a detailed characterization of this new scaffold was performed by testing its cytocompatibility, analyzing the surface topography, and understanding its thermal, physical and mechanical properties. The result showed that the PCLTF-GMPs has no critical cytotoxic effect. To confirm improvement, the surface properties were compared against the older version of PCLTF fabricated using salt porogen leaching. This PCLTF-GMPs scaffold showed no significant difference (unpaired t-test; p>0.05) in mechanical properties before and after gelatin leaching. However, it is mechanically weaker when compared to its predecessors. It has a high biodegradability rate of 16weeks. The pore size produced ranges from 40 to 300µm, and the RMS roughness is 613.7±236.9nm. These characteristics are condusive for osteoblast in-growth, as observed by the extension of filopodia across the macropores. Overall, this newly produced material has good thermal, physical and mechanical properties that complements its biocompatibility and ease of use.

The Management of Persistent Pain From a Branch of the Trifid Mandibular Canal due to Implant Impingement.

The mandibular canal is a conduit that allows the inferior alveolar neurovascular bundle to transverse the mandible to supply the dentition, jawbone, and soft tissue around the lower lip. It is now acknowledged that the mandibular canal is not a single canal but an anatomical structure with multiple branches and variations. Iatrogenic injury to branches of the mandibular canal that carry a neurovascular bundle has been reported to cause injury to the main canal as severe as if the main canal itself is traumatized. These injuries include bleeding, neurosensory disturbance, or the formation of traumatic neuroma, and so far, they have involved cases with the bifid mandibular canal. This current report presents a case of neurosensory disturbance that resulted from the impingement of a branch of a trifid mandibular canal during implant insertion. Its management included analgesics, reexamination, and reinserting a shorter implant.

Modified silk fibroin scaffolds with collagen/decellularized pulp for bone tissue engineering in cleft palate: Morphological structures and biofunctionalities.

Cleft palate is a congenital malformation that generates a maxillofacial bone defect around the mouth area. The creation of performance scaffolds for bone tissue engineering in cleft palate is an issue that was proposed in this research. Because of its good biocompatibility, high stability, and non-toxicity, silk fibroin was selected as the scaffold of choice in this research. Silk fibroin scaffolds were prepared by freeze-drying before immerging in a solution of collagen, decellularized pulp, and collagen/decellularized pulp. Then, the immersed scaffolds were freeze-dried. Structural organization in solution was observed by Atomic Force Microscope (AFM). The molecular organization of the solutions and crystal structure of the scaffolds were characterized by Fourier transform infrared (FT-IR) and X-ray diffraction (XRD), respectively. The weight increase of the modified scaffolds and the pore size were determined. The morphology was observed by a scanning electron microscope (SEM). Mechanical properties were tested. Biofunctionalities were considered by seeding osteoblasts in silk fibroin scaffolds before analysis of the cell proliferation, viability, total protein assay, and histological analysis. The results demonstrated that dendrite structure of the fibrils occurred in those solutions. Molecular organization of the components in solution arranged themselves into an irregular structure. The fibrils were deposited in the pores of the modified silk fibroin scaffolds. The modified scaffolds showed a beta-sheet structure. The morphological structure affected the mechanical properties of the silk fibroin scaffolds with and without modification. Following assessment of the biofunctionalities, the modified silk fibroin scaffolds could induce cell proliferation, viability, and total protein particularly in modified silk fibroin with collagen/decellularized pulp. Furthermore, the histological analysis indicated that the cells could adhere in modified silk fibroin scaffolds. Finally, it can be deduced that modified silk fibroin scaffolds with collagen/decellularized pulp had the performance for bone tissue engineering and a promise for cleft palate treatment.

Detection of host-specific immunogenic proteins in the saliva of patients with oral squamous cell carcinoma.

The main purpose of this article is to develop a new and reliable saliva-based clinical diagnostic method for the early detection of oral squamous cell carcinoma (OSCC). This study used an immunoproteomic approach which allowed the detection of immunogenic host proteins in patients' samples using pooled human antibodies. In an attempt to investigate potential biomarkers of OSCC, two-dimensional electrophoresis (2-DE) followed by immunoblotting of saliva from patients and controls were compared. The protein spots of interest were analyzed using 2-DE image analyzer and subsequently subjected to MALDI-TOF/TOF and then matched against NCBI database. The result showed that four protein clusters, namely Human Pancreatic Alpha-amylase (HPA), Human Salivary Amylase (sAA), keratin-10 (K-10), and Ga Module Complexed with Human Serum Albumin (GA-HSA), had exhibited immunoreactivity in western blot. The results are suggestive of the potential use of the differentially expressed saliva protein as tumor biomarkers for the detection of OSCC. However, further studies are recommended to validate this finding.

Development of a novel model for the investigation of implant-soft tissue interface.

BACKGROUND: In dental implant treatment, the long-term prognosis is dependent on the biologic seal formed by the soft tissue around the implant. The in vitro investigation of the implant-soft tissue interface is usually carried out using a monolayer cell-culture model that lacks a polarized-cell phenotype. This study developed a tissue-engineered three-dimensional oral mucosal model (3D OMM) to investigate the implant-soft tissue interface. METHODS: A 3D OMM was constructed using primary human oral keratinocytes and fibroblasts cultured on a skin-derived scaffold at an air-liquid interface. A titanium implant was inserted into the engineered oral mucosa and further cultured to establish epithelial attachment. The 3D OMM was characterized using basic histology and immunostaining for cytokeratin (CK) 10 and CK13. Histomorphometric analyses of the implant-soft tissue interface were carried out using a light-microscopy (LM) examination of ground sections and semi-thin sections as well as scanning electron microscopy (SEM). RESULTS: Immunohistochemistry analyses suggests that the engineered oral mucosa closely resembles the normal oral mucosa. The LM and SEM examinations reveal that the 3D OMM forms an epithelial attachment on the titanium surface. CONCLUSION: The 3D OMM provided mimicking peri-implant features as seen in an in vivo model and has the potential to be used as a relevant alternative model to assess implant-soft tissue interactions.