Collagen: The Superior Material for Full-Thickness Oral Mucosa Tissue Engineering.

BACKGROUND: Tissue engineering has significantly progressed in developing full-thickness oral mucosa constructs designed to replicate the natural oral mucosa. These constructs serve as valuable in vitro models for biocompatibility testing and oral disease modeling and hold clinical potential for replacing damaged or lost oral soft tissue. However, one of the major challenges in tissue engineering of the oral mucosa is the identification of an appropriate scaffold with optimal porosity, interconnected porous networks, biodegradability, and biocompatibility. These characteristics facilitate cell migration, nutrient delivery, and vascularization. Various biomaterials have been investigated for constructing tissue-engineered oral mucosa models; collagen has demonstrated superior outcomes compared with other materials. HIGHLIGHT: This review discusses the different types of tissue-engineered oral mucosa developed using various materials and includes articles published between January 2000 and December 2022 in PubMed and Google Scholar. The review focuses on the superiority of collagen-based scaffolds for tissue engineering of oral mucosa, explores in vitro applications, and discusses potential clinical applications. CONCLUSION: Among the various scaffold materials used for engineering the connective tissue of the oral mucosa, collagen-based scaffolds possess excellent biological properties, offering high-quality oral mucosa constructs and high resemblance to the native human oral mucosa in terms of histology and expression of various differentiation markers.

Evaluation of Mental Foramen and Mandibular Canal Course and Location in an Emirati Subpopulation.

INTRODUCTION AND AIMS: Violations of the mandibular canal (MC) and mental foramen (MF) and subsequent injuries to their neurovascular bundle have been reported after surgical and nonsurgical dental procedures. Besides using advanced technologies such as cone-beam computed tomography (CBCT), clinicians should be aware of the anatomy and location of MC and MF in different populations. This study aims to describe the morphologic characteristics of the MF, MC, and its intrabony location in relation to the apices of mandibular posterior teeth in an Emirati subpopulation using CBCT. METHODS: A total of 3700 CBCT scans were screened, and 154 scans that met the inclusion and exclusion criteria were randomly selected. The scans were assessed using 3-dimensional multiplanar imaging for the following structures: the location of MF and the MC course, its intrabony location, and its relationship to the apices of the mandibular posterior teeth. The data were analysed statistically using SPSS software. RESULTS: The MC ran lingually and inferiorly at the posterior region and became more buccal and superior towards the MF. The distal root of the mandibular second molar was found to be the closest root to the MC (2.06 ± 1.83 mm). Moreover, the most common location of the MF was distal to the contact area between the 2 premolars (0.83 ± 1.84 mm) with a significant negative correlation to age (with and increase in age, the MF moves distally). The distance between the root apices and the MC was statistically significantly affected by age (positive correlation) and gender (male patients had a greater distance). CONCLUSIONS: The common course of the MC is lingual and inferior posteriorly and becomes more buccal and superior towards the MF, which is located mostly between the mandibular first and second premolars. Furthermore, the distal root of the mandibular second molar is the closest to the MC and has a positive relationship with age.

Biological Evaluation of Oral Care Products Using 3D Tissue-Engineered In Vitro Models of Plaque-Induced Gingivitis.

BACKGROUND: The aim of this study was to investigate and visualize the anti-inflammatory and anti-bacterial effects of different oral care products using an infected and inflamed 3D tissue-engineered gingival mucosal model. METHODS: A 3D full-thickness oral mucosal model was engineered inside tissue culture inserts using collagen hydrogels populated with human gingival fibroblasts and THP-1 monocytes and layered with oral epithelial cell lines. Oral saliva bacteria were cultured and added to the surface of the models and inflammation was further simulated with lipopolysaccharide (LPS) of Escherichia coli. The 3D models were exposed to three different types of toothpastes, a chlorhexidine antiseptic mouthwash, different antibiotics, and a mechanical rinse with phosphate-buffered saline (PBS) prior to biological evaluation using the PrestoBlue tissue viability assay, histology, optical coherence tomography (OCT), confocal microscopy, and measurement of the release of the inflammatory markers IL-1ß, IL-6, and IL-8 with ELISA. RESULTS: Multiple-endpoint analyses of the infected oral mucosal models treated with different anti-bacterial agents showed consistent outcomes in terms of tissue viability, histology, OCT, and confocal microscopy findings. In terms of anti-inflammatory testings, the positive control group showed the highest level of inflammation compared with all other groups. Depending on the anti-bacterial and anti-inflammatory potential of the test groups, different levels of inflammation were observed in the test groups. CONCLUSIONS: The inflamed 3D oral mucosal model developed in this study has the potential to be used as a suitable in vitro model for testing the biocompatibility, anti-inflammatory, and anti-bacterial properties of oral care products including mouthwashes and toothpastes. The results of this study indicate that the chlorhexidine mouthwash has both anti-bacterial and cytotoxic effects on the 3D oral mucosal model. Hyaluronic-acid-containing toothpaste has significant anti-bacterial and anti-inflammatory effects on the 3D oral mucosal model.

Effect of Different Silane Coupling Agents on the Bond Strength between Hydrogen Peroxide-Etched Epoxy-Based- Fiber-Reinforced Post and Composite Resin Core.

The aim of the study was to evaluate the effect of various silane coupling agents on the micro-push-out bond strength between a hydrogen peroxide-etched epoxy-based fiber-reinforced post and composite resin core. Seventy-five cross-linked epoxy-based fiber-reinforced posts were etched with 24% hydrogen peroxide for 10 min. Then they were divided into five groups according to various silane coupling agents and bonded to a composite core. A Universal Testing Machine was utilized to evaluate the push-out bond strength. In addition, all groups' modes of failure were assessed. The push-out bond strength data in MPa were analyzed using ANOVA and a Tukey HSD post hoc test to reveal any difference between the groups. Results revealed that the application of a two-bottle silane coupling agent exhibited the highest bond strength, while the application of a one-bottle silane coupling agent demonstrated the lowest bond strength for a hydrogen peroxide-etched fiber post bonded to a composite core material, which was statistically significant (p < 0.05). The strongest association with the highest bond strength was found with the two-bottle silane coupling agent when compared to the one-bottle. The study highlighted that the application of a silane-coupling agent may affect the bond strength between composite and epoxy-based fiber-reinforced posts.

Development and Comparison of Conventional and 3D-Printed Laboratory Models of Maxillary Defects.

BACKGROUND: Recording accurate impressions from maxillary defects is a critical and challenging stage in the prosthetic rehabilitation of patients following maxillectomy surgery. The aim of this study was to develop and optimize conventional and 3D-printed laboratory models of maxillary defects and to compare conventional and digital impression techniques using these models. METHODS: Six different types of maxillary defect models were fabricated. A central palatal defect model was used to compare conventional silicon impressions with digital intra-oral scanning in terms of dimensional accuracy and total time taken to record the defect and produce a laboratory analogue. RESULTS: Digital workflow produced different results than the conventional technique in terms of defect size measurements which were statistically significant (p < 0.05). The time taken to record the arch and the defect using an intra-oral scanner was significantly less compared with the traditional impression method. However, there was no statistically significant difference between the two techniques in terms of the total time taken to fabricate a maxillary central defect model (p > 0.05). CONCLUSIONS: The laboratory models of different maxillary defects developed in this study have the potential to be used to compare conventional and digital workflow in prosthetic treatment procedures.

Molecular insights into the role of electronic cigarettes in oral carcinogenesis.

Electronic cigarette (EC) usage or vaping has seen a significant rise in recent years across various parts of the world. They have been publicized as a safe alternative to smoking; however, this is not supported strongly by robust research evidence. Toxicological analysis of EC liquid and aerosol has revealed presence of several toxicants with known carcinogenicity. Oral cavity is the primary site of exposure of both cigarette smoke and EC aerosol. Role of EC in oral cancer is not as well-researched as that of traditional smoking. However, several recent studies have shown that it can lead to a wide range of potentially carcinogenic molecular events in oral cells. This review delineates the oral carcinogenesis potential of ECs at the molecular level, providing a summary of the effects of EC usage on cancer therapy resistance, cancer stem cells (CSCs), immune evasion, and microbiome dysbiosis, all of which may lead to increased tumor malignancy and poorer patient prognosis. This review of literature indicates that ECs may not be as safe as they are perceived to be, however further research is needed to definitively determine their oncogenic potential.

3D Printing of Dental Prostheses: Current and Emerging Applications.

Revolutionary fabrication technologies such as three-dimensional (3D) printing to develop dental structures are expected to replace traditional methods due to their ability to establish constructs with the required mechanical properties and detailed structures. Three-dimensional printing, as an additive manufacturing approach, has the potential to rapidly fabricate complex dental prostheses by employing a bottom-up strategy in a layer-by-layer fashion. This new technology allows dentists to extend their degree of freedom in selecting, creating, and performing the required treatments. Three-dimensional printing has been narrowly employed in the fabrication of various kinds of prostheses and implants. There is still an on-demand production procedure that offers a reasonable method with superior efficiency to engineer multifaceted dental constructs. This review article aims to cover the most recent applications of 3D printing techniques in the manufacturing of dental prosthetics. More specifically, after describing various 3D printing techniques and their advantages/disadvantages, the applications of 3D printing in dental prostheses are elaborated in various examples in the literature. Different 3D printing techniques have the capability to use different materials, including thermoplastic polymers, ceramics, and metals with distinctive suitability for dental applications, which are discussed in this article. The relevant limitations and challenges that currently limit the efficacy of 3D printing in this field are also reviewed. This review article has employed five major scientific databases, including Google Scholar, PubMed, ScienceDirect, Web of Science, and Scopus, with appropriate keywords to find the most relevant literature in the subject of dental prostheses 3D printing.

Mechanical properties of the rabbit and human decellularized patches for well-tolerated/reinforced organ in cardiac tissue engineering.

Introduction: Natural decellularized patches have been developed as the therapeutic platform for the treatment of different diseases, especially cardiovascular disorders. Decellularized scaffolds (as both cell-seeded and cell-free patches) are broadly studied in heart tissue redevelopment in vivo and in vitro. The designed regenerative bio-scaffold must have desirable physicochemical properties including mechanical stiffness for load-bearing, and appropriate anatomical characteristics to mimic the native biological environment properly and facilitate tissue reconstruction. In this context, the current study was designed to investigate rabbit decellularized derma's similarity with human decellularized skin in terms of mechanical properties for cardiac tissue engineering application. Methods: Fifty two rabbit dermal specimens were provided and divided into two groups: the experimental (decellularized) group and the control (group). Similarly, twelve human skin specimens were divided into the experimental (decellularized) and control groups. Initially, the effect of decellularization on the mechanical performance of scaffolds was analyzed. Then, the mechanical strength of decellularized rabbit skin was compared to decellularized human derma by measuring the stress strain and Young's modulus of the samples. Results: The results showed that rabbit decellularized skin has a similar elastic range to human decellularized skin, despite being more elastic (P>0.05). In addition, after decellularization, both rabbit and human skin showed a non-significant decrease in elasticity (P>0.05). It is worth noting that the elasticity reduction in rabbit samples after skin decellularization was lower than in human samples. Conclusion: According to the results of this study and the similarities of rabbit decellularized derm to human skin and its advantages over it, along with the biological complexity of native cardiac ECM, this scaffold can be used as an alternative matrix for tissue-engineered cardiac patches.

Application of microneedle patches for drug delivery; doorstep to novel therapies.

In the past decade, microneedle-based drug delivery systems showed promising approaches to become suitable and alternative for hypodermic injections and can control agent delivery without side effects compared to conventional approaches. Despite these advantages, the procedure of microfabrication is facing some difficulties. For instance, drug loading method, stability of drugs, and retention time are subjects of debate. Besides, the application of novel refining fabrication methods, types of materials, and instruments are other issues that need further attention. Herein, we tried to summarize recent achievements in controllable drug delivery systems (microneedle patches) in vitro and in vivo settings. In addition, we discussed the influence of delivered drugs on the cellular mechanism and immunization molecular signaling pathways through the intradermal delivery route. Understanding the putative efficiency of microneedle patches in human medicine can help us develop and design sophisticated therapeutic modalities.

Development of a Clinically Relevant Index for Tooth Wear Treatment Needs.

BACKGROUND: This study aimed to develop a tooth wear classification system that combined the extent, severity, and aesthetic impact of tooth wear and correlated them with the most appropriate clinical management strategy. METHODS: Three hundred photographs were used to develop a classification tool that contained four levels of severity and aesthetic impact (0, 1, 2, and 3) in three age groups of patients. Ten examiners assessed and classified the cases using validated forms. Additionally, they selected the recommended treatment modality for each level. The analysis was conducted using a coefficient correlation test. RESULTS: The coefficient correlation for the severity was 0.81, 0.82 in the upper anterior and posterior segments, and 0.85 and 0.77 for the lower anterior and posterior segments, respectively. The aesthetic impact correlation coefficient was 0.84. Examiners had agreed that minor cases required monitoring or simple restorative interventions. The moderate-level cases had variety in the recommended management options depending on the aim of treatment. The severe level cases often required rehabilitation at an increased occlusal vertical dimension. CONCLUSION: Within the limitations of this preliminary study, a good agreement between the examiners was found using the provided tools. More strict criteria in the classification part of the tool can further improve the examiners' agreement.

Osteo-mucosal engineered construct: In situ adhesion of hard-soft tissues.

OBJECTIVES: The aim of this work was to combine engineered hard and soft tissue, adopting a new method for interfacial adhesion of osteo-mucosal construct. We hypothesized that the chemical procedure involved in this method not only adheres the components, but also improves the cell growth inside them. METHODS: 3D-printed functionally-graded porous hard-tissue scaffolds were characterized, functionalized by aminolysis and tyrosinase, and accommodated by human osteoblast cells. Introducing amino groups through aminolysis and inducing dopaquinones by tyrosinase can take part in the Michael additions to cause the adhesion. Subsequently, fully-differentiated engineered oral mucosa was formed directly on the surface of hard tissue. Constructs were assessed in term of morphology, structure, chemical composition, histology, and cytocompatibility. Interfacial adhesion was compared to a control group prepared by using a biological glue for the attachment of the soft and hard tissues. RESULTS: The data confirmed higher proliferation of osteoblast cells via aminolysis and improved osteoblast cells distribution and differentiation by incorporation of tyrosinase in collagen. There was evidence of multilayered, stratified epithelium on the osteo-mucosal model with viable fibroblasts and osteoblasts within the lamina propria and bone tissue layers. Our method of adhesion resulted in cohesive debonding within the engineered soft tissue; while in the control group, adhesive debonding and complete separation of the oral mucosa from the hard tissue was observed. Although the shear strength of the osteo-mucosal model (157.6 kDa ± 25.1) was slightly higher than that of the control group (149.4 kDa ± 23.1), there was no statistically significant difference between them (p > 0.05). However, the advantage of our in situ adhesion approach is the absence of a barrier like glue which can disrupt direct cellular communications between tissues. SIGNIFICANCE: This study provides a novel method of directly combining tissue-engineered human bone with oral mucosa, which has the potential to improve cell-ingrowth and tissue integration. This engineered tissue construct, after further optimization, can be used clinically as a graft material in various oral surgeries and can also be employed as an in vitro model to investigate many aspects of oral diseases and examine dental materials and oral health care products as a replacement of in vivo models.

3D Bioprinting for In Vitro Models of Oral Cancer: Toward Development and Validation.

The tumor microenvironment (TME) of oral carcinomas has highly complex contents and a dynamic nature which is difficult to study using oversimplified two-dimensional (2D) cell culture systems. By contrast, three dimensional (3D) in vitro models such as spheroids, organoids, and scaffold-based constructs have been able to replicate tumors three-dimensionality and have allowed a better understanding of the role of various microenvironmental cues in the initiation and progression of cancer. However, the heterogeneity of TME cannot be fully reproduced by these traditional tissue engineering strategies since they are unable to control the organization of multiple cell types in a complex architecture. 3D bioprinting is an emerging field that can be leveraged to produce biomimetic and complex tissue structures. Bioprinting allows for controllable and precise placement of multicomponent bioinks composed of multiple biomaterials, different types of cells, and soluble factors according to the natural compartments of the target tissue, aiming to reproduce the equivalent of the complex tissue. As such, 3D bioprinting provides a unique opportunity to fabricate in vitro tumor models with a complexity similar to that of the in vivo oral carcinoma. This will facilitate a thorough investigation of cellular physiology, cancer progression, and anti-cancer drug screening with unprecedented control and reproducibility. In this review, we discuss the role of 3D bioprinting in reconstituting oral cancer, the prospects of application to fill the literature gap, and the challenges that need to be addressed in order to exploit this emerging technology for future work in oral cancer research.

Characterisation of Microparticle Waste from Dental Resin-Based Composites.

Clinical applications of resin-based composite (RBC) generate environmental pollution in the form of microparticulate waste. METHODS: SEM, particle size and specific surface area analysis, FT-IR and potentiometric titrations were used to characterise microparticles arising from grinding commercial and control RBCs as a function of time, at time of generation and after 12 months ageing in water. The RBCs were tested in two states: (i) direct-placement materials polymerised to simulate routine clinical use and (ii) pre-polymerised CAD/CAM ingots milled using CAD/CAM technology. RESULTS: The maximum specific surface area of the direct-placement commercial RBC was seen after 360 s of agitation and was 1290 m2/kg compared with 1017 m2/kg for the control material. The median diameter of the direct-placement commercial RBC was 6.39 µm at 360 s agitation and 9.55 µm for the control material. FTIR analysis confirmed that microparticles were sufficiently unique to be identified after 12 months ageing and consistent alteration of the outermost surfaces of particles was observed. Protonation-deprotonation behaviour and the pH of zero proton charge (pHzpc) ≈ 5-6 indicated that the particles are negatively charged at neutral pH7. CONCLUSION: The large surface area of RBC microparticles allows elution of constituent monomers with potential environmental impacts. Characterisation of this waste is key to understanding potential mitigation strategies.

Phenolated alginate-collagen hydrogel induced chondrogenic capacity of human amniotic mesenchymal stem cells.

Horseradish peroxidase (HRP)-catalyzed hydrogels are considered to be an important platform for tissue engineering applications. In this study, we investigated the chondrogenic capacity of phenolated (1.2%) alginate-(0.5%) collagen hydrogel on human amniotic mesenchymal stem cells after 21 days. Using NMR, FTIR analyses, and SEM imaging, we studied the phenolation and structure of alginate-collagen hydrogel. For physicochemical evaluations, gelation time, mechanical properties, swelling, and degradation rate were assessed. The survival rate was monitored using the MTT assay and DAPI staining. Western blotting was performed to measure the chondrogenic differentiation of cells. NMR showed successful phenolation of the alginate-collagen hydrogel. FTIR exhibited the interaction between the functional groups of collagen with phenolated alginate. SEM showed the existence of collagen microfibrils in the alginate-collagen hydrogel. Compared to phenolated alginate, the addition of collagen increased hydrogel elasticity by 10%. Both swelling rate and biodegradability were reduced in the presence of collagen. We noted an increased survival rate in phenolated alginate-collagen compared to the control cells (p < 0.05). Western blotting revealed the increase of chondrocyte-associated proteins such as SOX9 and COL2A1 in phenolated-alginate-collagen hydrogels after 21 days. These data showed that phenolated alginate-collagen hydrogel is an appropriate 3 D substrate to induce chondrogenic capacity of human mesenchymal stem cells.

Simultaneous detection of monomers associated with resin-based dental composites using SPME and HPLC.

As resin-based composites (RBC) replace dental amalgam for environmental reasons, there is a requirement to understand the environmental impact of this alternative dental restorative material. In this study we standardize the simultaneous detection of five monomeric components associated with RBCs using high performance liquid chromatography (HPLC) coupled with solid-phase microextraction (SPME). Factors affecting method performance (detection wavelength, calibration conditions, method sensitivity/accuracy/precision, extraction time/efficiency) are evaluated using standard solutions containing the mixture of TEGDMA, UDMA, Bis-GMA, BPA and HEMA. Detection sensitivity and analytical efficiency of the method is optimized for these compounds using 200 nm detection wavelength, PDMS/DVB fiber and extraction time of 90 min. Analytical accuracy of the HPLC is >95% for all monomers, with precision of 2.3-5.1%. Detection limits under the conditions described are 25 µg/L for HEMA, BPA, UDMA, Bis-GMA, and 100 µg/L for TEGDMA. The extraction time is governed by the largest molecular weight compounds.

Development of a Radiographic Index for Periodontitis.

The use of radiographic indices is noticeably diminished due to the lack of simplicity and standardisation among the existing ones. The aim of this study was to introduce a radiographic index to aid clinicians in determining the extent and severity of interproximal alveolar bone loss (iABL), in relation to individual root lengths, among patients suffering from periodontitis. A retrospective analysis of 50 anonymised dental panoramic tomograms (DPTs) of patients with periodontitis was conducted. Visual interpretation of iABL was recorded by a single investigator and by 20 volunteering clinicians for the 'worst site' in each quintet. Results were compared to a gold standard quantification method. Intra-examiner and inter-examiner agreement were measured using the Kappa coefficient and the intra-class correlation coefficient, respectively. Validity was assessed using Cramér's V test. The mean intra-examiner agreement on the severity and pattern of iABL was 0.808 (K) and 0.802 (K), respectively. A stronger overall inter-examiner agreement was noted when the severity in contrast to the pattern of iABL and presence/absence of furcation involvement were analysed. The statistically significant total mean agreement values from this correlation coefficient were 0.892 and 0.739, respectively. A very strong association between all the visual interpretations carried out by all participants and the gold standard measurements was evident. Within the limitations of this study, the proposed radiographic index may serve as a simple, yet valid and reliable, adjunctive screening tool to further assist clinicians in determining the extent and severity of iABL in patients with periodontitis.

Tissue Engineering Strategies to Increase Osteochondral Regeneration of Stem Cells; a Close Look at Different Modalities.

The homeostasis of osteochondral tissue is tightly controlled by articular cartilage chondrocytes and underlying subchondral bone osteoblasts via different internal and external clues. As a correlate, the osteochondral region is frequently exposed to physical forces and mechanical pressure. On this basis, distinct sets of substrates and physicochemical properties of the surrounding matrix affect the regeneration capacity of chondrocytes and osteoblasts. Stem cells are touted as an alternative cell source for the alleviation of osteochondral diseases. These cells appropriately respond to the physicochemical properties of different biomaterials. This review aimed to address some of the essential factors which participate in the chondrogenic and osteogenic capacity of stem cells. Elements consisted of biomechanical forces, electrical fields, and biochemical and physical properties of the extracellular matrix are the major determinant of stem cell differentiation capacity. It is suggested that an additional certain mechanism related to signal-transduction pathways could also mediate the chondro-osteogenic differentiation of stem cells. The discovery of these clues can enable us to modulate the regeneration capacity of stem cells in osteochondral injuries and lead to the improvement of more operative approaches using tissue engineering modalities.

Fibroblast encapsulation in gelatin methacryloyl (GelMA) versus collagen hydrogel as substrates for oral mucosa tissue engineering.

PURPOSE: Over the past decades, a variety of biomaterials have been investigated in terms of their suitability for oral mucosa tissue engineering. The aim of this study was to compare collagen and GelMA hydrogels as connective tissue scaffolds for fibroblasts and as substrates for seeding and culture of oral epithelial keratinocyte cells. METHODS: Human primary oral fibroblast and keratinocyte cells were isolated from gingival biopsies. The mixture of fibroblasts with GelMA or collagen gel were aliquoted within six-well tissue culture plate inserts and cross-linked using visible light or reconstitution buffer/heat, respectively. The viability of fibroblasts in the hydrogels was investigated after one and three days of cultivation using the PrestoBlue assay. Following the addition and culture of oral keratinocytes onto the connective tissue constructs, the tissue-engineered oral mucosa was assessed histologically. RESULTS: The tissue viability assay shows that collagen hydrogels encapsulating fibroblasts displayed significantly higher cell viability than cell-laden GelMA constructs after 24 and 72 h (p < 0.05). A stratified and differentiated epithelium has formed on the surface of cell-laden collagen hydrogel but not on the surface of the GelMA-based substrate. CONCLUSION: Collagen-based scaffold offers superior biological properties compared to GelMA hydrogel in terms of oral fibroblast growth, as well as epithelial cell adhesion and differentiation. Therefore, collagen-based hydrogels remain the preferred choice for oral mucosa tissue engineering.

Implant Soft-Tissue Attachment Using 3D Oral Mucosal Models-A Pilot Study.

PURPOSE: The aim of this study was to investigate soft-tissue attachment to different metal, ceramic, and polymer implant surfaces using an inflamed, three-dimensional (3D), tissue-engineered, human oral mucosal model, as well as multiple-endpoint qualitative and quantitative biological approaches. METHODS: Normal human oral fibroblasts, OKF6/TERT-2 keratinocytes and THP-1 monocytes were cultured, and full-thickness, 3D oral mucosal models were engineered inside tissue culture inserts. Sand-blasted and acid-etched (SLA) and machined (M) titanium-zirconium alloy (TiZr; commercially known as Roxolid; Institut Straumann AG, Switzerland), ceramic (ZrO2), and polyether ether ketone (PEEK) rods (Ø 4 mm × 8 mm) were inserted into the center of tissue-engineered oral mucosa following a Ø 4mm punch biopsy. Inflammation was simulated with addition of the lipopolysaccharide (LPS) of Escherichia coli (E. coli) and tumor necrosis factor (TNF)-alpha to the culture medium. Implant soft-tissue attachment was assessed using histology, an implant pull-test with PrestoBlue assay, and scanning electron microscopy (SEM). RESULTS: Inflamed, full-thickness, 3D human oral mucosal models with inserted implants were successfully engineered and histologically characterized. The implant pull-test with PrestoBlue assay showed higher viability of the tissue that remained attached to the TiZr-SLA surface compared to the other test groups. This difference was statistically significant (p < 0.05). SEM analysis showed evidence of epithelial cell attachment on different implant surfaces. CONCLUSIONS: The inflamed, 3D, oral mucosal model has the potential to be used as a suitable in vitro test system for visualization and quantification of implant soft-tissue attachment. The results of our study indicate greater soft tissue attachment to TiZr-SLA compared to TiZr-M, ceramic, and PEEK surfaces.

Three-dimensional bio-printing and bone tissue engineering: technical innovations and potential applications in maxillofacial reconstructive surgery.

BACKGROUND: Bone grafting has been considered the gold standard for hard tissue reconstructive surgery and is widely used for large mandibular defect reconstruction. However, the midface encompasses delicate structures that are surrounded by a complex bone architecture, which makes bone grafting using traditional methods very challenging. Three-dimensional (3D) bioprinting is a developing technology that is derived from the evolution of additive manufacturing. It enables precise development of a scaffold from different available biomaterials that mimic the shape, size, and dimension of a defect without relying only on the surgeon's skills and capabilities, and subsequently, may enhance surgical outcomes and, in turn, patient satisfaction and quality of life. REVIEW: This review summarizes different biomaterial classes that can be used in 3D bioprinters as bioinks to fabricate bone scaffolds, including polymers, bioceramics, and composites. It also describes the advantages and limitations of the three currently used 3D bioprinting technologies: inkjet bioprinting, micro-extrusion, and laser-assisted bioprinting. CONCLUSIONS: Although 3D bioprinting technology is still in its infancy and requires further development and optimization both in biomaterials and techniques, it offers great promise and potential for facial reconstruction with improved outcome.