Green tea polyphenol epigallocatechin-3-gallate and cranberry proanthocyanidins act in synergy with cathelicidin (LL-37) to reduce the LPS-induced inflammatory response in a three-dimensional co-culture model of gingival epithelial cells and fibroblasts

Objectives: The human antimicrobial peptide cathelicidin (LL-37) possesses anti-inflammatory properties that may contribute to attenuating the inflammatory process associated with chronic periodontitis. Plant polyphenols, including those from cranberry and green tea, have been reported to reduce inflammatory cytokine secretion by host cells. In the present study, we hypothesized that A-type cranberry proanthocyanidins (AC-PACs) and green tea epigallocatechin-3-gallate (EGCG) act in synergy with LL-37 to reduce the secretion of inflammatory mediators by oral mucosal cells. Methods: A three-dimensional (3D) co-culture model of gingival epithelial cells and fibroblasts treated with non-cytotoxic concentrations of AC-PACs (25 and 50 mg/ml), EGCG (1 and 5 mg/ml), and LL-37 (0.1 and 0.2 mM) individually and in combination (AC-PACs + LL-37 and EGCG + LL-37) were stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS). Multiplex ELISA assays were used to quantify the secretion of 54 host factors, including chemokines, cytokines, growth factors, matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs). Results: LL-37, AC-PACs, and EGCG, individually or in combination, had no effect on the regulation of MMP and TIMP secretion but inhibited the secretion of several cytokines. ACPACs and LL-37 acted in synergy to reduce the secretion of CXC-chemokine ligand 1 (GRO-a), granulocyte colony-stimulating factor (G-CSF), and interleukin-6 (IL-6), and had an additive effect on reducing the secretion of interleukin-8 (IL-8), interferon-g inducible protein 10 (IP-10), and monocyte chemoattractant protein-1 (MCP-1) in response to LPS stimulation. EGCG and LL-37 acted in synergy to reduce the secretion of GRO-a, G-CSF, IL-6, IL-8, and IP-10, and had an additive effect on MCP-1 secretion. Conclusion: The combination of LL-37 and natural polyphenols from cranberry and green tea acted in synergy to reduce the secretion of several cytokines by an LPS-stimulated 3D coculture model of oral mucosal cells. Such combinations show promising results as potential adjunctive therapies for treating inflammatory periodontitis.

Synergistic Anti-Inflammatory Activity of the Antimicrobial Peptides Human Beta-Defensin-3 (hBD-3) and Cathelicidin (LL-37) in a Three-Dimensional Co-Culture Model of Gingival Epithelial Cells and Fibroblasts

Given the spread of antibiotic resistance in bacterial pathogens, antimicrobial peptides that can also modulate the immune response may be a novel approach for effectively controlling periodontal infections. In the present study, we used a three-dimensional (3D) co-culture model of gingival epithelial cells and fibroblasts stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS) to investigate the anti-inflammatory properties of human beta-defensin-3 (hBD-3) and cathelicidin (LL-37) and to determine whether these antimicrobial peptides can act in synergy. The 3D co-culture model composed of gingival fibroblasts embedded in a collagen matrix overlaid with gingival epithelial cells had a synergistic effect with respect to the secretion of IL-6 and IL-8 in response to LPS stimulation compared to fibroblasts and epithelial cells alone. The 3D co-culture model was stimulated with non-cytotoxic concentrations of hBD-3 (10 and 20 mu M) and LL-37 (0.1 and 0.2 mu M) individually and in combination in the presence of A. actinomycetemcomitans LPS. A multiplex ELISA assay was used to quantify the secretion of 41 different cytokines. hBD-3 and LL-37 acted in synergy to reduce the secretion of GRO-alpha, G-CSF, IP-10, IL-6, and MCP-1, but only had an additive effect on reducing the secretion of IL-8 in response to A. actinomycetemcomitans LPS stimulation. The present study showed that hBD-3 acted in synergy with LL-37 to reduce the secretion of cytokines by an LPS-stimulated 3D model of gingival mucosa. This combination of antimicrobial peptides thus shows promising potential as an adjunctive therapy for treating inflammatory periodontitis.

Isolation and characterization of Wharton's jelly-derived multipotent mesenchymal stromal cells obtained from bovine umbilical cord and maintained in a defined serum-free three-dimensional system

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Three-dimensional aspects of the lingual papillae and their connective tissue cores in the tongue of rats: a scanning electron microscope study

The aim of the present study was to describe the tridimensional morphological characteristics of the lingual papillae and their connective tissue cores (CTCs) in Sprague Dawley rats. Four types of papillae were reported on the dorsal surface. Filiform papillae were distributed on the tongue surface and after epithelial maceration a conic and multifilamentary shape of the CTCs was revealed. Fungiform papillae were reported on the rostral and middle regions covered by a squamous epithelium. After the removal of the epithelium, the shape of a volcano with the taste orifice at its top was noted. Foliate papillae were composed of five pairs of epithelial folds situated on the lateral-caudal margin of the tongue. After the removal of the epithelium, they were shown to be limited by thin laminar projections. The vallate papilla with an oval shape was present in the caudal region and delimited by an incomplete groove. The morphological characteristics of the lingual papillae of Sprague Dowley rats, three-dimensional SEM images, and the types of papillae on the dorsal surface were similar to those reported previously in other rodent mammals. The maceration technique revealed the details of extracellular matrix with varied shapes form of connective tissue cores.

Passivity Versus Unilateral Angular Misfit: Evaluation of Stress Distribution on Implant-Supported Single Crowns: Three-Dimensional Finite Element Analysis

The aim of this study was to evaluate the effect of unilateral angular misfit of 100 Km on stress distribution of implant-supported single crowns with ceramic veneering and gold framework by three-dimensional finite element analysis. Two three-dimensional models representing a maxillary section of premolar region were constructed: group 1 (control)-crown completely adapted to the implant and group 2-crown with unilateral angular misfit of 100 Km. A vertical force of 100 N was applied on 2 centric points of the crown. The von Mises stress was used as an analysis criterion. The stress values and distribution in the main maps (204.4 MPa for group 1 and 205.0 MPa for group 2) and in the other structures (aesthetic veneering, framework, retention screw, implant, and bone tissue) were similar for both groups. The highest stress values were observed between the first and second threads of the retention screw. Considering the bone tissue, the highest stress values were exhibited in the peri-implant cortical bone. The unilateral angular misfit of 100 Km did not influence the stress distribution on the implant-supported prosthesis under static loading.

Platform Switching: Biomechanical Evaluation Using Three-Dimensional Finite Element Analysis

Purpose: The objective of this study was to evaluate, using three-dimensional finite element analysis (3D FEA), the stress distribution in peri-implant bone tissue, implants, and prosthetic components of implant-supported single crowns with the use of the platform-switching concept. Materials and Methods: Three 3D finite element models were created to replicate an external-hexagonal implant system with peri-implant bone tissue in which three different implant-abutment configurations were represented. In the regular platform (RP) group, a regular 4.1-mm-diameter abutment (UCLA) was connected to regular 4.1-mm-diameter implant. The platform-switching (PS) group was simulated by the connection of a wide implant (5.0 mm diameter) to a regular 4.1-mm-diameter UCLA abutment. In the wide-platform (WP) group, a 5.0-mm-diameter UCLA abutment was connected to a 5.0-mm-diameter implant. An occlusal load of 100 N was applied either axially or obliquely on the models using ANSYS software. Results: Both the increase in implant diameter and the use of platform switching played roles in stress reduction. The PS group presented lower stress values than the RP and WP groups for bone and implant. In the peri-implant area, cortical bone exhibited a higher stress concentration than the trabecular bone in all models and both loading situations. Under oblique loading, higher intensity and greater distribution of stress were observed than under axial loading. Platform switching reduced von Mises (17.5% and 9.3% for axial and oblique loads, respectively), minimum (compressive) (19.4% for axial load and 21.9% for oblique load), and maximum (tensile) principal stress values (46.6% for axial load and 26.7% for oblique load) in the peri-implant bone tissue. Conclusion: Platform switching led to improved biomechanical stress distribution in peri-implant bone tissue. Oblique loads resulted in higher stress concentrations than axial loads for all models. Wide-diameter implants had a large influence in reducing stress values in the implant system. INT J ORAL MAXILLOFAC IMPLANTS 2011;26:482-491

Effect of Metal-Ceramic or All-Ceramic Superstructure Materials on Stress Distribution in a Single Implant-Supported Prosthesis: Three-Dimensional Finite Element Analysis

Purpose: This three-dimensional finite element analysis study evaluated the effect of different material combinations on stress distribution within metal-ceramic and all-ceramic single implant-supported prostheses. Materials and Methods: Three-dimensional finite element models reproducing a segment of the maxilla with a missing left first premolar were created. Five groups were established to represent different superstructure materials: GP, porcelain fused to gold alloy; GR, modified composite resin fused to gold alloy; TP, porcelain fused to titanium; TR, modified composite resin fused to titanium; and ZP, porcelain fused to zirconia. A 100-N vertical force was applied to the contact points of the crowns. All models were fixed in the superior region of bone tissue and in the mesial and distal faces of the maxilla section. Stress maps were generated by processing with finite element software. Results: Stress distribution and stress values of supporting bone were similar for the GP, GR, TP, and ZP models (1,574.3 MPa, 1,574.3 MPa, 1,574.3 MPa, and 1,574.2 MPa, respectively) and different for the TR model (1,838.3 MPa). The ZP model transferred less stress to the retention screw (785 MPa) than the other groups (939 MPa for GP, 961 MPa for GR, 1,010 MPa for TP, and 1,037 MPa for TR). Conclusion: The use of different materials to fabricate a superstructure for a single implant-supported prosthesis did not affect the stress distribution in the supporting bone. The retention screw received less stress when a combination of porcelain and zirconia was used. Int J Oral Maxillofac Implants 2011;26:1202-1209

Three-Dimensional Finite Element Analysis of Vertical and Angular Misfit in Implant-Supported Fixed Prostheses

Purpose: Three-dimensional finite element analysis was used to evaluate the effect of vertical and angular misfit in three-piece implant-supported screw-retained fixed prostheses on the biomechanical response in the peri-implant bone, implants, and prosthetic components. Materials and Methods: Four three-dimensional models were fabricated to represent a right posterior mandibular section with one implant in the region of the second premolar (2PM) and another in the region of the second molar (2M). The implants were splinted by a three-piece implant-supported metal-ceramic prosthesis and differed according to the type of misfit, as represented by four different models: Control = prosthesis with complete fit to the implants; UAM (unilateral angular misfit) = prosthesis presenting unilateral angular misfit of 100 pm in the mesial region of the 2M; UVM (unilateral vertical misfit) = prosthesis presenting unilateral vertical misfit of 100 pm in the mesial region of the 2M; and TVM (total vertical misfit) = prosthesis presenting total vertical misfit of 100 pm in the platform of the framework in the 2M. A vertical load of 400 N was distributed and applied on 12 centric points by the software Ansys, ie, a vertical load of 150 N was applied to each molar in the prosthesis and a vertical load of 100 N was applied at the 2PM. Results: The stress values and distribution in peri-implant bone tissue were similar for all groups. The models with misfit exhibited different distribution patterns and increased stress magnitude in comparison to the control. The highest stress values in group UAM were observed in the implant body and retention screw. The groups UVM and TVM exhibited high stress values in the platform of the framework and the implant hexagon, respectively. Conclusions: The three types of misfit influenced the magnitude and distribution of stresses. The influence of misfit on peri-implant bone tissue was modest. Each type of misfit increased the stress values in different regions of the system. INT J ORAL MAXILLOFAC IMPLANTS 2011;26:788-796

Comparison of different designs of implant-retained overdentures and fixed full-arch implant-supported prosthesis on stress distribution in edentulous mandible - A computed tomography-based three-dimensional finite element analysis

A finite element analysis was used to compare the effect of different designs of implant-retained overdentures and fixed full-arch implant-supported prosthesis on stress distribution in edentulous mandible. Four models of an human mandible were constructed. In the OR (O'ring) group, the mandible was restored with an overdenture retained by four unsplinted implants with O'ring attachment; in the BC (bar-clip) -C and BC groups, the mandibles were restored with overdentures retained by four splinted implants with bar-clip anchor associated or not with two distally placed cantilevers, respectively; in the FD (fixed denture) group, the mandible was restored with a fixed full-arch four-implant-supported prosthesis. Models were supported by the masticatory muscles and temporomandibular joints. A 100-N oblique load was applied on the left first molar. Von Mises (σvM), maximum (σmax) and minimum (σmin) principal stresses (in MPa) analyses were obtained. BC-C group exhibited the highest stress values (σvM=398.8, σmax=580.5 and σmin=-455.2) while FD group showed the lowest one (σvM=128.9, σmax=185.9 and σmin=-172.1). Within overdenture groups, the use of unsplinted implants reduced the stress level in the implant/prosthetic components (59.4% for σvM, 66.2% for σmax and 57.7% for σmin versus BC-C group) and supporting tissues (maximum stress reduction of 72% and 79.5% for σmax, and 15.7% and 85.7% for σmin on the cortical and trabecular bones, respectively). Cortical bone exhibited greater stress concentration than the trabecular bone for all groups. The use of fixed implant dentures and removable dentures retained by unsplinted implants to rehabilitate edentulous mandible reduced the stresses in the periimplant bone tissue, mucosa and implant/prosthetic components. © 2013 Elsevier Ltd.

A new fibrin sealant as a three-dimensional scaffold candidate for mesenchymal stem cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of Tapered and External Hexagon Connections on Bone Stresses Around Tilted Dental Implants: Three-Dimensional Finite Element Method With Statistical Analysis

Background: The purpose of this study is to analyze the tension distribution on bone tissue around implants with different angulations (0 degrees, 17 degrees, and 30 degrees) and connections (external hexagon and tapered) through the use of three-dimensional finite element and statistical analyses.Methods: Twelve different configurations of three-dimensional finite element models, including three inclinations of the implants (0 degrees, 17 degrees, and 30 degrees), two connections (an external hexagon and a tapered), and two load applications (axial and oblique), were simulated. The maximum principal stress values for cortical bone were measured at the mesial, distal, buccal, and lingual regions around the implant for each analyzed situation, totaling 48 groups. Loads of 200 and 100 N were applied at the occlusal surface in the axial and oblique directions, respectively. Maximum principal stress values were measured at the bone crest and statistically analyzed using analysis of variance. Stress patterns in the bone tissue around the implant were analyzed qualitatively.Results: The results demonstrated that under the oblique loading process, the external hexagon connection showed significantly higher stress concentrations in the bone tissue (P < 0.05) compared with the tapered connection. Moreover, the buccal and mesial regions of the cortical bone concentrated significantly higher stress (P < 0.005) to the external hexagon implant type. Under the oblique loading direction, the increased external hexagon implant angulation induced a significantly higher stress concentration (P = 0.045).Conclusions: The study results show that: 1) the oblique load was more damaging to bone tissue, mainly when associated with external hexagon implants; and 2) there was a higher stress concentration on the buccal region in comparison to all other regions under oblique load.

Straight and angulated abutments in platform switching: influence of loading on bone stress by three-dimensional finite element analysis

PURPOSE: In view of reports in the literature on the benefits achieved with the use of platform switching, described as the use of an implant with a larger diameter than the abutment diameter, the goal being to prevent the (previously) normal bone loss down to the first thread that occurs around most implants, thus enhancing soft tissue aesthetics and stability and the need for implant inclination due to bone anatomy in some cases, the aim of this study was to evaluate bone stress distribution on peri-implant bone, by using three-dimensional finite element analysis to simulate the influence of implants with different abutment angulations (0 and 15 degrees) in platform switching. METHODS: Four mathematical models of an implant-supported central incisor were created with varying abutment angulations: straight abutment (S1 and S2) and angulated abutment at 15 degrees (A1 and A2), submitted to 2 loading conditions (100 N): S1 and A1-oblique loading (45 degrees) and S2 and A2-axial loading, parallel to the long axis of the implant. Maximum (σmax) and minimum (σmin) principal stress values were obtained for cortical and trabecular bone. RESULTS: Models S1 and A1 showed higher σmax in cortical and trabecular bone when compared with S2 and A2. The highest σmax values (in MPa) in the cortical bone were found in S1 (28.5), followed by A1 (25.7), S2 (11.6), and A2 (5.15). For the trabecular bone, the highest σmax values were found in S1 (7.53), followed by A1 (2.87), S2 (2.85), and A2 (1.47). CONCLUSIONS: Implants with straight abutments generated the highest stress values in bone. In addition, this effect was potentiated when the load was applied obliquely.

Computed tomographic-dacryocystography (CT-DCG) of the normal canine nasolacrimal drainage system with three-dimensional reconstruction

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Three-Dimensional Semiautomatic Road Extraction From a High-Resolution Aerial Image by Dynamic-Programming Optimization in the Object Space

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)