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1.
Micromachines (Basel) ; 9(2)2018 Jan 27.
Article in English | MEDLINE | ID: mdl-30393322

ABSTRACT

Current commercially available barrier membranes for oral surgery have yet to achieve a perfect design. Existing materials used are either non-resorbable and require a second surgery for their extraction, or alternatively are resorbable but suffer from poor structural integrity or degrade into acidic by-products. Silk has the potential to overcome these issues and has yet to be made into a commercially available dental barrier membrane. Reactive inkjet printing (RIJ) has recently been demonstrated to be a suitable method for assembling silk in its regenerated silk fibroin (RSF) form into different constructs. This paper will establish the properties of RSF solutions for RIJ and the suitability of RIJ for the construction of RSF barrier membranes. Printed RSF films were characterised by their crystallinity and surface properties, which were shown to be controllable via RIJ. RSF films degraded in either phosphate buffered saline or protease XIV solutions had degradation rates related to RSF crystallinity. RSF films were also printed with the inclusion of nano-hydroxyapatite (nHA). As reactive inkjet printing could control RSF crystallinity and hence its degradation rate, as well as offering the ability to incorporate bioactive nHA inclusions, reactive inkjet printing is deemed a suitable alternative method for RSF processing and the production of dental barrier membranes.

2.
Tissue Cell ; 52: 71-77, 2018 Jun.
Article in English | MEDLINE | ID: mdl-29857831

ABSTRACT

Recent advances in three-dimensional printing technology have led to a rapid expansion of its applications in tissue engineering. The present study was designed to develop and characterize an in vitro multi-layered human alveolar bone, based on a 3D printed scaffold, combined with tissue engineered oral mucosal model. The objective was to incorporate oral squamous cell carcinoma (OSCC) cell line spheroids to the 3D model at different anatomical levels to represent different stages of oral cancer. Histological evaluation of the 3D tissue model revealed a tri-layered structure consisting of distinct epithelial, connective tissue, and bone layers; replicating normal oral tissue architecture. The mucosal part showed a well-differentiated stratified oral squamous epithelium similar to that of the native tissue counterpart, as demonstrated by immunohistochemistry for cytokeratin 13 and 14. Histological assessment of the cancerous models demonstrated OSCC spheroids at three depths including supra-epithelial level, sub-epithelial level, and deep in the connective tissue-bone interface. The 3D tissue engineered composite model closely simulated the native oral hard and soft tissues and has the potential to be used as a valuable in vitro model for the investigation of bone invasion of oral cancer and for the evaluation of novel diagnostic or therapeutic approaches to manage OSCC in the future.


Subject(s)
Carcinoma, Squamous Cell/pathology , Head and Neck Neoplasms/pathology , Models, Anatomic , Mouth Neoplasms/pathology , Printing, Three-Dimensional , Tissue Engineering/methods , Alveolar Process/pathology , Humans , Spheroids, Cellular , Squamous Cell Carcinoma of Head and Neck , Tissue Scaffolds , Tumor Cells, Cultured
3.
Oncotarget ; 9(26): 18277-18295, 2018 Apr 06.
Article in English | MEDLINE | ID: mdl-29719605

ABSTRACT

Bone loss resulting from degenerative diseases and trauma is a significant clinical burden which is likely to grow exponentially with the aging population. In a number of conditions where pre-formed materials are clinically inappropriate an injectable bone forming hydrogel could be beneficial. The development of an injectable hydrogel to stimulate bone repair and regeneration would have broad clinical impact and economic benefit in a variety of orthopedic clinical applications. We have previously reported the development of a Laponite® crosslinked pNIPAM-co-DMAc (L-pNIPAM-co-DMAc) hydrogel delivery system, loaded with hydroxyapatite nanoparticles (HAPna), which was capable of inducing osteogenic differentiation of mesenchymal stem cells (MSCs) without the need for additional growth factors in vitro. However to enable progression towards clinical acceptability, biocompatibility and efficacy of the L-pNIPAM-co-DMAc hydrogel to induce bone repair in vivo must be determined. Biocompatibility was evaluated by subcutaneous implantation for 6 weeks in rats, and efficacy to augment bone repair was evaluated within a rat femur defect model for 4 weeks. No inflammatory reactions, organ toxicity or systemic toxicity were observed. In young male rats where hydrogel was injected, defect healing was less effective than sham operated controls when rat MSCs were incorporated. Enhanced bone healing was observed however, in aged exbreeder female rats where acellular hydrogel was injected, with increased deposition of collagen type I and Runx2. Integration of the hydrogel with surrounding bone was observed without the need for delivered MSCs; native cell infiltration was also seen and bone formation was observed within all hydrogel systems investigated. This hydrogel can be delivered directly into the target site, is biocompatible, promotes increased bone formation and facilitates migration of cells to promote integration with surrounding bone, for safe and efficacious bone repair.

4.
Tissue Eng Part C Methods ; 24(2): 99-107, 2018 02.
Article in English | MEDLINE | ID: mdl-29092692

ABSTRACT

Advances in tissue engineering have permitted assembly of multilayered composite tissue constructs for potential applications in the treatment of combined hard and soft tissue defects and as an alternative in vitro test model to animal experimental systems. The aim of this study was to develop and characterize a novel three-dimensional combined human alveolar bone and gingival mucosal model based on primary cells isolated from the oral tissues. Bone component of the model was engineered by seeding primary human alveolar osteoblasts into a hydroxyapatite/tricalcium phosphate scaffold and culturing in a spinner bioreactor. The engineered bone was then laminated, using an adhesive tissue sealant, with tissue-engineered gingival mucosa consisting of air/liquid interface-cultured normal human gingival keratinocytes on oral fibroblast-populated collagen gel scaffold. Histological characterization revealed a structure consisting of established epithelial, connective tissue and bone layers closely comparable to normal oral tissue architecture. The mucosal component demonstrated a mature epithelium undergoing terminal differentiation similar to that characteristic of native buccal mucosa, as confirmed using cytokeratin 13 and cytokeratin 14 immunohistochemistry. Ultrastructural analysis confirmed the presence of desmosomes and hemidesmosomes in the epithelial layer, a continuous basement membrane, and newly synthesized collagen in the connective tissue layer. Quantitative polymerase chain reaction (qPCR) assessment of osteogenesis-related gene expression showed a higher expression of genes encoded collagen I (COL1) and osteonectin (ON) compared with osteocalcin (OC), osteopontin (OP), and alkaline phosphatase (ALP). Enzyme-linked immunosorbent assay quantification of COL1, ON, and OC confirmed a pattern of secretion, which paralleled the model's gene expression profile. We demonstrate in this study that, replicating the anatomical setting between oral mucosa and the underlying alveolar bone is feasible and the developed model showed characteristics similar to those of normal tissue counterparts. This trilayered model therefore offers great scope as an advanced and anatomically representative tissue-engineered alternative to animal models.


Subject(s)
Alveolar Process/cytology , Bone Regeneration , Fibroblasts/cytology , Gingiva/cytology , Mouth Mucosa/cytology , Osteoblasts/cytology , Tissue Engineering/methods , Alveolar Process/metabolism , Biomarkers/metabolism , Cells, Cultured , Fibroblasts/metabolism , Gingiva/metabolism , Humans , Mouth Mucosa/metabolism , Osteoblasts/metabolism , Tissue Scaffolds
5.
J Biomed Mater Res A ; 104(12): 3157-3163, 2016 12.
Article in English | MEDLINE | ID: mdl-27502461

ABSTRACT

Tissue engineering is increasingly being recognized as a new approach that could alleviate the burden of tissue damage currently managed with transplants or synthetic devices. Making this novel approach available in the future for patients who would potentially benefit is largely dependent on understanding and addressing all those factors that impede the translation of this technology to the clinic. Cell-associated factors in particular raise many challenges, including those related to cell sources, up- and downstream techniques, preservation, and the creation of in vitro microenvironments that enable cells to grow and function as far as possible as they would in vivo. This article highlights the main confounding issues associated with cells in tissue engineering and how these issues may hinder the advancement of therapeutic tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 3157-3163, 2016.


Subject(s)
Stem Cells/cytology , Tissue Engineering/methods , Animals , Humans , Preservation, Biological/methods , Stem Cells/metabolism , Tissue and Organ Procurement
6.
J Mater Sci Mater Med ; 27(4): 65, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26883949

ABSTRACT

Tissue engineering of bone and oral mucosa have been extensively studied independently. The aim of this study was to develop and investigate a novel combination of bone and oral mucosa in a single 3D in vitro composite tissue mimicking the natural structure of alveolar bone with an overlying oral mucosa. Rat osteosarcoma (ROS) cells were seeded into a hydroxyapatite/tri-calcium phosphate scaffold and bone constructs were cultured in a spinner bioreactor for 3 months. An engineered oral mucosa was fabricated by air/liquid interface culture of immortalized OKF6/TERET-2 oral keratinocytes on collagen gel-embedded fibroblasts. EOM was incorporated into the engineered bone using a tissue adhesive and further cultured prior to qualitative and quantitative assessments. Presto Blue assay revealed that ROS cells remained vital throughout the experiment. The histological and scanning electron microscope examinations showed that the cells proliferated and densely populated the scaffold construct. Micro computed tomography (micro-CT) scanning revealed an increase in closed porosity and a decrease in open and total porosity at the end of the culture period. Histological examination of bone-oral mucosa model showed a relatively differentiated parakeratinized epithelium, evenly distributed fibroblasts in the connective tissue layer and widely spread ROS cells within the bone scaffold. The feasibility of fabricating a novel bone-oral mucosa model using cell lines is demonstrated. Generating human 'normal' cell-based models with further characterization is required to optimize the model for in vitro and in vivo applications.


Subject(s)
Bone Development/physiology , Bone and Bones/physiology , Mouth Mucosa/physiology , Tissue Engineering/methods , Tissue Scaffolds , Animals , Cell Line, Tumor , Humans , Keratinocytes/physiology , Microscopy, Electron, Scanning , Osteosarcoma , Rats , X-Ray Microtomography
7.
J R Soc Interface ; 9(77): 3528-38, 2012 Dec 07.
Article in English | MEDLINE | ID: mdl-22915635

ABSTRACT

For dental implants, it is vital that an initial soft tissue seal is achieved as this helps to stabilize and preserve the peri-implant tissues during the restorative stages following placement. The study of the implant-soft tissue interface is usually undertaken in animal models. We have developed an in vitro three-dimensional tissue-engineered oral mucosal model (3D OMM), which lends itself to the study of the implant-soft tissue interface as it has been shown that cells from the three-dimensional OMM attach onto titanium (Ti) surfaces forming a biological seal (BS). This study compares the quality of the BS achieved using the three-dimensional OMM for four types of Ti surfaces: polished, machined, sandblasted and anodized (TiUnite). The BS was evaluated quantitatively by permeability and cell attachment tests. Tritiated water (HTO) was used as the tracing agent for the permeability test. At the end of the permeability test, the Ti discs were removed from the three-dimensional OMM and an Alamar Blue assay was used for the measurement of residual cells attached to the Ti discs. The penetration of the HTO through the BS for the four types of Ti surfaces was not significantly different, and there was no significant difference in the viability of residual cells that attached to the Ti surfaces. The BS of the tissue-engineered oral mucosa around the four types of Ti surface topographies was not significantly different.


Subject(s)
Cell Adhesion , Dental Implants , Models, Biological , Mouth Mucosa/physiology , Cells, Cultured , Humans , Permeability , Surface Properties , Tissue Engineering , Titanium
8.
J Mater Sci Mater Med ; 23(10): 2521-9, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22752884

ABSTRACT

Potassium fluorrichterite (KNaCaMg(5)Si(8)O(22)F(2)) glass-ceramics were modified by either increasing the concentration of calcium (GC5) or by the addition of P(2)O(5) (GP2). Rods (2 × 4 mm) of stoichiometric fluorrichterite (GST), modified compositions (GC5 and GP2) and 45S5 bioglass, which was used as the reference material, were prepared using a conventional lost-wax technique. Osteoconductivity was investigated by implantation into healing defects in the midshaft of rabbit femora. Specimens were harvested at 4 and 12 weeks following implantation and tissue response was investigated using computed microtomography (µCT) and histological analyses. The results showed greatest bone to implant contact in the 45S5 bioglass reference material at 4 and 12 weeks following implantation, however, GST, GC5 and GP2 all showed direct bone tissue contact with evidence of new bone formation and cell proliferation along the implant surface into the medullary space. There was no evidence of bone necrosis or fibrous tissue encapsulation around the test specimens. Of the modified potassium fluorrichterite compositions, GP2 showed the greatest promise as a bone substitute material due to its osteoconductive potential and superior mechanical properties.


Subject(s)
Ceramics/chemistry , Glass/chemistry , Magnesium Silicates/chemistry , Animals , Male , Rabbits , Tomography, X-Ray Computed
9.
Int J Dent ; 2012: 396316, 2012.
Article in English | MEDLINE | ID: mdl-22505899

ABSTRACT

The aim of this study was to assess the osteoconductive and osteogenic properties of processed bovine dentin using a robust rabbit calvarial defect model. In total, 16 New Zealand White rabbits were operated to create three circular defects in the calvaria. One defect was left unfilled, one filled with collected autogenous bone, and the third defect was filled with the dentin-based bone substitute. Following surgery and after a healing period of either 1 or 6 weeks, a CT scan was obtained. Following sacrificing, the tissues were processed for histological examination. The CT data showed the density in the area grafted with the dentin-based material was higher than the surrounding bone and the areas grafted with autologous bone after 1 week and 6 weeks of healing. The area left unfilled remained an empty defect after 1 week and 6 weeks. Histological examination of the defects filled with the dentin product after 6 weeks showed soft tissue encapsulation around the dentin particles. It can be concluded that the rabbit calvarial model used in this study is a robust model for the assessment of bone materials. Bovine dentin is a biostable material; however, it may not be suitable for repairing large 4-wall defects.

10.
J Biomed Mater Res A ; 100(2): 269-77, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22045611

ABSTRACT

A three dimensional tissue-engineered human oral mucosal model (3D OMM) used in the investigation of implant-soft tissue interface was recently reported. The aim of this study was to examine the ultrastructural features of soft tissue attachment to various titanium (Ti) implant surfaces based on the 3D OMM. Two techniques, that is, focus ion beam (FIB) and electropolishing techniques were used to prepare specimens for transmission electron microscopic (TEM) analysis of the interface. The 3D OM consisting of both epithelial and connective tissue layers was constructed by co-culturing human oral keratinocytes and fibroblasts onto an acellular dermis scaffold. Four types of Ti surface topographies were tested: polished, machined (turned), sandblasted, and TiUnite. The specimens were then processed for TEM examination using FIB (Ti remained) and electropolishing (Ti removed) techniques. The FIB sections showed some artifact and lack of details of ultrastructural features. In contrast, the ultrathin sections prepared from the electropolishing technique showed a residual Ti oxide layer, which preserved the details for intact ultrastructural interface analysis. There was evidence of hemidesmosome-like structures at the interface on the four types of Ti surfaces, which suggests that the tissue-engineered oral mucosa formed epithelial attachments on the Ti surfaces.


Subject(s)
Dental Implants , Imaging, Three-Dimensional , Mouth Mucosa/ultrastructure , Tissue Engineering , Humans , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Models, Biological , Mouth Mucosa/drug effects , Titanium/pharmacology
11.
J Mater Sci Mater Med ; 22(9): 2065-70, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21706217

ABSTRACT

Potassium fluorrichterite (KNaCaMg(5)Si(8)O(22)F(2)) glass-ceramics were modified by either increasing the concentration of calcium in the glass (GC5), or by the addition of P(2)O(5) to produce potassium fluorrichterite-fluorapatite (GP2). The solubility of the stoichiometric composition (GST), GC5 and GP2 were measured using the standard test described in ISO 6872:1995 (Dental Ceramics). Ion release profiles were determined for Si, Ca, Mg, Na, K and P using inductively coupled plasma mass spectrometry and fluoride ion (F(-)) concentration was measured using an ion-selective electrode. The cytotoxicity of all compositions was assessed using cultured rat osteosarcoma cells (ROS, 17/2.8). Cell response was qualitatively assessed using scanning electron microscopy (SEM) and quantitatively using the Alamar blue assay. GST was the least soluble and also released the lowest concentration of ions following immersion in water. Of the modified compositions, GC5 demonstrated intermediate solubility but the greatest ion release while GP2 exhibited the highest solubility. This was most likely due to GC5 having the greatest proportion of residual glass following crystallisation. The mass loss exhibited by GP2 may have been due in part to the partial disintegration of the surface of specimens during solubility testing. SEM demonstrated that all compositions supported the growth of healthy ROS cells on their surfaces, and this data was further supported by the quantitative Alamar blue assay.


Subject(s)
Biocompatible Materials , Ceramics , Fluorine Compounds/chemistry , Glass , Potassium Compounds/chemistry , Animals , Cell Line, Tumor , In Vitro Techniques , Ion-Selective Electrodes , Microscopy, Electron, Scanning , Rats , Solubility
12.
J Periodontol ; 80(5): 769-75, 2009 May.
Article in English | MEDLINE | ID: mdl-19405830

ABSTRACT

BACKGROUND: The biologic safety profile of oral health care products is often assumed on the basis of simplistic test models such as monolayer cell culture systems. We developed and characterized a tissue-engineered human oral mucosal model, which was proven to represent a potentially more informative and more clinically relevant alternative for the biologic assessment of mouthwashes. The aim of this study was to evaluate the biologic effects of alcohol-containing mouthwashes on an engineered human oral mucosal model. METHODS: Three-dimensional (3D) models were engineered by the air/liquid interface culture technique using human oral fibroblasts and keratinocytes. The models were exposed to phosphate buffered saline (negative control), triethylene glycol dimethacrylate (positive control), cola, and three types of alcohol-containing mouthwashes. The biologic response was recorded using basic histology; a cell proliferation assay; 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tissue-viability assay; transmission electron microscopy (TEM) analysis; and the measurement of release of interleukin (IL)-1beta by enzyme-linked immunosorbent assay. RESULTS: Statistical analysis showed that there was no significant difference in tissue viability among the mouthwashes, cola, and negative control groups. However, exposure to the positive control significantly reduced the tissue viability and caused severe cytotoxic epithelial damage as confirmed by histology and TEM analysis. A significant increase of IL-1beta release was observed with the positive control and, to a lesser extent, with two of the tested mouthrinses. CONCLUSIONS: The 3D human oral mucosal model can be a suitable model for the biologic testing of mouthwashes. The alcohol-containing mouthwashes tested in this study do not cause significant cytotoxic damage and may slightly stimulate IL-1beta release.


Subject(s)
Mouth Mucosa/drug effects , Mouthwashes/toxicity , Tissue Engineering/methods , Tissue Survival/drug effects , Cell Proliferation , Coloring Agents , Drug Combinations , Ethanol/toxicity , Humans , Interleukin-1beta/biosynthesis , Microscopy, Electron, Transmission , Models, Biological , Mouth Mucosa/metabolism , Salicylates/toxicity , Terpenes/toxicity , Tetrazolium Salts , Thiazoles
13.
J Dent ; 36(5): 331-6, 2008 May.
Article in English | MEDLINE | ID: mdl-18359139

ABSTRACT

OBJECTIVE: Tissue-engineered human oral mucosal models have been developed for biocompatibility assessment of biomaterials. The aim of this study was to evaluate the biological effects of three different composite resin systems on a three-dimensional human oral mucosal model. METHODS: Full-thickness oral mucosal models were engineered by air/liquid interface culture of a human oral keratinocyte cell line on a lamina propria composed of oral fibroblasts seeded into a porous scaffold. The surface of the tissue models was exposed to three types of experimental composite resins: a TEGDMA-based, a UDMA-based, and a BisGMA/TEGDMA (80:20)-based composite resin for 24h. The response of the engineered oral mucosa to the test materials was assessed using routine histology, the Alamar Blue tissue viability assay and IL-1beta release measured by ELISA. RESULTS: Compared to the other materials tested, the TEGDMA-based composite resin caused significant damage to the oral mucosal model. Statistical analysis by one-way ANOVA followed by Tukey's analysis showed that there was a significant decrease in the viability of tissue models after 24h exposure to TEGDMA-based composite resin. Also exposure to TEGDMA-based composite resin significantly increased the amount of IL-1beta released from the oral mucosal model. CONCLUSION: The 3D human oral mucosal model has the potential to be a more relevant and more informative model than monolayer cell culture systems for biocompatibility testing of dental materials. The results obtained from multiple-endpoint analysis of the oral mucosal model indicate significant mucotoxicity of high TEGDMA-containing composite resins.


Subject(s)
Composite Resins/toxicity , Dental Materials/toxicity , Mouth Mucosa/drug effects , Tissue Engineering , Biocompatible Materials/toxicity , Bisphenol A-Glycidyl Methacrylate/toxicity , Cell Survival/drug effects , Cells, Cultured , Fibroblasts/drug effects , Fibroblasts/pathology , Humans , Interleukin-1beta/analysis , Keratinocytes/drug effects , Keratinocytes/pathology , Materials Testing , Methacrylates/toxicity , Mouth Mucosa/pathology , Polyethylene Glycols/toxicity , Polymethacrylic Acids/toxicity , Polyurethanes/toxicity , Time Factors , Tissue Scaffolds , Tissue Survival/drug effects
14.
J Mater Sci Mater Med ; 19(2): 839-46, 2008 Feb.
Article in English | MEDLINE | ID: mdl-17665105

ABSTRACT

Modified fluorcanasite glass-ceramics were produced by controlled two stage heat-treatment of as-cast glasses. Castability was determined using a spiral castability test and the lost-wax method. Specimens were cast into moulds formed from gypsum and phosphate bonded investments to observe their effect on the casting process, surface roughness, surface composition and biocompatibility. Both gypsum and phosphate bonded investments could be successfully used for the lost-wax casting of fluorcanasite glasses. Although the stoichiometric glass composition had the highest castability, all modified compositions showed good relative castability. X-ray diffraction showed similar bulk crystallisation for each glass, irrespective of the investment material. However, differences in surface crystallisation were detected when different investment materials were used. Gypsum bonded investment discs showed slightly improved in vitro biocompatibility than equivalent phosphate bonded investment discs under the conditions used.


Subject(s)
Ceramics/chemistry , Glass/chemistry , Silicates/chemistry , Animals , Biocompatible Materials/chemistry , Bone Substitutes/chemistry , Cell Line, Tumor , Rats , Surface Properties , X-Ray Diffraction
15.
Dent Mater ; 23(1): 40-4, 2007 Jan.
Article in English | MEDLINE | ID: mdl-16426672

ABSTRACT

OBJECTIVES: The aim of this study was to evaluate and compare the biological effects of three resin monomers on three human gingival fibroblast (HGF) cell lines and immortalised human keratinocytes. METHODS: Primary HGFs and HaCaT keratinocytes were cultured for 24h and grown to sub-confluent monolayers. Resin monomers were dissolved in dimethyl sulphoxide (DMSO) and diluted with culture medium. Cultures were exposed to different concentrations of monomers (10(-2) to 10mM) for 24h. Cell viability measured by Alamar Blue assay, and cell culture supernatant was examined for the presence of human interlukin-1beta (IL-1beta) using sandwich enzyme-linked immunosorbant assay (ELISA). TC50 values were calculated from fitted dose-response curves. RESULTS: All monomers showed toxic effects on the HGFs and HaCaT cells and inhibited chemical reduction of Alamar Blue in high concentrations. Statistical analysis of TC50 values by one-way ANOVA followed by Tukey's analysis showed that there is a significant difference in TC50 values between the cell lines (p<0.05), although the rank order of monomer toxicity remained the same for different cell lines. None of these monomers-induced IL-1beta release from HGFs and HaCaT cells. SIGNIFICANCE: Dental resin monomers are toxic to human gingival fibroblasts and HaCaT keratinocytes. However, they cannot induce IL-1beta release from these cells by themselves. Alamar Blue assay is a sensitive method for the evaluation of cytotoxicity and it can detect different sensitivities of different cell lines to the resin monomers.


Subject(s)
Composite Resins/toxicity , Dental Materials/toxicity , Fibroblasts/drug effects , Gingiva/drug effects , Keratinocytes/drug effects , Skin/drug effects , Bisphenol A-Glycidyl Methacrylate/toxicity , Cell Line , Cell Survival/drug effects , Culture Media , Dose-Response Relationship, Drug , Gingiva/cytology , Humans , Indicators and Reagents , Inflammation Mediators/analysis , Interleukin-1beta/analysis , Materials Testing , Methacrylates/toxicity , Oxazines , Polyethylene Glycols/toxicity , Polymethacrylic Acids/toxicity , Polyurethanes/toxicity , Skin/cytology , Time Factors , Xanthenes
16.
Tissue Eng ; 12(3): 569-77, 2006 Mar.
Article in English | MEDLINE | ID: mdl-16579690

ABSTRACT

Together, the chondrocyte and its pericellular matrix have been collectively termed the chondron. Current opinion is that the pericellular matrix has both protective and signalling functions between chondrocyte and extracellular matrix. Formation of a native chondrocyte pericellular matrix or chondron structure might therefore be advantageous when tissue engineering a functional hyaline cartilage construct. The presence of chondrons has not been previously described in cartilage engineered on a scaffold. In this paper, we describe a modified immunochemical method to detect collagen VI, a key molecular marker for the pericellular matrix, and an investigation of type VI collagen distribution in engineered hyaline cartilage constructs. Cartilage constructs were engineered from adult human or bovine hyaline chondrocytes cultured on sponge or nonwoven fiber based HYAFF 11 scaffolds. Type VI collagen was detected in all constructs, but a distinctive, high-density, chondron-like distribution of collagen VI was present only in constructs exhibiting additional features of hyaline cartilage engineered using nonwoven HYAFF 11. Chondron structures were localized in areas of the extracellular matrix displaying strong collagen II and GAG staining of constructs where type II collagen composed a high percentage (over 65%) of the total collagen.


Subject(s)
Cartilage/metabolism , Collagen Type VI/metabolism , Tissue Engineering/methods , Animals , Biocompatible Materials , Cattle , Chondrocytes/metabolism , Collagen Type II/metabolism , Extracellular Matrix/metabolism , Glycosaminoglycans/metabolism , Humans , Hyaluronic Acid/analogs & derivatives , Immunohistochemistry , Materials Testing , Polymers
17.
Dent Mater ; 21(6): 551-6, 2005 Jun.
Article in English | MEDLINE | ID: mdl-15904698

ABSTRACT

OBJECTIVE: To assess the effect of varying CaF(2) on the chemical solubility of apatite-mullite glass-ceramic (G-C) materials in both the glassy and crystallized states. METHODS: Apatite-mullite forming glasses used in this study are ionomer cement derivatives based on the general formula (4.5SiO(2)-3Al(2)O(3)-1.5P(2)O(5)-3CaO-XCaF(2)). Six glass formulations were produced where X=0.5, 1, 1.5, 2, 2.5 and 3, and called HG1-6, respectively. Batches were melted in covered silliminite crucibles in a furnace overnight at 1050 degrees C, then at 1450 degrees C for 2h, before quenching in water. The six glass compositions were analyzed using differential thermal analysis (DTA), X-ray diffraction (XRD) and X-ray fluorescence spectrometry (XRF). Thirty discs (2mm thick and 12 mm diameter) were produced per glass using the lost wax casting technique. Ten were left as cast and 10 heat treated to either apatite or apatite-mullite. Solubility testing was carried out according to International Standard BS EN ISO 6872 1999 and the mass difference in solubility calculated as mug/cm(2). A lithium disilicate G-C system was used as a control material. RESULTS: All compositions formed glasses and on heat treatment could form apatite and apatite-mullite. The as-cast glass samples were the most soluble followed by the apatite samples. The apatite-mullite G-C was significantly less soluble than the other two phases (p<0.05) for all six compositions. The control material was significantly less soluble than all the HG glass-ceramic compositions for every phase (p<0.05). Decreasing the CaF(2) content (3-0.5 mol%) led to a decrease in solubility, without affecting the ability of the material to form apatite and apatite-mullite phases. SIGNIFICANCE: Increasing the CaF(2) content increases the chemical solubility for the glass, apatite G-C and apatite-mullite G-C phases. The solubility values obtained show that all the compositions, as cast and heat treated would be suitable for use as core ceramics.


Subject(s)
Ceramics/chemistry , Dental Porcelain/chemistry , Aluminum Silicates/chemistry , Analysis of Variance , Apatites/chemistry , Calcium Fluoride/chemistry , Crystallization , Crystallography, X-Ray , Differential Thermal Analysis , Glass Ionomer Cements/chemistry , Materials Testing , Solubility , Spectrometry, X-Ray Emission
18.
Int J Oral Maxillofac Implants ; 18(3): 377-82, 2003.
Article in English | MEDLINE | ID: mdl-12814312

ABSTRACT

PURPOSE: The aim of this study was to report the outcome of sinus augmentation surgery with autogenous bone grafting in routine dental implant practice. MATERIALS AND METHODS: Twenty-seven sinus augmentation procedures were undertaken on 18 consecutive patients (mean age 43.7 years). The mandibular symphysis was used as the donor site for 11 patients. The iliac crest was used as a donor site for 7 bilateral cases. RESULTS: Six patients had implants placed at the time of grafting: the other 13 had a mean bone graft consolidation period of 24.7 weeks (range 9 to 39 weeks) before implants were placed. One patient who had a repeat procedure had both immediate and delayed techniques. A total of 79 Brånemark System Mk II implants were placed in grafted bone (and 2 Mk IV implants were placed in a patient who had to have a repeat procedure) and proceeded to occlusal loading. After a mean follow-up period of 162 weeks (range 76 to 288 weeks), 16 implants failed to integrate in grafted bone, representing an 80.25% survival rate. Fourteen patients proceeded to the planned prosthesis, 3 patients had a compromised treatment plan, and 1 patient was restored conventionally. This represents 94% of patients who were rehabilitated. DISCUSSION AND CONCLUSION: The sinus augmentation procedure using autogenous bone grafting can Increase bone volume to allow implant placement where there is insufficient bone. The survival of implants in the grafted bone, as measured by integration and successful loading, was reduced compared to implants placed in normal maxillary bone. Infection during the healing of the grafted site reduces the success of subsequent implant osseointegration.


Subject(s)
Alveolar Ridge Augmentation/methods , Bone Transplantation , Dental Implants , Maxilla/surgery , Maxillary Sinus/surgery , Adolescent , Adult , Aged , Bone Transplantation/methods , Dental Abutments , Dental Implantation, Endosseous/methods , Dental Restoration Failure , Female , Follow-Up Studies , Guided Tissue Regeneration, Periodontal , Humans , Male , Middle Aged , Osseointegration , Patient Care Planning , Survival Analysis , Transplantation, Autologous , Treatment Outcome
19.
Int J Oral Maxillofac Implants ; 18(2): 238-41, 2003.
Article in English | MEDLINE | ID: mdl-12705302

ABSTRACT

PURPOSE: Loss of alveolar bone in the anterior maxilla may preclude implant placement or compromise positioning and thus diminish the final esthetic result of the restoration. Bone augmentation can overcome such difficulties but may affect osseointegration. The aim of this study was to report the outcome of buccal onlay bone grafting in the anterior maxilla in routine dental implant practice. MATERIALS AND METHODS: Seventeen consecutive patients (12 men and 5 women, mean age 31.4 years) received autogenous bone grafts from the mandibular symphysis to the anterior maxilla. A total of 35 Brånemark System MK II implants were placed in grafted bone. RESULTS: Fifteen patients had a mean period of graft consolidation of 19.7 weeks (range 13 to 32 weeks). Two patients had simultaneous graft and implant placement; 1 implant failed to Integrate in this group. This represents a survival rate of 97.1% of implants in functional loading after a mean follow-up period of 153.6 weeks from occlusal loading (range 74 to 283 weeks). DISCUSSION AND CONCLUSION: Mandibular block onlay grafts appear to be a predictable method for augmenting the width of the anterior maxilla prior to implant placement.


Subject(s)
Alveolar Ridge Augmentation/methods , Bone Transplantation , Dental Implantation, Endosseous/methods , Dental Implants , Maxilla/surgery , Adolescent , Adult , Aged , Dental Prosthesis Retention , Dental Prosthesis, Implant-Supported , Dental Restoration Failure , Female , Humans , Male , Middle Aged , Osseointegration , Treatment Outcome
20.
Int. j. oral maxillofac. implants ; 13(1): 44-51, 1998. ilus
Article in English | BBO - Dentistry | ID: biblio-851036

ABSTRACT

The healing of standardized bone defects grafted with either particulate ionomeric or hydroxyapatite bone substitutes was compared in the mandibular ramus of 30 Sprague-Dawley rats. The possible additional response achieved when combining these materials with a guided bone regeneration (GBR) technique was also evaluated. Three groups of 10 animals received either no implant material or ionomeric or hydroxyapatite bone substitute in defects in the right ramus. The left mandibular defects received the same treatment, except that the operation site was covered by a membrane (GBR technique). Half of the animals were sacrificed at 4 and 10 weeks following surgery, and the inflammatory response at the implant site and the amount of new bone formed in the defects were determined histomorphometrically. Defects implanted with ionometric bone substitute exhibited more bone formation (4 weeks = 3.19 ± 0.38 mm², 10 weeks = 5.35 ± 0.26 mm²) than both defects that received no treatment (4 weeks = 0.88 ± 0.35 mm², 10 weeks = 2.1 ± 0.49 mm²), membrane alone (4 weeks = 1.21 ± 0.005 mm² or hydroxyapatite bone substitute (4 weeks = 1.41 ± 0.46 mm², 10 weeks = 3.34 ± 0,41 mm²) at 4 weeks (P<=.01) and at 10 weeks (P<=.05). The use of a GBR technique did not increase the amount of bone formed, compared to the use of bone substitutes alone. Hydroxyapatite and ionomeric bone substitutes used alone were more effective in inducing repair of the defects than was GBR membrane alone. The use of hydroxyapatite was associated with a greater inflammatory reaction (P<=.01) than was ionomer in this model


Subject(s)
Animals , Rats , Bone Regeneration , Glass Ionomer Cements/analysis , Glass Ionomer Cements/therapeutic use , Hydroxyapatites/administration & dosage , Hydroxyapatites/analysis , Hydroxyapatites/therapeutic use , Mandible/anatomy & histology , Mandible/chemistry
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