Ian M. Brook

Development of biodegradable electrospun scaffolds for dermal replacement

Our objective is to develop a synthetic biodegradable replacement dermal substitute for tissue engineering of skin and oral mucosa. Our in vivo criteria were that candidate scaffolds should allow surrounding cells to migrate fully into the scaffolds, enabling vasculogenesis and remodelling without invoking a chronic inflammatory response. We examined a total of six experimental electrospun polymer scaffolds: (1) poly-l-lactide (PLLA); (2) PLLA+10% oligolactide; (3) PLLA+rhodamine and (4-6) three poly(d,l)-lactide-co-glycolide (PLGA) random multiblock copolymers, with decreasing lactide/glycolide mole fractions (85:15, 75:25 and 50:50). These were evaluated for degradation in vitro up to 108 days and in vivo in adult male Wistar rats from 4 weeks to 12 months. In vivo, all scaffolds permitted good cellular penetration, with no adverse inflammatory response outside the scaffold margin and with no capsule formation around the periphery. The breakdown rate for each scaffold in vitro versus in vivo was similar, and an increase in the ratio of polyglycolide to polylactide correlated with an increase in breakdown rate, as expected. Scaffolds of PLLA were stable in vivo even after 12 months whereas scaffolds fabricated from PLGA 85:15 and 75:25 revealed a 50% loss of mass after 4 and 3 months, respectively. In vitro PLGA 85:15 and 75:25 scaffolds were able to support keratinocyte, fibroblast and endothelial cell growth and extracellular matrix production, with evidence of new collagen production after 7 days. In conclusion, the data supports the development of PLGA 85:15 and 75:25 electrospun polymer scaffolds as potential degradable biomaterials for dermal replacement.

Tissue-engineered Oral Mucosa: a Review of the Scientific Literature

Tissue-engineered oral mucosal equivalents have been developed for clinical applications and also for in vitro studies of biocompatibility, mucosal irritation, disease, and other basic oral biology phenomena. This paper reviews different tissue-engineering strategies used for the production of human oral mucosal equivalents, their relative advantages and drawbacks, and their applications. Techniques used for skin tissue engineering that may possibly be used for in vitro reconstruction of human oral mucosa are also discussed.

CHARACTERISATION OF THE ULTRASTRUCTURE OF GLASS-IONOMER (POLY-ALKENOATE) CEMENT

Set glass-ionomer cements were sectioned with a diamond knife and examined in the transmission electron microscope. Their appearance was in accordance with the theoretical structure of these materials, close examination revealing glass particles surrounded by a siliceous layer set in a hydrogel matrix. The elemental composition of each region was determined by X-ray microanalysis (energy dispersive). The results of microanalysis supported the ultrastructural observations, with ions that originated from the glass particles being detected throughout the matrix of the set cement. It was suggested that the mobility of these ions in the matrix phase was important in determining the biocompatibility and adhesive properties of glass-ionomer cements

Tissue-engineered Oral Mucosa

Advances in tissue engineering have permitted the three-dimensional (3D) reconstruction of human oral mucosa for various in vivo and in vitro applications. Tissue-engineered oral mucosa have been further optimized in recent years for clinical applications as a suitable graft material for intra-oral and extra-oral repair and treatment of soft-tissue defects. Novel 3D in vitro models of oral diseases such as cancer, Candida, and bacterial invasion have been developed as alternatives to animal models for investigation of disease phenomena, their progression, and treatment, including evaluation of drug delivery systems. The introduction of 3D oral mucosal reconstructs has had a significant impact on the approaches to biocompatibility evaluation of dental materials and oral healthcare products as well as the study of implant-soft tissue interfaces. This review article discusses the recent advances in tissue engineering and applications of tissue-engineered human oral mucosa.

Preliminary investigation of novel bone graft substitutes based on strontium–calcium–zinc–silicate glasses

Bone graft procedures typically require surgeons to harvest bone from a second site on a given patient (Autograft) before repairing a bone defect. However, this results in increased surgical time, excessive blood loss and a significant increase in pain. In this context a synthetic bone graft with excellent histocompatibility, built in antibacterial efficacy and the ability to regenerate healthy tissue in place of diseased tissue would be a significant step forward relative to current state of the art philosophies. We developed a range of calcium–strontium–zinc–silicate glass based bone grafts and characterised their structure and physical properties, then evaluated their in vitro cytotoxicity and in vivo biocompatibility using standardised models from the literature. A graft (designated BT109) of composition 0.28SrO/0.32ZnO/0.40 SiO2 (mol fraction) was the best performing formulation in vitro shown to induce extremely mild cytopathic effects (cell viability up to 95%) in comparison with the commercially available bone graft Novabone® (cell viability of up to 72%). Supplementary to this, the grafts were examined using the standard rat femur healing model on healthy Wister rats. All grafts were shown to be equally well tolerated in bone tissue and new bone was seen in close apposition to implanted particles with no evidence of an inflammatory response within bone. Complimentary to this BT109 was implanted into the femurs of ovariectomized rats to monitor the response of osteoporotic tissue to the bone grafts. The results from this experiment indicate that the novel grafts perform equally well in osteoporotic tissue as in healthy tissue, which is encouraging given that bone response to implants is usually diminished in ovariectomized rats. In conclusion these materials exhibit significant potential as synthetic bone grafts to warrant further investigation and optimisation.

In vitro interaction between primary bone organ cultures, glass-ionomer cements and hydroxyapatite/tricalcium phosphate ceramics.

Primary organ cultures derived from neonate rat calvaria were maintained for 2 wk and used to study in vitro response of osteoblast and periosteal cells to the component and composite forms of three different glass-ionomer (polyalkenoic) cements, comparing them to densely sintered hydroxyapatite and tricalcium phosphate ceramics. Qualitative analysis by scanning and transmission electron microscopy revealed that osteoblasts colonized all the solid test materials, although there was a less favourable response to materials with a rough surface topography and to unset and fluoride-containing glasses. On solid materials migrated cells maintained their tessellated morphology and exhibited numerous micro-appendages anchoring them to the surface of the test materials. A collagen-containing extracellular matrix was elaborated on to the ceramics and set glass-ionomer cements, except for one (AquaCem). Mineralization of the extracellular matrix was seen adjacent to hydroxyapatite and tricalcium phosphate ceramics, that adjacent to the latter morphologically resembling bone.

Crystallization modifies osteoconductivity in an apatite-mullite glass-ceramic.

The response to implantation of novel apatite glass–ceramics was evaluated using a weight bearing in vivo bone implant model. Five novel glasses with varying calcium to phosphate ratios were cast as short rods and heat-treated to crystallize principally apatite. One glass ceramic had an apatite stoichiometry (Ca : P=1.67); three were phosphate-rich and one calcium-rich. One of the phosphate-rich glasses was also tested in its glassy state to determine the effect of crystallization on the biological response. Rods were implanted into the midshaft of rat femurs and left for 28 days. The femurs were then harvested and processed for scanning electron microscopy, energy dispersive X-ray microanalysis and conventional histology as ground and polished sections. Four of the materials exhibited evidence of osseointegration and osteoconduction. However, there was a marked inflammatory response to one of the phosphate-rich glass–ceramics, and to the non-crystallized glass. Crystallization of the latter significantly improved the bone tissue response. The glass–ceramic with an apatite stoichiometry elicited the most favorable response and merited further study as an osteoconductive bone substitute in maxillofacial and orthopedic surgery.

Self-reinforced polyglycolic acid membrane: a bioresorbable material for orbital floor repair. Initial clinical report

A self-reinforced polyglycolic acid membrane has been used successfully to repair 15 orbital floor fractures in 12 consecutive patients. As polyglycolic acid is absorbable it does not cause the complications of long-term infection and migration associated with non-absorbable bioinert alloplastic repair materials.

Biocompatibility of Resin-based Dental Materials

Oral and mucosal adverse reactions to resin-based dental materials have been reported. Numerous studies have examined the biocompatibility of restorative dental materials and their components, and a wide range of test systems for the evaluation of the biological effects of these materials have been developed. This article reviews the biological aspects of resin-based dental materials and discusses the conventional as well as the new techniques used for biocompatibility assessment of dental materials.

Biologic Assessment of Antiseptic Mouthwashes Using a Three-Dimensional Human Oral Mucosal Model

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.