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1.
J Mech Behav Biomed Mater ; 97: 330-338, 2019 09.
Article in English | MEDLINE | ID: mdl-31153114

ABSTRACT

OBJECTIVES: To determine the effect of heat on flexural strength (FS), maximum strain (MS), storage modulus (SM), tan delta (TD) and chemical changes through micro-Raman spectroscopy of dentine exposed to 2.5% NaOCl or saline. METHOD: ology: Dentine bars were randomly allocated to 8 test groups. Half (groups 2,4,6,8) were treated with NaOCl for 20 min; the rest (groups 1,3,5,7) remained in saline. FS/MS were measured in groups 1-4 (n = 15) (3/4 were also heated to 200 °C & re-hydrated in saline). Micro-Raman spectroscopy was performed on bars from groups 1-4. SM/TD were measured in 5-8: in 5/6 (n = 10), repeated after heating (200 °C), then following re-hydration; in 7/8 (n = 3) after heating to 25-185 °C. RESULTS: Increase in MS on heat and FS/MS on heat + NaOCl was not significant (P > 0.05). SM increased (P = 0.06) after heat treatment but reduced to initial state after rehydration (P = 0.03). TD did not change (P = 0.4) after heat (200 °C) treatment but rehydration increased it compared with pre-treatment state (P = 0.001). For dentine bars pre-treated with NaOCl, SM did not change (P = 0.6) after heat (200 °C) treatment or rehydration but TD significantly increased (P = 0.02) upon re-hydration compared with pre- (P=0.007), or post- (P = 0.03) heat-treatment states. SM and TD varied between 25-185 °C with no consistent trend amongst the NaOCl pre-treated bars. Micro-Raman only detected chemical changes following NaOCl treatment in the mineral phase. CONCLUSIONS: Exposure of dentine bars to heat and NaOCl produced only moderate changes to quasi-static but marked changes to viscoelastic properties, which may be explained by chemical alterations.


Subject(s)
Carbonates/chemistry , Dentin/chemistry , Hot Temperature , Sodium Hypochlorite/pharmacology , Cuspid/drug effects , Cuspid/pathology , Elasticity , Humans , Incisor/drug effects , Incisor/pathology , Molar/drug effects , Molar/pathology , Spectrum Analysis, Raman , Stress, Mechanical , Viscosity
2.
J Mech Behav Biomed Mater ; 50: 277-89, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26164218

ABSTRACT

Bone substitute composite materials with poly(L-lactide-co-glycolide) (PLGA) matrices and four different bioactive fillers: CaCO3, hydroxyapatite (HA), 45S5 Bioglass(®) (45S5 BG), and ICIE4 bioactive glass (a lower sodium glass than 45S5 BG) were produced via melt blending, extrusion and moulding. The viscoelastic, mechanical and thermal properties, and the molecular weight of the matrix were measured. Thermogravimetric analysis evaluated the effect of filler composition on the thermal degradation of the matrix. Bioactive glasses caused premature degradation of the matrix during processing, whereas CaCO3 or HA did not. All composites, except those with 45S5 BG, had similar mechanical strength and were stiffer than PLGA alone in compression, whilst all had a lower tensile strength. Dynamic mechanical analysis demonstrated an increased storage modulus (E') in the composites (other than the 45S5 BG filled PLGA). The effect of water uptake and early degradation was investigated by short-term in vitro aging in simulated body fluid, which indicated enhanced water uptake over the neat polymer; bioactive glass had the greatest water uptake, causing matrix plasticization. These results enable a direct comparison between bioactive filler type in poly(α-hydroxyester) composites, and have implications when selecting a composite material for eventual application in bone substitution.


Subject(s)
Bone Substitutes/chemistry , Lactic Acid/chemistry , Mechanical Phenomena , Polyglycolic Acid/chemistry , Temperature , Biomimetic Materials/chemistry , Body Fluids , Materials Testing , Molecular Weight , Polylactic Acid-Polyglycolic Acid Copolymer , Time Factors
3.
Bone ; 71: 244-56, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25460184

ABSTRACT

Bone cell culture systems are essential tools for the study of the molecular mechanisms regulating extracellular matrix mineralization. MC3T3-E1 osteoblast cell cultures are the most commonly used in vitro model of bone matrix mineralization. Despite the widespread use of this cell line to study biomineralization, there is as yet no systematic characterization of the mineral phase produced in these cultures. Here we provide a comprehensive, multi-technique biophysical characterization of this cell culture mineral and extracellular matrix, and compare it to mouse bone and synthetic apatite mineral standards, to determine the suitability of MC3T3-E1 cultures for biomineralization studies. Elemental compositional analysis by energy-dispersive X-ray spectroscopy (EDS) showed calcium and phosphorus, and trace amounts of sodium and magnesium, in both biological samples. X-ray diffraction (XRD) on resin-embedded intact cultures demonstrated that similar to 1-month-old mouse bone, apatite crystals grew with preferential orientations along the (100), (101) and (111) mineral planes indicative of guided biogenic growth as opposed to dystrophic calcification. XRD of crystals isolated from the cultures revealed that the mineral phase was poorly crystalline hydroxyapatite with 10 to 20nm-sized nanocrystallites. Consistent with the XRD observations, electron diffraction patterns indicated that culture mineral had low crystallinity typical of biological apatites. Fourier-transform infrared spectroscopy (FTIR) confirmed apatitic carbonate and phosphate within the biological samples. With all techniques utilized, cell culture mineral and mouse bone mineral were remarkably similar. Scanning (SEM) and transmission (TEM) electron microscopy showed that the cultures had a dense fibrillar collagen matrix with small, 100nm-sized, collagen fibril-associated mineralization foci which coalesced to form larger mineral aggregates, and where mineralized sites showed the accumulation of the mineral-binding protein osteopontin. Light microscopy, confocal microscopy and three-dimensional reconstructions showed that some cells had dendritic processes and became embedded within the mineral in an osteocyte-like manner. In conclusion, we have documented characteristics of the mineral and matrix phases of MC3T3-E1 osteoblast cultures, and have determined that the structural and compositional properties of the mineral are highly similar to that of mouse bone.


Subject(s)
Bone and Bones/physiology , Bone and Bones/ultrastructure , Calcification, Physiologic , Extracellular Matrix/metabolism , Osteoblasts/physiology , Osteoblasts/ultrastructure , Animals , Cells, Cultured , Mice , Minerals/metabolism , Spectrometry, X-Ray Emission , Spectroscopy, Fourier Transform Infrared , Vibration , X-Ray Diffraction
4.
J Mater Chem B ; 2(33): 5339-5343, 2014 Sep 07.
Article in English | MEDLINE | ID: mdl-32261754

ABSTRACT

Incorporation of anionic fibroin derived polypeptides into dense collagen gels provided a dynamic, three-dimensional, tissue-equivalent matrix together with biochemical cues that resembled the role of the bone morphogenic growth factors commonly used to promote osteogenic differentiation of mesenchymal stem cells.

5.
Acta Biomater ; 9(12): 9360-9, 2013 Dec.
Article in English | MEDLINE | ID: mdl-23896567

ABSTRACT

Cell-based therapies such as autologous chondrocyte implantation require in vitro cell expansion. However, standard culture techniques require cell passaging, leading to dedifferentiation into a fibroblast-like cell type. Primary chondrocytes grown on continuously expanding culture dishes (CE culture) limits passaging and protects against dedifferentiation. The authors tested whether CE culture chondrocytes were advantageous for producing mechanically competent cartilage matrix when three-dimensionally seeded in dense collagen gels. Primary chondrocytes, grown either in CE culture or passaged twice on static silicone dishes (SS culture; comparable to standard methods), were seeded in dense collagen gels and cultured for 3 weeks in the absence of exogenous chondrogenic growth factors. Compared with gels seeded with SS culture chondrocytes, CE chondrocyte-seeded gels had significantly higher chondrogenic gene expression after 2 and 3 weeks in culture, correlating with significantly higher aggrecan and type II collagen protein accumulation. There was no obvious difference in glycosaminoglycan content from either culture condition, yet CE chondrocyte-seeded gels were significantly thicker and had a significantly higher dynamic compressive modulus than SS chondrocyte-seeded gels after 3 weeks. Chondrocytes grown in CE culture and seeded in dense collagen gels produce more cartilaginous matrix with superior mechanical properties, making them more suitable than SS cultured cells for tissue engineering applications.


Subject(s)
Cartilage/physiology , Cell Culture Techniques/methods , Chondrocytes/cytology , Collagen Type II/pharmacology , Gels/chemistry , Aggrecans/metabolism , Animals , Cartilage/drug effects , Cattle , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , Chondrocytes/drug effects , Chondrocytes/metabolism , DNA/metabolism , Extracellular Matrix/drug effects , Extracellular Matrix/metabolism , Fluorescent Antibody Technique , Glycosaminoglycans/metabolism , Hydrogel, Polyethylene Glycol Dimethacrylate , Phenotype , Rats , Tissue Scaffolds
6.
J Dent Res ; 92(7): 648-54, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23632809

ABSTRACT

While advances in biomineralization have been made in recent years, unanswered questions persist on bone- and tooth-cell differentiation, on outside-in signaling from the extracellular matrix, and on the link between protein expression and mineral deposition. In the present study, we validate the use of a bioengineered three-dimensional (3D) dense collagen hydrogel scaffold as a cell-culture model to explore these questions. Dental pulp progenitor/stem cells from human exfoliated deciduous teeth (SHEDs) were seeded into an extracellular matrix-like collagen gel whose fibrillar density was increased through plastic compression. SHED viability, morphology, and metabolic activity, as well as scaffold mineralization, were investigated over 24 days in culture. Additionally, measurements of alkaline phosphatase enzymatic activity, together with immunoblotting for mineralized tissue cell markers ALPL (tissue-non-specific alkaline phosphatase), DMP1 (dentin matrix protein 1), and OPN (osteopontin), demonstrated osteo/odontogenic cell differentiation in the dense collagen scaffolds coincident with mineralization. Analyses of the mineral phase by electron microscopy, including electron diffraction and energy-dispersive x-ray spectroscopy, combined with Fourier-transform infrared spectroscopy and biochemical analyses, were consistent with the formation of apatitic mineral that was frequently aligned along collagen fibrils. In conclusion, use of a 3D dense collagen scaffold promoted SHED osteo/odontogenic cell differentiation and mineralization.


Subject(s)
Calcification, Physiologic/physiology , Dental Pulp/cytology , Fibrillar Collagens , Hydrogel, Polyethylene Glycol Dimethacrylate , Stem Cells/physiology , Tissue Scaffolds , Alkaline Phosphatase/analysis , Apatites/analysis , Biomarkers/analysis , Cell Culture Techniques , Cell Differentiation/physiology , Cell Shape/physiology , Cell Survival/physiology , Child , Child, Preschool , Extracellular Matrix/chemistry , Extracellular Matrix Proteins/analysis , Fibrillar Collagens/chemistry , Gels , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Odontogenesis/physiology , Osteogenesis/physiology , Osteopontin/analysis , Phosphoproteins/analysis , Pressure , Time Factors , Tissue Engineering/instrumentation , Tissue Scaffolds/chemistry , Tooth, Deciduous/cytology
7.
Acta Biomater ; 8(4): 1616-26, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22248526

ABSTRACT

Reinforcing biodegradable polymers with phosphate-based glass fibres (PGF) is of interest for bone repair and regeneration. In addition to increasing the mechanical properties, PGF can also release bioinorganics, as they are water soluble, a property that may be controllably translated into a fully degradable composite. Herein, the effect of Si and Fe on the solubility of calcium-containing phosphate-based glasses (PG) in the system (50P(2)O(5)-40CaO-(10-x)SiO(2)-xFe(2)O(3), where x=0, 5 and 10 mol.%) were investigated. On replacing SiO(2) with Fe(2)O(3), there was an increase in the glass transition temperature and density of the PG, suggesting greater crosslinking of the phosphate chains. This significantly reduced the dissolution rates of degradation and ion release. Two PG formulations, 50P(2)O(5)-40CaO-10Fe(2)O(3) (Fe10) and 50P(2)O(5)-40CaO-5Fe(2)O(3)-5SiO(2) (Fe5Si5), were melt drawn into fibres and randomly incorporated into polycaprolactone (PCL). Initially, the flexural strength and modulus significantly increased with PGF incorporation. In deionized water, PCL-Fe(5)Si(5) displayed a significantly greater weight loss and ion release compared with PCL-Fe10. In simulated body fluid, brushite was formed only on the surface of PCL-Fe(5)Si(5). Dynamic mechanical analysis in phosphate buffered saline (PBS) at 37°C revealed that the PCL-Fe10 storage modulus (E') was unchanged up to day 7, whereas the onset of PCL-Fe(5)Si(5)E' decrease occurred at day 4. At longer-term ageing in PBS, PCL-Fe(5)Si(5) flexural strength and modulus decreased significantly. MC3T3-E1 preosteoblasts seeded onto PCL-PGF grew up to day 7 in culture. PGF can be used to control the properties of biodegradable composites for potential application as bone fracture fixation devices.


Subject(s)
Bone Substitutes/pharmacology , Calcium Phosphates/pharmacology , Glass/chemistry , Iron/pharmacology , Polyesters/pharmacology , Silicon/pharmacology , Animals , Body Fluids/chemistry , Cell Adhesion/drug effects , Cell Survival/drug effects , Differential Thermal Analysis , Hydrogen-Ion Concentration/drug effects , Ions , Materials Testing , Mice , Microscopy, Electron, Scanning , Osteoblasts/cytology , Osteoblasts/drug effects , Temperature , Time Factors , Water/chemistry
8.
Acta Biomater ; 7(2): 829-40, 2011 Feb.
Article in English | MEDLINE | ID: mdl-20849987

ABSTRACT

The long-term (600days) in vitro degradation of highly porous poly(D,L-lactide) (PDLLA)/Bioglass-filled composite foams developed for bone tissue engineering scaffolds has been investigated in simulated body fluid (SBF). Foams of ∼93% porosity were produced by thermally induced phase separation (TIPS). The degradation profile for foams of neat PDLLA and the influence of Bioglass addition were comprehensively assessed in terms of changes in dimensional stability, pore morphology, weight loss, molecular weight and mechanical properties (dry and wet states). It is shown that the degradation process proceeded in several stages: (a) a quasi-stable stage, where water absorption and plasticization occurred together with weight loss due to Bioglass particle loss and dissolution, resulting in decreased wet mechanical properties; (b) a stage showing a slight increase in the wet mechanical properties and a moderate decrease in dimensions, with the properties remaining moderately constant until the onset of significant weight loss, whilst molecular weight continued to decrease; (c) an end stage of massive weight loss, disruption of the pore structure and the formation of blisters and embrittlement of the scaffold (evident on handling). The findings from this long-term in vitro degradation investigation underpin studies that have been and continue to be performed on highly porous poly(α-hydroxyesters) scaffolds filled with bioactive glasses for bone tissue engineering applications.


Subject(s)
Body Fluids/chemistry , Body Fluids/cytology , Bone and Bones/drug effects , Ceramics/pharmacology , Polyesters/pharmacology , Tissue Scaffolds/chemistry , Absorption/drug effects , Biocompatible Materials/pharmacology , Elastic Modulus/drug effects , Hydrogen-Ion Concentration/drug effects , Microscopy, Electron, Scanning , Molecular Weight , Porosity/drug effects , Time Factors , Water
9.
J Biomed Mater Res B Appl Biomater ; 93(2): 394-400, 2010 May.
Article in English | MEDLINE | ID: mdl-20119947

ABSTRACT

Local delivery of antibiotics may provide the advantage of reducing the potential side effects associated with their systemic administration. This study assessed, in vitro, the antimicrobial efficacy of tetracycline hydrochloride (TCH) adsorbed onto Bio-Oss bone grafts against a range of pathogenic bacteria. Various levels of TCH were adsorbed onto Bio-Oss granules by immersing in TCH aqueous solutions of different initial concentrations for 48 h at room temperature. TCH release was assessed in phosphate buffered saline at 37 degrees C, and its antimicrobial efficacy, up to 96 h, was tested against two Gram-negative bacteria associated with periodontal diseases: Aggregatibacter (formerly Actinobacillus) actinomycetemcomitans, and Porphyromonas gingivalis, and one Gram-positive bacterium associated with soft-tissue and bone infections: Staphylococcus aureus. The range of TCH concentrations studied was also assessed for cytotoxicity against osteoblast-like human osteosarcoma cell lines. The amount of TCH adsorbed and released from Bio-Oss was concentration dependent. All TCH adsorbed Bio-Oss resulted in a reduction of A. actinomycetemcomitans, P. gingivalis, and S. aureus and higher concentrations were generally more effective in reducing or eliminating bacterial growth. The proliferation of HOS cells was not substantially reduced except for the maximum concentration of TCH. In addition to its osteoconductive role, TCH adsorbed Bio-Oss could also be functional in negating systemically antibiotic prophylactic treatment in the prevention of implant or biomaterial related infections.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacteria/growth & development , Bacterial Infections/prevention & control , Bone Substitutes/pharmacology , Drug Delivery Systems , Minerals/pharmacology , Tetracycline/pharmacology , Bone Regeneration/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Humans
10.
Bone ; 43(2): 377-385, 2008 Aug.
Article in English | MEDLINE | ID: mdl-18502716

ABSTRACT

Bone is distinguished from other tissues by its mechanical properties, in particular stiffness. However, we know little of how osteoblasts react to the stiffness of their microenvironment; in this study we describe their response to a dense (>10 wt.%) collagenous 3D environment. Primary pre-osteoblasts were seeded within a novel form of native collagen, dense collagen, and cultured for up to 14 days in the presence and absence of osteogenic supplements: analysis was via Q-PCR, histology, fluorescent in situ zymography, MMP loss-of-function and tensile testing. Differentiation as measured through the up-regulation of Bsp (247-fold), Alp (14.2-fold), Col1A1 (4.5-fold), Mmp-13 (8.0-fold) and Runx2 (1.2-fold) transcripts was greatly accelerated compared to 2D plastic at 7 and 14 days in the same medium. The scale of this enhancement was confirmed through the use of growth factor stimulation on 2D via the addition of BMP-6 and the Hedgehog agonist purmorphamine. In concert, these molecules were capable of the same level of osteo-induction (measured by Bsp and Alp expression) as the dense collagen alone. Mineralisation was initially localised to remodelled pericellular regions, but by 14 days embedded cells were discernible within regions of apatite (confirmed by MicroRaman). Tensile testing of the matrices showed that this had resulted in a significant increase in Young's modulus at low strain values, consistent with a stiffening of the matrix. To determine the need for matrix remodelling in the mineralisation event the broad spectrum MMP Inhibitor Ilomastat was used. It was found that in its presence mineralisation could still occur (though serum-specific) and the apoptosis associated with MMP inhibition in hydrated collagen gels was abrogated. Analysis of gene expression indicated that this was due to the up-regulation of Mmp-13 in the presence of Ilomastat in dense collagen (400-fold), demonstrating a powerful feedback loop and a potential mechanism for the rescue from apoptosis. Osteoid-like matrix (dense collagen) is therefore a potent stimulant of osteoblast differentiation in vitro and provides an environment that enables survival and differentiation in the presence of MMP inhibition.


Subject(s)
Apoptosis , Cell Differentiation , Collagen/metabolism , Matrix Metalloproteinase Inhibitors , Osteogenesis , Animals , Apoptosis/drug effects , Cell Differentiation/drug effects , Cryoultramicrotomy , Culture Media , Enzyme Inhibitors/pharmacology , Gene Expression Regulation/drug effects , Mice , Mice, Inbred ICR , Osteoblasts/cytology , Osteoblasts/drug effects , Osteoblasts/enzymology , Osteogenesis/drug effects
11.
J Biomed Mater Res A ; 86(4): 987-95, 2008 Sep 15.
Article in English | MEDLINE | ID: mdl-18067165

ABSTRACT

A highly interconnected porous scaffold made from 45S5 Bioglass was fabricated by the polymer replica technique and surface functionalized for protein immobilization. Subsequently rat-tail collagen type I was immobilized on the scaffolds. The protein and ion release rates were determined by UV-vis spectroscopy and ion chromatography, respectively, and the impact on hydroxyapatite (HA) formation on the scaffolds upon immersion in SBF was evaluated. It was discovered that the surface functionalization enhanced the stability of the collagen attachment and stability against the increment of pH in a biological environment, resulting in similar collagen release kinetics in solutions of different pH values. Without the surface modification, collagen release was considerably expedited by the increment of pH in a surrounding solution. It was also found that the collagen immobilization does not effect the formation of carbonated HA on the scaffold surface. The stable collagen attachment to the functionalized scaffold makes this approach potentially suitable for improving cell attachment and thus for enhancing the application potential of the scaffold in tissue engineering.


Subject(s)
Ceramics/metabolism , Collagen/metabolism , Tissue Scaffolds , Animals , Glass , Hydrogen-Ion Concentration , Kinetics , Metals , Microscopy, Electron, Scanning , Porosity , Rats , Surface Properties , Time Factors , X-Ray Diffraction
12.
Acta Biomater ; 3(4): 563-72, 2007 Jul.
Article in English | MEDLINE | ID: mdl-17270508

ABSTRACT

There is currently a need to expand the range of graft materials available to orthopaedic surgeons. This study investigated the effect of ternary phosphate-based glass (PBG) compositions on the behaviour of osteoblast and osteoblast-like cells. PBGs of the formula (in mol.%) P(2)O(5)(50)-CaO(50-X)-Na(2)O(X), where X is either 2, 4, 6, 8 or 10, were produced and their influence on the proliferation, differentiation and death in vitro of adult human bone marrow stromal cells (hBMSCs) and human fetal osteoblast 1.19 (HFOB 1.19) cells were assessed. Tissue culture plastic (TCP) and hydroxyapatite (HA) were used as controls. Exposure to PBGs in culture inhibited cell adhesion and proliferation and increased cell death in both cell types studied. There was no significant difference in percentage cell death between the PBGs, which was significantly greater than the controls. However, compared with other PBGs, a greater number of cells were found on the 48mol.% CaO which may have been due to either increased adherence or proliferation, or both. This composition was capable of supporting osteogenic proliferation and early differentiation, and supports the notion that chemical modification of the glass could lead to a more biologically compatible substrate with the potential to support osteogenic grafting. Realisation of this potential should lead to the development of novel grafting strategies for the treatment of problematic bone defects.


Subject(s)
Glass/chemistry , Osteoblasts/cytology , Osteoblasts/physiology , Phosphates/chemistry , Stromal Cells/cytology , Stromal Cells/physiology , Adult , Bone Marrow Cells/cytology , Cell Adhesion , Cell Culture Techniques , Cell Death , Cell Differentiation , Cell Survival , Cells, Cultured , Durapatite/chemistry , Fetus , Humans , Osteoblasts/metabolism , Osteoblasts/ultrastructure , Phosphates/metabolism , Plastics/chemistry , Stromal Cells/metabolism , Stromal Cells/ultrastructure
13.
J Biomed Mater Res B Appl Biomater ; 80(2): 322-31, 2007 Feb.
Article in English | MEDLINE | ID: mdl-16838353

ABSTRACT

Phosphate glass (PG) of the composition 0.46(CaO)-0.04(Na(2)O)-0.5(P(2)O(5)) was used as filler in poly-L-lactic acid (PLA) foams developed as degradable scaffolds for bone tissue engineering. The effect of PG on PLA was assessed both in bulk and porous composite foams. Composites with various PG content (0, 5, 10, and 20 wt %) were melt-extruded, and either compression-molded or foamed through supercritical CO(2). Dynamic mechanical analysis on the bulk composites showed that incorporating 20 wt % PG resulted in a significant increase in storage modulus. Aging studies in deionized water in terms of weight loss, pH change, and ion release inferred that the degradation was due to PG dissolution, and dependent on the amount of glass in the composites. Foaming was only possible for composites containing 5 and 10 wt % PG, as an increase in PG increased the foam densities; however, the level of porosity was maintained above 75%. PLA-T(g) in the foams was higher than those obtained for the bulk. Compressive moduli showed no significant reinforcement with glass incorporation in either expansion direction, indicating no anisotropy. Biocompatibility showed that proliferation of human fetal bone cells was more rapid for PLA compared to PLA-PG foams. However, the proliferation rate of PLA-PG foams were similar to those obtained for foams of PLA with either hydroxyapatite or beta-tricalcium phosphate.


Subject(s)
Biocompatible Materials , Glass , Lactic Acid , Polymers , Tissue Engineering/methods , Biocompatible Materials/chemistry , Biomechanical Phenomena , Cells, Cultured , Drug Stability , Glass/chemistry , Humans , Lactic Acid/chemistry , Materials Testing , Microscopy, Electron, Scanning , Osteoblasts/cytology , Particle Size , Phosphates/chemistry , Polyesters , Polymers/chemistry , Thermodynamics , Time Factors
14.
J Mater Sci Mater Med ; 17(11): 979-87, 2006 Nov.
Article in English | MEDLINE | ID: mdl-17122908

ABSTRACT

The first and foremost function of a tissue engineering scaffold is its role as a substrate for cell attachment, and their subsequent growth and proliferation. However, cells do not attach directly to the culture substrate; rather they bind to proteins that are adsorbed to the scaffold's surface. Like standard tissue culture plates, tissue engineering scaffolds can be chemically treated to couple proteins without losing the conformational functionality; a process called surface functionalization. In this work, novel highly porous 45S5 Bioglass-based scaffolds have been functionalized applying 3-AminoPropyl-TriethoxySilane (APTS) and glutaraldehyde (GA) without the use of organic solvents. The efficiency and stability of the surface modification was assessed by X-ray photoemission spectroscopy (XPS). The bioactivity of the functionalized scaffolds was investigated using simulated body fluid (SBF) and characterized by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD). It was found that the aqueous heat-treatment applied at 80 degrees C for 4 hrs during the surface functionalization procedure accelerated the structural transition of the crystalline Na2Ca2Si3O9 phase, present in the original scaffold structure as a result of the sintering process used for fabrication, to an amorphous phase during SBF immersion. The surface functionalized scaffolds exhibited an accelerated crystalline hydroxyapatite layer formation upon immersion in SBF caused by ion leaching and the increased surface roughness induced during the heat treatment step. The possible mechanisms behind this phenomenon are discussed.


Subject(s)
Biocompatible Materials , Ceramics , Glass , Materials Testing , Biocompatible Materials/chemistry , Ceramics/chemistry , Glass/chemistry , Microscopy, Electron, Scanning , Spectrometry, X-Ray Emission , X-Ray Diffraction
15.
J Biomed Mater Res B Appl Biomater ; 79(2): 360-4, 2006 Nov.
Article in English | MEDLINE | ID: mdl-16680683

ABSTRACT

The setting behavior of a brushite-forming cement (beta-tricalcium phosphate/mono calcium monophosphate) was investigated using an indentation technique (the Gillmore needles method) and isothermal differential scanning calorimetry (DSC). The two objectives of the study were to investigate whether DSC could be used to real-time monitor a fast-setting calcium phosphate cement (CPC) and to determine if it is possible to correlate DSC results directly with conventional setting-time measurements. Best-fit linear correlation analysis revealed that both the initial and final setting time (T(i) and T(f)) measured by indentation were strongly correlated to the maximum heat flow measured with DSC. It seems therefore possible to predict the setting times, usually achieved with user dependent indentation methods, of this specific fast setting CPC on the basis of objective DSC measurements. The drawbacks of DSC, however, are its overall complexity and expense and the fact that only exothermal reactions can be investigated in comparison to the Gillmore needles method, furthermore, it is not possible to monitor the complete reaction as the first 2 or 3 min are lost due to sample preparation and apparatus set up.


Subject(s)
Bone Cements/chemistry , Calcium Phosphates/chemistry , Calorimetry, Differential Scanning
16.
J Biomater Appl ; 20(1): 65-80, 2005 Jul.
Article in English | MEDLINE | ID: mdl-15972364

ABSTRACT

This article reports on the use of ion chromatography (IC) to investigate extensively the release profiles of both cations and anions and characterize the relationship between composition and degradation for a ternary-based Na(2)O-CaO-P(2)O(5) glass system developed as biomaterials. Studies are carried out on glasses with the formula 45P(2)O(5)-55(xCaO-Na(2)O) in deionized water, where x = 30, 35, and 40 mol%, using a cumulative release method, where the solution is changed at regular intervals. Degradation behavior is linear with time where the degradation rate shows an initial decrease with increasing CaO content. This rate then increases with a further addition of CaO. Cation release profiles follow similar trends to the degradation rates. Anion release profiles show a decrease for the PO(4) and linear polyphosphate (P(2)O(7) and P(3)O(10)) species with increasing CaO content. This decrease is attributed to the cross-linking of the Ca(2+) ions. In contrast, the cyclic P(3)O(9) anion exhibits the highest amount of anionic release, which demonstrates similar trends to the cations. These release patterns suggest that the cyclic P(3)O(9) species dominate the degradation rates. The proposed mode of degradation is a hydrolysis reaction, with the cyclic metaphosphate undergoing acid/base catalysis. The pH remains constant for the 30 and 35 mol% CaO glasses, and drops to about 5.5 for the 40 mol% composition. By using a response factor, it is possible to semiquantitatively analyze the additional peaks observed in the chromatograms. Suggestions are also put forward as to the identity of some of these unidentified peaks.


Subject(s)
Absorbable Implants , Biocompatible Materials/chemistry , Glass/chemistry , Materials Testing , Phosphates/chemistry , Phosphorus Compounds/chemistry , Anions , Biocompatible Materials/analysis , Cations , Diffusion
17.
Acta Biomater ; 1(5): 553-63, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16701835

ABSTRACT

Phosphate-based glass fibres (PGF) have the unique characteristic of being completely soluble in an aqueous environment, releasing bioactive and biocompatible ions. They have been proposed as tissue engineering scaffolds for craniofacial skeletal muscle regeneration, where myoblasts are seeded directly onto the fibres. Studies have shown that these cells have a preference in their initial attachment to fibres of certain composition and size, which in turn control the rate of degradation. This study investigated the relationship between the surface properties, degradation properties and ion release (cationic and anionic species) by altering the chemical composition of the PGF. Iron oxide (Fe2O3) was incorporated into glasses containing P2O5 (50 mol%), CaO (30 mol%) and Na2O (20 mol%). Six glass compositions with Fe2O3 ranging from 0 to 5 mol% by replacing the equivalent Na2O mol% were investigated. Contact angle measurements showed that polar interactions occurring on the glass surfaces diminished with increasing Fe2O3 content. This behaviour was reflected in the estimated surface energies of the glasses, where the overall surface energy decreased with increasing Fe2O3 content due to the decrease in polar or acid/base component. The incorporation of up to 5 mol% Fe2O3 into PGF resulted in a significant reduction in the degradation rate (by two orders of magnitude), which can be related to the formation of more hydration resistant P-O-Fe bonds. However, the degradation rate increased with decreasing fibre diameter (comparing average diameters of 31.6 +/- 6.5 microm versus 13.1 +/- 1.3 microm) for a given mass of fibre, and this is related to the surface area to volume ratio. Taken together the results suggest that fibres with the larger diameters and containing 3-5 mol% Fe2O3 could initially be a more durable scaffold than ones with 1 or 2 mol% Fe2O3 for initial cell attachment.


Subject(s)
Absorbable Implants , Biocompatible Materials/chemistry , Glass/chemistry , Iron/chemistry , Phosphates/chemistry , Tissue Engineering/methods , Diffusion , Ions , Materials Testing , Molecular Weight , Particle Size , Surface Properties
18.
Acta Biomater ; 1(6): 643-52, 2005 Nov.
Article in English | MEDLINE | ID: mdl-16701845

ABSTRACT

This study developed highly porous degradable composites as potential scaffolds for bone tissue engineering. These scaffolds consisted of poly-D,L-lactic acid filled with 2 and 15 vol.% of 45S5 Bioglass particles and were produced via thermally induced solid-liquid phase separation and subsequent solvent sublimation. The scaffolds had a bimodal and anisotropic pore structure, with tubular macro-pores of approximately 100 microm in diameter, and with interconnected micro-pores of approximately 10-50 microm in diameter. Quasi-static and thermal dynamic mechanical analysis carried out in compression along with thermogravimetric analysis was used to investigate the effect of Bioglass on the properties of the foams. Quasi-static compression testing demonstrated mechanical anisotropy concomitant with the direction of the macro-pores. An analytical modelling approach was applied, which demonstrated that the presence of Bioglass did not significantly alter the porous architecture of these foams and reflected the mechanical anisotropy which was congruent with the scanning electron microscopy investigation. This study found that the Ishai-Cohen and Gibson-Ashby models can be combined to predict the compressive modulus of the composite foams. The modulus and density of these complex foams are related by a power-law function with an exponent between 2 and 3.


Subject(s)
Bone Substitutes/chemistry , Ceramics/chemistry , Models, Chemical , Polyesters/chemistry , Tissue Engineering/methods , Anisotropy , Ceramics/analysis , Compressive Strength , Computer Simulation , Elasticity , Glass , Materials Testing , Molecular Weight , Particle Size , Polyesters/analysis , Porosity
19.
Biomaterials ; 26(15): 2247-54, 2005 May.
Article in English | MEDLINE | ID: mdl-15585226

ABSTRACT

Phosphate-based glass fibres (PGF) of the general formula Na(2)O-CaO-P(2)O(5) are degradable in an aqueous environment, and therefore can function as antibacterial delivery systems through the inclusion of ions such as copper. In this study, PGF with varying amounts of copper oxide (CuO) were developed for potential uses in wound healing applications. PGF with 0, 1, 5 and 10 mol% CuO were produced with different diameters and characterised in terms of structural and antibacterial properties. The effect of CuO and fibre pulling speed on the glass properties were investigated using rapid differential scanning calorimetry, differential thermal analysis and X-ray diffraction. The effect of two fibre diameters on short-term (3 h) attachment and killing against Staphylococcus epidermidis were investigated and were related to their rate of degradation in deionised water, as well as copper ion release measured using ion chromatography. Thermal analysis showed that there was a significant increase in the PGF glass transition temperature as the CuO content increased. There was a significant decrease in the rate of degradation with increasing CuO content and an increase in fibre diameter. Over 6 h, both the amount and rate of copper ions released increased with CuO content, as well as a reduction in fibre diameter thus increasing the surface area to volume ratio. There was a decrease in the number of viable staphylococci both attached to the CuO-containing fibres and in the surrounding environment.


Subject(s)
Absorbable Implants , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Copper/administration & dosage , Copper/chemistry , Glass/chemistry , Staphylococcus epidermidis/drug effects , Anti-Bacterial Agents/administration & dosage , Cell Adhesion/drug effects , Cell Survival/drug effects , Drug Implants/administration & dosage , Drug Implants/chemistry , Materials Testing , Particle Size , Phosphates/chemistry , Staphylococcus epidermidis/cytology , Transition Temperature
20.
J Biomater Sci Polym Ed ; 15(6): 727-39, 2004.
Article in English | MEDLINE | ID: mdl-15255522

ABSTRACT

Denture soft lining materials are compliant cushions used at the oral tissue-denture interface. They are generally required to have sufficient compliance to redistribute mastication load, as well as an adequate modulus for long-term dimensional stability and control over the water uptake. This study investigated the effect of using silane treated fumed silica as a filler on the properties of experimental soft lining materials based on blends of isoprene-styrene (SIS) block co-polymer and mixtures of methyl methacrylate (MMA) and 1,6-hexandiol dimethacrylate (HDMA). The overall elastomer/monomer ratios were maintained, whereas the monomers ranged from 10 to 60% HDMA. The silica filler level was maintained at 10 wt% with respect to SIS. The properties investigated were the dynamic mechanical parameters of storage modulus (E') and tan delta as a function of temperature and the quasi-static mechanical parameters of ultimate tensile strength (UTS) and elongation to break (Eb) as well as absorption properties that were carried out in water and saline. In both unfilled and filled systems there was an increase in E' and a decrease in tan delta with an increase in HDMA. Silica addition tended to increase E' and substantially reduce tan delta in materials with less than 20% HDMA. UTS decreased with filler and HDMA content, however, Eb was greater for filled systems. Generally, in the long term, the water uptake decreased with increasing HDMA content and E'. The silanated silica further reduced the water uptake, indicating a cross-linking effect, thus increasing the restraining force on droplet growth. The uptake in saline was significantly reduced indicating an osmotically controlled uptake process.


Subject(s)
Biocompatible Materials/chemistry , Dental Implants , Elastomers/chemistry , Silicon Dioxide/chemistry , Diffusion , Methacrylates/chemistry , Methylmethacrylate/chemistry , Osmosis , Polymers/chemistry , Sodium Chloride/chemistry , Temperature , Tensile Strength , Time Factors , Water/chemistry
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