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An innovative approach was developed to engineer a multi-layered chitosan scaffold for osteochondral defect repair. A combination of freeze drying and porogen-leaching out methods produced a porous, bioresorbable scaffold with a distinct gradient of pore size (mean = 160–275 μm). Incorporation of 70 wt% nano-hydroxyapatite (nHA) provided additional strength to the bone-like layer. The scaffold showed instantaneous mechanical recovery under compressive loading and did not delaminate under tensile loading. The scaffold supported the attachment and proliferation of human mesenchymal stem cells (MSCs), with typical adherent cell morphology found on the bone layer compared to a rounded cell morphology on the chondrogenic layer. Osteogenic and chondrogenic differentiation of MSCs preferentially occurred in selected layers of the scaffold in vitro, driven by the distinct pore gradient and material composition. This scaffold is a suitable candidate for minimally invasive arthroscopic delivery in the clinic with potential to regenerate damaged cartilage and bone.

Hypothesis The accretion of ice on component surfaces often causes severe impacts or accidents in modern industries. Applying icephobic surface is considered as an effective solution to minimise the hazards. However, the durability of the current icephobic surface and coatings for long-term service remains a great challenge. Therefore, it is indeed to develop new durable material structures with great icephobic performance. Experiments A new design concept of combining robust porous metallic skeletons and icephobic filling was proposed. Nickel/polydimethylsiloxane (PDMS) two-phase layer was prepared using porous Ni foam skeletons impregnated with PDMS as filling material by a two-step method. Findings Good icephobicity and mechanical durability have been verified. Under external force, micro-cracks could easily initiate at the ice/solid interface due to the small surface cavities and the difference of local elastic modulus between the ice and PDMS, which would promote the ice fracture and thus lead to low ice adhesion strength. The surface morphology and icephobicity almost remain unchanged after water-sand erosion, showing greatly improved mechanical durability. By combining the advantages of the mechanical durability of porous Ni skeleton and the icephobicity of PDMS matrix, the Ni foam/PDMS two-phase layer demonstrates great potentials for ice protection with long-term service time.

In recent years, it has been found that small weight percent additions of silicon to HA can be used to enhance the initial response between bone tissue and HA. A large amount of research has been concerned with bulk materials, however, only recently has the attention moved to the use of these doped materials as coatings. This paper focusses on the development of a co-RF and pulsed DC magnetron sputtering methodology to produce a high percentage Si containing HA (SiHA) thin films (from 1.8 to 13.4 wt.%, one of the highest recorded in the literature to date). As deposited thin films were found to be amorphous, but crystallised at different annealing temperatures employed, dependent on silicon content, which also lowered surface energy profiles destabilising the films. X-ray photoelectron spectroscopy (XPS) was used to explore the structure of silicon within the films which were found to be in a polymeric (SiO2, Q4) state. However, after annealing, the films transformed to a SiO44&minus, Q0, state, indicating that silicon had substituted into the HA lattice at higher concentrations than previously reported. A loss of hydroxyl groups and the maintenance of a single-phase HA crystal structure further provided evidence for silicon substitution. Furthermore, a human osteoblast cell (HOB) model was used to explore the in vitro cellular response. The cells appeared to prefer the HA surfaces compared to SiHA surfaces, which was thought to be due to the higher solubility of SiHA surfaces inhibiting protein mediated cell attachment. The extent of this effect was found to be dependent on film crystallinity and silicon content.

A major challenge in orthopedics is the repair of large non-union bone fractures. A promising therapy for this indication is the use of biodegradable bioinspired biomaterials that stabilize the fracture site, relieve pain and initiate bone formation and healing. This study uses a multidisciplinary evaluation strategy to assess immunogenicity, allergenicity, bone responses and physicochemical properties of a novel biomaterial scaffold. Two-photon stereolithography generated personalized custom-built scaffolds with a repeating 3D structure of Schwarz Primitive minimal surface unit cell with a specific pore size of ?400 ?m from three different methacrylated poly(D,L-lactide-co-?-caprolactone) copolymers with lactide to caprolactone monomer ratios of 16?:?4, 18?:?2 and 9?:?1. Using in vitro and in vivo assays for bone responses, immunological reactions and degradation dynamics, we found that copolymer composition influenced the scaffold physicochemical and biological properties. The scaffolds with the fastest degradation rate correlated with adverse cellular effects and mechanical stiffness correlated with in vitro osteoblast mineralization. The physicochemical properties also correlated with in vivo bone healing and immune responses. Overall these observations provide compelling support for these scaffolds for bone repair and illustrate the effectiveness of a promising multidisciplinary strategy with great potential for the preclinical evaluation of biomaterials.

Icephobicity of materials has received intensive attention in recent years due to the increasing requirement of ice protection in aerospace, wind energy and power lines. However, the influencing factors of material icephobicity have not been well identified. In this work, the effect of surface gaseous adsorption on icing behaviour of materials was investigated for the first time. Ni-Cu-P coatings with different surface morphologies were fabricated and used as the objects of the study. Environmental scanning electron microscopy (ESEM) was utilized to observe the water condensation and ice formation on the coatings. X-ray photoelectron spectroscopy (XPS) was employed to analyse the variations of surface adsorption. Droplets icing time and ice adhesion strength of the coatings were also studied. The results showed that the icing time of water droplets on the Ni-Cu-P coatings increased significantly, and the ice adhesion strength decreased sharply with the spontaneous surface adsorption of gaseous species (mainly hydrocarbon groups) in air. The adsorbed hydrocarbon species would promote the formation of air pockets between the ice-coating interface, which could effectively reduce the interfacial contact of the formed ice with the coating. When the adsorbed hydrocarbon species were removed by plasma cleaning, water droplets tended to have more direct contacts with the coatings prior to icing, leading to the formation of interlocked ice and significantly increased the ice adhesion on the surface. The variation of surface icephobicity can also be attributed to the changes of surface energy due to the surface adsorption. The results indicated that the surface gaseous adsorption in air played an important role in determining the surface icing behaviour and the icephobicity of the materials.

Human mesenchymal stem cells (MSCs) show promise for musculoskeletal repair applications. Animal-derived serum is extensively used for MSC culture as a source of nutrients, extracellular matrix proteins and growth factors. However, the routine use of fetal calf serum (FCS) is not innocuous due to its animal antigens and ill-defined composition, driving the development of alternatives protocols. The present study sought to reduce exposure to FCS via the transient use of human serum. Transient exposure to animal serum had previously proved successful for the osteogenic differentiation of MSCs but had not yet been tested with alternative serum sources. Here, human serum was used to support the proliferation of MSCs, which retained surface marker expression and presented higher alkaline phosphatase activity than those in FCS-based medium. Addition of osteogenic supplements supported strong mineralisation over a 3-week treatment. When limiting serum exposure to the first five days of treatment, MSCs achieved higher differentiation with human serum than with FCS. Finally, human serum analysis revealed significantly higher levels of osteogenic components such as alkaline phosphatase and 25-Hydroxyvitamin D, consistent with the enhanced osteogenic effect. These results indicate that human serum used at the start of the culture offers an efficient replacement for continuous FCS treatment and could enable short-term exposure to patient-derived serum in the future.

Multiphase hybrids were fabricated from poly(d,l-lactide-co-ɛ-caprolactone) (PLCL) copolymer scaffolds impregnated with silk-elastin-like recombinamers (SELRs) hydrogel containing 2 wt% hydroxyapatite nanoparticles (nHA). The PLCL scaffolds, triply-periodic minimal surface geometry, were manufactured using two-photon stereolithography. In vitro degradation studies were conducted on PLCL scaffolds in inflamed tissue mimic media (pH ~ 4.5–6.5) or phosphate buffered saline (PBS) at 37 °C. Compression test revealed instant shape recovery of PLCL scaffolds after compression to 70% strain, ideal for arthroscopic delivery. Degradation of these scaffolds was accelerated in acidic media, where mass loss and compressive properties at day 56 were about 2–6 times lower than the scaffolds degraded in PBS. No significant difference was seen in the compressive properties between PLCL scaffolds and the hybrids due to the order of magnitude difference between the hydrogels and the PLCL scaffolds. Moreover, degradation properties of the hybrids did not significantly change by inclusion of SELR+/−nHA nanocomposite hydrogels. The hybrids lost approximately 40% and 84% of their initial weight and mechanical properties, respectively after 112 days of degradation. Cytotoxicity assessment revealed no cytotoxic effects of PLCL or PLCL-SELR+/−2%nHA scaffolds on bone marrow-derived human Mesenchymal Stem Cells. These findings highlight the potential of these hybrid constructs for bone and cartilage repair. Keywords: Poly(d,l-lactide-co-ɛ-caprolactone) copolymer, Multiphase hybrids, In vitro degradation, Mechanical recovery, human Mesenchymal Stem Cells

ICIE16 and 13-93 bioactive glasses have been proposed as alternative chemically stable compositions in physiological fluid keeping bioactivity comparable to Bioglass®. ICIE16 and 13-93 bioactive glasses coatings were produced via an emerging suspension high-velocity oxy-fuel (SHVOF) thermal spraying technique. Suspensions of ICIE16 and 13-93 with 10 wt% solid loading in isopropanol (IPA) and water were used to produce coatings on AISI304 stainless steel using a flame power of 50 kW and 75 kW. For both glass formulations, the coatings deposited at a lower flame power were more porous, less hard, and less rough (~6% porous and 242 HV) than the coatings obtained at a higher flame power (~4% porosity and 300 HV). ICIE16 coatings showed more dissolution in SBF (simulated body fluid) than the 13-93 coatings. Moreover, the 13-93 glass coating sprayed at 75 kW showed the highest stability in SBF since only 2% of the coating was resorbed in SBF after 7 days of immersion and revealed apatite precipitation after 7 days. In-vitro cell tests, using MG63 cells, showed good cell attachment and proliferation on the surfaces of the coatings, revealing good cytocompatibility. The 13-93 coating sprayed at 75 kW revealed the highest cell proliferation after 7 days of incubation. This can be attributed to the higher surface roughness of the coating (Ra = 6.5 ± 0.6 μm).

Hydrophobic metallic coatings have attracted increasing interest in the recent years due to their excellent mechanical durability. But the wetting behavior and hydrophobic mechanism of metallic coatings are far from clear. In this work, Ni-Cu-P ternary coatings with hierarchical structure were prepared on 304 stainless steel by electrodeposition method. The surface morphologies, phase compositions, and wetting behavior were studied systemically. Time-dependent wetting behaviour of Ni-Cu-P coatings had been clearly observed, and the surface of the as-deposited coatings changed from hydrophilic state to hydrophobic state after aging in ambient air. The related surface wetting mechanism was investigated with the assistance of plasma cleaning to study the possible surface adsorption contributing to the time-dependent wetting behavior. The variations of the surface species were analyzed using X-ray photoelectron spectroscopy (XPS), showing the composition change of both carbon and the oxygen. The atomic ratio of hydrocarbon on the Ni-Cu-P coating first increased from 78.7% to 86.5% when stored in ambient air and then decreased from 82.3% to 65.9% after the plasma cleaning treatment; while the variation of oxygen content was an opposite trend. The results indicated that the observed time-dependent wettability was a combined result of the adsorption of airborne hydrocarbon and the change of lattice oxygen on the coating surface.

Phosphate‐based glasses (PBGs) are ideal materials for regenerative medicine strategies because their composition, degradation rates, and ion release profiles can easily be controlled. Strontium has previously been found to simultaneously affect bone resorption and deposition. Therefore, by combining the inherent properties of resorbable PBG and therapeutic activity of strontium, these glasses could be used as a delivery device of therapeutic factors for the treatment of orthopaedic diseases such as osteoporosis. This study shows the cytocompatibility and osteogenic potential of PBGs where CaO is gradually replaced by SrO in the near invert glass system 40P2O5·(16‐x)CaO·20Na2O·24MgO·xSrO (x = 0, 4, 8, 12, and 16 mol%). Direct seeding of MG63 cells onto glass discs showed no significant difference in cell metabolic activity and DNA amount measurement across the different formulations studied. Cell attachment and spreading was confirmed via scanning electron microscopy (SEM) imaging at Days 3 and 14. Alkaline phosphatase (ALP) activity was similarly maintained across the glass compositions. Follow‐on studies explored the effect of each glass composition in microsphere conformation (size: 63‐125 μm) on human mesenchymal stem cells (hMSCs) in 3D cultures, and analysis of cell metabolic activity and ALP activity showed no significant differences at Day 14 over the compositional range investigated, in line with the observations from MG63 cell culture studies. Environmental SEM and live cell imaging at Day 14 of hMSCs seeded on the microspheres showed cell attachment and colonisation of the microsphere surfaces, confirming these formulations as promising candidates for regenerative medicine strategies addressing compromised musculoskeletal/orthopaedic diseases.

A single solvent based film casting process for fabricating porous polymer films was developed in this study. The porous film was produced by mixing concentrated polylactic acid/ chloroform solution (20 wt%) and fresh chloroform solvent followed by film casting. The average pore sizes of the films produced were seen to increase from 2.1 (±0.1) µm to 6.4 (±0.2) µm with increasing ratio of concentrated PLA solution and fresh solvent from 1:2 to 1:4. Functional groups of PLA after casting into porous film were confirmed via Fourier transform infrared (FTIR) spectroscopy analysis. Cytocompatibility studies (via Alamar Blue assessment) utilising MG-63 cells on the porous PLA films revealed an increase in cell metabolic activity up to 8 days post-seeding. In addition, these direct cell culture studies showed that the porous membranes supported cell adhesion and growth not only on the surface but also through the porous structures of the membrane, highlighting the suitability of these porous films in tissue engineering applications.

Low moduli cell culture substrates can be used to apply dynamic mechanical strain to cells, by surface deformation. Understanding the surface interaction with cells is critical to improving cell adhesion and normal growth. A medical grade polyurethane (PU), Chronoflex AL 80A, was modified by oxygen plasma etching and characterised by X-ray photoelectron spectroscopy. Etching resulted in increased cross-linking at the isocyanate bond and formation of new oxygen moieties. The model, derived from patent data and XPS data of the unetched PU, indicated that the additional oxygen was likely to be hydroxyl and carbonyl groups. Etched membranes enhanced protein adhesion, resulting in full surface coverage compared to unetched PU. The etched PU supported cell adhesion and spreading, while the unetched PU was not conducive to monolayer formation.

Completely resorbable composites are an attractive alternative for metallic bone-fracture fixation devices. However, failure of their interfacial integrity within aqueous environments, which can lead to a rapid loss of overall mechanical properties, has been reported in the literature. In this study coupling agents were investigated for phosphate glass fibre reinforced poly(lactic acid) composites. Three coupling agents with varying wettability were employed to improve initial mechanical properties and their retention in vitro via improvement of the interfacial bond between polymer matrix and fibres. Coupling agents were grafted onto the glass fibres by dip-coating in coupling agent solution at optimised concentrations. Three-aminopropyltriethoxy silane and sorbitol ended PLA oligomer treatments improved the initial flexural properties (27% strength with APS and 17% modulus via SPLA treatment) of the composites and 3-aminopropyltriethoxy silane and hexamethylene diisocyanate (HDI) treatments also decreased the loss of flexural strength and modulus during degradation. HDI treated samples retained 57.2% and 64.7% of their initial strength and modulus, respectively compared to control where only 34% of initial strength and 52% of initial modulus was retained after 28 days of degradation in PBS solution. Initial improvements in flexural properties were associated with improved shear bond strength at the interface due to covalent bonding between the glass fibres and polymer matrix provided by the coupling agents. Delay in mechanical property loss with degradation was suggested to be due to the hydrophobicity at the interface, which could have hindered the interfacial integrity loss and consequently loss of mechanical integrity of the composites. All coupling agent treated and control composites were tested for cytocompatibility using a primary human osteoblast cell line. A comparable response to the control, in terms of cell adhesion, proliferation and differentiation was observed supporting the use of these agents within implantable devices.

Bioreactors can be used for mechanical conditioning and to investigate the mechanobiology of cells in vitro. In this study a polyurethane (PU), Chronoflex AL, was evaluated for use as a flexible cell culture substrate in a novel bioreactor capable of imparting cyclic uniaxial tensile strain to cells. PU membranes were plasma etched, across a range of operating parameters, in oxygen. Contact angle analysis and X-ray photoelectron spectroscopy showed increases in wettability and surface oxygen were related to both etching power and duration. Atomic force microscopy demonstrated that surface roughness decreased after etching at 20 W but was increased at higher powers. The etching parameters, 20 W 40 s, produced membranes with high surface oxygen content (21%), a contact angle of 66° ± 7° and reduced topographical features. Etching and protein conditioning membranes facilitated attachment, and growth to confluence within 3 days, of MG-63 osteoblasts. After 2 days with uniaxial strain (1%, 30 cycles/min, 1500 cycles/day), cellular alignment was observed perpendicular to the principal strain axis, and found to increase after 24 h. The results indicate that the membrane supports culture and strain transmission to adhered cells. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 2356–2364, 2014.

In the UK, it is now recognised that 1 in 2 people born after 1960 will develop some form of cancer during their lifetime. Diagnosing patients whilst in the early stages drastically improves their chances of survival but up until now the gold standard for cancer detection is via a lengthy excision biopsy procedure, which relies on the skill of a histopathologist. Evidently, the need for a faster solution is paramount. The mid-infrared (MIR) spectral region covers the wavelengths 3-25 μm and characteristic vibrational spectra unique to each molecular type. Subtle changes in the specific spectral response within this region are indicative of changes within the cells relative to normal cells, signifying the presence or absence of a disease. Our goal is to carry out disease diagnosis in vivo. Reaching these wavelengths has previously presented difficulties as conventional MIR blackbody light sources are weak and optical fibers for transmitting MIR light to/from tissue in vivo can be limited by strong material absorption such as silica glass >2.4 μm and tellurite, and heavy metal fluoride, >4.75 μm. However, chalcogenide glasses have been shown to transmit MIR light out to 25 μm. This paper reports on a glass composition in the Ge-Sb-Se system and its suitability as an optical fiber for the transmission of MIR to and from tissue samples, enabling in vivo mapping for an immediate diagnostic response- a technique termed ‘optical biopsy’.

© 2016 IOP Publishing Ltd. The manufacture of 3D scaffolds with specific controlled porous architecture, defined microstructure and an adjustable degradation profile was achieved using two-photon polymerization (TPP) with a size of 2 × 4 × 2 mm3. Scaffolds made from poly(D,L-lactide-co-ε-caprolactone) copolymer with varying lactic acid (LA) and ε -caprolactone (CL) ratios (LC16:4, 18:2 and 9:1) were generated via ring-opening-polymerization and photoactivation. The reactivity was quantified using photo-DSC, yielding a double bond conversion ranging from 70% to 90%. The pore sizes for all LC scaffolds were see 300 μm and throat sizes varied from 152 to 177 μm. In vitro degradation was conducted at different temperatures; 37, 50 and 65°C. Change in compressive properties immersed at 37°C over time was also measured. Variations in thermal, degradation and mechanical properties of the LC scaffolds were related to the LA/CL ratio. Scaffold LC16:4 showed significantly lower glass transition temperature (T g) (4.8°C) in comparison with the LC 18:2 and 9:1 (see 32°C). Rates of mass loss for the LC16:4 scaffolds at all temperatures were significantly lower than that for LC18:2 and 9:1. The degradation activation energies for scaffold materials ranged from 82.7 to 94.9 kJ mol-1. A prediction for degradation time was applied through a correlation between long-term degradation studies at 37°C and short-term studies at elevated temperatures (50 and 65°C) using the half-life of mass loss (Time (M1/2)) parameter. However, the initial compressive moduli for LC18:2 and 9:1 scaffolds were 7 to 14 times higher than LC16:4 (see 0.27) which was suggested to be due to its higher CL content (20%). All scaffolds showed a gradual loss in their compressive strength and modulus over time as a result of progressive mass loss over time. The manufacturing process utilized and the scaffolds produced have potential for use in tissue engineering and regenerative medicine applications.

In this study three chemical agents Amino-propyl-triethoxy-silane (APS), sorbitol ended PLA oligomer (SPLA) and Hexamethylene diisocyanate (HDI) were identified to be used as coupling agents to react with the phosphate glass fibre (PGF) reinforcement and the polylactic acid (PLA) polymer matrix of the composite. Composites were prepared with short chopped strand fibres (l = 20 mm, ϕ = 20 µm) in a random arrangement within PLA matrix. Improved, initial composite flexural strength (~20 MPa) was observed for APS treated fibres, which was suggested to be due to enhanced bonding between the fibres and polymer matrix. Both APS and HDI treated fibres were suggested to be covalently linked with the PLA matrix. The hydrophobicity induced by these coupling agents (HDI, APS) helped to resist hydrolysis of the interface and thus retained their mechanical properties for an extended period of time as compared to non-treated control. Approximately 70% of initial strength and 65% of initial modulus was retained by HDI treated fibre composites in contrast to the control, where only ~50% of strength and modulus was retained after 28 days of immersion in PBS at 37 °C. All coupling agent treated and control composites demonstrated good cytocompatibility which was comparable to the tissue culture polystyrene (TCP) control, supporting the use of these materials as coupling agent’s within medical implant devices.

Eight different chemicals were investigated as potential candidate coupling agents for phosphate glass fibre reinforced polylactic acid composites. Evidence of reaction of the coupling agents with phosphate glass and their effect on surface wettability and glass degradation were studied along with their principle role of improving the interface between glass reinforcement and polymer matrix. It was found that, with an optimal amount of coupling agent on the surface of the glass/polymer, interfacial shear strength improved by a factor of 5. Evidence of covalent bonding between agent and glass was found for three of the coupling agents investigated, namely: 3-aminopropyltriethoxysilane; etidronic acid and hexamethylene diisocyanate. These three coupling agents also improved the interfacial shear strength and increased the hydrophobicity of the glass surface. It is expected that this would provide an improvement in the macroscopic properties of full-scale composites fabricated from the same materials which may also help to retain these properties for the desired length of time by retarding the breakdown of the fibre/matrix interface within these composites.

Polymers prepared from polylactic acid (PLA) have found a multitude of uses as medical devices. For a material that degrades, the main advantage is that an implant would not necessitate a second surgical event for removal. In this study, fibers produced from a quaternary phosphate-based glass (PBG) in the system 50P2O5-40CaO-5Na2O-5Fe2O3 were used to reinforce PLA polymer. The purpose of this study was to assess the effect of screw holes in a range of PBG-reinforced PLA composites with varying fiber layup and volume fraction. The flexural properties obtained showed that the strength and modulus values increased with increasing fiber volume fraction; from 96 MPa to 320 MPa for strength and between 4 GPa and 24 GPa for modulus. Furthermore, utilizing a larger number of thinner unidirectional (UD) fiber prepreg layers provided a significant increase in mechanical properties, which was attributed to enhanced wet out and thus better fiber dispersion during production. The effect of gamma sterilization via flexural tests showed no statistically significant difference between the sterilized and nonsterilized samples, with the exception of the modulus values for samples with screw holes. Degradation profiles revealed that samples with screw holes degraded faster than those without screw holes due to an increased surface area for the plates with screw holes in PBS up to 30 days. Scanning electron microscope (SEM) analysis revealed fiber pullout before and after degradation. Compared with various fiber impregnation samples, with 25% volume fraction, 8 thinner unidirectional prepreg stacked samples had the shortest fiber pull-out lengths in comparison to the other samples investigated.

Novel composite scaffolds were produced using long continuous bidirectional fibers embedded in an electrospun matrix, with the aim of using them in soft tissue engineering applications. The fibers are of polydioxanone and the matrix of polylactic acid. The novel manufacturing method consists of direct electrospinning performed on both sides of a collector that supports the already arranged fibers. The scaffolds were tested in vitro using 3T3 mouse fibroblasts as-obtained or functionalized with biotin or poly (dopamine). Functionalization did not significantly affect cells attachment, metabolic activity, or proliferation, but poly (dopamine) was proven to be effective in inducing hydrophilicity to the surface.

A range of nanocomposites were prepared using cellulose nanowhiskers (CNWs) and poly(lactic acid) (PLA) via a solvent casting process. Acid hydrolysis process was used to produce CNWs from bleached cotton. Structural morphology and surface topography of the CNWs and nanocomposites were examined using transmission (TEM) and scanning electron microscopy. TEM images revealed rod-like whiskers in the nano-scale region which were dispersed within the PLA matrix. The presence of the functional groups of CNWs and PLA were confirmed via FTIR analysis. Tensile tests were conducted on thin films and the nanocomposites containing 1 wt% CNWs showed a 34 and 31% increase in tensile strength and modulus, respectively, compared to pure PLA. The dynamic mechanical analysis showed that the tensile storage modulus also increased in the visco-elastic temperature region with increasing CNWs content in the nanocomposites. Thermogravimetric analysis showed that all the materials investigated were thermally stable from room temperature to 210 °C. A positive effect of CNWs on the crystal nucleation of PLA polymer in the nanocomposites was observed using differential scanning calorimetry and X-ray diffraction analysis. The degradation profiles of the nanocomposites in deionised water over 1 week revealed a mass loss of 1.5–5.6% at alternate temperatures (25, 37 and 50 °C) and at the same conditions the swelling ratio and water uptake were seen to increase with CNWs content in the nanocomposites, which was strongly influenced by the presence of crystalline CNWs.

We report results from an initial small animal study designed to provide information on the biocompatibility of a novel biodegradable composite designed for craniomaxillofacial reconstruction. Rat calvarium was chosen as a clinically analogous model, which allowed comparison between experimental groups (PCL alone, PCL/phosphate glass, or PCL/bioglass implants) and control groups (empty defects or bone grafted defects). All animals recovered well from surgery and no clinical complications were observed. Histological assessment indicated a lack of inflammatory response. The amount of new bone formation at the dural aspect of the implant was statistically significantly higher in the PCL/phosphate glass group than the other experimental groups. This study confirms, in a clinically analogous model, the promise of the novel PCL/phosphate glass composite material. Work is planned toward manufacturing scale up and clinical trials. (C) 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 96A: 230-238, 2011.

Two chitosan-alginate gel systems in the form of membranes were produced and evaluated. The first membrane was produced by a novel gel system formed after blending N-(methylsulfonic acid) chitosan with ammonium alginate (CAG1) and the second was an N-(methylsulfonic acid) chitosan-sodium alginate blend cross-linked with glutaraldehyde and calcium chloride (CAG2). The cytocompatibility and hemocompatibility of the gels were examined by assessing the cell viability of 3T3 Swiss mouse fibroblasts, whole blood hemolysis, and platelet activation. Cell viability was not significantly different by exposure to these gels compared to the controls. Both gel types had minimal effect on hemolysis of whole heparinized rabbit blood after 1-h exposure. Further platelet activation by the surfaces was also minimal. These results indicate that these novel gels merit further investigation for blood contact applications.

Recently, phosphate-based glass (PBG) fibers have been used to reinforce the biodegradable polymers polycaprolactone and polylactic acid, in order to fabricate materials suitable for use as resorbable bone fracture fixation devices. However, the PBG fibers investigated tended to degrade too quickly for application. Therefore, more durable PBG formulations were sought with emphasis remaining firmly placed on their biocompatibility. In this study, four invert PBG formulations (in the system P2O5—CaO—MgO—Na 2O) were produced with fixed phosphate and calcium content at 40 and 25 mol%, respectively. MgO was added at 10—30 mol% in place of Na 2O and the maximum divalent cation to phosphate ratio obtained was 1.375. Thermal analyses showed a linear increase in Tg with increasing MgO content. This was proposed to be due to an increase in the cross-link density of the glass network, which also improved the chemical durability of the glass. EDX analyses were also conducted to verify the final composition of the glass. XRD analyses confirmed the amorphous nature of the glasses investigated. Rapid quenching of the Mg30 glass revealed a degree of surface crystallization, which was shown to be a CaMgP2O7 phase. The degradation rates of the glasses investigated decreased with increasing MgO content. The decrease in rate seen was almost two orders of magnitude (a ×50 difference was seen between glass Mg0 and Mg30). The cytocompatibility studies of the formulations investigated showed good cellular response over time for up to 14 days. Statistical analysis revealed that the formulations investigated gave a response comparable to the tissue culture plastic control. It is suggested that invert PBG provide degradation profiles and the cytocompatibility response desired to make these glasses useful for bone repair applications.

Increased magnitude of biomaterial surface roughness and micromachined-grooved surfaces has both been shown to stimulate osteoblast activity, but have not been compared in the same study quantitatively. A series of titanium alloy (Ti6Al4V) samples were prepared using simple machining techniques to undertake such a comparison. Samples were either grit blasted (Gb) or shot peened (Sp) to give random discontinuities, or silicon carbide ground (SiC) to produce ordered grooves. These were compared with micropolished samples (Mp). The samples were coated with a 1 mum continuous coating of hydroxyapatite to remove differences in surface chemistry. Human osteoblast-like cells were seeded onto the materials and metabolic activity, proliferation, alkaline phosphatase activity, and osteocalcin production assessed. Cell responses were highly dependent on the substrate that they were cultured on. Cells cultured on the smooth and ordered (Mp and SiC, respectively) samples had higher metabolic activity and a more elongated morphology than those cultured on the randomly structured Gb or Sp samples. Over 21 days, cell metabolic activity peaked relative to the control between 7 and 14 days on the Mp sample, and between 14 and 21 days on the Gb, Sp, and SiC samples. In common with other researchers, we note that micron scale topography may have potential for influencing osseointegration. More importantly, as the magnitude of the discontinuities on SiC, Gb, and Sp were similar, the differences in cell responses does not appear to lie with the size of the features, but whether the features showed an ordered or disordered structure.

Any biomaterial implanted within the human body is influenced by the interactions that take place between its surface and the surrounding biological milieu. These interactions are known to influence the tissue interface dynamic, and thus act to emphasize the need to study cell-surface interactions as part of any biomaterial design process. The work described here investigates the relationship between human osteoblast attachment, spreading and focal contact formation on selected surfaces using immunostaining and digital image processing for vinculin, a key focal adhesion component. Our observations show that a relationship exists between levels of cell attachment, the degree of vinculin-associated plaque formation and biocompatibility. It also suggests that cell adhesion is not indicative of how supportive a substrate is to cell spreading, and that cell spreading does not correlate with focal contact formation.

There has been little information to date based on the potential health effects and hazards associated with the inhalation of carbon nanofibrous materials by workers despite their growing use in industry. This study examines the in vitro effects of a range of nanofibres and nanotubes for their ability to stimulate the release of the pro-inflammatory cytokine TNF-α and reactive oxygen species (ROS) from monocytic cells. Also assessed were the toxic effect of the nanomaterials on the cells and the phagocytic ability of the cells after exposure. Our studies showed that the cellular response varied with fibre morphology and state of aggregation; long, straight, well-dispersed nanofilaments produced significantly more TNF-α and ROS in monocytic cells compared with highly curved and entangled materials. We also demonstrated that monocytic cell phagocytic ability was reduced after exposure to all of the nanotubes used in this study. Microscopic examination of the cells after treatment with the nanotubes showed ‘frustrated phagocytosis’, The frustrated phagocytosis suggests that clearance of nanotubes from the lungs by macrophages may be impaired. There was no evidence of a toxic effect at any of the doses or time points used. These considerations may have important consequences for workers exposed to these nanomaterials.

A common phenomenon in tissue engineering is rapid tissue formation on the outer edge of the scaffold which restricts cell penetration and nutrient exchange to the scaffold centre, resulting in a necrotic core. To address this problem, we generated scaffolds with both random and anisotropic open porous architectures to enhance cell and subsequent tissue infiltration throughout the scaffold for applications in bone and cartilage engineering. Hydroxyapatite (HA) and poly(D,L-lactic acid) (P(DL)LA) scaffolds with random open porosity were manufactured, using modified slip-casting and by supercritical fluid processing respectively, and subsequently characterised. An array of porous aligned channels (400 microm) was incorporated into both scaffold types and cell (human osteoblast sarcoma, for HA scaffolds; ovine meniscal fibrochondrocytes, for P(DL)LA scaffolds) and tissue infiltration into these modified scaffolds was assessed in vitro (cell penetration) and in vivo (tissue infiltration; HA scaffolds only). Scaffolds were shown to have an extensive random, open porous structure with an average porosity of 85%. Enhanced cell and tissue penetration was observed both in vitro and in vivo demonstrating that scaffold design alone can influence cell and tissue infiltration into the centre of tissue engineering scaffolds.

Three series of phosphate glasses were produced by melting together sodium phosphate salt (NaH 2 PO 4 ) and the phosphate salts of either calcium (CaHPO 4 ), magnesium (MgHPO 4 · 3H 2 O) or iron (FePO 4 · 2H 2 O) in a 5% gold/95% platinum crucible at 1200 °C. The glass compositions were confirmed by EDX and XRD analysis. Glass transition temperature ( T g ), density and durability in water were determined for all the compositions. Maximum metal oxide contents before devitrification were between 55% and 59% for CaO + Na 2 O and 59% and 62% for MgO + Na 2 O. The normalized equivalent for Fe 2 O 3 + Na 2 O was between 55% and 61%. Density values for the glasses lay between 2.49 and 2.75 g cm −3 . T g s lay between 295 °C and 470 °C. Degradation rates in deionized water at 37 °C lay between 0.03 g cm −2 h −1 for Na phosphate glasses and 9 × 10 −6 g cm −2 h −1 for Ca phosphate glasses, 3 × 10 −6 g cm −2 h −1 for Mg phosphate glasses and −6 g cm −2 h −1 for Fe phosphate glasses. The effect of metal addition on properties goes as Fe > Mg > Ca for degradation rates and T g and Fe > Mg ≈ Ca for density. The change in properties with metal addition was seen to be linear for Fe and Ca additions but not with Mg addition. This is in agreement with the anomalous behavior of magnesium phosphate glasses.

There is a clinical need for synthetic scaffolds that will promote bone regeneration. Important factors include obtaining an optimal porosity and size of interconnecting windows whilst maintaining scaffold mechanical strength, enabling complete penetration of cells and nutrients throughout the scaffold, preventing the formation of necrotic tissue in the centre of the scaffold. To address this we investigated varying slip deflocculation in order to control the resulting porosity, pore size and interconnecting window size whilst maintaining mechanical strength. Hydroxyapatite (HA) porous ceramics were prepared using a modified slip casting process. Rheological measurements of the HA slips were used to identify deflocculation conditions which resulted in changes in the cell and window sizes of the resulting ceramics. Sintered ceramics were characterised by X-ray diffraction (XRD) and scanning electron microscopy (SEM). Pore and window size distribution was determined by SEM. XRD analysis confirmed that the crystal structure remained HA after the sintering process. SEM showed that HA porous ceramics presented a highly interconnected porous network with average pore sizes ranging from 391+/-39 to 495+/-25 microm. The average window size varied from 73+/-5 to 135+/-7 microm. Pore diameters obtained were controllable in the range 200-500 microm. Window sizes were in the range 30-250 microm. The use of dispersant concentration allows pore and window size to be modified whilst maintaining control over porosity demonstrated by a porosity of 85% for seven different dispersant concentrations. The advantage of this approach allows the correlation between the rheological conditions of the slip and the resultant sintered ceramic properties. In particular, optimising the ceramic strength by controlling the agglomeration during the casting process.

There is a clinical need for synthetic scaffolds that promote bone regeneration. A common problem encountered when using scaffolds in tissue engineering is the rapid formation of tissue on the outer edge of the scaffold whilst the tissue in the centre becomes necrotic. To address this, the scaffold design should improve nutrient and cell transfer to the scaffold centre. In this study, hydroxyapatite scaffolds with random, open porosity (average pore size of 282+/-11microm, average interconnecting window size of 72+/-4microm) were manufactured using a modified slip-casting methodology with a single aligned channel inserted into the centre. By varying the aligned channel diameter, a series of scaffolds with channel diameters ranging from 170 to 421microm were produced. These scaffolds were seeded with human osteosarcoma (HOS TE85) cells and cultured for 8 days. Analysis of cell penetration into the aligned channels revealed that cell coverage increased with increasing channel diameter; from 22+/-3% in the 170microm diameter channel to 38+/-6% coverage in the 421microm channel. Cell penetration into the middle section of the 421microm diameter channel (average cell area coverage 121x10(3)+/-32x10(3)microm(2)) was significantly greater than that observed within the 170microm channel (average cell area coverage 26x10(3)+/-6x10(3)microm(2)). In addition, the data presented demonstrates that the minimum channel (or pore) diameter required for cell penetration into such scaffolds is approximately 80microm. These results will direct the development of scaffolds with aligned macroarchitecture for tissue engineering bone.

The degradation profiles of six sodium iron phosphate glass formulations have been investigated using simple dissolution trials in deionized water. The glasses were produced from the appropriate phosphate salts by melting at 1200°C in 5% Au/95% Pt crucibles. Dissolution rates varied from 0.2 gcm−2h−1 for the 1% Fe glass to essentially zero over the 6-week test period for the 15% Fe and 20% Fe glasses. The overall degradation rate was found to vary according to the approximate relation: rate = 1.3e−0.79x gcm−2h−1, where x is the percentage iron content of the glass. Glasses with 10% or greater iron content were observed to maintain a constant density over the course of the tests and thus appeared to degrade from the surface and not the bulk. In vitro cell response tests were conducted on the glasses using macrophages and primary craniofacial osteoblasts. These tests were performed on the glasses with 10% or greater iron content because glasses with lower iron content degraded too quickly. Confocal microscopy revealed a rounded macrophage morphology and IL-1β production was low, suggesting little macrophage activation. However, a significant level of peroxide production was observed. Osteoblasts were observed to attach to the glass surfaces and spread, exhibiting a similar cytosketetal organization to the cells on the Thermanox controls, with a high level of F-actin organization. On balance, the 15% Fe glass performed slightly better than the 20% Fe glass in these assays. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res 71A: 283–291, 2004

Changes in the thermal properties of sodium phosphate glasses during melt production have been investigated using Pt/Au and fused alumina crucibles. Glasses were produced from NaH2PO4 as a starting material, providing an intrinsic Na2O:P2O5 ratio of 1:1 and giving an O/P = 3, that is, a metaphosphate. In Pt/Au crucibles, glass transition temperatures rose to a plateau value of 295°C at a rate determined by melt temperature. No contamination of the glass by platinum or gold was detected or indicated in the results. Ea for the reaction was found to be 66.4 kJ mol−1. In fused alumina crucibles, glass transition temperatures rose to over 450°C, with these values showing some convergence at higher furnace temperatures. Extensive erosion of the alumina crucibles was observed. The amount of alumina incorporation within the glasses correlated well with the rise in glass transition temperature up to a maximum of 15.5 mol % Al2O3 content. Al2O3 incorporation above this value caused a reduction in the value of the Tg. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 71B: 22–29, 2004

We have developed a new bone replacement material based on polycaprolactone (PCL), which can act as a suitable matrix for monomer transfer molding of degradable composites. A boron trifluoride catalyst with glycerol additive was used to produce PCL with a degradation rate that can be altered by treatment with fluoride ions. The effect of cations on the degradation of the polymer and macrophage cell responses are discussed. We found that treatment with fluoride ions reduced the degradation rate. No significant difference between these three fluorides was observed although a general trend was seen where KF-treated PCL appeared to degrade slower than NaF-treated PCL which was slower than NH4F-treated PCL. Variation in solubilities of the salts was observed where the K+ cation had the highest solubility and the Na+ cation had the lowest solubility, which suggests that NaF was able to degrade the polymer more efficiently than the other fluorides. No significant macrophage activation was observed after culture on the polymer surfaces as determined by peroxide and IL-1β release, whereas some activation occurred after culture in degradation products. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res 69A: 17–25, 2004

There is no ideal material for craniofacial bone repair at present. The aim of this study was to test the biocompatibility of polycaprolactone (PCL) synthesised by a novel method allowing control of molecular weight and degradation rate, with regard to it being used as matrix for a biodegradable composite for craniofacial bone repair. Human primary craniofacial cells were used, isolated from paediatric skull after surgery. Cell responses were analysed using various assays and antibody staining. Cells attached and spread on the PCL in a similar manner to the Thermanox controls as shown by phalloidin staining of F-actin. Cells maintained the osteoblast phenotype as demonstrated by alkaline phosphatase assay and antibody staining throughout the time points studied, up to 28 days. Cells proliferated on the PCL as shown by a DNA assay. Collagen-1 staining showed extensive production of a collagen-1 containing extracellular matrix, which was also shown to be mineralised by alizarin red staining. Short-term (up to 48 h) attachment studies and long-term (up to 28 days) expression of markers of the osteoblast phenotype have been demonstrated on the PCL. This new method of synthesising PCL shows biocompatibility characteristics that give it potential to be used for craniofacial bone repair.

Protein adsorption and adhesion of primary human osteoblasts on chemically patterned, metal-oxide-based surfaces comprising combinations of titanium, aluminium, vanadium and niobium were investigated. Single metal samples with a homogeneous surface and bimetal samples with a surface pattern produced by photolithographic techniques were used. The physical and chemical properties of the samples have been extensively characterised and are presented in a companion paper. Here, we describe their properties in terms of cell responses during the initial 24h of cell culture. Regarding the cell number and activity there was no significant difference between any of the single metal surfaces. However the morphology of cells on vanadium surfaces became spindle-like. In contrast to the behaviour on single metal samples, cells exhibited a pronounced reaction on bimetallic surfaces that contained aluminium. Cells tended to stay away from aluminium, which was the least favoured metal in all two-metal combinations. An initial cell alignment relative to the pattern geometry was detectable after 2h and was fully developed after 18h of incubation. The organisation of f-actin and microtubules as well as the localisation of vinculin were all more pronounced on non-aluminium regions. We hypothesised that the differences in cell response could be associated with differences in the adsorption of serum proteins onto the various metal oxides. Protein adsorption experiments were performed using microscopy in conjunction with immunofluorescent stains. They indicated that both fibronectin and albumin adsorption were significantly greater on the non-aluminium regions, suggesting that differences in cellular response correlate with a modulation of the concentration of serum proteins on the surface.

Physical, chemical and biological properties of calcium phosphate coatings fabricated by a pulse laser deposition method at room temperature (RT PLD) have been studied. In vitro evaluation of RT PLD coatings on bioresorbable polymers (Poly-e-caprolactone and Poly-L-lactide) have been carried out. It was shown that both polymers support osteoblast growth, with increased cell activity, alkaline phosphatase activity and total protein content on those surfaces that have been coated. The advantages of RT PLD coatings in biomaterials surface optimization are discussed.

A recently commercialised hydroxyapatite electrochemically assisted chemical deposition technique (BoneMaster) has been shown to induce increased bone apposition; whether this response is caused by the surface topography or chemistry is unknown. An in-vitro examination using human osteoblast-like cells was performed on a series of BoneMaster-coated surfaces. The chemistry was separated from the topography using a thin gold coating; Thermanox coverslips were used as a control. BoneMaster surfaces showed significantly greater alkaline phosphatase activity and osteocalcin production compared with controls; however, no difference was found between the gold-coated and uncoated BoneMaster samples, indicating topography is the main contributing factor.

Pulsed laser ablation is a new method for deposition of thin layers of hydroxyapatite (HA) on to biomaterial surfaces. In this paper, we report activity and morphology of osteoblasts grown on HA surfaces fabricated using different laser conditions. Two sets of films were deposited from dense HA targets, at three different laser fluences: 3, 6 and 9 Jcm(-2). One set of the surfaces was annealed at 575 degrees C to increase the crystallinity of the deposited films. Primary human osteoblasts were seeded onto the material surfaces and cytoskeletal actin organisation was examined using confocal laser scanning microscopy. The annealed surfaces supported greater cell attachment and more defined cytoskeletal actin organisation. Cell activity, measured using the alamar Blue assay, was also found to be significantly higher on the annealed samples. In addition, our results show distinct trends that correlate with the laser fluence used for deposition. The cell activity increases with increasing fluence. This pattern was repeated for alkaline phosphatase production by the cells. Differences in cell spreading were apparent which were correlated with the fluence used to deposit the HA. The optimum surface for initial attachment and spreading of osteoblasts was one of the HA films deposited using 9 J cm(-2) laser fluence and subsequently annealed at 575 degrees C.

Murine 3T3 fibroblasts and primary human osteoblasts have been cultured on self-assembled monolayers (SAMs) formed by the adsorption of alkanethiols of varying terminal group and alkyl chain length on gold. It has been found that the interactions of both cell types with SAMs are influenced by surprisingly subtle changes in adsorbate structure. Both the nature of the adsorbate terminal group and the length of the alkyl chain have a significant influence on cellular attachment and growth. For both cell types, attachment and growth on the carboxylic acid terminated SAMs was extensive and was accompanied by a high degree of cell spreading. In contrast, attachment and growth on long-chain methyl terminated SAMs were poor. Attachment and growth on short-chain methyl terminated SAMs were intermediate, with reasonable numbers of attached cells being observed and some degree of cell spreading. Sharply contrasting behaviour was observed for the two cell types when cultured on hydroxy terminated SAMs. It was found that fibroblasts attached poorly to these surfaces, while osteoblasts attached and grew nearly as successfully on short-chain hydroxy terminated SAMs as on carboxylic acid terminated SAMs. Osteoblasts attached less well to long-chain hydroxy terminated SAMs, but better than to methyl terminated SAMs.

The initial attachment of human bone-derived cells (HBDC) to several polymer systems has been studied in vitro. A novel polymer system based on poly(ethyl methacrylate) polymer and tetrahydrofurfuryl methacrylate monomer (PEMA/THFMA) was compared with a variant in which THFMA was replaced by 2-hydroxyethyl methacrylate (PEMA/HEMA). Tissue culture polystyrene (TCPS) and polystyrene (PS) were used as reference materials. The ability of the substrates to adsorb the attachment glycoproteins fibronectin (Fn) and vitronectin (Vn) from serum and the subsequent effect on radiolabeled HBDC attachment were examined. Initial cell attachment from the medium containing 10% (v/v) serum was highest on TCPS; on PEMA/THFMA and PEMA/HEMA substrates it was about 25% of this level, and on PS it was only 10% of that on TCPS. Attachment of HBDC to all substrates was dependent on the presence of Vn, which, unlike Fn, was able to adsorb in the face of competition from other serum components. Both Vn and Fn were able to support cell attachment when precoated onto all substrates. In comparison to TCPS, PEMA/THFMA did not show enhanced adsorption of either Fn or Vn from serum, and this was reflected in the level of cell attachment. Interestingly, the potency of preadsorbed Fn for cell attachment was much higher on this substrate than on any other: 45 ng/cm2 Fn when adsorbed to PEMA/THFMA gave a level of cell attachment 1.6-fold higher than the same density of Fn on PS or TCPS. The maximum Fn surface density achieved on HEMA/PEMA was 16 ng/cm2. Cells on PEMA/THFMA showed typical clustering of the α5 β1 Fn receptor, but this was not evident in cells attached to PEMA/HEMA even when precoated with Fn. This study indicates that the initial attachment of HBDC to all substrates was Vn dependent. It also indicates that on PEMA/THFMA the favorable presentation of subsequently adsorbed Fn may assist matrix assembly. © 1999 John Wiley & Sons, Inc. J Biomed Mater Res, 44, 1–11, 1999.

Primary human osteoblasts were cultured on self-assembled monolayers (SAMs) of alkylthiols on gold with carboxylic acid and methyl termini, and the kinetics of cell attachment and proliferation were measured. Over 90 min approximately twice as many cells attached to carboxylic-acid-terminated monolayers as attached to methyl-terminated monolayers. After 24 h the number of cells attached to carboxylic-acid-terminated monolayers was ten times that attached to the methyl-terminated monolayers. Cell morphology and cytoskeletal actin organization also were found to be different. Osteoblasts were cultured on SAMs that were patterned by photolithographic techniques. Cells attached almost exclusively to carboxylic-acid-functionalized areas of the patterned surfaces, leaving methyl-functionalized regions bare. The patterns strongly influenced the morphology of the attached cells. After 24 h, cells were observed to bridge between carboxylic-acid-terminated regions separated by 75 μm, methyl-terminated regions but not those separated by 150 μm methyl-terminated regions. After 6 days in culture osteoblasts formed multilayers on the carboxylic-acid-terminated regions of the pattern. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 41, 431–442, 1998.

A range of ‘bioartificial’ collagen/poly(vinyl alcohol) blends have been produced, cast as films and cross-linked using either glutaraldehyde or a dehydrothermal treatment (DHT). Films were used as substrates for the culture of osteoblast-like cells. The attachment, adhesion and proliferative responses of these cells to the range of films were examined using proliferation assays, light, electron and confocal microscopy. There was an inverse relationship between collagen content of gluataraldehyde cross-linked films and the extent of cell proliferation on them. A cytotoxicity assay demonstrated no toxic effect related to increasing collagen content. The greatest differences in cell responses observed were associated with the choice of cross-linking method. Films cross-linked with glutaraldehyde showed variation related to collagen content in cell adhesion, proliferation and morphology. Such differences were not apparent with the DHT cross-linked films. Collagen/PVA ‘bioartificial’ films can be dehydrothermally cross-linked to increase biological stability and reduce water solubility. The method of cross-linking employed is the greater influence in determining osteoblast compatibility with these materials. The DHT cross-linking method is a preferable alternative to the use of glutaraldehyde. Collagen/PVA bioartificial films cross-linked by the DHT method have shown potential for biocompatibility with osteoblasts.

A novel polymer system comprising poly(ethyl methacrylate) (PEMA) powder gelled with hydroxyethylmethacrylate (HEMA), n-butylmethacrylate (nBM) monomer mixtures has been produced. The monomers were combined in different ratios to vary the relative hydrophobicity of the system. Surface and bulk properties of this copolymer system were investigated. Surfaces were relatively featureless with little variation due to composition. Contact angles ranged from 76 to 83°. Equilibrium water content of the polymers was directly related to the mole fraction of HEMA content. The uptake of water in the earlier stages was proportional to t 1 2 , consistent with a diffusion process; the slope of this plot enabled diffusion coefficients to be measured. The maximum equilibrium water content was 16%. Water uptake was reduced in phosphate-buffered saline, but addition of bovine serum albumin did not affect water uptake. Desorption was also linear on a t 1 2 plot in the early stages. There was a direct relationship between water uptake and loss. The polymer system was capable of releasing albumin; the amount of albumin released was inversely related to the HEMA content of the system. The relationship of the properties of the polymer system to biological interactions and potential applications are discussed.

Fully resorbable phosphate glass fiber reinforced polymer composites have shown real potential for replacing some of the existing metallic bone fracture fixation devices. However, some of these composites have not provided suitable mechanical strength profiles over the required healing period for bone. Typically, it has been seen that these composites can lose up to 50% or more of their strength within the first week of degradation. Functionalizing the glass surface to promote polymer adhesion or to introduce hydrophobicity at the glass surface could potentially introduce control over the mechanical properties of the composite and their retention. In this study eight chemical agents namely, Glycerol 2-phosphate disodium salt; 3-phosphonopropionic acid; 3-aminopropyltriethoxy silane; etidronic acid; hexamethylene diisocyanate; sorbitol/sodium ended PLA oligomers and amino phosphonic acid, were selected to functionalise the bulk phosphate glass surface. Selected chemical agents had one functional group (-OH or O C N) to react with the glass and another functionality (either -OH, NH2, or Na) to react with the polymer matrix and/or produce hydrophobicity at the fiber surface. Bulk phosphate glass surface-treated with the above agents were assessed for the cytotoxicity of degradation products cell-material interaction in short- and long-term direct cytocompatibility studies. Results obtained from these cytocompatibility studies (using human osteosarcoma (MG63) and primary human osteoblast cell lines) revealed no cytotoxicity from the degradation products and a response comparable to controls in terms of cell functions (attachment, viability, metabolic activity, proliferation, and differentiation) and morphology.

In this study eight different phosphate-based glass compositions were prepared by melt-quenching: four in the (P2O5)45-(CaO)16-(Na2O)15-x -(MgO)24-(B2O3) x system and four in the system (P2O5)50-(CaO)16-(Na2O)10-x -(MgO)24-(B2O3) x , where x = 0,1, 5 and 10 mol%. The effect of B2O3 addition on the thermal properties, density, molar volume, dissolution rates, and cytocompatibility were studied for both glass systems. Addition of B2O3 increased the glass transition (T(g)), crystallisation (T(c)), melting (T(m)), Liquidus (T(L)) and dilatometric softening (T(d)) temperature and molar volume (V(m)). The thermal expansion coefficient (α) and density (ρ) were seen to decrease. An assessment of the thermal stability of the glasses was made in terms of their processing window (crystallisation onset, T(c,ons) minus glass transition temperature, T(g)), and an increase in the processing window was observed with increasing B2O3 content. Degradation studies of the glasses revealed that the rates decreased with increasing B2O3 content and a decrease in degradation rates was also observed as the P2O5 content reduced from 50 to 45 mol%. MG63 osteoblast-like cells cultured in direct contact with the glass samples for 14 days revealed comparative data to the positive control for the cell metabolic activity, proliferation, ALP activity, and morphology for glasses containing up to 5 mol% of B2O3.

Hydroxyapatite (HA), heat-treated hydroxyapatite (HAH), and fluorapatite (FA) coatings on titanium were loaded with human growth hormone (GH), and the subsequent release was monitored in vitro. The amount of GH released was significantly increased from the HA coating that had received a post-plasma-spraying heat treatment prior to incorporation of the growth hormone. Scanning electron microscopy was used to assess the surfaces of the ceramic coatings prior and postincubation with GH. Surface changes were observed on the HA and HAH coatings but not on the FA coatings after incubation with GH. Osteoblast-like cells were grown on the coatings and maintained in culture. Scanning electron microscopy was used to study the morphology of the cells and the interaction of the cells with the ceramic coatings. We used thymidine uptake and DNA content to determine the relative rates of cell division on the different coatings; the optimum rate of cell proliferation was observed on the HAH coating loaded with 0.1 IU/mL GH. © 1995 John Wiley & Sons, Inc.

The knowledge of mechanical properties of root cell walls is vital to understand how these properties interact with relevant genetic and physiological processes to bring about growth. Expansion of cell walls is an essential component of growth, and the regulation of cell wall expansion is one of the ways in which the mechanics of growth is controlled, managed and directed. In this study, the inherent surface mechanical properties of living Arabidopsis thaliana whole-root epidermal cells were studied at the nanoscale using the technique of atomic force microscopy (AFM). A novel methodology was successfully developed to adapt AFM to live plant roots. Force-Indentation (F-I) experiments were conducted to investigate the mechanical properties along the length of the root. F-I curves for epidermal cells of roots were also generated by varying turgor pressure. The F-I curves displayed a variety of features due to the heterogeneity of the surface. Hysteresis is observed. Application of conventional models to living biological systems such as cell walls in nanometer regimes tends to increase error margins to a large extent. Hence information from the F-I curves were used in a preliminary semiquantitative analysis to infer material properties and calculate two parameters. The work done in the loading and unloading phases (hysteresis) of the force measurements were determined separately and were expressed in terms of "Index of Plasticity" (η), which characterized the elasticity properties of roots as a viscoelastic response. Scaling approaches were used to find the ratio of hardness to reduced modulus (H/E(*)).