Jonny J. Blaker

In vitro attachment of staphylococcus epidermidis to surgical sutures with and without Ag-containing bioactive glass coating

The ability of a silver-doped bioactive glass (AgBG) coating to prevent bacterial colonization on surgical sutures was investigated in vitro. Bioactive glass powders, in the form of 45S5 Bioglass® and AgBG, were used to coat Mersilk® sutures using an optimized ‘in house’ slurry-dipping process. In vitro experiments were carried out using Staphylococcus epidermidis under both batch and flow conditions. While the traditional batch culture testing was used to determine the number of viable cells adhered to the surface, the flow-cell was used to visualize attachment and detachment over time. Under batch conditions of up to 180 min, statistically significant differences were observed in the colony forming units (CFU) per suture for both the coated and uncoated Mersilk® sutures. The results showed that the AgBG coating had the greatest effect on limiting bacterial attachment (8 102 CFU) when compared to the 45S5 Bioglass® coating (3.2 103 CFU) and the uncoated Mersilk® (1.2 104 CFU). Also under flow conditions differences were seen between the coated and uncoated sutures. Therefore, this preliminary study has demonstrated the quantification and visualization of bacterial attachment onto sutures in order to compare the antibacterial properties of Ag-containing bioactive glass coatings. The bactericidal properties imparted by Ag-containing glass open new opportunities for use of the composite sutures in wound healing and body wall repair.

Phase behavior of medium and high internal phase water-in-oil emulsions stabilized solely by hydrophobized bacterial cellulose nanofibrils.

Water-in-oil emulsions stabilized solely by bacterial cellulose nanofibers (BCNs), which were hydrophobized by esterification with organic acids of various chain lengths (acetic acid, C2-; hexanoic acid, C6-; dodecanoic acid, C12-), were produced and characterized. When using freeze-dried C6-BCN and C12-BCN, only a maximum water volume fraction (ϕw) of 60% could be stabilized, while no emulsion was obtained for C2-BCN. However, the maximum ϕw increased to 71%, 81%, and 77% for C2-BCN, C6-BCN, and C12-BCN, respectively, 150 h after the initial emulsification, thereby creating high internal phase water-in-toluene emulsions. The observed time-dependent behavior of these emulsions is consistent with the disentanglement and dispersion of freeze-dried modified BCN bundles into individual nanofibers with time. These emulsions exhibited catastrophic phase separation when ϕw was increased, as opposed to catastrophic phase inversion observed for other Pickering emulsions.

Thermal Characterizations of Silver-containing Bioactive Glass-coated Sutures:

This study utilized and compared a number of thermal analysis methods to characterize the thermal properties of commercial sutures with and without antimicrobial coatings of silver-doped bioactive glass (AgBG) interlocking particulates. The effect of a slurry dipping technique used to coat resorbable Vicryl® (polyglactin 910) and non-resorbable Mersilk® surgical sutures with AgBG was investigated using conventional differential scanning calorimetry (DSC), high speed calorimetry (or HYPERDSC™), and modulated temperature DSC (MTDSC). These methods were compared in terms of their ability to resolve the thermal transitions of the types of suture materials. Differential thermal analysis (DTA) and thermogravimetric analysis (TGA) were used to verify the thermal degradation temperatures of these materials and to quantify the AgBG coatings on the sutures. The use of complementary thermal analysis techniques enabled the understanding of the effect of the AgBG coating technique on the morphological properties of the sutures. The slurry dipping technique had no significant effect on the thermal transitions of both types of materials. The use of high speed calorimetry through DSC offered better resolution for the transitions that appeared to be weak through conventional heating regimes, and was able to separate broad double transitions. Furthermore, it was shown not to compromise either the melting temperature or the enthalpy of melting. Therefore this method allows for the accurate determination of thermal transitions through much shorter experimental times thus allowing for an increased sample throughput. The combined DTA and TGA indicated that a greater AgBG coating was obtained in the case of the Mersilk® sutures.

Enhancing the Hydrophilicity and Cell Attachment of 3D Printed PCL/Graphene Scaffolds for Bone Tissue Engineering

Scaffolds are physical substrates for cell attachment, proliferation, and differentiation, ultimately leading to the regeneration of tissues. They must be designed according to specific biomechanical requirements, i.e., certain standards in terms of mechanical properties, surface characteristics, porosity, degradability, and biocompatibility. The optimal design of a scaffold for a specific tissue strongly depends on both materials and manufacturing processes, as well as surface treatment. Polymeric scaffolds reinforced with electro-active particles could play a key role in tissue engineering by modulating cell proliferation and differentiation. This paper investigates the use of an extrusion-based additive manufacturing system to produce poly(ε-caprolactone) (PCL)/pristine graphene scaffolds for bone tissue applications and the influence of chemical surface modification on their biological behaviour. Scaffolds with the same architecture but different concentrations of pristine graphene were evaluated from surface property and biological points of view. Results show that the addition of pristine graphene had a positive impact on cell viability and proliferation, and that surface modification leads to improved cell response.

Nacre-nanomimetics: Strong, Stiff, and Plastic

The bricks and mortar in the classic structure of nacre have characteristic geometry, aspect ratios and relative proportions; these key parameters can be retained while scaling down the absolute length scale by more than 1 order of magnitude. The results shed light on fundamental scaling behavior and provide new opportunities for high performance, yet ductile, lightweight nanocomposites. Reproducing the toughening mechanisms of nacre at smaller length scales allows a greater volume of interface per unit volume while simultaneously increasing the intrinsic properties of the inorganic constituents. Layer-by-layer (LbL) assembly of poly(sodium 4-styrenesulfonate) (PSS) polyelectrolyte and well-defined [Mg2Al(OH)6]CO3.nH2O layered double hydroxide (LDH) platelets produces a dense, oriented, high inorganic content (∼90 wt %) nanostructure resembling natural nacre, but at a shorter length scale. The smaller building blocks enable the (self-) assembly of a higher quality nanostructure than conventional mimics, leading to improved mechanical properties, matching those of natural nacre, while allowing for substantial plastic deformation. Both strain hardening and crack deflection mechanisms were observed in situ by scanning electron microscopy (SEM) during nanoindentation. The best properties emerge from an ordered nanostructure, generated using regular platelets, with narrow size dispersion.

Characterisation of 'wet' polymer surfaces for tissue engineering applications: Are flat surfaces a suitable model for complex structures?

Abstract Tissue engineering scaffolds are 3D constructs that simulate the growth environment in vivo. The present work aims to address the question of whether thin films, i.e., flat surfaces, are a suitable model for more complex 3D structures? With this in mind a complete study of the morphology and surface chemistry of poly(D,Llactide) (PDLLA) substrates, fabricated into two different structures, is presented. The polymer structures studied include a 3D, porous, foam-like scaffold prepared by the thermally induced phase separation (TIPS) method and flat polymer thin films made by solvent casting. Based on the maximum bubble point test, a new method to assess the wettability of wet pore wall surfaces inside highly porous 3D structures was developed and tested. The maximum pore diameter determined using the maximum bubble point test for the total wetting liquids was confirmed through image analysis of scanning electron micrographs. The method allows the determination of the contact angle between the wet pore wall and a contacting liquid. The captive bubble method was employed to characterise the wettability of flat polymer films in contact with water. Both structures were further characterised using zeta- (ζ-) potential measurements to assess the surface chemistry of the polymer. The results demonstrate that PDLLA contains acidic functional groups and is hydrophobic. In order to evaluate the sensitivity of the test methods, the polymer surfaces were modified by protein adsorption using fibronectin and collagen. ζ-Potential and wettability measurements show that proteins indeed adsorb on virgin PDLLA surfaces. Protein adsorption causes the wettability of the PDLLA for water to improve. Our results strongly indicate that flat surfaces are not a suitable model for surfaces in complex 3D structures such as highly porous tissue engineering scaffolds. Such scaffolds must be characterised as a 3D system.

Morphological, mechanical and biological assessment of PCL/pristine graphene scaffolds for bone regeneration

Scaffolds are physical substrates for cell attachment, proliferation, and differentiation, ultimately leading to the regeneration of tissues. They must be designed according to specific biomechanical requirements such as mechanical properties, surface characteristics, biodegradability, biocompatibility, and porosity. The optimal design of a scaffold for a specific tissue strongly depends on both materials and manufacturing processes. Polymeric scaffolds reinforced with electroactive particles could play a key role in tissue engineering by modulating cell proliferation and differentia-tion. This paper investigates the use of an extrusion additive manufacturing system to produce PCL/pristine graphene scaffolds for bone tissue applications. PCL/pristine graphene blends were prepared using a melt blending process. Scaffolds with regular and reproducible architecture were produced with different concentrations of pristine graphene. Scaffolds were evaluated from morphological, mechanical, and biological view. The results suggest that the addition of pristine graphene improves the mechanical performance of the scaffolds, reduces the hydrophobicity, and improves cell viability and proliferation.

Hybrid sol–gel inorganic/gelatin porous fibres via solution blow spinning

Hybrid sol–gel inorganic–organic fibres offer great potential in tissue engineering and regenerative medicine. A significant challenge is to process them using scalable technologies into useful scaffolds that provide control over fibre diameter, morphology, mechanical properties, ion release, degradation and cell response. In this work we develop formulations that are amenable to processing via solution blow spinning (SBS), a rapid technique using simple equipment to spray nano-/micro-fibres without any electric fields. The technique is extended to produce porous class I and II hybrid fibres using cryogenic SBS, with formulations developed based on tetraethyl orthosilicate/gelatin that are relatively facile to lyophilise. The formulations developed here take advantage of the reversible thermally activated conformation change of gelatin in aqueous solutions from random coil to triple helix to enable viscosity tuning and therefore fibre spinning. Gelatin is functionalised with (3-glycidyloxypropyl)trimethoxysilane to produce class II hybrids which exhibit controllable time- and temperature-dependent viscosity profiles which can be tuned for spinning into highly porous fibres.

Polymer-Ceramic Composite Scaffolds: The Effect of Hydroxyapatite and β-tri-Calcium Phosphate

The design of bioactive scaffolds with improved mechanical and biological properties is an important topic of research. This paper investigates the use of polymer-ceramic composite scaffolds for bone tissue engineering. Different ceramic materials (hydroxyapatite (HA) and β-tri-calcium phosphate (TCP)) were mixed with poly-ε-caprolactone (PCL). Scaffolds with different material compositions were produced using an extrusion-based additive manufacturing system. The produced scaffolds were physically and chemically assessed, considering mechanical, wettability, scanning electron microscopy and thermal gravimetric tests. Cell viability, attachment and proliferation tests were performed using human adipose derived stem cells (hADSCs). Results show that scaffolds containing HA present better biological properties and TCP scaffolds present improved mechanical properties. It was also possible to observe that the addition of ceramic particles had no effect on the wettability of the scaffolds.

A comparative study of the effects of different bioactive fillers in PLGA matrix composites and their suitability as bone substitute materials: A thermo-mechanical and in vitro investigation

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.