Development and microwave characterization of selected biocompatible materials and conducting polymers

The 20th century witnessed the extensive use of microwaves in industrial, scientific and medical fields. The major hindrance to many developments in the ISM field is the lack of knowledge about the effect of microwaves on materials used in various applications. The study of the interaction of microwaves with materials demanded the knowledge of the dielectric properties of these materials. However, the dielectric properties of many of these materials are still unknown or less studied. This thesis is an effort to shed light into the dielectric properties of some materials which are used in medical, scientific and industrial fields. Microwave phantoms are those materials used in microwave simulation applications. Effort has been taken to develop and characterize low cost, eco-friendly phantoms from Biomaterials and Bioceramics. The interaction of microwaves with living tissues paved way to the development of materials for electromagnetic shielding. Materials with good conductivity/absorption properties could be used for EMI shielding applications. Conducting polymer materials are developed and characterized in this context. The materials which are developed and analyzed in this thesis are Biomaterials, Bioceramics and Conducting polymers. The use of materials of biological origin in scientific and medical applications provides an eco-friendly pathway. The microwave characterization of the materials were done using cavity material perturbation method. Low cost and ecofriendly biomaterial films were developed from Arrowroot and Chitosan. The developed films could be used in applications such as microwave phantom material, capsule material in pharmaceutical applications, trans-dermal patch material and eco-friendly Band-Aids. Bioceramics with better bioresorption and biocompatibility were synthesized. Bioceramics such as Hydroxyapatite, Beta tricalcium phosphate and Biphasic Calcium Phosphate were studied. The prepared bioceramics could be used as phantom material representing Collagen, Bone marrow, Human abdominal wall fat and Human chest fat. Conducting polymers- based on Polyaniline, are developed and characterized. The developed materials can be used in electromagnetic shielding applications such as in anechoic chambers, transmission cables etc

Biological response of pulps submitted to different capping materials

Pulp capping is a procedure that comprises adequate protection of the pulp tissue exposed to the oral environment, aiming at the preservation of its vitality and functions. This study evaluated the response of the dental pulps of dog teeth to capping with mineral trioxide aggregate (MTA) or calcium hydroxide P.A. For that purpose, 37 teeth were divided into two groups, according to the capping material employed. Two dogs were anesthetized and, after placement of a rubber dam, their pulps were exposed in a standardized manner and protected with the experimental capping materials. The cavities were then sealed with resin-modified glass ionomer cement and restored with composite resin. After sixty days, the animals were killed and the specimens were processed in order to be analyzed with optic microscopy. It was observed that MTA presented a higher success rate compared to calcium hydroxide, presenting a lower occurrence of infection and pulp necrosis.

Biocompatibility of resin-based dental materials applied as liners in deep cavities prepared in human teeth

Background: Since only a few data have been published concerning the effects of resinous dental materials on the pulp-dentin complex, the aim of this study was to evaluate the biocompatibility of resin-based materials applied as liners in deep cavities prepared in duman teeth. Methods: After preparing class V cavities, the following dental materials were applied on the axial walls: group 1, Vitrebond™ (VIT; 3M ESPE); group 2, Ultra-Blend® Plus™ (UBP; Untradent); and group 3, Clearfil™ SE Bond (CSEB; Kuraray). In group 4 (control), the hard-setting calcium hydroxide cement Dycal (CH; Caulk/Dentsply) was used. The teeth extracted at 7 days or between 30 and 85 days after the clinical procedures were processed for histological evaluation. Results: For all the experimental and control groups, most of specimens exhibited no pulpal response or slight inflammatory reaction associated with slight tissue disorganization at 7-day period. Moderate inflammatory pulpal response occurred only in one tooth (RDT = 262 μm) of group 3 in which transdentinal diffusion of resin components was observed. Conclusion: The resin-based dental cements VIT and UBP as well as the bonding agent CSEB presented acceptable biocompatibility when applied in deep cavities prepared in sound human teeth. © 2006 Wiley Periodicals, Inc.

Ureasil-polyether hybrid film-forming materials

The objectives of this work were to study the suitability and highlight the advantages of the use of cross-linked ureasil-polyether hybrid matrices as film-forming systems. The results revealed that ureasil-polyethers are excellent film-forming systems due to specific properties, such as their biocompatibility, their cosmetic attractiveness for being able to form thin and transparent films, their short drying time to form films and their excellent bioadhesion compared to the commercial products known as strong adhesives. Rheological measurements have demonstrated the ability of these hybrid matrices to form a film in only a few seconds and Water Vapor Transmitting Rate (WVTR) showed adequate semi-occlusive properties suggesting that these films could be used as skin and wound protectors. Both the high skin bioadhesion and non-cytotoxic character seems to be improved by the presence of multiple amine groups in the hybrid molecules. © 2012 Elsevier B.V.

Cytotoxicity testing of burn wound dressings, ointments and creams : a method using polycarbonate cell culture inserts on a cell culture system

We have developed a method to test the cytotoxicity of wound dressings, ointments, creams and gels used in our Burn Centre, by placing them on a permeable Nunc Polycarbonate cell culture insert, incubated with a monolayer of cells (HaCaTs and primary human keratinocytes). METHODS: We performed two different methods to determine the relative toxicity to cells. (1) Photo visualisation: The dressings or compounds were positioned on the insert's membrane which was placed onto the monolayer tissue culture plate. After 24 h the surviving adherent cells were stained with Toluidine Blue and photos of the plates were taken. The acellular area of non-adherent dead cells which had been washed off with buffer was measured as a percentage of the total area of the plate. (2) Cell count of surviving cells: After 24 h incubation with the test material, the remaining cells were detached with trypsin, spun down and counted in a Haemocytometer with Trypan Blue, which differentiates between live and dead cells. RESULTS: Seventeen products were tested. The least cytotoxic products were Melolite, White soft Paraffin and Chlorsig1% Ointment. Some cytotoxicity was shown with Jelonet, Mepitel((R)), PolyMem((R)), DuoDerm((R)) and Xeroform. The most cytotoxic products included those which contained silver or Chlorhexidine and Paraffin Cream a moisturizer which contains the preservative Chlorocresol. CONCLUSION: This in vitro cell culture insert method allows testing of agents without direct cell contact. It is easy and quick to perform, and should help the clinician to determine the relative cytotoxicity of various dressings and the optimal dressing for each individual wound.

Computer simulation of scaffold degradation

Scaffolds are porous biocompatible materials with suitable microarchitectures that are designed to allow for cell adhesion, growth and proliferation. They are used in combination with cells in regenerative medicine to promote tissue regeneration by means of a controlled deposition of natural extracellular matrix by the hosted cells therein. This healing process is in many cases accompanied by scaffold degradation up to its total disappearance when the scaffold is made of a biodegradable material. This work presents a computational model that simulates the degradation of scaffolds. The model works with three-dimensional microstructures, which have been previously discretised into small cubic homogeneous elements, called voxels. The model simulates the evolution of the degradation of the scaffold using a Monte Carlo algorithm, which takes into account the curvature of the surface of the fibres. The simulation results obtained in this study are in good agreement with empirical degradation measurements performed by mass loss on scaffolds after exposure to an etching alkaline solution.

Single-cell force spectroscopy, an emerging tool to quantify cell adhesion to biomaterials

Cell adhesion receptors play a central role in sensing and integrating signals provided by the cellular environment. Thus, understanding adhesive interactions at the cell-biomaterial interface is essential to improve the design of implants that should emulate certain characteristics of the cell's natural environment. Numerous cell adhesion assays have been developed; among these, atomic force microscopy-based single-cell force spectroscopy (AFM-SCFS) provides a versatile tool to quantify cell adhesion at physiological conditions. Here we discuss how AFM-SCFS can be used to quantify the adhesion of living cells to biomaterials and give examples of using AFM-SCFS in tissue engineering and regenerative medicine. We anticipate that in the near future, AFM-SCFS will be established in the biomaterial field as an important technique to quantify cell-biomaterial interactions and thereby will contribute to the optimization of implants, scaffolds, and medical devices.

Biomimicking natural nanopatterned topology by 3D laser lithography

Natural nanopatterned surfaces (nNPS) present on insect wings have demonstrated bactericidal activity [1, 2]. Fabricated nanopatterned surfaces (fNPS) derived by characterization of these wings have also shown superior bactericidal activity [2]. However bactericidal NPS topologies vary in both geometry and chemical characteristics of the individual features in different insects and fabricated surfaces, rendering it difficult to ascertain the optimum geometrical parameters underling bactericidal activity. This situation calls for the adaptation of new and emerging techniques, which are capable of fabricating and characterising comparable structures to nNPS from biocompatible materials. In this research, CAD drawn nNPS representing an area of 10 μm x10 μm was fabricated on a fused silica glass by Nanoscribe photonic professional GT 3D laser lithography system using two photon polymerization lithography. The glass was cleaned with acetone and isopropyl alcohol thrice and a drop of IP-DIP photoresist from Nanoscribe GmbH was cast onto the glass slide prior to patterning. Photosensitive IP-DIP resist was polymerized with high precision to make the surface nanopatterns using a 780 nm wavelength laser. Both moving-beam fixedsample (MBFS) and fixed-beam moving-sample (FBMS) fabrication approaches were tested during the fabrication process to determine the best approach for the precise fabrication of the required nanotopological pattern. Laser power was also optimized to fabricate the required fNPS, where this was changed from 3mW to 10mW to determine the optimum laser power for the polymerization of the photoresist for fabricating FNPS...

Synthesis and characterization of novel poly(ester carbonate)s based on pentaerythritol

Novel poly(ester carbonate)s were synthesized by the ring-opening polymerization Of L-lactide and functionalized carbonate monomer 9-phenyl-2,4,8,10-tetraoxaspiro[5,5]undecan-3-one derived from pentaerythritol with diethyl zinc as an initiator. H-1 NMR analysis revealed that the carbonate content in the copolymer was almost equal to that in the feed. DSC results indicated that T-g of the copolymer increased with increasing carbonate content in the copolymer. Moreover, the protecting benzylidene groups in the copolymer poly(L-lactide-co-9-phenyl-2,4,8,10-tetraoxaspiro[5,5]undecan-3-one) were removed by hydrogenation with palladium hydroxide on activated charcoal as a catalyst to give a functional copolymer, poly(L-lactide-co-2,2-dihydroxylmethyl-propylene carbonate), containing pendant primary hydroxyl groups. Complete deprotection was confirmed by H-1 NMR and FTIR spectroscopy. The in vitro degradation rate of the deprotected copolymers was faster than that of the protected copolymers in the presence of proteinase K. The cell morphology and viability on a copolymer film evaluated with ECV-304 cells showed that poly(ester carbonate)s derived from pentaerythritol are good biocompatible materials suitable for biomedical applications.

Beyond simple imaging with energetic beams – nanopatterning, correlative microscopy and statistical analysis of inclusions

Electron microscopy (EM) has advanced in an exponential way since the first transmission electron microscope (TEM) was built in the 1930’s. The urge to ‘see’ things is an essential part of human nature (talk of ‘seeing is believing’) and apart from scanning tunnel microscopes which give information about the surface, EM is the only imaging technology capable of really visualising atomic structures in depth down to single atoms. With the development of nanotechnology the demand to image and analyse small things has become even greater and electron microscopes have found their way from highly delicate and sophisticated research grade instruments to key-turn and even bench-top instruments for everyday use in every materials research lab on the planet. The semiconductor industry is as dependent on the use of EM as life sciences and pharmaceutical industry. With this generalisation of use for imaging, the need to deploy advanced uses of EM has become more and more apparent. The combination of several coinciding beams (electron, ion and even light) to create DualBeam or TripleBeam instruments for instance enhances the usefulness from pure imaging to manipulating on the nanoscale. And when it comes to the analytic power of EM with the many ways the highly energetic electrons and ions interact with the matter in the specimen there is a plethora of niches which evolved during the last two decades, specialising in every kind of analysis that can be thought of and combined with EM. In the course of this study the emphasis was placed on the application of these advanced analytical EM techniques in the context of multiscale and multimodal microscopy – multiscale meaning across length scales from micrometres or larger to nanometres, multimodal meaning numerous techniques applied to the same sample volume in a correlative manner. In order to demonstrate the breadth and potential of the multiscale and multimodal concept an integration of it was attempted in two areas: I) Biocompatible materials using polycrystalline stainless steel and II) Semiconductors using thin multiferroic films. I) The motivation to use stainless steel (316L medical grade) comes from the potential modulation of endothelial cell growth which can have a big impact on the improvement of cardio-vascular stents – which are mainly made of 316L – through nano-texturing of the stent surface by focused ion beam (FIB) lithography. Patterning with FIB has never been reported before in connection with stents and cell growth and in order to gain a better understanding of the beam-substrate interaction during patterning a correlative microscopy approach was used to illuminate the patterning process from many possible angles. Electron backscattering diffraction (EBSD) was used to analyse the crystallographic structure, FIB was used for the patterning and simultaneously visualising the crystal structure as part of the monitoring process, scanning electron microscopy (SEM) and atomic force microscopy (AFM) were employed to analyse the topography and the final step being 3D visualisation through serial FIB/SEM sectioning. II) The motivation for the use of thin multiferroic films stems from the ever-growing demand for increased data storage at lesser and lesser energy consumption. The Aurivillius phase material used in this study has a high potential in this area. Yet it is necessary to show clearly that the film is really multiferroic and no second phase inclusions are present even at very low concentrations – ~0.1vol% could already be problematic. Thus, in this study a technique was developed to analyse ultra-low density inclusions in thin multiferroic films down to concentrations of 0.01%. The goal achieved was a complete structural and compositional analysis of the films which required identification of second phase inclusions (through elemental analysis EDX(Energy Dispersive X-ray)), localise them (employing 72 hour EDX mapping in the SEM), isolate them for the TEM (using FIB) and give an upper confidence limit of 99.5% to the influence of the inclusions on the magnetic behaviour of the main phase (statistical analysis).

Functional properties of cell-seeded three-dimensionally woven poly(epsilon-caprolactone) scaffolds for cartilage tissue engineering.

Articular cartilage possesses complex mechanical properties that provide healthy joints the ability to bear repeated loads and maintain smooth articulating surfaces over an entire lifetime. In this study, we utilized a fiber-reinforced composite scaffold designed to mimic the anisotropic, nonlinear, and viscoelastic biomechanical characteristics of native cartilage as the basis for developing functional tissue-engineered constructs. Three-dimensionally woven poly(epsilon-caprolactone) (PCL) scaffolds were encapsulated with a fibrin hydrogel, seeded with human adipose-derived stem cells, and cultured for 28 days in chondrogenic culture conditions. Biomechanical testing showed that PCL-based constructs exhibited baseline compressive and shear properties similar to those of native cartilage and maintained these properties throughout the culture period, while supporting the synthesis of a collagen-rich extracellular matrix. Further, constructs displayed an equilibrium coefficient of friction similar to that of native articular cartilage (mu(eq) approximately 0.1-0.3) over the prescribed culture period. Our findings show that three-dimensionally woven PCL-fibrin composite scaffolds can be produced with cartilage-like mechanical properties, and that these engineered properties can be maintained in culture while seeded stem cells regenerate a new, functional tissue construct.

Injectable laminin-functionalized hydrogel for nucleus pulposus regeneration.

Cell delivery to the pathological intervertebral disc (IVD) has significant therapeutic potential for enhancing IVD regeneration. The development of injectable biomaterials that retain delivered cells, promote cell survival, and maintain or promote an NP cell phenotype in vivo remains a significant challenge. Previous studies have demonstrated NP cell - laminin interactions in the nucleus pulposus (NP) region of the IVD that promote cell attachment and biosynthesis. These findings suggest that incorporating laminin ligands into carriers for cell delivery may be beneficial for promoting NP cell survival and phenotype. Here, an injectable, laminin-111 functionalized poly(ethylene glycol) (PEG-LM111) hydrogel was developed as a biomaterial carrier for cell delivery to the IVD. We evaluated the mechanical properties of the PEG-LM111 hydrogel, and its ability to retain delivered cells in the IVD space. Gelation occurred in approximately 20 min without an initiator, with dynamic shear moduli in the range of 0.9-1.4 kPa. Primary NP cell retention in cultured IVD explants was significantly higher over 14 days when cells were delivered within a PEG-LM111 carrier, as compared to cells in liquid suspension. Together, these results suggest this injectable laminin-functionalized biomaterial may be an easy to use carrier for delivering cells to the IVD.

Experimental and numerical study of the cold crucible melting process

The cold crucible, or induction skull melting process as is otherwise known, has the potential to produce high purity melts of a range of difficult to melt materials, including Ti–Al and Ti6Al4V alloys for Aerospace, Ti–Ta and other biocompatible materials for surgical implants, silicon for photovoltaic and electronic applications, etc. A water cooled AC coil surrounds the crucible causing induction currents to melt the alloy and partially suspend it against gravity away from water-cooled surfaces. Strong stirring takes place in the melt due to the induced electromagnetic Lorentz forces and very high temperatures are attainable under the right conditions (i.e., provided contact with water cooled walls is minimised). In a joint numerical and experimental research programme, various aspects of the design and operation of this process are investigated to increase our understanding of the physical mechanisms involved and to maximise process efficiency. A combination of FV and Spectral CFD techniques are used at Greenwich to tackle this problem numerically, with the experimental work taking place at Birmingham University. Results of this study, presented here, highlight the influence of turbulence and free surface behaviour on attained superheat and also discuss coil design variations and dual frequency options that may lead to winning crucible designs.

Experimental and numerical study of the cold crucible melting process

The cold crucible, or induction skull melting process as is otherwise known, has the potential to produce high purity melts of a range of difficult to melt materials, including Ti–Al and Ti6Al4V alloys for Aerospace, Ti–Ta and other biocompatible materials for surgical implants, silicon for photovoltaic and electronic applications, etc. A water cooled AC coil surrounds the crucible causing induction currents to melt the alloy and partially suspend it against gravity away from water-cooled surfaces. Strong stirring takes place in the melt due to the induced electromagnetic Lorentz forces and very high temperatures are attainable under the right conditions (i.e., provided contact with water cooled walls is minimised). In a joint numerical and experimental research programme, various aspects of the design and operation of this process are investigated to increase our understanding of the physical mechanisms involved and to maximise process efficiency. A combination of FV and Spectral CFD techniques are used at Greenwich to tackle this problem numerically, with the experimental work taking place at Birmingham University. Results of this study, presented here, highlight the influence of turbulence and free surface behaviour on attained superheat and also discuss coil design variations and dual frequency options that may lead to winning crucible designs.

Integrated biomedical device for blood preparation

The separation of red blood cells from plasma flowing in microchannels is possible by bio-physical effects such as an axial migration effect and Zweifach-Fung bifurcation law. In the present study, subchannels are placed alongside a main channel to collect cells and plasma separately. The addition of a constriction in the main microchannel creates a local high shear force region, forcing the cells to migrate and concentrate towards the centre of the channel. The resulting lab-on-a-chip was manufactured using biocompatible materials. Purity efficiency was measured for mussel and human blood suspensions as different parameters including flow rate and geometries of parent and daughter channels were varied.