Cells, surfaces and adhesion

This thesis concerns cell adhesion to polymer surfaces with an experimental emphasis on hydrogels. The thesis begins with a review of the literature and a synthesis of recent evidence to describe the process of cell adhesion in a given situation. The importance of understanding integrin-adhesion protein interactions and adhesion protein-surface interactions is emphasised. The experimental chapters describe three areas of investigation. Firstly, in vitro cell culture techniques are used to explore a variety of surfaces including polyethylene glycol methacrylate (PEGMA) substituted hydrogels, sequence distribution modified hydrogels and worn contact lenses. Cell adhesion to PEGMA substituted gels is found to decrease with increases in polyethylene oxide chain length and correlations are made between sequence distribution and adhesion. Worn contact lenses are investigated for their cell adhesion properties in the presence of antibodies to specific adhesion proteins, demonstrating the presence of vitronectin and fibronectin on the lenses. The second experimental chapter addresses divalent cation regulation of integrin mediated cell adhesion. Several cell types and various cations are used. Zinc, previously not regarded as an important cation in the process, is found to inhibit 3T3 cell adhesion to vitronectin that is promoted by other divalent cations. The final experimental chapter concerns cell adhesion and growth on macroporous hydrogels. A variety of freeze-thaw formed porous gels are investiated and found generally to promote cell growth rate.Interpenetrating networkbased gels (IPN) are made porous by elution of dextrin particles of varying size and loading density. These materials provide the basis for synthetic cartilage. Cartilage cells (chondrocytes) plated onto the surface of the porous IPN materials maintain a rounded shape and hence phenotypic function when a critical pore size and density is achieved. In this way, a prospective implant, made porous at the perpendicular edges contacting natural cartilage can be both mechanically stabilised and encourage the maintenance of normal matrix production at the tissue interface.

Fluctuations and correlations in sandpiles and interfaces with boundary pinning

Interfaces are studied in an inhomogeneous critical state where boundary pinning is compensated with a ramped force. Sandpiles driven off the self-organized critical point provide an example of this ensemble in the Edwards-Wilkinson (EW) model of kinetic roughening. A crossover from quenched to thermal noise violates spatial and temporal translational invariances. The bulk temporal correlation functions have the effective exponents β1D∼0.88±0.03 and β2D∼0.52±0.05, while at the boundaries βb,1D/2D∼0.47±0.05. The bulk β1D is shown to be reproduced in a randomly kicked thermal EW model.

Fluctuations and correlations in sandpiles and interfaces with boundary pinning

Interfaces are studied in an inhomogeneous critical state where boundary pinning is compensated with a ramped force. Sandpiles driven off the self-organized critical point provide an example of this ensemble in the Edwards-Wilkinson (EW) model of kinetic roughening. A crossover from quenched to thermal noise violates spatial and temporal translational invariances. The bulk temporal correlation functions have the effective exponents β1D∼0.88±0.03 and β2D∼0.52±0.05, while at the boundaries βb,1D/2D∼0.47±0.05. The bulk β1D is shown to be reproduced in a randomly kicked thermal EW model.

Nanoscratch and nanowear testing of TiN coatings on M42 steel

Constant load, progressive load and multipass nanoscratch (nanowear) tests were carried out on 500 and 1500 nm TiN coatings on M42 steel chosen as model systems. The influences of film thickness, coating roughness, scratch direction relative to the grinding grooves on the critical load in the progressive load test and number of cycles to failure in the wear test have been determined. Progress towards the development of a suitable methodology for determining the scratch hardness from nanoscratch tests is discussed. © 2011 W. S. Maney & Son Ltd.

Humans assume a mixture of diffuse and point-source lighting when viewing sinusoidal shading patterns

Observers perceive sinusoidal shading patterns as being due to sinusoidally corrugated surfaces, and perceive surface peaks to be offset from luminance maxima by between zero and 1/4 wavelength. This offset varies with grating orientation. Physically, the shading profile of a sinusoidal surface will be approximately sinusoidal, with the same spatial frequency as the surface, only when: (A) it is lit suitably obliquely by a point source, or (B) the light source is diffuse and hemispherical--the 'dark is deep' rule applies. For A, surface peaks will be offset by 1/4 wavelength from the luminance maxima; for B, this offset will be zero. As the sum of two same-frequency sinusoids with different phases is a sinusoid of intermediate phase, our results suggest that observers assume a mixture of two light sources whose relative strength varies with grating orientation. The perceived surface offsets imply that gratings close to horizontal are taken to be lit by a point source; those close to vertical by a diffuse source. [Supported by EPSRC grants to AJS and MAG].

The role of oxidation in the protection of sliding metal surfaces

This study is concerned with the mechanisms of growth and wear of protective oxide films formed under various tribological conditions. In the study three different tribological systems are examined in each of which oxidational wear is the dominant equilibrium mode. These are an unlubricated steel on steel system sliding at low and elevated temperatures, a boundary lubricated aluminium bronze on steel system and an unlubricated reciprocating sliding 9% Cr steel system operated at elevated temperature, in an atmosphere of carbon dioxide. The results of mechanical measurements of wear and friction are presented for a range of conditions of load, speed and temper.ature for the systems, together with the results of extensive examinations of the surfaces and sub­ surfaces by various physical methods of analysis. The major part of the thesis, however, is devoted to the development and application of surface models and theoretical quantative expressions in order to explain the observed oxidational wear phenomena. In this work, the mechanisms of formation of load bearing ox ide plateaux are described and are found to be dependent on system geometry and environment. The relative importance of ''in contact" and "out of contact" oxidation is identified together with growth rate constants appropriate to the two situations. Hypotheses are presented to explain the mechanisms of removal of plateaux to form wear debris. The latter hypotheses include the effects of cyclic stressing and dislocation accumulation, together with effects associated with the kinetics of growth and physical properties of the various oxides. The proposed surf ace mode1s have led to the develop­ ment of quantitative expressions for contact temperature, unlubricated wear rates, boundary lubricated wear rates and the wear of rna ter ial during the transition from severe to mild wear. In general theoretical predictions from these expressions are in very good agreement with experimental values.

The reponse of lichen growth to transplantation to rock surfaces of different aspect

Thalli of four saxicolous lichens on slate rock fragments were transplanted from rock surafces to horizontal boards and then to south-east-facing and north-west-facing rock surfaces. The radial growth rate of Physcia orbicularis and Parmelia conspersa thalli declined after transplatation to north-west-facing rock surfaces and was unchanged after transplantation to south-east-facing rock surfaces cmpared with growth rates on the boards. The radial growth rate of P. glabratula ssp. fuliginosa thalli declined after transplantation to south-east-facing rock surfaces and was unchanged after transplantation to north-west-facing rock surfaces compared with grwoth rate on the boards. The radial growth rate of P. saxatilis thalli was similar on the horizontal boards, south-east-facing and north-west-facing rock surfaces. These results are dsicussed in relation to the aspect distribution of the four lichens in South Gwynedd, Wales.

Optical properties of some surfaces for solar energy application

The preThe present work is a study of the optical properties of some surfaces, in order to determine their applications in solar energy utilisation. An attempt has been made to investigate and measure the optical properties of two systems of surface moderately selective surfaces like thermally grown oxide of titanium, titanium oxide en aluminium and thermally grown oxides of stainless steel; and, selective surfaces of five different coloured stainless at (INCO surfaces) and of black nickel foil. A calorimetric instrument based on the steady state method for measuring directly the total emittance has been designed. Chapter 1 is an introductory survey of selective surface. It also includes a brief review of various preparation techniques in use since 1955. Chapter 2 investigates the theory of selective surfaces, defining their optical properties and their figures of merit. It also outlines the method of computing the optical properties (i.e. absorptance, a, and emittance, a) which have been adopted for the present work. Chapter 3 describes the measuring technique and the modes of operation of the equipment used in the experimental work carried out. Chapter 4 gives the results of the experimental work to measure the optical properties, the life testing and chemical composition of the surfaces under study. Chapter 5 deals with the experimentation leading to the design of a calorimetric instrument for measuring the total emmitance directly. Chapter 6 presents concluding remarks about the outcome of the present work and some suggestions for further work. sent work is a study of the optical properties of some surfaces, in order to determine their applications in solar energy utilisation. An attempt has been made to investigate and measure the optical properties of two systems of surface moderately selective surfaces like thermally grown oxide of titanium, titanium oxide en aluminium and thermally grown oxides of stainless steel; and, selective surfaces of five different coloured stainless at (INCO surfaces) and of black nickel foil. A calorimetric instrument based on the steady state method for measuring directly the total emittance has been designed. Chapter 1 is an introductory survey of selective surface. It also includes a brief review of various preparation techniques in use since 1955. Chapter 2 investigates the theory of selective surfaces, defining their optical properties and their figures of merit. It also outlines the method of computing the optical properties (i.e. absorptance, a, and emittance, a) which have been adopted for the present work. Chapter 3 describes the measuring technique and the modes of operation of the equipment used in the experimental work carried out. Chapter 4 gives the results of the experimental work to measure the optical properties, the life testing and chemical composition of the surfaces under study. Chapter 5 deals with the experimentation leading to the design of a calorimetric instrument for measuring the total emmitance directly. Chapter 6 presents concluding remarks about the outcome of the present work and some suggestions for further work.

The biology of the lichen Rhizocarpon geographicum: symbionts, growth, ecology, and lichenometry

Rhizocarpon geographicum (L.) DC. is one of the most widely distributed species of crustose lichens. This unusual organism comprises yellow-green ‘areolae’ that contain the algal symbiont which develop and grow on the surface of a non-lichenized, fungal ‘hypothallus’ that extends beyond the margin of the areolae to form a marginal ring. This species grows exceptionally slowly with annual radial growth rates (RGR) as low as 0.07 mm yr-1 and its considerable longevity has been exploited by geologists in the development of methods of dating the age of exposure of rock surfaces and glacial moraines (‘lichenometry’). Recent research has established some aspects of the basic biology of this important and interesting organism. This chapter describes the general structure of R. geographicum, how the areolae and hypothallus develop, why the lichen grows so slowly, the growth rate-size curve, and some aspects of the ecology of R. geographicum including whether the lichen can inhibit the growth of its neighbours by chemical means (‘allelopathy’). Finally, the importance of R. geographicum in direct and indirect lichenometry is reviewed.

Digluconate and isopropyl alcohol biocide formulation

Effective surface disinfection is a fundamental infection control strategy within healthcare. This study assessed the antimicrobial efficacy of novel biocide formulations comprising 5% and 2% eucalyptus oil (EO) combined with 2% chlorhexidine digluconate (CHG) and 70% isopropyl alcohol (IPA) contained within a wipe. The efficacy of this novel antimicrobial formulation to remove and eliminate methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli and Candida albicans from steel surfaces was investigated. Adpression studies of pre-contaminated wipes were also utilised to assess their potential to induce cross-contamination between hard surfaces. Furthermore, the bactericidal nature of the EO-formulation was established in addition to time-kill. The EO-containing formulations demonstrated bactericidal antimicrobial efficacy against all microorganisms and did not induce surface cross-contamination. There was no significant difference (p < 0.05) between the 5% and 2% EO formulations in their ability to remove microorganisms from steel surfaces, however both significantly (p < 0.05) removed more than the control formulations. Microbial biofilms were eliminated within 10 min (p < 0.05) when exposed to the EO formulations. Our novel EO-formulation demonstrated rapid antimicrobial efficacy for potential disinfection and elimination of microbial biofilms from hard surfaces and may therefore be a useful adjunct to current infection control strategies currently employed within healthcare facilities.

Controlled growth of poly(2-(diethylamino)ethyl methacrylate) brushes via atom transfer radical polymerisation on planar silicon surfaces

Progress in making pH-responsive polyelectrolyte brushes with a range of different grafting densities is reported. Polymer brushes of poly(2-(diethylamino)ethyl methacrylate) were synthesised via atom transfer radical polymerisation on silicon wafers using a 'grafted from' approach. The [11-(2-bromo-2-methyl) propionyloxy]undecyl trichlorosilane initiator was covalently attached to the silicon via silylation, from which the brushes were grown using a catalytic system of copper(I) chloride and pentamethyldiethylenetriamine in tetrahydrofuran at 80°C. X-ray reflectivity was used to assess the initiator surfaces and an upper limit on the grafting density of the polymer was determined. The quality of the brushes produced was analysed using ellipsometry and atomic force microscopy, which is also discussed.

Lichen growth and lichenometry

Lichenometry is one of the most widely used methods available for dating the surface age of various substrata including rock surfaces, boulders, walls, and archaeological remains. It depends on the assumption that if the lag time before colonisation of a substratum by a lichen is known and lichen age can be estimated, then a minimum date can be obtained by measuring the diameter (or another property related to size) of the largest lichen at the site. Lichen age can be determined by variety of methods including calibrating lichen size against surfaces of known age (‘indirect lichenometry’), by constructing a growth rate-size curve from direct measurement of lichen growth (‘direct lichenometry’), using radio-carbon (RC) dating, and from lichen ‘growth rings’. This chapter describes: (1) lichen growth rates and longevity, (2) methods of estimating lichen age, (3) the methodology of lichenometry and (4) applications of lichenometry. Despite its limitations, lichenometry is likely to continue to play an important role in dating a variety of surfaces and also in providing data that contribute to the debate regarding global warming and climate change.

Lichenometric dating (lichenometry) and the biology of the lichen genus rhizocarpon:challenges and future directions

Lichenometric dating (lichenometry) involves the use of lichen measurements to estimate the age of exposure of various substrata. Because of low radial growth rates and considerable longevity, species of the crustose lichen genus Rhizocarpon have been the most useful in lichenometry. The primary assumption of lichenometry is that colonization, growth and mortality of Rhizocarpon are similar on surfaces of known and unknown age so that the largest thalli present on the respective faces are of comparable age. This review describes the current state of knowledge regarding the biology of Rhizocarpon and considers two main questions: (1) to what extent does existing knowledge support this assumption; and (2) what further biological observations would be useful both to test its validity and to improve the accuracy of lichenometric dates? A review of the Rhizocarpon literature identified gaps in knowledge regarding early development, the growth rate/size curve, mortality, regeneration, competitive effects, colonization, and succession on rock surfaces. The data suggest that these processes may not be comparable on different rock surfaces, especially in regions where growth rates and thallus turnover are high. In addition, several variables could differ between rock surfaces and influence maximum thallus size, including rate and timing of colonization, radial growth rates, environmental differences, thallus fusion, allelopathy, thallus mortality, colonization and competition. Comparative measurements of these variables on surfaces of known and unknown age may help to determine whether the basic assumptions of lichenometry are valid. Ultimately, it may be possible to take these differences into account when interpreting estimated dates.

Simultaneous optimization of colloidal stability and interfacial charge transfer efficiency in photocatalytic Pt/CdS nanocrystals

Colloidal stability and efficient interfacial charge transfer in semiconductor nanocrystals are of great importance for photocatalytic applications in aqueous solution since they provide long-term functionality and high photocatalytic activity, respectively. However, colloidal stability and interfacial charge transfer efficiency are difficult to optimize simultaneously since the ligand layer often acts as both a shell stabilizing the nanocrystals in colloidal suspension and a barrier reducing the efficiency of interfacial charge transfer. Here, we show that, for cysteine-coated, Pt-decorated CdS nanocrystals and Na2SO3 as hole scavenger, triethanolamine (TEOA) replaces the original cysteine ligands in situ and prolongs the highly efficient and steady H2 evolution period by more than a factor of 10. It is shown that Na2SO3 is consumed during H2 generation while TEOA makes no significant contribution to the H2 generation. An apparent quantum yield of 31.5%, a turnover frequency of 0.11 H2/Pt/s, and an interfacial charge transfer rate faster than 0.3 ps were achieved in the TEOA stabilized system. The short length, branched structure and weak binding of TEOA to CdS as well as sufficient free TEOA in the solution are the keys to enhancing colloidal stability and maintaining efficient interfacial charge transfer at the same time. Additionally, TEOA is commercially available and cheap, and we anticipate that this approach can be widely applied in many photocatalytic applications involving colloidal nanocrystals.