Cellular interaction with novel biomaterials

The objective of this thesis is to report the behaviour of mammalian cells with biocompatible synthetic polymers with potential for applications to the human body. Composite hydrogel materials were tested as possible keratoprosthetic devices. It was found that surface topography is an important consideration, pores, channels and fibres exposed on the surface of the hydrogels tested can have significant effects on the extent of cell adheson and proliferation. It is recommended that the core component is fabricated out of one of the following to provide a non cell adhesive base; A8, A11, A13, A22, A23. The haptic periphery fabricated out of one of the following would provide a cell adhesive composite; A16, A30, A33, A37, A38, A42, A43, A44. The presence of vitronectin in the ocular tissue appears to lead to higher cell adhesion to the posterior surface of a contact lens when compared to the anterior surface. Group IV contact lenses adhere more cells than Group II contact lenses - this may indicate that more protein (including vitronectin) is able to adhere to the contact lens due to the Group IV contact lenses high water content and ionic hydrogel matrix. Artificial lung surfactant analogues were found to be non cytotoxic but also decreased cell proliferation when tested at higher concentrations. Poly(lysine ethyl ester adipamide) [PLETESA] had the most favourable response on cell proliferation and commercial styrene/maleic anhydride (pMA/STY sp2) the most pronounced inhibitory response. The mode of action that decreases cell proliferation appears to be through membrane destabilization. Tissue culture well plates coated with PLETESA allowed cells to adhere in a concentration dependent manner, multilaminar liposomes possibly of PLETESA were observed in solution in PLETESA coated wells. Polyhydroxybutryate (PHB) and polyhydroxyvalerate (PHV) blends that contained hydroxyapatite were found to be the most cell adhesive material of those materials tested. The blends that were most susceptible to degradation adhered the most cells in initial stages of degradation. The initial slight increase in cell adhesion may be due to the increased rugosity of the material. As the degradation continued the number of cells adhering to the samples decreased, this may indicate that the polarity was inhibitory to cell adhesion during the later stages of degradation.

Ocular biomaterials and the anterior eye

Hydrogels, water swollen polymer matrices, have been utilised in many biomedical applications, as there is the potential to manipulate the properties for a given application by changing the chemical structure of the constituent monomers The eye provides an excellent site to examne the interaction between a synthetic material and a complex biological fluid without invasive surgery. There is a need for the development of new synthetic hydrogels for use in the anterior eye, Three applications of hydrogels in the eye were considered in this thesis. For some patients, the only hope of any visual improvement lies in the use of an artificial cornea, or keratoprosthesis, Preliminary investigations of a series of simple homogeneous hydrogel copolymers revealed that the mechanical properties required to withstand surgery and in eye stresses, were not achieved This lead to work on the development of semi-interpenetrating polymer networks based on the aforementioned copolymers, Manufacture of the device and cell response were also studied. Lasers have been employed in ocular surgery to correct refractive defects. If an irregular surface is ablated, an irregular surface is obtained. A hydrogel system was investigated that could be applied to the eye prior to ablation to create a smooth surface. Factors that may influence ablation rate were explored, Soft contact lenses can be used as a probe to study the interaction between synthetic materials and the biological constituents of tears. This has lead to the development of many sensitive analytical techniques for protein and lipid deposition, one of which is fluorescence spectrophotometry. Various commercially available soft contact lenses were worn for different periods of time and then analysed for protein and lipid deposition using fluorescence spectrophotometry, The influence of water content, degree of ionicity and the lens material on the level and type of deposition was investigated.

Cellular responses to potential biomaterials

The aim of this study was to systematically investigate the factors considered to be responsible for anchorage-dependent cell behaviour to determine which, if any, of these factors exerts greater influence. An efficient means of doing so is the in vitro fibroblast cell culture model. The interaction of fibroblasts with novel substrata gives information about how a biological system reacts to a foreign material. The may ultimately lead to the development of improved biomaterials. This interdisciplinary study combines the elements of surface characterisation and biological testing to determine the nature of the biomaterial/host interface. Polarity and surface charge were found to have an important influence on fibroblast adhesion to hydrogel polymers, by virtue of their water-structuring effects. The same factors were found to affect cell adhesion on undegraded PHB-HV copolymers and their blends with polysaccharides. On degraded PHB-HV copolymers, the degradation process itself played the greatest role in influencing cell response. Increasing surface charge and mechanical instability in these polymers inhibited cell adhesion. Based on the observations of hydrogels and PHB-copolymers a novel material, gel-spun PHB was designed for use as a wound scaffold. In vitro tests using human and mammalian fibroblasts accentuated the importance of polarity and surface charge in determining cellular response. The overall view of cellular behaviour on a broad spectrum of materials highlighted the effects that polarity and surface charge have on water-structuring, and how this affects interfacial conversion. In degradable systems, mechanical stability also plays an inportant role in determining anchorage-dependent cell behaviour.

Melt processable biomaterials for degradable surgical fixation devices

There are currently few biomaterials which combine controlled degradation rates with ease of melt processability. There are however, many applications ranging from surgical fixation devices to drug delivery systems which require such combination properties. The work in this thesis is an attempt to increase the availability of such materials. Polyhydroxybutyrate-polyhydroxyvalerate copolymers are a new class of potentially biodegradable materials, although little quantitative data relating to their in vitro and in vivo degradation behaviour exists. The hydrolytic degradation of these copolymers has been examined in vitro under conditions ranging from `physiological' to extremes of pH and elevated temperature. Progress of the degradation process was monitored by weight loss and water uptake measurement, x-ray diffractometry, optical and electron microscopy, together with changes in molecular weight by gel permeation chromatography. The extent to which the degradation mechanism could be modified by forming blends with polysaccharides and polycaprolactone was also investigated. Influence of the valerate content, molecular weight, crystallinity, together with the physical form of the sample, the pH and the temperature of the aqueous medium on the hydrolytic degradation was investigated. Its progress was characterised by an initial increase in the wet weight, with concurrent decrease in the dry weight as the amorphous regions of the polymer are eroded, thereby producing an increase in matrix porosity. With the polysaccharide blends, this initial rate is dramatically affected, and erosion of the polysaccharide from the matrix markedly increases the internal porosity which leads to the eventual collapse of the matrix, a process which occurs, but less rapidly, in the degradation of the unblended polyhydroxybutyrate-polyhydroxyvalerate copolymers. Surface energy measurement and goniophotometry proved potentially useful in monitoring the early stages of the degradation, where surface rather than bulk processes predominate and are characterised by little weight loss.

Sulphonated biomaterials as glycosaminoglycan mimics in wound healing

This chapter considers the available evidence and underlying physicochemical principles that support the proposition that a biomimetic wound dressing based on glycosaminoglycan models offers a potential means of influencing wound bioactivity. Available evidence showing advantages in wound healing for experimental proteoglycanbased dressing materials is described, together with an overview of the bioactive role of sulphated macromolecules. This leads to an assessment of the analogies between the sulphonate group and the sulphate group and an explanation of their unique water binding behaviour. The available information suggests the desirability of an integrated physicochemical, biochemical and biological approach to the design and synthesis of new wound healing biomaterials.

The ageing ocular surface:challenges for biomaterials design and function

Changing demographics and in particular an increasingly ageing population, in combination with improved longevity, will have a major impact on changing the face of human diseases and likewise the demand for appropriate biomaterials. The ocular surface is a multifaceted system that combines to create a unique mucosal surface, which includes the cornea, conjunctiva, sclera and lids of the eye. Physical parameters such as the eyelids and eyelashes, combined with the numerous secretory glands that produce the complex tear film, act together to protect and maintain the cornea. Unfortunately an ageing tear film and lacrimal functional unit can lead to impairment of this magnificently orchestrated structure. No single mechanism or modification is responsible but, whatever the cause, the consequence is a reduction in tear stability. An uncompromised tear film is fundamental to a healthy ocular surface. In the face of progressively changing demographics and consequent requirements for medical intervention and medical device developments, it is important to understand what effects the ageing process has on these anterior ocular structures.

The development of biomaterials for contact lens applications:effects of wear modality on materials design

The successful design of polymers for contact lens applications depends on the ability to provide a balance of properties appropriate to the ocular environment. Principal relevant aspects of the anterior eye are the tear film, eyelid and cornea, which govern the requirements for surface properties, modulus and oxygen permeability, respectively. Permeability requirements and the developing view of the needs of the cornea, in terms of oxygen consumption and the particular roles of fluorine and silicon in the design of silicone hydrogels, which have proved to be the most successful family of materials for this demanding application, are discussed. The contact lens field is complicated by the fact that contact lenses are used in a range of wear modalities, the extremes of which can conveniently be classified as lenses that are disposed of at the end of a single period of daily wear and those used for 30. days of successive day-and-night periods, frequently referred to as extended or continuous wear. As silicone hydrogels developed, in the decade following their launch there has been a progressive trend in properties taking both modulus and water content closer to those of conventional hydrogels. This is particularly evident in the family of daily disposable contact lenses that have appeared since 2008.