Supra-assembly of siliceous vesicles

We report a new approach to produce macroporous (110 nm in diameter) ordered siliceous foams (MOSF) by using block copolymers as templates in the absence of any organic cosolvent. The fine three-dimensional honeycomb structure of MOSF was determined by electron tomography. A formation mechanism of MOSF that spans from the atomic to macroscopic scale is proposed, which involves the cooperative self-assembly of unilamellar vesicles followed by the supra-assembly of vesicles. The fusion of soft vesicles finally leads to MOSF with well-ordered and defined honeycomb structures.

Solution properties of star and linear poly(N-isopropylacrylamide)

The LCST transitions of novel N-isopropylacrylamide ( NIPAM) star polymers, prepared using the four-armed RAFT agent pentaerythritoltetrakis(3-(S-benzyltrithiocarbonyl) propionate) (PTBTP) and their hydrolyzed linear arms were studied using H-1 NMR, PFG-NMR, and DLS. The aim was to determine the effect of polymer architecture and the presence of end groups derived from RAFT agents on the LCST. The LCST transitions of star PNIPAM were significantly depressed by the presence of the hydrophobic star core and possibly the benzyl end groups. The effect was molecular weight dependent and diminished once the number of repeating units per arm >= 70. The linear PNIPAM exhibited an LCST of 35 degrees C, regardless of molecular weight; the presence of both hydrophilic and hydrophobic end groups after hydrolysis from the star core was suggested to cancel effects on the LCST. A significant decrease in R-H was observed below the LCST for star and linear PNIPAM and was attributed to the formation of n-clusters. Application of a scaling law to the linear PNIPAM data indicated the cluster size n = 6. Tethering to the hydrophobic star core appeared to inhibit n-cluster formation in the lowest molecular weight stars; this may be due to enhanced stretching of the polymer chains, or the presence of larger numbers of n-clusters at temperatures below those measured.

A blank slate? Layer-by-layer deposition of hyaluronic acid and chitosan onto various surfaces

Although poly(alpha-hydroxy esters), especially the PLGA family of lactic acid/glycolic acid copolymers, have many properties which make them promising materials for tissue engineering, the inherent chemistry of surfaces made from these particular polymers is problematic. In vivo, they promote a strong foreign-body response as a result of nonspecific adsorption and denaturation of serum proteins, which generally results in the formation of a nonfunctional fibrous capsule. Surface modification post-production of the scaffolds is an often-utilized approach to solving this problem, conceptually allowing the formation of a scaffold with mechanical properties defined by the bulk material and molecular-level interactions defined by the modified surface properties. A promising concept is the so-called blank slate: essentially a surface that is rendered resistant to nonspecific protein adsorption but can be readily activated to covalently bind bio-functional molecules such as extracellular matrix proteins, growth factors or polysaccharides. This study focuses on the use of the quartz crystal microbalance (QCM) to follow the layer-by-layer (LbL) electrostatic deposition of high molecular weight hyaluronic acid and chitosan onto PLGA surfaces rendered positively charged by aminolysis, to form a robust, protein-resistant coating. We further show that this surface may be further functionalized via the covalent attachment of collagen IV, which may then be used as a template for the self-assembly of basement membrane components from dilute Matrigel. The response of NIH-3T3 fibroblasts to these surfaces was also followed and shown to closely parallel the results observed in the QCM.

Synthesis of soluble phosphate polymers by RAFT and their in vitro mineralization

Soluble linear (non-cross-linked) poly(monoacryloxyethyl phosphate) (PMAEP) and poly(2-(methacryloyloxy)ethyl phosphate) (PMOEP) were successfully synthesized through reversible addition-fragmentation chain transfer (RAFT)-mediated polymerization and by keeping the molecular weight below 20 K. Above this molecular weight, insoluble (cross-linked) polymers were observed, postulated to be due to residual diene (cross-linkable) monomers formed during purification of the monomers, MOEP and MAEP. Block copolymers consisting of PMAEP or PMOEP and poly(2-(acetoacetoxy) ethyl methacrylate) (PAAEMA) were successfully prepared and were immobilized on aminated slides. Simulated body fluid studies revealed that calcium phosphate (CaP) minerals formed on both the soluble polymers and the cross-linked gels were very similar. Both the PMAEP polymers and the PMOEP gel showed a CaP layer most probably brushite or monetite based on the Ca/P ratios. A secondary CaP mineral growth with a typical hydroxyapatite (HAP) globular morphology was found on the PMOEP gel. The soluble PMOEP film formed carbonated HAP according to Fourier transform infrared (FTIR) spectroscopy. Block copolymers attached to aminated slides showed only patchy mineralization, possibly due to the ionic interaction of negatively charged phosphate groups and protonated amines.

Effect of the average soft-segment length on the morphology and properties of segmented polyurethane nanocomposites

Two organically modified layered silicates (with small and large diameters) were incorporated into three segmented polyurethanes with various degrees of microphase separation. Microphase separation increased with the molecular weight of the poly(hexamethylene oxide) soft segment. The molecular weight of the soft segment did not influence the amount of polyurethane intercalating the interlayer spacing. Small-angle neutron scattering and differential scanning calorimetry data indicated that the layered silicates did not affect the microphase morphology of any host polymer, regardless of the particle diameter. The stiffness enhancement on filler addition increased as the microphase separation of the polyurethane decreased, presumably because a greater number of urethane linkages were available to interact with the filler. For comparison, the small nanofiller was introduced into a polyurethane with a poly(tetramethylene oxide) soft segment, and a significant increase in the tensile strength and a sharper upturn in the stress-strain curve resulted. No such improvement occurred in the host polymers with poly(hexamethylene oxide) soft segments. It is proposed that the nanocomposite containing the more hydrophilic and mobile poly(tetramethylene oxide) soft segment is capable of greater secondary bonding between the polyurethane chains and the organosilicate surface, resulting in improved stress transfer to the filler and reduced molecular slippage. (c) 2006 Wiley Periodicals, Inc.

Zwitterionic and charge-balanced polyampholyte copolymer hydrogels

Zwitterionic copolymers were synthesised from N,N-dimethyl-N-(2- acryloylethyl)-N-(3-sulfopropyl) ammonium betaine (SPDA) and 2-hydroxyethyl methacrylate (HEMA) produce a series of polyzwitterion hydrogels. For the synthesis of the charge-balanced copolymer hydrogels, two cationic monomers were selected: 2-(diethylamino) ethyl methacrylate (DMAEMA) and 3-(dimethylamino) propyl methacrylamide (DMAPMA), and an anionic monomer; 2-acrylamido-2- methylpropane sulphonic acid (AMPS). Two series of charge-balanced copolymers were synthesized from stoichiometrically equivalent ratios of DMAEMA or DMAPMA and AMPS with HEMA as a termonomer. All synthesized copolymers produced clear and cohesive hydrogels. The zwitterionic and charge-balanced copolymers displayed similar equilibrium water contents together with similar mechanical and surface energy properties. The swelling of the zwitterionic and the charge-balanced copolymers shows some features of antipolyelectrolyte behavior.

Variations of local heat transfer coefficient in piped flow of viscous liquids

Measurements were carried out to determine local coefficients of heat transfer in short lengths of horizontal pipe, and in the region of an discontinuity in pipe diameter. Laminar, transitional and turbulent flow regimes were investigated, and mixtures of propylene glycol and water were used in the experiments to give a range of viscous fluids. Theoretical and empirical analyses were implemented to find how the fundamental mechanism of forced convection was modified by the secondary effects of free convection, temperature dependent viscosity, and viscous dissipation. From experiments with the short tube it was possible to determine simple empirical relationships describing the axial distribution of the local 1usselt number and its dependence on the Reynolds and Prandtl numbers. Small corrections were made to account for the secondary effects mentioned above. Two different entrance configurations were investigated to demonstrate how conditions upstream could influence the heat transfer coefficients measured downstream In experiments with a sudden contraction in pipe diameter the distribution of local 1u3se1t number depended on the Prandtl number of the fluid in a complicated way. Graphical data is presented describing this dependence for a range of fluids indicating how the local Nusselt number varied with the diameter-ratio. Ratios up to 3.34:1 were considered. With a sudden divergence in pipe diameter, it was possible to derive the axial distribution of the local Nusse1t number for a range of Reynolds and Prandtl numbers in a similar way to the convergence experiments. Difficulty was encountered in explaining some of the measurements obtained at low Reynolds numbers, and flow visualization techniques wore used to determine the complex flow patterns which could lead to the anomalous results mentioned. Tests were carried out with divergences up to 1:3.34 to find the way in which the local Nusselt number varied with the diameter ratio, and a few experiments were carried out with very large ratios up .to 14.4. A limited amount of theoretical analysis of the 'divergence' system was carried out to substantiate certain explanations of the heat transfer mechanisms postulated.

Polymer-phospholipid complexes for the solubilisation and delivery of drugs to the eye

Aim: Topical application of ophthalmic drugs is very inefficient; contact lenses used as drug delivery devices could minimize the drug loss and side effects. Styrene-maleic acid copolymers (PSMA) can form polymer-phospholipid complexes with dipalmitoyl phosphatidylcholine (DMPC) in the form of nanometric vesicles, which can easily solubilise hydrophobic drugs. They can be dispersed on very thin contact lens coatings to immobilize the drug on their surface. Methods: Two types of complexes stable at different pH values (5 and 7 respectively) where synthesized and loaded with drugs of different hydrophilicities during their formation process. The drug release was studied in vitro and compared to the free drug. Results: The mean sizes of the complexes obtained by light scattering were 50 nm and 450 nm respectively with low polydispersities. However, they were affected by the drugs load and release. An increase was observed in the duration of the release in the case of hydrophobic drugs, from days to weeks, avoiding initial “burst” and with a lesser amount of total drug released due to the interaction of the drug with the phospholipid core. The size and charge of the different drugs and the complexes nature also affected the release profile. Conclusions: Polymer-phospholipid complexes in the form of nanoparticles can be used to solubilise and release hydrophobic drugs in a controlled way. The drug load and release can be optimised to reach therapeutic values in the eye.

Functional polymers for biomedical application : synthesis and applications

Aromatic and aliphatic diacid chlorides were used to condense naturally occurring diamino acids and their esterified derivatives. It was anticipated the resulting functional polyamides would biodegrade to physiologically acceptable compounds and show pH dependant solubility could be used for biomedical applications ranging from enteric coatings to hydrosoluble drug delivery vehicles capable of targeting areas of low physiological pH. With these applications in mind the polymers were characterised by infra red spectroscopy, gel permeation chromatography and in the case of aqueous soluble polymers by potentiometric titration. Thin films of poly (lysine ethyl ester isophthalamide) plasticised with poly (caprolactone) were cast from DMSO/chloroform solutions and their mechanical properties measured on a Hounsfield Hti tensiometer. Interfacial synthesis was investigated as a synthetic route for the production of linear functional polyamides. High molecular weight polymer was obtained only when esterified diamino acids were condensed with aromatic diacid chlorides. The method was unsuitable for the production of copolymers of free and esterified amino acids with a diacid chloride. A novel miscible mixed solvent single phase reaction was investigated for production of copolymers of esterified and non-esterified amino acids with diacid chlorides. Aliphatic diacid chlorides were unsuitable for condensing diamino acids using this technique because of high rates of hydrolysis. The technique gave high molecular weight homopolymers from esterified diamino acids and aromatic diacid chlorides.

Synthesis and characterisation of hydrogel polymers for medical applications

A fundamental if poorly understood problem that hydrogels display is the tendency of these contact lens materials to dehydrate, causing certain complications of the corneal epithelium. However, recent studies have indicated that the evaporation rate of water from different hydrogel lenses is the same and the severity of conditions such as corneal staining is controlled by the states of water in the material. A study was therefore undertaken which concluded that increased corneal desiccating staining occurred as the proportion of water existing in the bound state decreased. The possibility of using dehydrated hydrogels as packaging materials with desiccating properties has also been investigated. As hydrogels have a high affinity for water they have adequate ability to function as a moisture scavenger in an enclosed atmosphere. It was concluded that this ability is maximised by a high total water content and an increase in the proportion of this water existing in the bound state for the material when it is fully hydrated. N-vinyl pyrrolidone has a low reactivity in vinyl polymerisation reactions which results in polymers with local domains of the same chemical type which can lead to deposition. As contact lenses comprising of this monomer are susceptible to deposition, a monomer with a higher reactivity in vinyl polymerisations is acryloylmorpholine and its incorporation in favour of NVP is encouraged. Unfortunately a large proportion of high EWC hydrogels are mechanically weak and attempts to increase this property by increasing hydrophobicity or cross-linking results in a decrease in EWC. Monomers with the potential to carry a positive charge were incorporated into a high EWC, AMO-HEMA copolymer and the physical properties were investigated. Although EWC increased, mechanical properties decreased only slightly. Therefore simultaneous incorporation of a positively charged monomer and a negatively charged monomer was investigated. The resulting copolymers showed increased water content and increased initial modulus. A technique for measuring the coefficient of friction of contact lenses during lubrication has been developed.

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.

Synthesis and evaluation of novel polymer modifiers for biodegradable polymers

The effects of ester plasticizers and copolymers on the mechanical properties of the natural biodegradable polymers, poly(3-hydroxybutyrate) [PHB] and poly(lactic acid) [PLA] have been studied after subjecting to melt processing conditions. Ester plasticizers were synthesized from citric, tartaric and maleic acids using various alcohols. A variety of PLA copolymers have also been prepared from poly(ethylene glycol) derivatives using stannous octanoate catalysed ring opening polymerisations of DL-lactide. A novel PLA star copolymer was also prepared from an ethoxylated pentaerythritol. The structures of these copolymers were determined by NMR spectroscopy. The plasticizing effect of the synthesised additives at various concentrations was determined. While certain additives were capable of improving the mechanical properties of PLA, none were effective in PHB. Moreover, it was found that certain combinations of additives exhibited synergistic effects. Possible mechanisms are discussed. Biotic and abiotic degradation studies showed that the plasticizers (esters and copolymers) did not inhibit the biodegradability of PHB or PLA in compost at 60°C. Simple toxicity tests carried out on compost extract and its ability to support the growth of cress seeds was established. PLA was found to be susceptible to limited thermal degradation under melt processing conditions. Conventional phenolic antioxidants showed no significant effect on this process, suggesting that degradation was not predominantly a free radical process. PLA also underwent photo-oxidative degradation with UV light and the process could be accelerated in the presence of a photoactivator such as iron (III) diisononyl dithiocarbamate. The mechanisms for the above processes are discussed. Finally, selected compounds were prepared on a pilot plant scale. Extruded and blown films were prepared containing these additives with conventional polymer processing equipment. The mechanical properties were similar to those obtained with laboratory produced compression moulded films.

Cellular interactions with hydrogels

An efficient means of evaluating potential biomaterials is to use the in vitro fibroblast cell culture model. However, the chemistry which influences cell adhesion on polymer substrates is poorly understood. The work in this thesis aims to rationalise several theories of current opinion and introduce new chemical techniques that may predict cellular behaviour. The keratoprosthesis is a typical example of the need to be able to manipulate cell adhesion of materials since both adhesive and non adhesive sections are needed for proper integration and optical function. Calcein AM/ethidium homodimer-1 and DAPI assays were carried out using 3T3 and EKl.BR cells. Poly(HEMA) was found to be the most cell adhesive hydrogel tested. The reactivity of monomers and the resulting sequence distribution were found to affect surface properties and this may explain the poor levels of cell adhesion seen on NVP/MMA copolymers. Surface free energy is shown to be dependent on the polar and non polar groups present along the backbone chain of the polymers. Dehydrated and hydrated contact angle measurements show the effect of rotation of surface groups around the backbone chain. This effect is most apparent on hydrogels containing methacrylic acid. Dynamic contact angle measurements confirm sequence distribution irregularities and demonstrate the mobility of surface groups. Incorporation of NVI or DEAEMA into the hydrogels does not affect the mobility of the surface groups despite their bulkiness. Foetal calf serum was used for the first time as a test solution in an attempt to mimic a biological environment during surface experiments. A Vroman effect may be present, and may involve different surface proteins for each material tested. This interdisciplinary study combines surface characterisation and biological testing to further the knowledge of the biomaterial/host interface. Surface chemistry techniques appear to be insufficiently sensitive to predict cellular behaviour. The degree of ionisation of hydrogels containing ionic groups depends on the nature of the functional groups as well as the concentration and this is an important parameter to consider when comparing charged materials.

Novel hydrogels for keratoprosthesis

Hydrogels may be described as cross~linked hydrophilic polymers that swell but do not dissolve in water. They 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. This project is focused on the development of novel hydrogels for keratoprosthesis (KPro). The most commonly used KPro model consists of a tansparent central stem witb a porous peripheral skirt. Clear poly (methyl methacrylate) (PMMA) core material used in the Strampelli KPros prosthesis has not been the cause of failure found in other core and skirt prostheses. However, epithelialization of this kind of solid, rigid optic material is clearly impossible. The approach to the development of a hydrogeJ for potential KPro use adopted in this work is to develop soft core material to mimic the properties of the natural cornea by incorporating some hydrophilic monomers such as N, N-dimethyacrylamide (NNSMA) N~vinyl pyrrolidone (NVP) and acryloylmorpholine (AMO) with methyl methactylate (MMA). Most of these materials have been used in other ophthalmic applications, such as contact lens. However, an unavoidable limitation of simple .MMA copolymers as conventional hydrogels is poor mechanical strength. The hydrogel for use in this application must be able to withstand the stresses involved during the surgical procedure involved with KPro surgery and the in situ stresses such as the deforming force of the eyelid during the blink cycle. Thus, semi-interpenetrating polymer networks (SIPNs) based on ester polyurethane, AMO, NVP and NNDMA were examined in this work and were found to have much improved mechanical properties at water contents between 40% and 70%. Polyethylene glycol monomethacrylate (PEG MA) was successfully incorporated in order to modulate protein deposition and cell adhesion. Porous peripheral skirts were fabricated using different types of porosigen. The water content mechanical properties, surface properties and cell response of these various materials have been investigated in this thesis. These studies demonstrated that simple hydrogel SIPNs which show isotropic mechanical behaviour, are not ideal KPro materials since they do not mimic the anisotropic behaviour of natural cornea. The final stage of the work has concentrated on the study of hydrogels reinforced with mesh materials. They offer a promising approach to making a hydrogel that is very flexible but strong under tension, thereby having mechanical properties closer to the natural cornea than has been previously possible.

The synthesis of novel biodegradable polyesters and copolyesters from anhydrosulfites

This thesis was concerned primarily with the synthesis and the ring-opening polymerisation of anhydrosulfites (1,3,2-dioxa-thiolan-4-one-2-oxides), and secondly with the copolymerisation of anhydrosulfites with -caprolactone. The polyesters and copolyesters synthesised are of considerable interest in medical applications and also for use as environmental friendly packaging. A range of anhydrosulfites were prepared according to an established method. Aliphatic anhydrosulfites were obtained with a level of purity satisfactory for polymerisation whereas aromatic anhydrosulfites decomposed during distillation and purification by chromatographic techniques. Aliphatic anhydrosulfites with a substituent, such as methyl, isopropyl, n-butyl and isobutyl were studied by NMR spectroscopy. Analysis of these spectra revealed that the five-membered anhydrosulfite ring was puckered and that when the substituent was bulky, rotations about the alkyl chains were restricted. A wide range of anionic initiators may be used to initiate anhydrosulfites. Lithium alkyls turned out to be more successful than alkali metal alkoxides and amides. The molecular weights were found to depend on the basicity of the initiator, the monomer-to-initiator ratio, the nature of the solvent and the polymerisation temperature. The molecular weight M0 of poly(L-lactic acid) ranged from (0.5 to 6)x104. Highly crystalline and purely isotactic poly(lactic acid) was synthesised from L-lactic acid anhydrosulfite (L-LAAS) whereas DL-LAAS led to an amorphous polymer with randomly distributed D-and L-lactic units. This indicated that this polymerisation was not stereoselective. However, the bulkiness of the substituent in the anhydrosulfites molecule was found to influence the stereoselectivity of the polymerisation, thus polyesters with isobutyl or n-butyl pendant group were preferentially isotactic. Block-copolymers of ε-caprolactone and several anhydrosulfites were successfully produced. Block-copolymers of LAAS with ε-caprolactone were also synthesised, but the incorporation of caprolactone units was rather small. In contrast, random copolymerisation of LAAS and ε-caprolactone led to polymers with blocky structures similar to those obtained in the block-copolymerisation of LAAS with  ε-caprolactone.