GRAFT COPOLYMERIZATION OF p-ACRYLOYLOXYBENZOIC ACID AND p-METHACRYLOYLOXYBENZOIC ACID ONTO ISOTACTIC POLYPROPYLENE The monomers, p-acryloyloxybenzoic acid (ABA) and p-methacryloyloxybenzoic acid (MBA) were synthesized by condensation reaction of corresponding acid chlorides with p-hydroxybenzoic acid in alkaline medium. The polymerization of the monomers were studied by several techniques. Polyacryloyloxybenzoic acid (PABA) was obtained by &amp / #947 / -radiation induced, solution and bulk melt polymerization by initiation of dicumyl peroxide (DCP). Polymethacryloyloxybenzoic acid (PMBA) could be obtained by only bulk melt polymerization. The graft copolymerization of the monomers onto isotactic polypropylene, IPP, was successfully carried out only with bulk melt polymerization. The IPP used in the graft copolymerization was firstly subjected to &amp / #947 / -radiation to create active sites for grafting. The graft copolymerization of the monomers, ABA and MBA onto IPP were initiated by these active sites. The grafting was studied at constant concentration of the monomers in the reaction mixture (50%) and at different reaction temperatures (170, 185, 202, 215 and 225&deg / C). The maximum grafting of PABA and PMBA were found to be 33.1% and 33.9% in the graft copolymers, respectively. The maximum grafting was reached in shorter times at higher temperatures, and it also increased with the increase of concentration of the monomers in the reaction medium. The graft copolymerization was also carried out by using initiator, DCP at 170&deg / C, however, the grafting extent was lower due to the homopolymerization of the monomers and the direct reactions between peroxides of initiator and peroxides on IPP. The graft copolymers were characterized by several techniques, DSC, WAX, TG/IR, MS, SEM and mechanical testing. The formation of both crystalline forms of (&amp / #61537 / 1 and &amp / #61537 / 2) were observed in the products obtained at 170&deg / C. The graft copolymerization of ABA did not have any significance on the formation of both forms of &amp / #61537 / form, while MBA lead to increase in &amp / #61537 / 2 form. The &amp / #61538 / crystalline modification formed in PABA-g-PP products obtained at 185&deg / C and at higher temperature and also in the second run of DSC studies after fast cooling. &amp / #61538 / form was not observed in graft copolymers of PMBA The decomposition mechanism of PABA, PMBA and the graft coproducts were studied by mass spectrometry and TG/IR. The polymers degraded predominantly by decomposition of side groups giving phenol, benzoic acid, hydroxybenzoic acid, carbondioxide and cyclodiene mainly. The mechanical properties of the graft copolymers showed an improvement particularly in tensile strength and modulus. The maximum tensile strength and modulus of PABA-g-PP were found as 41.1 and 881 MPa, and the values of PMBA-g-PP were measured as 35.9 and 721 MPa, respectively. These values were 28.1 and 486 MPa for irradiated IPP and 33.9 and 632 MPa for virgin IPP, respectively. The copolymerization of ABA did not alter the impact properties of the graft copolymer, while a slight decrease was observed in PMBA-g-PP samples. The tensile and impact fractured surface of the graft copolymers studied by scanning electron microscopy showed homogeneous structure. The brittle nature with some extent of ductility was seen in the samples.

No description available.

Tissue engineering (TE) approaches have emerged as an alternative to traditional tissue and organ replacements. The aim of this work was to contribute to the understanding of the effects of cell-material and endothelial cell (EC) paracrine signaling on cell responses using poly(ethylene glycol) diacrylate (PEGDA) hydrogels as a material platform. Three TE applications were explored. First, the effect of glycosaminoglycan (GAG) identity was evaluated for vocal fold restoration. Second, the influence of GAG identity was explored and a novel approach for stable endothelialization was developed for vascular graft applications. Finally, EC paracrine signaling in the presence of cyclic stretch, and hydrophobicity and inorganic content were studied for osteogenic applications. In terms of vocal fold restoration, it was found that vocal fold fibroblast (VFF) phenotype and extracellular matrix (ECM) production were impacted by GAG identity. VFF phenotype was preserved in long-term cultured hydrogels containing high molecular weight hyaluronan (HAHMW). Furthermore, collagen I deposition, fibronectin production and smooth muscle alpha-actin (SM-alpha-actin) expression in PEG-HA, PEG-chondroitin sulfate C and PEG- heparan sulfate (HS) gels suggest that CSC and HS may be undesirable for vocal fold implants. Regarding vascular graft applications, the impact of GAG identity on smooth muscle cell (SMC) foam cell formation was explored. Results support the increasing body of literature that suggests a critical role for dermatan sulfate (DS)-bearing proteoglycans in early atherosclerosis. In addition, an approach for fabricating bi-layered tissue engineering vascular grafts (TEVGs) with stable endothelialization was validated using PEGDA as an intercellular "cementing" agent between adjacent endothelial cells (ECs). Finally, mesenchymal stem cell (MSC) differentiation toward osteogenic like cells was evaluated. ECM and cell phenotypic data showed that elevated scaffold inorganic content and hydrophobicity were indeed correlated with increased osteogenic differentiation. Moreover, the present results suggest that EC paracrine signaling enhances MSC osteogenesis in the presence of cyclic stretch.

No description available.

Atherosclerosis, the leading cause of mortality in Western societies, affects large elastic arteries, causing focal deposition of proliferative inflammatory and lipid-laden cells within the artery. Several risk factors have been causally implicated in the ???reaction to injury??? hypothesis first described by Ross in 1969. The ???injury??? sustained by endothelial cells may be either mechanical or chemical. Environmental factors have a role in the production of chemical agents that are injurious to the endothelium. Mechanical stresses such as wall tensile stress are proportional to systemic blood pressure and pulse pressure. Essentially, these systemic pressures are fairly evenly distributed throughout the circulation. However, atherosclerotic lesions characteristically occur at focal sites within the human vasculature; at or near bifurcations, within the ostia of branch arteries and at regions of marked or complex curvature, where local haemodynamic abnormalities occur. The most discussed haemodynamic factor seems to be low or highly oscillating wall shear stress which exists on the outer wall of bifurcations and on the inner aspect of curving vessels. The magnitude of these haemodynamic forces may not be great but the subtleties of their variable spatial distribution may help to explain the multifocal distribution of atherosclerotic plaques. With the altered haemodynamics there is endothelial injury and phenotypic changes in the endothelium result, which in turn lead to endothelial cell dysfunction. These haemodynamic variables are difficult to measure directly in vivo. In this work a novel model is developed utilising human autologous vein bypass grafts as a surrogate vessel for the observation of pathological structural changes in response to altered haemodynamics. The influence of haemodynamic factors (such as wall shear stress) in the remodeling of the vein graft wall and the pathogenesis of Myointimal Hyperplasia (MIH) and resultant wall thickening in femoral bypass grafts is analysed. The haemodynamic determinants of MIH (which have been established in many animal models) are similar to those implicated in atherosclerosis. The accelerated responses of the vein (Intimal hyperplasia develops much more rapidly than atherosclerotic lesions in native vessels) make it an ideal model to expediently examine the hypothesised relationships prospectively in an in vivo setting. Furthermore, the utilisation of in vivo data acquired from non-invasive diagnostic methods (such as Magnetic Resonance Angiography (MRA) and Duplex ultrasound) combined with the application of state-of-the-art Computational Fluid Dynamic (CFD) techniques makes the model essentially non-invasive. The following hypotheses are examined: 1) regions of Low shear and High tensile stress should develop disproportionately greater wall thickening, 2) regions of greater oscillatory blood flow should develop greater wall thickening, and 3) regions of lower wall shear should undergo inward (or negative) remodelling and result in a reduction in vessel calibre. The conclusions reached are that abnormal haemodynamic forces, namely low Time-averaged Wall Shear Stress, are associated with subsequent wall thickening. These positive findings have great relevance to the understanding of vein graft MIH and atherosclerosis. It was also evident that with non-invasive data and CFD techniques, some of the important haemodynamic factors are realistically quantifiable (albeit indirectly). The detection of parameters known to be causal in the development of graft intimal hyperplasia or other vascular pathology may improve ability to predict clinical problems. From a surgical perspective this might be employed to facilitate selection of at-risk grafts for more focused postoperative surveillance and reintervention. On a broader stage the utilisation of such analyses may be useful in predicting individuals at greater risk of developing atherosclerotic deposits, disease progression, and the likelihood of clinical events such as heart attack, stroke and threat of limb loss.

Immune responses to antigens presented at skin, or other epithelial surfaces such as the cervix, are important for the clearance of viral infections, such as human papillomavirus (HPV) that infect epithelial cells [13]. Elucidation of the components of an effective immune response to antigens presented in this manner will potentially aid in design of immune modulatory techniques or therapeutic vaccine strategies to treat conditions such as cervical cancer. This thesis addresses the role of CD4+ T lymphocytes in immune responses to antigens presented in skin. CD4+ T cells have a well established role in the priming of CD8+ T cells, such that priming without help results in defective CD8+ T cell memory response [15]. The role of CD4+ T cells in the immune response subsequent to priming is less well delineated [15, 16]. Murine skin grafting is a model of antigen presented at an epithelial surface. The model used in this thesis utilises grafts transgenically expressing neo-antigens (human growth hormone=hGH, ovalbumin=OVA) under the control of a keratin promoter (K14 or K5) in the graft. The corresponding mice are termed K14hGH and K5mOVA. With hGH as the antigen, rejection of such skin grafts were shown to require CD4+ T cells [1]. The most surprising finding was that this requirement for CD4+ T cells was maintained even in an antigen-experienced host (in the recall immune response to hGH). CD4+ T cells are required by graft-primed recipients to reject hGH-expressing grafts, but are not required to reject grafts expressing alternative antigens such as OVA. In an adoptive transfer model into lymphopaenic hosts, when high numbers of CD8+ T cells were transferred, any addition of CD4+ T cells was superfluous. However, with low numbers of OVA-specific CD8+ T cells, the addition of CD4+ T cells resulted in a significantly faster rate of K5mOVA skin graft rejection. This helper enhancement of K5mOVA skin graft rejection is maintained, even 7 when CD8+ T cells were previously activated to a memory phenotype prior to transfer, indicating that CD4+ T cells do have effects after CTL priming in vivo. The requirement for CD4+ T cells in the rejection of C57.K14hGH grafts is abrogated by the addition of a local inflammatory stimulus (TLR7 agonist, imiquimod). This is a local rather than systemic effect, suggesting an influence on trafficking or local effector function. Administration of agonist anti-CD40 antibody also partially abrogates the need for CD4+ T cells in rejection of C57.K14hGH grafts by primed hosts. Although CD40 has a well established role in priming of naïve CTL responses, our findings indicate that CD40 can alter events after priming, and suggests a possible mechanism for the role of CD4+ T cells in this system. With these data, we speculate that CD4+ T cells may provide help by altering the state of APC cross-presenting antigen to experienced CD8+ T cells, and that this can be substituted by TLR or CD40 mediated activation of APC. The result may be an increased number of effector CD8+ T cells, as we demonstrate that high numbers of antigen-specific CD8+ T cells can abrogate this effect.

No description available.

The replacement of missing teeth by osseointegrated dental implants is a commonly utilised treatment option in dentistry. However, successful treatment outcomes are dependent on sufficient bone quantity in the proposed surgical site for implant placement (Buser et al., 2004). Surgical augmentation of bone defects is commonly performed prior to or during implant placement. Bone augmentation procedures of the maxillary sinus or guided bone regeneration (GBR) procedures of alveolar ridge defects have utilised a variety of bone graft materials in block or particulate form, either alone or in combination with resorbable or non-resorbable barrier membranes. Objective: The aim of this study was to determine whether Hydroxyapatite β-Tricalcium Phosphate (HA-TCP) either alone or combined with Enamel Matrix Derivative (EMD) or recombinant human Platelet Derived Growth Factor-BB (rhPDGF-BB) is osteoinductive when implanted into a nonosseous site. Methods: Twenty CD-1 adult male mice underwent intramuscular implantation into both hindlimbs of an empty gelatine capsule or a gelatine capsule containing one of the following: 10 mg of uncoated particulate HA-TCP, (Straumann Bone Ceramic®, HA-TCP), EMD coated HA-TCP, (Emdogain®, HATCP + EMD) or rhPDGF-BB coated HA-TCP (HA-TCP + PDGF). Ten animals were sacrificed at four and eight weeks with five specimens from each group retrieved at each time point. The area of graft placement was radiographed and after graft retrieval, a semi-quantitative histological examination was performed with the aim of assessing the inflammatory changes, reparative processes and osteoinduction within the graft site. Results: At both 4 and 8 weeks, histological analysis failed to demonstrate any osteoinductive activity in any of the specimens from the three experimental groups. A minimal chronic inflammatory response and foreign body reaction was seen in the experimental groups which reduced over time. The particles were embedded within fibrous connective tissue and were encapsulated by a dense cellular layer consisting of active fibroblasts and occasional macrophages with the thickness of this layer decreasing over time. At 4 weeks, a greater density of the fibrous connective tissue was demonstrated in the HA-TCP + EMD group (P<0.001) while a greater thickness in the capsule thickness was seen in the HA-TCP group (P=0.022) although no differences were seen after 8 weeks. Greater neovascularisation was seen in the HA-TCP + PDGF group after 8 weeks (P=0.043) while greater amounts of adipose tissue surrounding the particles were detected in the HA-TCP + PDGF group at 4 weeks (P=0.002) and in the HA-TCP + EMD group at eight weeks (P=0.002). Conclusions: The results of this study suggest that the use of commercially available HA-TCP alone or in combination with EMD or rhPDGF-BB is biocompatible but not osteoinductive in the murine model. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1522641 / Thesis (D.Clin.Dent.) -- University of Adelaide, School of Dentistry, 2010

Immune responses to antigens presented at skin, or other epithelial surfaces such as the cervix, are important for the clearance of viral infections, such as human papillomavirus (HPV) that infect epithelial cells [13]. Elucidation of the components of an effective immune response to antigens presented in this manner will potentially aid in design of immune modulatory techniques or therapeutic vaccine strategies to treat conditions such as cervical cancer. This thesis addresses the role of CD4+ T lymphocytes in immune responses to antigens presented in skin. CD4+ T cells have a well established role in the priming of CD8+ T cells, such that priming without help results in defective CD8+ T cell memory response [15]. The role of CD4+ T cells in the immune response subsequent to priming is less well delineated [15, 16]. Murine skin grafting is a model of antigen presented at an epithelial surface. The model used in this thesis utilises grafts transgenically expressing neo-antigens (human growth hormone=hGH, ovalbumin=OVA) under the control of a keratin promoter (K14 or K5) in the graft. The corresponding mice are termed K14hGH and K5mOVA. With hGH as the antigen, rejection of such skin grafts were shown to require CD4+ T cells [1]. The most surprising finding was that this requirement for CD4+ T cells was maintained even in an antigen-experienced host (in the recall immune response to hGH). CD4+ T cells are required by graft-primed recipients to reject hGH-expressing grafts, but are not required to reject grafts expressing alternative antigens such as OVA. In an adoptive transfer model into lymphopaenic hosts, when high numbers of CD8+ T cells were transferred, any addition of CD4+ T cells was superfluous. However, with low numbers of OVA-specific CD8+ T cells, the addition of CD4+ T cells resulted in a significantly faster rate of K5mOVA skin graft rejection. This helper enhancement of K5mOVA skin graft rejection is maintained, even 7 when CD8+ T cells were previously activated to a memory phenotype prior to transfer, indicating that CD4+ T cells do have effects after CTL priming in vivo. The requirement for CD4+ T cells in the rejection of C57.K14hGH grafts is abrogated by the addition of a local inflammatory stimulus (TLR7 agonist, imiquimod). This is a local rather than systemic effect, suggesting an influence on trafficking or local effector function. Administration of agonist anti-CD40 antibody also partially abrogates the need for CD4+ T cells in rejection of C57.K14hGH grafts by primed hosts. Although CD40 has a well established role in priming of naïve CTL responses, our findings indicate that CD40 can alter events after priming, and suggests a possible mechanism for the role of CD4+ T cells in this system. With these data, we speculate that CD4+ T cells may provide help by altering the state of APC cross-presenting antigen to experienced CD8+ T cells, and that this can be substituted by TLR or CD40 mediated activation of APC. The result may be an increased number of effector CD8+ T cells, as we demonstrate that high numbers of antigen-specific CD8+ T cells can abrogate this effect.

Immune responses to antigens presented at skin, or other epithelial surfaces such as the cervix, are important for the clearance of viral infections, such as human papillomavirus (HPV) that infect epithelial cells [13]. Elucidation of the components of an effective immune response to antigens presented in this manner will potentially aid in design of immune modulatory techniques or therapeutic vaccine strategies to treat conditions such as cervical cancer. This thesis addresses the role of CD4+ T lymphocytes in immune responses to antigens presented in skin. CD4+ T cells have a well established role in the priming of CD8+ T cells, such that priming without help results in defective CD8+ T cell memory response [15]. The role of CD4+ T cells in the immune response subsequent to priming is less well delineated [15, 16]. Murine skin grafting is a model of antigen presented at an epithelial surface. The model used in this thesis utilises grafts transgenically expressing neo-antigens (human growth hormone=hGH, ovalbumin=OVA) under the control of a keratin promoter (K14 or K5) in the graft. The corresponding mice are termed K14hGH and K5mOVA. With hGH as the antigen, rejection of such skin grafts were shown to require CD4+ T cells [1]. The most surprising finding was that this requirement for CD4+ T cells was maintained even in an antigen-experienced host (in the recall immune response to hGH). CD4+ T cells are required by graft-primed recipients to reject hGH-expressing grafts, but are not required to reject grafts expressing alternative antigens such as OVA. In an adoptive transfer model into lymphopaenic hosts, when high numbers of CD8+ T cells were transferred, any addition of CD4+ T cells was superfluous. However, with low numbers of OVA-specific CD8+ T cells, the addition of CD4+ T cells resulted in a significantly faster rate of K5mOVA skin graft rejection. This helper enhancement of K5mOVA skin graft rejection is maintained, even 7 when CD8+ T cells were previously activated to a memory phenotype prior to transfer, indicating that CD4+ T cells do have effects after CTL priming in vivo. The requirement for CD4+ T cells in the rejection of C57.K14hGH grafts is abrogated by the addition of a local inflammatory stimulus (TLR7 agonist, imiquimod). This is a local rather than systemic effect, suggesting an influence on trafficking or local effector function. Administration of agonist anti-CD40 antibody also partially abrogates the need for CD4+ T cells in rejection of C57.K14hGH grafts by primed hosts. Although CD40 has a well established role in priming of naïve CTL responses, our findings indicate that CD40 can alter events after priming, and suggests a possible mechanism for the role of CD4+ T cells in this system. With these data, we speculate that CD4+ T cells may provide help by altering the state of APC cross-presenting antigen to experienced CD8+ T cells, and that this can be substituted by TLR or CD40 mediated activation of APC. The result may be an increased number of effector CD8+ T cells, as we demonstrate that high numbers of antigen-specific CD8+ T cells can abrogate this effect.