The role of microRNA 21 in the development of in-stent restenosis

Coronary heart disease is a major cause of morbidity and mortality worldwide. Percutaneous coronary intervention (PCI) has become the most widely used method of coronary artery revascularisation. The use of stents to hold open atherosclerosis induced arterial narrowing has significantly reduced elastic recoil and acute vessel occlusion following balloon angioplasty. However, bare metal stents have been associated with in-stent restenosis attributed to vascular smooth muscle cell (VSMC) hyperplasia and excessive neointimal formation. The resultant luminal renarrowing may manifest clinically with the return of symptoms such as chest pain or shortness of breath. The development of drug eluting stents has significantly reduced the incidence of in-stent restenosis (ISR). Unfortunately the antiproliferative medications used not only inhibit VSMC proliferation but also re-endothelialisation of the stented vessel. In addition, the drug impregnated polymer coating has been associated with a chronic inflammatory response within the vessel wall predisposing patients to stent thrombosis. Thus the identification of novel therapies which promote vessel healing without excessive proliferative or inflammatory response may improve long term outcome and reduce the need for repeated revascularisation. MicroRNAs (miRs) are short (18-25 nucleotide) non-coding RNAs acting to regulate gene expression. By binding to the 3’untranslated region of mRNA they act to fine tune gene expression either by mRNA degradation or translational repression. Originally identified in coordinating tissue development microRNAs have also been shown to play important roles coordinating the inflammatory response and in numerous cardiovascular diseases. MiR-21 has been identified in human atherosclerotic plaques, arteriosclerosis obliterans and abdominal aortic aneurysms. In addition, its up regulation has been documented in preclinical models of vascular injury. This study sought to identify the role of miR-21 in the development of ISR. Utilising a small animal model of stenting and in vitro techniques, we sought to investigate its influence upon VSMC and immune cell response following stenting. 19 The refinement of a murine stenting model within the Baker laboratory and the electrochemical dissolution of the metal stent from within harvested vascular tissues significantly improved the ability to perform detailed histological analysis. In addition, identification of miRNAs using in situ hybridisation was achieved for the first time within stented tissue. Neointimal formation and ISR was significantly reduced in mice in which miR-21 had been genetically deleted. In addition, neointimal composition was found to be altered in miR-21 KO mice with reductions in VSMC and elastin content demonstrated. Importantly, no difference in re-endothelialisation was observed. In vitro analysis demonstrated that VSMCs from miR-21 KO mice had both reduced proliferative and migratory capacity following platelet derived growth factor stimulation. Molecular analysis revealed that these differences may, at least in part, be due to de-repression of programmed cell death 4 (PDCD4). PDCD4 is a known miR-21 target within VSMCs implicated in the suppression of proliferation and promotion of apoptosis. Unfortunately, initial attempts at antimiR mediated knockdown of miR-21 in vivo, failed to produce a similar change in the suppression of ISR. Furthermore, a significant alteration in macrophage polarisation state within the neointima of miR-21 WT and KO mice was noted. Immunohistochemical staining revealed a preponderance of anti-inflammatory M2 macrophages in KO mice. Analysis of bone marrow derived macrophages from miR-21 KO mice demonstrated an increased level of the peroxisome proliferation activating receptor-γ (PPARγ) which facilitates M2 polarisation. Importantly, significant alterations in numerous pro-inflammatory cytokines, which also have mitogenic effects, were also found following genetic deletion of miR-21. In Summary, this is the first study to look at miRs in the development of ISR. MiR-21 plays an important role in the development of ISR by influencing the proliferative response of VSMCs and modulating the immune response following stent deployment. Further attempts to modulate miR-21 expression following PCI may reduce ISR and the need for repeat revascularisation while also reducing the risk of stent thrombosis.

Mid-infrared photonic sensors based on metamaterial structures

In this work three different metallic metamaterials (MMs) structures such as asymmetric split ring resonators (A-SRRs), dipole and split H-shaped (ASHs) structures that support plasmonic resonances have been developed. The aim of the work involves the optimization of photonic sensor based on plasmonic resonances and surface enhanced infrared absorption (SEIRA) from the MM structures. The MMs structures were designed to tune their plasmonic resonance peaks in the mid-infrared region. The plasmonic resonance peaks produced are highly dependent on the structural dimension and polarisation of the electromagnetic (EM) source. The ASH structure particularly has the ability to produce the plasmonic resonance peak with dual polarisation of the EM source. The double resonance peaks produced due to the asymmetric nature of the structures were optimized by varying the fundamental parameters of the design. These peaks occur due to hybridization of the individual elements of the MMs structure. The presence of a dip known as a trapped mode in between the double plasmonic peaks helps to narrow the resonances. A periodicity greater than twice the length and diameter of the metallic structure was applied to produce narrow resonances for the designed MMs. A nanoscale gap in each structure that broadens the trapped mode to narrow the plasmonic resonances was also used. A thickness of 100 nm gold was used to experimentally produce a high quality factor of 18 in the mid-infrared region. The optimised plasmonic resonance peaks was used for detection of an analyte, 17β-estradiol. 17β-estradiol is mostly responsible for the development of human sex organs and can be found naturally in the environment through human excreta. SEIRA was the method applied to the analysis of the analyte. The work is important in the monitoring of human biology and in water treatment. Applying this method to the developed nano-engineered structures, enhancement factors of 10^5 and a sensitivity of 2791 nm/RIU was obtained. With this high sensitivity a figure of merit (FOM) of 9 was also achieved from the sensors. The experiments were verified using numerical simulations where the vibrational resonances of the C-H stretch from 17β-estradiol were modelled. Lastly, A-SRRs and ASH on waveguides were also designed and evaluated. These patterns are to be use as basis for future work.