Collagen fibre orientation and dispersion govern ultimate tensile strength, stiffness and the fatigue performance of bovine pericardium.

The durability of bovine pericardium leaflets employed in bioprosthetic heart valves (BHVs) can significantly limit the longevity of heart valve prostheses. Collagen fibres are the dominant load bearing component of bovine pericardium, however fibre architecture within leaflet geometries is not explicitly controlled in the manufacture of commercial devices. Thus, the purpose of this study was to ascertain the influence of pre-determined collagen fibre orientation and dispersion on the mechanical performance of bovine pericardium. Three tissue groups were tested in uniaxial tension: cross-fibre tissue (XD); highly dispersed fibre-orientations (HD); or preferred-fibre tissue (PD). Both the XD and PD tissue were tested under cyclic loading at 1.5 Hz and a stress range of 2.7 MPa. The results of the static tensile experiments illustrated that collagen fibre orientation and degree of alignment significantly influenced the material's response, whereby, there was a statistically significant decrease in material properties between the XD groups and both the PD and HD groups for ultimate tensile strength and stiffness (p < 0.01). Furthermore, HD tissue had a stiffness of approximately 58% of the PD group, and XD tissue had a stiffness of approximately 18% of the PD group. The dynamic behaviour of the XD and PD groups was extremely distinct; for example a Weibull analysis indicated that the 50% probability of failure in specimens with fibres orientated perpendicular (XD) to the loading direction occurred at 375 cycles. Due to this failure, XD specimens survived on average less than 20% of the cycles completed by those in which fibres were aligned along the loading direction (PD). The results from this study indicate that fibre architecture is a significant factor in determining static strength and fatigue life in bovine pericardium, and thus must be incorporated in the design process to improve future device durability.

The use of small angle light scattering in assessing strain induced collagen degradation in arterial tissue ex vivo.

Collagen is the predominant load bearing component in many soft tissues including arterial tissue and is therefore critical in determining the mechanical integrity of such tissues. Degradation of collagen fibres is hypothesized to be a strain dependent process whereby the rate of degradation is affected by the magnitude of strain applied to the collagen fibres. The aim of this study is to investigate the ability of small angle light scattering (SALS) imaging to identify strain dependent degradation of collagen fibres in arterial tissue ex vivo, and determine whether a strain induced protection mechanism exists in arterial tissue as observed in pure collagen and other collagenous tissues. SALS was used in combination with histological and second harmonic generation (SHG) analysis to determine the collagen fibre architecture in arterial tissue subjected to strain directed degradation. SALS alignment analysis identified statistically significant differences in fibre alignment depending on the strain magnitude applied to the tissue. These results were also observed using histology and SHG. Our findings suggest a strain protection mechanism may exist for arterial collagen at intermediate strain magnitudes between 0% and 25%. These findings may have implications for the onset and progression of arterial disease where changes in the mechanical environment of arterial tissue may lead to changes in the collagen degradation rate.

Strain mediated enzymatic degradation of arterial tissue: Insights into the role of the non-collagenous tissue matrix and collagen crimp.

Collagen fibre remodelling is a strain dependent process which is stimulated by the degradation of existing collagen. To date, literature has focussed on strain dependent degradation of pure collagen or structurally simple collagenous tissues, often overlooking degradation within more complex, heterogenous soft tissues. The aim of this study is to identify, for the first time, the strain dependent degradation behaviour and mechanical factors influencing collagen degradation in arterial tissue using a combined experimental and numerical approach. To achieve this, structural analysis was carried out using small angle light scattering to determine the fibre level response due to strain induced degradation. Next, strain dependent degradation rates were determined from stress relaxation experiments in the presence of crude and purified collagenase to determine the tissue level degradation response. Finally, a 1D theoretical model was developed, incorporating matrix stiffness and a gradient of collagen fibre crimp to decouple the mechanism behind strain dependent arterial degradation. SALS structural analysis identified a strain mediated degradation response in arterial tissue at the fibre level not dissimilar to that found in literature for pure collagen. Interestingly, two distinctly different strain mediated degradation responses were identified experimentally at the tissue level, not seen in other collagenous tissues. Our model was able to accurately predict these experimental findings, but only once the load bearing matrix, its degradation response and the gradient of collagen fibre crimp across the arterial wall were incorporated. These findings highlight the critical role that the various tissue constituents play in the degradation response of arterial tissue. STATEMENT OF SIGNIFICANCE: Collagen fibre architecture is the dominant load bearing component of arterial tissue. Remodelling of this architecture is a strain dependent process stimulated by the degradation of existing collagen. Despite this, degradation of arterial tissue and in particular, arterial collagen, is not fully understood or studied. In the current study, we identified for the first time, the strain dependent degradation response of arterial tissue, which has not been observed in other collagenous tissues in literature. We hypothesised that this unique degradation response was due to the complex structure observed in arterial tissue. Based on this hypothesis, we developed a novel numerical model capable of explaining this unique degradation response which may provide critical insights into disease development and aid in the design of interventional medical devices.

Collagen fibre characterisation in arterial tissue under load using SALS.

The collagen fibre architecture of arterial tissue is known to play a key role in its resultant mechanical behaviour, while maladaptive remodelling of this architecture may be linked to disease. Many of the techniques currently used to analyse collagen fibre architecture require time consuming tissue preparation procedures and are destructive in nature. The aim of this study is to fully explore Small Angle Light Scattering (SALS) as a means to non-destructively assess collagen fibre architecture in arterial tissue and subsequently gain insights into load induced reorientation. The optimised configuration of the SALS system for arterial tissue was determined using quantitative comparisons to histological analyses of porcine carotid artery as its basis. Once established, layer specific fibre orientation and the influence of tissue loading was determined for thin sections of carotid artery using SALS. This process was subsequently repeated for intact carotid artery layers. A single family of circumferentially orientated collagen fibres were found in the intima (- 0.1 ± 1.4° (5.5°)) and media (- 1.7 ± 1.9° (4.7°)) while two perpendicular families of fibres were identified in the adventitia (- 6.4 ± 0.7° (37.7°)) and (118.3 ± 2.7 (39.9°)). An increase in fibre alignment in response to a 20% circumferential strain was also identified using SALS, characterised by an increase in scattered light eccentricity. RESULTS: determined using SALS agreed with those found using traditional destructive techniques, however SALS has the important benefits of allowing vessel layers to remain intact, and has a fast processing time. SALS unique ability to identify load induced reorganisation in intact arterial layers offers an efficient means to gain crucial insights into arterial disease and its development over time.

Analysis of wind-driven ambient noise in a shallow water environment with a sandy seabed.

On the New Jersey continental shelf ambient sound levels were recorded during tropical storm Ernesto that produced wind speeds up to 40 knots in early September 2006. The seabed at the position of the acoustic measurements can be approximately described as coarse sand. Differences between the ambient noise levels for the New Jersey shelf measurements and deep water reference measurements are modeled using both normal mode and ray methods. The analysis is consistent with a nonlinear frequency dependent seabed attenuation for the New Jersey site.

Isolation of human and mouse HMG2a cDNAs: evidence for an HMG2a-specific 3' untranslated region.

We have isolated cDNAs of the human gene for high mobility group protein HMG2a, using the method of direct cDNA selection. The gene maps to chromosome band Xq28, and is located within 40 kb from marker DXS1684, at a distance of 5.4 Mb from the telomere. The deduced human HMG2a protein sequence has a length of 199 amino acids and is 97% identical to the sequence of chicken HMG2a. The 3' untranslated regions of the HMG2a gene in both species are highly homologous (87% identical nucleotides), and are even more conserved than the coding sequences (84% identical nucleotides). In addition, a partial cDNA sequence of the putative HMG2a gene from mouse was identified. The 3' untranslated regions from human and mouse are 90% identical. We conclude that the 3' untranslated sequences have been under strong selective pressure during evolution. Whereas expression of the chicken HMG2a gene has previously been demonstrated in liver of newly hatched chicken, the human HMG2a gene is transcribed mainly in placenta.

A gene in human chromosome band Xq28 (GABRE) defines a putative new subunit class of the GABAA neurotransmitter receptor.

We have isolated and sequenced a novel human gene (GABRE) of the GABAA neurotransmitter receptor family. A cDNA sequence of the gene coding for a 506 amino acid protein was identified, representing a member of a putative new class (epsilon) of the GABAA receptor. The gene is transcribed at least at low level in several different tissues, with the highest levels being detected in adult heart and placenta. Alternative splicing of GABRE transcripts isolated from different tissues was observed at multiple positions of the gene, yielding an unusually complex variety of cDNA variants. The structure of the 5' region of most cDNAs is compatible with expression of protein sequence epsilon only in adult brain, whereas in other tissues, the majority of transcripts code for truncated protein sequences. The GABRE gene extends over 14 kb and is clustered together with the alpha 3 and the putative beta 4 GABAA receptor subunit genes in an approximately 0.8-Mb interval in chromosome band Xq28, located in the candidate regions of two different neurologic diseases. Based on features of conservation of protein sequences, gene structure, and genomic organization of GABAA receptor gene clusters, we propose that the epsilon and gamma subunit genes have a common ancestor and that GABAA receptor gene clusters in the human genome have diverged by multiple duplication events of an ancestral gene cluster containing one each alpha, beta, and gamma/epsilon precursor gene.

Association of primary sclerosing cholangitis with HLA-B8.

The frequency of HLA antigens was studied in 25 patients with primary sclerosing cholangitis and compared with a control group of 562 kidney donors. Fourteen patients also had ulcerative colitis. A significant increase in the frequency of HLA-B8 (60%) was found in the primary sclerosing cholangitis patients compared with controls (25%) (p less than 0.001). HLA-B8 was found in eight patients with ulcerative colitis. The frequency of HLA-B12 was significantly decreased (8%) compared with controls (30%) (p less than 0.02). Piecemeal necrosis was observed on liver histology in 66% of HLA-B8 positive and 50% of HLA-B8 negative patients. Low titres of serum autoantibodies were frequently found in the primary sclerosing cholangitis group but did not correspond to the presence of HLA-B8. Raised serum concentrations of IgM and IgG were not related to HLA-B8. This study has shown that in patients with primary sclerosing cholangitis there exists a disease susceptibility gene closely associated with the B locus of the major histocompatibility complex which may be modified by other factors such as ulcerative colitis. Patients with ulcerative colitis and HLA-B8 may be particularly liable to develop primary sclerosing cholangitis.