Simultaneous Upregulation of Elastolytic and Elastogenic Factors Are Necessary for Regulated Collateral Diameter Expansion

Background: During arteriogenesis, outward remodeling of the arterial wall expands luminal diameter to produce increased conductance in developing collaterals. We have previously shown that diameter expansion without loss of internal elastic lamina (IEL) integrity requires both degradation of elastic fibers and LOX-mediated repair. The aim of this study was to investigate the expression of genes involved in remodeling of the extracellular matrix (ECM) using a model of arteriogenesis.Methods: Sprague-Dawley rats underwent femoral artery ligation with distal arteriovenous fistula (FAL + AVF) placement. Profunda femoral arteries (PFA) were harvested for analysis at various time points. Serum desmosine, an amino acid found exclusively in elastin, was evaluated with enzyme-linked immunosorbent assay (ELISA) as a marker of tissue elastolysis. Tissue mRNA isolated from FAL + AVF exposed PFAs was compared to the contralateral sham-operated using qPCR. HCAECs were cultured under low shear stress (8 dyn·s/cm2) for 24 h and then exposed to high shear stress (40 dyn·s/cm2) for 2–6 h. Primers used included FBN-1, FBN-2, Timp-2, LOX-1, Trop-E, Cath-K, Cath-S, MMP-2, MMP-9, FBLN-4, and FBLN-5 and were normalized to GAPDH. mRNA fold changes were quantified using the 2-ΔΔCq method. Comparisons between time points were made with non-parametric ANOVA analysis with Bonferroni adjustment.Results: PFAs showed IEL reorganization during arteriogenesis. Serum desmosine levels are significantly elevated at 2 days and one week, with a return to baseline thereafter (p < 0.01). Expression of ECM structural proteins (FBN-1, FBN-2, FBLN-4, FBLN-5, Tropoelastin, TIMP-2, LOX-1) and elastolytic proteins (MMP-2, MMP-9, Cathepsin S, Cathepsin K) exhibited an early peak (p < 0.05) relative to sham PFAs. After two weeks, expression returned to baseline. HCAECs demonstrated upregulation of FBN-2, FBLN-5, LOX-1 and Trop-E at 4 h of high shear stress, as well as elastolytic protein MMP-2.Conclusions: Elastin degradation begins early in arteriogenesis and is mediated by local upregulation of elastolytic genes. Elastolysis appears to be simultaneously balanced by production of elastic fiber components which may facilitate stabilization of the IEL. Endothelial cells are central to initiation of arteriogenesis and begin ECM remodeling in response to altered shear stress.

Enhancements in Fracturing Fluid Engineering Reawakens a North Sea Giant

The Valhall field, operated by AkerBP, has been a major hub in the North Sea, on stream for thirty-eight years and recently passed one billion barrels of oil produced. The field requires stimulation for economical production. Mechanically strong formations are acid stimulated, while weaker formations require large tip-screenout design proppant fractures. Fracture deployment methods on Valhall have remained relatively unchanged since the nineties and are currently referred to as "conventional". Those consist in a sequence of placing a proppant frac, cleaning out the well with coiled tubing, opening a sleeve or shooting perforations, then coil pulling out of hole pumping the proppant frac. For the past few years, AkerBP and their service partners have worked on qualifying an adapted version of the annular coiled tubing fracturing practice for the offshore infrastructure - a first for the industry, which has been a strategic priority for the operator as it significantly reduces execution time and accelerates production. As with all technology trials, the implementation of this practice on Valhall had to begin on a learning curve through various forms of challenges. Whilst investigating the cause and frequency of premature screenouts during the initial implementation of annular fracturing, the team decided to challenge the conventional standards for fluid testing and quality control. Carefully engineered adjustments were made with regards to high shear testing conditions, temperature modelling, and mixing sequences, these did not only identify the root cause for the unexpected screenouts, but also helped create the current blueprint for engineering a robust fluid. Since the deployment of the redefined recipe, adjusted testing procedures and changes made to the stimulation vessel, there have not been any cases of fluid induced screenouts during the executions. The fewer types of additives now required for the recipe have lowered the cost of treatments and the lower gel loading leads to reduced damage in the fractures, thereby contributing to enhanced production over the lifetime of the wells. This paper describes the investigation, findings and the resulting changes made to the fluid formulation and quality control procedures to accommodate for high shear and dynamic wellbore temperature conditions. It discusses the rationale behind the "reality" testing model and, proves that significant value is created from investing time in thoroughly understanding fluid behaviour in the lab, prior to pumping it on large-scale capital-intensive operations. The study demonstrated that there is always value in innovating or challenging pre-conceived practices, and the learnings from this investigation significantly improved the track record for annular fracturing on Valhall, redefined fluid engineering for the North Sea and will inform future annular fracturing deployments on other offshore assets around the world.

Mechanical Deformation of Peripapillary Retina in Response to Acute Intraocular Pressure Elevation

Elevated intraocular pressure (IOP) may cause mechanical injuries to the optic nerve head (ONH) and the peripapillary tissues in glaucoma. Previous studies have reported the mechanical deformation of the ONH and the peripapillary sclera (PPS) at elevated IOP. The deformation of the peripapillary retina (PPR) has not been well-characterized. Here we applied high-frequency ultrasound elastography to map and quantify PPR deformation, and compared PPR, PPS and ONH deformation in the same eye. Whole globe inflation was performed in ten human donor eyes. High-frequency ultrasound scans of the posterior eye were acquired while IOP was raised from 5 to 30 mmHg. A correlation-based ultrasound speckle tracking algorithm was used to compute pressure-induced displacements within the scanned tissue cross-sections. Radial, tangential, and shear strains were calculated for the PPR, PPS, and ONH regions. In PPR, shear was significantly larger in magnitude than radial and tangential strains. Strain maps showed localized high shear and high tangential strains in PPR. In comparison to PPS and ONH, PPR had greater shear and a similar level of tangential strain. Surprisingly, PPR radial compression was minimal and significantly smaller than that in PPS. These results provide new insights into PPR deformation in response of IOP elevation, suggesting that shear rather than compression was likely the primary mode of IOP-induced mechanical insult in PPR. High shear, especially localized high shear, may contribute to the mechanical damage of this tissue in glaucoma.

SIPA in 10 milliseconds: VWF tentacles agglomerate and capture platelets under high shear

Shear-Induced Platelet Aggregation (SIPA) occurs under elevated shear rates (~10000 s-1) found in stenotic coronary and carotid arteries. The pathologically high-shear environment can lead to occlusive thrombosis by SIPA from the interaction of nonactivated platelets and von Willebrand factor (VWF) via glycoprotein Ib (GPIb)-A1 binding. This process under high shear rates is difficult to visualize experimentally with concurrent molecular- and cellular-resolutions. To understand this fast bonding, we employ a validated multiscale in-silico model incorporating measured molecular kinetics and a thrombosis-on-a-chip device to delineate the flow-mediated biophysics of VWF and platelets assembly into mural micro-thrombi. We show that SIPA begins with VWF elongation, followed by agglomeration of platelets in the flow by soluble VWF entanglement before mural capture of the agglomerate by immobilized VWF. The entire SIPA process occurs on the order of 10 ms with the agglomerate travelling a lag distance of a few hundred microns before capture, matching in vitro results. Increasing soluble VWF concentration by ~20x in silico leads to a 2~3x increase in SIPA rates, matching the increase in occlusion rates found in vitro. The morphology of mural aggregates is primarily controlled by VWF molecular weight (length), where normal-length VWF leads to cluster or elongated aggregates and ultra-long VWF leads to loose aggregates seen by others' experiments. Finally, we present phase diagrams of SIPA which provides biomechanistic rationales for a variety of thrombotic and hemostatic events in terms of platelet agglomeration and capture.