Blood vessels usually have poor local contrast, and the application of existing edge detection algorithms yield results which are not satisfactory. An operator for feature extraction based on the optical and spatial properties of objects to be recognized is introduced. The gray-level profile of the cross section of a blood vessel is approximated by a Gaussian-shaped curve. The concept of matched filter detection of signals is used to detect piecewise linear segments of blood vessels in these images. Twelve different templates that are used to search for vessel segments along all possible directions are constructed. Various issues related to the implementation of these matched filters are discussed. The results are compared to those obtained with other methods. >

/pdf/detection-of-blood-vessels-in-retinal-images-using-two-2lxvrugvz5.pdf

Detection of blood vessels in retinal images using two-dimensional matched filters

Single cardiomyocytes contain myofibrils that harbor the sarcomere-based contractile machinery of the myocardium. Cardiomyocytes differentiated from human pluripotent stem cells (hPSC-CMs) have potential as an in vitro model of heart activity. However, their fetal-like misalignment of myofibrils limits their usefulness for modeling contractile activity. We analyzed the effects of cell shape and substrate stiffness on the shortening and movement of labeled sarcomeres and the translation of sarcomere activity to mechanical output (contractility) in live engineered hPSC-CMs. Single hPSC-CMs were cultured on polyacrylamide substrates of physiological stiffness (10 kPa), and Matrigel micropatterns were used to generate physiological shapes (2,000-µm(2) rectangles with length:width aspect ratios of 5:1-7:1) and a mature alignment of myofibrils. Translation of sarcomere shortening to mechanical output was highest in 7:1 hPSC-CMs. Increased substrate stiffness and applied overstretch induced myofibril defects in 7:1 hPSC-CMs and decreased mechanical output. Inhibitors of nonmuscle myosin activity repressed the assembly of myofibrils, showing that subcellular tension drives the improved contractile activity in these engineered hPSC-CMs. Other factors associated with improved contractility were axially directed calcium flow, systematic mitochondrial distribution, more mature electrophysiology, and evidence of transverse-tubule formation. These findings support the potential of these engineered hPSC-CMs as powerful models for studying myocardial contractility at the cellular level.

https://www.pnas.org/content/pnas/112/41/12705.full.pdf

Contractility of single cardiomyocytes differentiated from pluripotent stem cells depends on physiological shape and substrate stiffness

The differentiation of fibroblasts to contractile myofibroblasts, which is characterized by de novo expression of alpha-smooth muscle actin (alpha-SMA), is crucial for wound healing and a hallmark of tissue scarring and fibrosis. These processes often follow inflammatory events, particularly in soft tissues such as skin, lung and liver. Although inflammatory cells and damaged epithelium can release transforming growth factor beta1 (TGF-beta1), which largely mediates myofibroblast differentiation, the biophysical environment of inflammation and tissue regeneration, namely increased interstitial flow owing to vessel hyperpermeability and/or angiogenesis, may also play a role. We demonstrate that low levels of interstitial (3D) flow induce fibroblast-to-myofibroblast differentiation as well as collagen alignment and fibroblast proliferation, all in the absence of exogenous mediators. These effects were associated with TGF-beta1 induction, and could be eliminated with TGF-beta1 blocking antibodies. Furthermore, alpha1beta1 integrin was seen to play an important role in the specific response to flow, as its inhibition prevented fibroblast differentiation and subsequent collagen alignment but did not block their ability to contract the gel in a separate floating gel assay. This study suggests that the biophysical environment that often precedes fibrosis, such as swelling, increased microvascular permeability and increased lymphatic drainage--all which involve interstitial fluid flow--may itself play an important role in fibrogenesis.

/pdf/interstitial-fluid-flow-induces-myofibroblast-27i2dhegri.pdf

Interstitial fluid flow induces myofibroblast differentiation and collagen alignment in vitro

The planar fibrous connective tissues of the body are composed of a dense extracellular network of collagen and elastin fibers embedded in a ground matrix, and thus can be thought of as biocomposites. Thus, the quantification of fiber architecture is an important step in developing an understanding of the mechanics of planar tissues in health and disease. We have used small angle light scattering (SALS) to map the gross fiber orientation of several soft membrane connective tissues. However, the device and analysis methods used in these studies required extensive manual intervention and were unsuitable for largescale fiber architectural mapping studies. We have developed an improved SALS device that allows for rapid data acquisition, automated high spatial resolution specimen positioning, and new analysis methods suitable for large-scale mapping studies. Extensive validation experiments revealed that the SALS device can accurately measure fiber orientation for up to a tissue thickness of at least 500 μm to an angular resolution of∼1o and a spatial resolution of±254 μm. To demonstrate the new device’s capabilities, structural measurements from porcine aortic valve leaflets are presented. Results indicate that the new SALS device provides an accurate method for rapid quantification of the gross fiber structure of planar connective tissues.

A small angle light scattering device for planar connective tissue microstructural analysis

Investigates a measure of "dominant perceived orientation" that has been developed to match the output of a human study involving 40 subjects. The results of this measure are compared with humans analyzing seven "teaser" images to test its effectiveness for finding perceptually dominant orientations. The use of low-level orientation is then applied to a "quick search" problem important in image database applications. Since both pigeons and humans are able to perform coarse classification of certain kinds of scenes, e.g., city from country, without taking time or brain-power to solve the image understanding problem, the authors conjecture that the collective behavior of low-level textural features such as orientation may be doing most of the work. The authors demonstrate a simple test of global multiscale orientation for quickly searching a database of vacation photos for likely "city/suburb" shots. The orientation features achieve agreement with human classification in 91 out of 98 of the scenes.

Texture orientation for sorting photos "at a glance"

Collagen fibers and their component fibrils make up the protenaceous " backbone" of most tissues and provide the majority of their resistance to tensile loading. Spatial orientation of collagen fibrils is an important factor in determining tissue properties. This is particularly true in ligament tissue, since ligaments must be loose enough to allow joints to move but tight enough to prevent joint surfaces from separating. A method is presented here to reproducibly quantify this collagen arrangement, which should be useful in studies on ligament healing and growth.

Hieu Nguyen

Papers

A Fourier Domain Directional Filterng Method for Analysis of Collagen Alignment in Ligaments