Extracellular cross-striated banded structures in human connective tissue.

Extracellular cross-striated banded structures have been found in the connective tissues of a variety of organs. Rarely have they been found in conditions such as: hyperplastic parathyroid, follicular adenoma of the thyroid, metastatic hemangiopericytoma in the lung, sarcoidosis of lymph node, and two biopsies of nerve as reported here. The shape and size of the structure varies, as well as its periodicity, which is usually in the range of 1000 to 15000 A. An unusual exception to this general rule is the banded structure in the follicular adenoma of the thyroid where the periodicity is 270 A. These structures are quite different from native collagen or experimentally reconstituted long-space collagen, both by staining reaction with phosphotungstic acid or uranyl acetate and lead nitrate, and in their ultrastructural morphology.     

A new method of measuring the cross-sectional area of connective tissue structures

There appears to be no generally accepted method of measuring in-situ the cross-sectional area of connective tissues, particularly small ones, before mechanical testing. An instrument has therefore been devised to measure the cross-sectional area of one such tissue, the rabbit medial collateral ligament, directly and nondestructively. However, the methodology is general and could be applied to other tissues with appropriate changes in detail. The concept employed in the instrument is to measure the thickness of the tissue as a function of position along the width of the tissue. The plot obtained of thickness versus width position is integrated to provide the cross-sectional area. This area is accurate to within 5 percent, depending mainly on alignment of the instrument and pre-load of the ligament. Results on the mid-substance of the rabbit medial collateral ligaments are repeatable and reproducible. Values of maximum width and thickness are less variable than those obtained with a vernier caliper. The measured area is considerably less than that estimated assuming rectangular cross-section and slightly less than that estimated on the assumption of elliptical cross-section.     

Paravascular connective tissue structures of the veins of the pelvic cavity in man

The data on structure of the paravasal connective tissue formations in some parietal pelvic veins, in the utero-vaginal plexus veins, in the fatty tissue space veins of the rectum, of the vesicular and prostatic plexuses are presented, as well as those on connections between the fascial vaginae for the veins in the human subabdominal part of the pelvis and its fascial nodes. Theoretical interpretation of gaping of the pelvic fundal venous vessels is presented. Participation of the paravasal connective tissue formations in organization of the fascial nodes of the human small pelvis is stated.     

Finite element model of subsynovial connective tissue deformation due to tendon excursion in the human carpal tunnel

Carpal tunnel syndrome (CTS) is a nerve entrapment disease, which has been extensively studied by the engineering and medical community. Although the direct cause is unknown, in vivo and in vitro medical research has shown that tendon excursion creates microtears in the subsynovial connective tissue (SSCT) surrounding the tendon in the carpal tunnel. One proposed mechanism for the SSCT injury is shearing, which is believed to cause fibrosis of the SSCT. Few studies have reported quantitative observations of SSCT response to mechanical loading. Our proposed model is a 2-D section that consists of an FDS tendon, interstitial SSCT and adjacent stationary tendons. We believe that developing this model will allow the most complete quantitative observations of SSCT response to mechanical loading reported thus far. Boundary conditions were applied to the FEA model to simulate single finger flexion. A velocity was applied to the FDS tendon in the model to match loading conditions of the documented cadaver wrist kinematics studies. The cadaveric and FEA displacement results were compared to investigate the magnitude of stiffness required for the SSCT section of the model. The relative motions between the model and cadavers matched more closely than the absolute displacements. Since cadaveric models do not allow identification of the SSCT layers, an FEA model will help determine the displacement and stress experienced by each SSCT layer. Thus, we believe this conceptual model is a first step in understanding how the SSCT layers are recruited during tendon excursion.     

Fibroblast cytoskeletal remodeling contributes to connective tissue tension

The visco-elastic behavior of connective tissue is generally attributed to the material properties of the extracellular matrix rather than cellular activity. We have previously shown that fibroblasts within areolar connective tissue exhibit dynamic cytoskeletal remodeling within minutes in response to tissue stretch ex vivo and in vivo. Here, we tested the hypothesis that fibroblasts, through this cytoskeletal remodeling, actively contribute to the visco-elastic behavior of the whole tissue. We measured significantly increased tissue tension when cellular function was broadly inhibited by sodium azide and when cytoskeletal dynamics were compromised by disrupting microtubules (with colchicine) or actomyosin contractility (via Rho kinase inhibition). These treatments led to a decrease in cell body cross-sectional area and cell field perimeter (obtained by joining the end of all of a fibroblast's processes). Suppressing lamellipodia formation by inhibiting Rac-1 decreased cell body cross-sectional area but did not affect cell field perimeter or tissue tension. Thus, by changing shape, fibroblasts can dynamically modulate the visco-elastic behavior of areolar connective tissue through Rho-dependent cytoskeletal mechanisms. These results have broad implications for our understanding of the dynamic interplay of forces between fibroblasts and their surrounding matrix, as well as for the neural, vascular, and immune cell populations residing within connective tissue.     

Hydrogen peroxide is a novel inducer of connective tissue growth factor.

Connective tissue growth factor (CTGF) has recently been described as a fibrogenic factor and is greatly induced by various extracellular stimuli, such as transforming growth factor-beta (TGF-beta), dexamethasone, and serotonin. CTGF induces collagen type I and fibronectin, and the deposition of such molecules leads to fibrotic disease in many tissues. Intracellular reactive oxygen species (ROS) are generated by extracellular stress conditions and are produced as by-products of cellular metabolism. Imbalanced cellular redox status is a potent pathogenic factor that leads to various degenerative diseases, including tissue fibrosis. Since CTGF is believed to play a crucial role in fibrotic disease formation in many tissues, we examined the role of ROS in CTGF gene expression in human lens epithelial cell line B3. The results showed that CTGF was induced by reactive oxygen species such as hydrogen peroxide and hydroxyl radicals. Next, we examined whether CTGF induction by ROS is via newly synthesized TGF-beta. The results showed that ROS directly induced CTGF mRNA not via the increased TGF-beta synthesis or activation. Next, we treated AG490, which is the well-known inhibitor of Janus kinase (JAK), with hydrogen peroxide. AG490 abrogated the CTGF induction by ROS in a dose-dependent manner. The results suggest that JAK-2/-3 seems to be involved in the enhanced CTGF mRNA expression by hydrogen peroxide. In this report, we present that hydrogen peroxide is a novel inducer of CTGF gene expression and that JAK-2/-3 activation seems to play a role in CTGF induction.