Chondrocyte viability is higher after prolonged storage at 37 degrees C than at 4 degrees C for osteochondral grafts.

Background: Osteochondral allografts are currently stored at 4 degrees C for 2 to 6 weeks before implantation. At 4 degrees C, chondrocyte viability, especially in the superficial zone, deteriorates starting at 2 weeks. Alternative storage conditions could maintain chondrocyte viability beyond 2 weeks, and thereby facilitate increased graft availability and enhanced graft quality.

Articular cartilage tensile integrity: modulation by matrix depletion is maturation-dependent.

Articular cartilage function depends on the molecular composition and structure of its extracellular matrix (ECM). The collagen network (CN) provides cartilage with tensile integrity, but must also remodel during growth. Such remodeling may depend on matrix molecules interacting with the CN to modulate the tensile behavior of cartilage.

Cartilage growth and remodeling: modulation of balance between proteoglycan and collagen network in vitro with beta-aminopropionitrile.

Objective: To examine the effect of beta-aminopropionitrile (BAPN), an inhibitor of lysyl oxidase, on growth and remodeling of immature articular cartilage in vitro.

Design: Immature bovine articular cartilage explants from the superficial and middle layers were cultured for 13 days in serum-containing medium with or without BAPN. Variations in tissue size, accumulation of proteoglycan and collagen (COL), and tensile mechanical properties were assessed.

Differential regulation of proteoglycan 4 metabolism in cartilage by IL-1alpha, IGF-I, and TGF-beta1.

Objectives: To determine (1) if interleukin-1 alpha (IL-1alpha), insulin like growth factor I (IGF-I), and transforming growth factor-beta 1 (TGF-beta1) regulate proteoglycan 4 (PRG4) metabolism in articular cartilage, in terms of chondrocytes expressing PRG4 and PRG4 bound at the articular surface, and (2) if these features of cartilage PRG4 metabolism correlate with its secretion.

Methods: Articular cartilage explants were harvested and cultured for 6 days with or without 10% fetal bovine serum (FBS), alone, or with the addition of 10ng/ml IL-1alpha, 300ng/ml IGF-I, or 10ng/ml TGF-beta1. PRG4 expression by chondrocytes in the cartilage disks was assessed by immunohistochemistry (IHC).

PRG4 exchange between the articular cartilage surface and synovial fluid.

The boundary lubrication function of articular cartilage is mediated in part by proteoglycan 4 (PRG4) molecules, found both in synovial fluid (SF) and bound to the articular cartilage surface. Currently the mechanism by which PRG4 binds to the articular surface is not well understood. The objectives of this study were to determine (1) the effect of bathing fluid contents on PRG4 concentration at the articular surface ([PRG4](cart)), and (2) whether native PRG4 can be removed from the surface and subsequently repleted with PRG4 from synovial fluid.

Directional shear flow and Rho activation prevent the endothelial cell apoptosis induced by micropatterned anisotropic geometry.

To study the roles of anisotropic cell morphology and directionality of mechanical force in apoptosis, the spreading of human umbilical vein endothelial cells (HUVECs) was constrained by growing on micropatterned (MP) strips of fibronectin (FN, 20 microg/cm2) with widths of 15, 30, and 60 microm on silicone membrane. Cells on 30- and 60-microm strips, like cells on a nonpatterned (NP) surface coated with FN, showed clear actin stress fibers with anchoring spots of phosphorylated focal adhesion kinase (p-FAK) and no significant apoptosis. On 15-microm strips, cells had few stress fibers, no p-FAK, and significant apoptosis.

Effect of synovial fluid on boundary lubrication of articular cartilage.

Objectives: The lubrication of articulating cartilage surfaces in joints occurs through several distinct modes. In the boundary mode of lubrication, load is supported by surface-to-surface contact, a feature that makes this mode particularly important for maintenance of the normally pristine articular surface. A boundary mode of lubrication is indicated by a kinetic friction coefficient being invariant with factors that influence formation of a fluid film, including sliding velocity and axial load.

Regulation of mTOR by mechanically induced signaling events in skeletal muscle.

Mechanical stimuli play a major role in the regulation of skeletal muscle mass, and the maintenance of muscle mass contributes significantly to disease prevention and the quality of life. Although a link between mechanical stimuli and the regulation of muscle mass has been recognized for decades, the mechanisms involved in converting mechanical information into the molecular events that control this process have not been defined. Nevertheless, significant advancements are being made in this field, and it has recently been established that signaling through a rapamycin-sensitive pathway is necessary for mechanically induced growth of skeletal muscle.

The role of phospholipase D and phosphatidic acid in the mechanical activation of mTOR signaling in skeletal muscle.

Signaling by the mammalian target of rapamycin (mTOR) has been reported to be necessary for mechanical load-induced growth of skeletal muscle. The mechanisms involved in the mechanical activation of mTOR signaling are not known, but several studies indicate that a unique [phosphotidylinositol-3-kinase (PI3K)- and nutrient-independent] mechanism is involved. In this study, we have demonstrated that a regulatory pathway for mTOR signaling that involves phospholipase D (PLD) and the lipid second messenger phosphatidic acid (PA) plays a critical role in the mechanical activation of mTOR signaling.

Mechanical stimuli and nutrients regulate rapamycin-sensitive signaling through distinct mechanisms in skeletal muscle.

The mammalian target of rapamycin (mTOR) has been identified as a growth factor and nutrient-sensitive molecule that controls the translational machinery and cell growth. Rapamycin-sensitive (RS) signaling events have also been shown to be necessary for mechanical load-induced growth of skeletal muscle, but the mechanisms involved in the mechanical activation of RS signaling are not known. The finding that mechanical stimuli induce nutrient uptake in skeletal muscle raises the possibility that mechanically induced RS signaling is mediated via a nutrient-dependent mechanism.

Molecular basis of mechanical modulation of endothelial cell migration.

Vascular endothelial cells (ECs) play important roles in the regulation of vascular functions. Loss of endothelial integrity can lead to vascular diseases such as stenosis resulting from atherosclerosis. The migration of ECs into wounded area in the vessel wall is required for the restoration of its integrity and functions.

Site- and exercise-related variation in structure and function of cartilage from equine distal metacarpal condyle.

Objective: Determine (1) the site-associated response of articular cartilage of the equine distal metacarpal condyle to training at a young age as assessed by changes in indentation stiffness and alterations in cartilage structure and composition, and (2) relationships between indentation stiffness and indices of cartilage structure and composition.

Method: Experimental animals (n=6) were trained on a track (increasing exercise to 1km/day by 5 months); controls (n=6) were pasture-reared. Animals were euthanized at 18 months and four osteochondral samples were harvested per metacarpal condyle from dorsal-medial, dorsal-lateral, palmar-medial, and palmar-lateral aspects.

Mechanical characterization of native and tissue-engineered cartilage.

Cartilage functions as a low-friction, wear-resistant, load-bearing tissue. During a normal gait cycle, one cartilage surface rolls and slides against another, all the while being loaded and unloaded. The durability of the tissue also makes for an impressive material to study.

In vitro physical stimulation of tissue-engineered and native cartilage.

Because of the limited availability of donor cartilage for resurfacing defects in articular surfaces, there is tremendous interest in the in vitro bioengineering of cartilage replacements for clinical applications. However, attaining mechanical properties in engineered cartilaginous constructs that approach those of native cartilage has not been previously achieved when constructs are cultured under free-swelling conditions. One approach toward stimulating the development of constructs that are mechanically more robust is to expose them to physical environments that are similar, in certain ways, to those encountered by native cartilage.

Growth of immature articular cartilage in vitro: correlated variation in tensile biomechanical and collagen network properties.

Articular cartilage biochemical composition and mechanical properties evolve during in utero and in vivo growth, with marked differences between fetus, newborn, and young adult. The objectives of this study were to test whether in vitro growth of bovine fetal and newborn calf articular cartilage explants resulted in changes in biochemical and tensile properties during up to 6 weeks of free-swelling culture in serum-supplemented medium. During this culture period, both fetal and calf cartilage grew markedly in size, increasing in dry and wet mass by 150-270%.

Tensile mechanical properties of bovine articular cartilage: variations with growth and relationships to collagen network components.

One approach to repairing articular defects is to regenerate cartilage by recapitulating the changes that occur during fetal and postnatal growth into adulthood, and to thereby restore functional biomechanical properties, especially those of the normally strong superficial region. The objectives of this study were (1) to characterize and compare tensile biomechanical properties of the superficial region of articular cartilage of the patellofemoral groove (PFG) and femoral condyle (FC) from bovine animals over a range of growth stages (third-trimester fetal, 1-3 week-old calf, and adult), and (2) to determine if these properties were correlated with collagen network components. With growth from the fetus to the adult, the equilibrium and dynamic tensile moduli and strength of cartilage samples increased by an average of 391-1060%, while the strain at the failure decreased by 43%.

Adhesion of perichondrial cells to a polylactic acid scaffold.

The number of chondrogenic cells available locally is an important factor in the repair process for cartilage defects. Previous studies demonstrated that the number of transplanted rabbit perichondrial cells (PC) remaining in a cartilage defect in vivo, after being carried into the site in a polylactic acid (PLA) scaffold, declined markedly within two days. This study examined the ability of in vitro culture of PC/PLA constructs to enhance subsequent biomechanical stability of the cells and the matrix content in an in vitro screening assay.

Development-associated differences in integrative cartilage repair: roles of biosynthesis and matrix.

A recurring problem in tissue transplantation therapies for articular cartilage defects is the lack of integration between the implant and the host cartilage. Previous studies have shown that in vitro integration between explants of calf cartilage is markedly higher than that between fetal cartilage, despite similarly high levels of deposition of newly synthesized collagen. The aim of this study was to determine if cellular biosynthesis and extracellular matrix each contribute to these development-associated differences in integrative repair in vitro.

Interplay between integrins and FLK-1 in shear stress-induced signaling.

Blood flow can modulate vascular cell functions. We studied interactions between integrins and Flk-1 in transducing the mechanical shear stress due to flow. This application of a step shear stress caused Flk-1.

Induction of apoptosis in vascular smooth muscle cells by mechanical stretch.

We studied the response of porcine vascular smooth muscle cells (PVSMCs) to cyclic sinusoidal stretch at a frequency of 1 Hz. Cyclic stretch with an area change of 25% caused an increase in PVSMC apoptosis, which was accompanied by sustained activation of c-Jun NH(2)-terminal kinases (JNK) and the mitogen-activated protein kinase p38. Cyclic stretch with an area change of 7% had no such effect.

Shear stress activation of SREBP1 in endothelial cells is mediated by integrins.

We investigated the effect of shear stress on the sterol regulatory element-binding protein 1 (SREBP1) in vascular endothelial cells (ECs) and the mechanotransduction mechanism involved. Application of a shear stress (12 dyn/cm(2)) caused the proteolytic cleavage of SREBP1 and the ensuing translocation of its transcription factor domain into the nucleus. As a result, shear stress increased the mRNAs encoding the low density lipoprotein receptor (LDLR), as well as the binding of (125)I-LDL.

Similarity of stress distribution in bone for various implant surface roughness heights of similar form.

Background: Surface roughness effects on osseointegration can be considered from two viewpoints: purely mechanical effects of stresses attributable to roughness and cell and molecular response to surface roughness.

Purpose: The goal of this study is to provide a theoretical basis to understand the effects of surface roughness size on the osseointegration of implants. The emphasis is primarily on the purely mechanical effects.

Interaction of force-fitting and surface roughness of implants.

Background: Increased surface roughness may increase installation torque and thus appear to increase the initial stability of an implant. However, it is not immediately clear if the increased torque is attributable to an increase in the effective diameter of the implant or to increased resistance of the bone because of the greater roughness.

Purpose: Force-fitting stresses arise when an implant is placed into a predrilled hole of smaller-diameter in bone.

Development of hemorheology: perspective in instrumentation development.

New viscometers for blood viscometry, improved intravital microscope, and related instruments, which are capable of measuring important rheological factors for microcirculatory research were developed and applied for hemorheological studies. As the results, four major determinants of the suspension viscosity were determined and the role of suspension viscosity of the blood as a function of microcirculatory flow was clarified. Meanwhile, the use of fluorescence microscopy and digitized video microscopic techniques has allowed the investigation of the structure and function of cells at the level of the single intact living cell.