Telocytes in human heart valves

Valve interstitial cells (VICs) are responsible for maintaining the structural integrity and dynamic behaviour of the valve. Telocytes (TCs), a peculiar type of interstitial cells, have been recently identified by Popescu's group in epicardium, myocardium and endocardium (visit www.telocytes.com ). The presence of TCs has been identified in atria, ventricles and many other tissues and organ, but not yet in heart valves. We used transmission electron microscopy and immunofluorescence methods (double labelling for CD34 and c‐kit, or vimentin, or PDGF Receptor‐β) to provide evidence for the existence of TCs in human heart valves, including mitral valve, tricuspid valve and aortic valve. TCs are found in both apex and base of heart valves, with a similar density of 27–28 cells/mm² in mitral valve, tricuspid valve and aortic valve. Since TCs are known for the participation in regeneration or repair biological processes, it remains to be determined how TCs contributes to the valve attempts to re‐establish normal structure and function following injury, especially a complex junction was found between TCs and a putative stem (progenitor) cell.     

Mechanical aspects of the development of artificial heart valves

The development of an antithrombogenic coating permits a hybrid design for artificial heart valves. A substrate material optimized for its application is coated to meet the electrochemical requirements of improved hemocompatibility. But future progress in artificial heart valves requires an improvement in design as well as of the material. The basis of both aspects is the determination of such fundamental mechanical properties as the elasticity and plasticity of the valve ring and the deformation and fraction behaviour of the occluder. Analytical and numerical calculations of various different models result in different requirements for the substrate of ring and occluder. A combination of high elastic temper and low resistance to flow requires a ring material with a Young's modulus of 40 GPa or more, and a 0.2% proof stress to (Young's modulus)2/3 ratio of 0.3 MPa1/3. The best occluder materials should have a Young's modulus of more than 50 GPa and a flexural strength of at least 800 MPa. On the basis of these criteria, a heart valve consisting of a TiA15Fe2,5 ring and occluders made partially stabilized zirconia is introduced.     

Explaining embryological development: Should integration be the goal?

Two approaches to an integration of evolution and development are often distinguished, one “neo-Darwinian” and the other “structuralist”. Should these approaches in turn be integrated? Kelly Smith recently stated that we need a “more complete” theory of biological order, suggesting integration as the ideal. In response to him, I argue that a recognition of different types of scientific questions and causal explanation is more urgent. Do we understand development when we know the crucial factors in the process of differentiation, or rather when we know the laws that govern the transformations of fields? Without a recognition of these different explanatory ideals, “integration” is likely to have the character of annexation.     

Molecules mediating cell-ECM and cell-cell communication in human heart valves

The specific phenotype of different tissues depends on the interactions of cells with neighboring cells and the surrounding extracellular matrix, which is mediated by cell adhesion receptors including integrins, immunoglobulin family members, syndecans, and selectins. The aim of this study was to investigate the adhesion profile of native human valve interstitial cells (ICs) in situ and in vitro by analyzing these adhesion receptors. Flow cytometry and immunocytochemistry was used to quantify the expression of the specific receptors on ICs cultured from all human cardiac valves, and immunohistochemistry were used to profile their distribution pattern in valve tissue sections. The valve leaflets and cultured ICs from all valves expressed alpha1, alpha2, alpha3, alpha4, and alpha5 integrins to varying degrees and percentages with very little expression of alpha6 and alphaV. Valve leaflet ICs from all valves, expressed predominantly beta1 integrin but no beta3 or beta4 integrin. Syndecan-1 and Syndecan-4 were not detected. Intercellular adhesion molecule-1 was weakly detected, whereas vascular adhesion molecule-1 was barely detectable and E-selectin was not detected. This study has delineated the identity of some of the integrins synthesized and expressed by human valve ICs and the specificity of adhesion molecules with which the valve ICs interact with the extracellular matrix and mediate intercellular interactions. This pattern of expression of cell surface adhesion molecules may be considered as a basis for a fingerprint on which to base future cell alternatives and would provide useful information for valve tissue engineering.     

Atomic force microscopy and FT-IR spectroscopy investigations of human heart valves

Human aortic, mitral, tricuspid and pulmonary heart valves were investigated by the contact mode atomic force microscopy (AFM) in air, and using FT-IR spectroscopy in the frequency range 950-4000 cm(-1). Heart valves were collected post mortem from 65-78 years old patients who died from non-cardiac diseases. All of the examined valves showed considerable heterogeneity in the surface topography of collagen fibrils as well as in their organization on the tissue surface. The AFM images revealed areas with significantly different spatial organization of the collagen fibril bundles. We observed zones with multidirectional, stacked collagen fibrils as well as areas of thin fibrils packed regularly, densely and "in phase". The majority of the collagen fibrils reproduced the typical transverse D-banding pattern, with the band interval varying in rather wide range of 70-90 nm. Using AFM imaging, objects that correspond to some pathological states of heart valves at their early stages, i.e. some forms of mineral deposits, were observed. The FT-IR spectra allowed us to recognize main components, i.e. collagen and elastin, in di.erent layers (ventricularis, fibrosa) of the valve leaflets as well as they gave also support for the presence of mineral deposits on the valve surface. The presented results showed, that the AFM imaging and FT-IR spectroscopy can be applied as a complementary methods for structural characterization of heart valves at the molecular and supramolecular levels.     

Collagen composition of normal and myxomatous human mitral heart valves.

The collagens were studied in 13 normal and 19 myxomatous human mitral valves. The collagens of the valve were completely solubilized by using a method consisting of guanidinium chloride extraction, limited pepsin digestions and CNBr cleavage of the residue. The normal valves contained 74% type I, 24% type III and 2% type V collagen. The type I and type III collagens had similar solubility patterns, although only type I collagen was detected in the guanidinium chloride extract. Type V collagen was only detected in the first pepsin extract. The type I and III collagens had higher contents of hydroxylysine than did the same collagens from age-matched dermis. The two-dimensional electrophoretic 'maps' of CNBr-cleavage peptides showed low recoveries of the C-terminal alpha 1(I) CB6 and alpha 1(III) CB9 peptides, which are involved in forming intermolecular cross-linkages. Most of the reducible cross-linkages were present in large-Mr peptide complexes, and these complexes were shown by labelling with 125I to include the tyrosine-containing alpha 1(I) CB6 peptide. The myxomatous valves contained 67% type I, 31% type III and 2% type V collagens. There was a significant increase in the concentration of each type of collagen, which consisted of a 9% increase of type I collagen, a 53% increase of type III collagen and a 25% increase of type V collagen. The contents of hydroxylysine in type I and III collagens and the electrophoretic 'maps' of the CNBr-cleavage peptides involved in cross-linkages did not differ significantly from the results obtained from the normal valves. The biochemical findings suggest that there is an increased production of collagen, in particular type III collagen, and glycosaminoglycan as well as a proliferation of cells as part of a repair process in the myxomatous valves.     

The effect of liquid nitrogen submersion on cryopreserved human heart valves

Cryopreservation and storage of human heart valves have become an accepted means of maintaining a usable supply of heart valves for outflow track reconstructive surgery. Valves are typically stored at the vapor phase temperature of liquid nitrogen, -130 degrees C and below, to reduce the chance of recrystallization within the tissues. Concern over the effects of submersion of the valves in liquid nitrogen, i.e., plunging to -196 degrees C, prompted this study. Cryopreserved valves were plunged into liquid nitrogen, held for 5 min, and then processed (thawed) by standardized protocols. The thawed valves were then assessed using scanning electron microscopy and the more traditional histology at the light microscope level. Cuspal tissues plunged into liquid nitrogen appear to have numerous microfractures over both surfaces of the tissue, penetrating into the collagen/proteoglycan matrix. Control cryopreserved valves do not exhibit these microfractures. Histologically, the submerged valves appear normal. The clinical use of valves which have been submerged in liquid nitrogen is discussed.     

The appearance of ganglioside during embryological development of the frog

Embryological stages after fertilization were examined for the appearance of gangliosides. In Rana pipiens, gangliosides were found to appear first at the late gastrula, a stage well before any morphological appearance of a nervous system. Furthermore, at this stage and at a later stage, hatching, only two of the four major gangliosides were detected, namely, monosialoganglioside G_M1 and disialoganglioside G_D1a.     

Vascular-nervous relationships in the valves of the heart]

Method of silver nitrate impregnation was used in order to study 50 preparations of not-changed atrioventricular valves of the heart of domestic bulls and 30 preparations of the same valves of adult humans. It has been shown that in heart valves there are certain relationships between striated muscle fibres, blood vessels and nerve elements. The nerve structures of the valves are represented by nerve bundles of different thickness. In their composition there are comparatively thin non-myelinated and thicker myelinated fibres. Towards the free edge of cusps the nerve bundles become thinner and the nerve trunks give off separate thin nerve fibres disposed along the vessels of a capillary type and in some places getting around them. In certain portions of cusps the nerve bundles, some of which have zigzag sinuosity, cross blood vessels in different directions. In man the major mass of blood vessels and nerve elements are disposed near the base of the valve cusps, accompanying the muscle fibre bundles penetrating from the base side. In the bull heart valves an amount of blood vessels and nerve elements is found in considerable portions of the cusps not connected with muscle fibres.     

Sculpting the heart: Cellular mechanisms shaping valves and trabeculae

The transformation of the heart from a simple tube to a complex organ requires the orchestration of several morphogenetic processes. Two structures critical for cardiac function, the cardiac valves and the trabecular network, are formed through extensive tissue morphogenesis—endocardial cell migration, deadhesion and differentiation into fibroblast-like cells during valve formation, and cardiomyocyte delamination and apico-basal depolarization during trabeculation. Here, we review current knowledge of how these specialized structures acquire their shape by focusing on the underlying cellular behaviors and molecular mechanisms, highlighting findings from in vivo models and briefly discussing the recent advances in cardiac cell culture and organoids.