Apoptosis and Calcification

Calcification in necrosis has long been known. Of the tissue components, the cells are most vulnerable. Nevertheless, little attention has been paid to the role of cell death in calcification. This review attempts to update the mechanism of calcification with an emphasis on the role of apoptosis in calcification. A brief review on the basic sciences relevant to calcification is followed by a discussion of abnormal Ca2+ and Pi homeostasis in cell injury and apoptosis. Concomitant increases in Ca2+ and Pi in blebs (and matrix vesicles) formed by apoptotic and/or necrotic cells are apparently the primary mechanism of calcification. In addition, membranous cellular degradation products (CDP) resulting from cell disintegration in toto frequently serve as the nidus of calcification. Published data on physiological calcification are compared with findings in various dystrophic calcinoses. This led to the conclusion that apoptosis most likely underlies the mechanism of both physiological and pathological calcifications. It is concluded that calcification is an important function of apoptosis. The mechanism of calcification by CDP and morphology of the resultant calcific deposits are complex

Determinants of Weddellite Formation: Chondroitin Sulfates and Citrate Determine Weddellite Formation In Vitro

In synthetic urine (SU), addition of oxalate tends to form monohydrates of calcium oxalate. However, addition of oxalate to natural urine preferably forms calcium oxalate dihydrate (COD). Urine apparently contains a determinant for COD formation. To identify the determinant, the effects of pH, temperature, oxalate, calcium, urate, citrate, magnesium, sulfate and chondroitin sulfates (CS) on calcium oxalate crystal formation were studied. Lower temperatures, higher oxalate concentrations and higher pH favored COD formation in a SU. Mixed CS in the presence of citrate were the most decisive determinant of COD formation. Substitution of CS for agar and gelatin produced similar results, indicating that the colloidal effect of the macromolecules deter-mines COD formation. Identification of the determinants led to a simple, reproducible method of COD formation in SU without natural urine. Addition of strontium to SU resulted in dodecahedral bipyramids. Inter-penetration twinning of bi pyramids occur within seconds of the crystal formation

Calcification of Liposomes and Red Cell Ghosts In Vitro and In Vivo

To study the role of membranous phospholipids in calcification , liposomes made of phosphatidylserine and red blood cell ghosts (RBCG) prepared from rat and canine blood were incubated in minimal essential medium (MEM) with 2 .5 mM (mmol/l) calcium and 1.3 mM phosphate, pH 7.4 (MEM-2.5), and homologous (rat) or autologous (canine) plasma filtrate (serum) at 37°C for up to 1 week. Calcification was determined by electron probe microanalysis , electron diffraction, and depletions of calcium and phosphate from MEM-2.5 and sera. Liposomes and rat-RBCG incubated in MEM-2.5 and serum calcified in a week, whereas isolated collagen and elastin did not. Liposomes and rat-RBCG implanted in rat peritoneal cavities calcified in 4 weeks. Calcification of both liposomes and RBCG under identical conditions suggests that phospholipids in membrane may play a role in calcification. Canine RBCG incubated in MEM-2.5 and serum began to calcify on day one and grew heavier with further incubations . Calcification of.RBCG in autologous serum indicates that calcification is prevented by normal red cells. The capacity to prevent calcification by red cells is apparently abolished by removal, during RBCG preparation, of the cytoplasmic content

The Growth of Fetal Human Sensory Ganglion Neurons in Culture: A Scanning Electron Microscopic Study

Sensory neurons of 8-week human fetal dorsal root ganglia were dissociated into single cells by trypsinization and cultured on coverslips for 4 months, in either serum-containing or serum-free chemically defined media. At different times the cultures were fixed and prepared for scanning electron microscopy. Fetal sensory neurons in culture regenerated axons within 24 hours which were characterized by axonal growth cones at their tips and the neuronal perikarya assumed spherical or a bonnet-like morphology

Survival and Growth of Adult Human Oligodendrocytes in Culture: Scanning Electron Microscopy

The oligodendrocytes in culture serve as an important model for the study of demyelination diseases. We have previously originated a method of isolating human oligodendrocytes. In order to establish their three dimensional morphology, scanning electron microscopy of the cultured oligodendrocytes was performed. The oligodendrocytes bulk isolated from adult human brain were maintained in culture for more than 2 months. At the various periods in culture, the cells were studied by scanning electron microscopy and immunofluorescence staining using marker antibodies for the identification of oligodendrocytes. The three dimensional organization and the surface morphology of the cultured oligodendrocytes were investigated. They displayed an extensive network of the cell processes and characteristic surface morphology

Calcification in Aging Canine Aortic Valve

Aging changes of aortic valves are thought to underlie the mechanism of calcification, which leads to calcific aortic stenosis in humans. The study of calcification in the aging valvular connective tissue has been hindered by the lack of a suitable animal model. In search of the model, canine aortic valves demonstrated age changes including calcification remarkably similar to those in humans. The mechanism of calcification was studied in the aortic valves of aged Beagles by electron microscopy. Fibroblasts in the canine aortic valves showed the most prominent age changes. The cells accumulated numerous residual bodies and appeared to disintegrate. The resultant membranous cellular degradation products which sequestered in the extracellular space were the nidi of calcification. It appeared that the membrane of cell debris played an important role in calcification. Canine aortic valve is an ideal model for the study of calcification in relation to aging of the valvular connective tissue