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Biomedical subjects

M Silbermann

Publications and source records attributed to M Silbermann.

At least 109 records · Page 6Linked to original sources
PubMed1986

A tissue culture system supporting cartilage cell differentiation, extracellular mineralization, and subsequent bone formation, using mouse condylar progenitor cells.

Undifferentiated progenitor cells of mandibular condyles of neonatal mice were kept in a tissue culture system for up to 9 days. After 2 days in culture, new chondroblasts developed within the explants, whereas the peripheries of the latter were occupied by undifferentiated cells undergoing mitosis. By 4 days in culture, many of the cartilage cells showed signs of hypertrophy, while the matrix revealed positive reactivity for type II collagen and matrix metachromasia. The process of maturation of cartilage cells appeared to have reached its final stages after 6 days in culture, at a time when the initial loci of matrix mineralization could be easily identified. Concomitantly, peripheral areas bordering the cartilaginous core, as well as portions of the cartilage, reacted positively for type I collagen and fibronectin. Light microscopy examination showed signs of new bone formation after 9 days in culture. The extracellular matrix at the upper portion of the explant, facing the medium-air interface, reacted intensely with antibodies against type I collagen and fibronectin, but not with antibodies against type II collagen. Further, the newly formed osteoid was found to have undergone mineralization, thus forming an expanded layer of new bony tissue. A close spatial association was found between mature, mineralized cartilage and new bone. Hence, we hereby introduce a new in vitro system serving as an experimental model for studies related to the differentiation of progenitor cells into chondroblasts, which in turn promote ossification.

Animals↗
PubMed1986

Structural and metabolic changes characterizing the aging of various cartilages in mice.

It is well documented that various organs and tissues in the body show a differential, non-homogeneous pattern of development and aging. The present study evaluates some of the morphological and metabolic changes characterizing the aging of different types of cartilages in normal male mice. Representatives of hyaline, fibrous, articular and elastic types of cartilage were obtained from animals ranging from 1 week to 1 year of age. Our determinations included the following: total body and organ weights, proteins and DNA concentrations, [3H]leucine and [35S]sulfate uptake. The biochemical assays were accompanied by morphological examinations of corresponding tissue specimens. This investigation clearly indicates that the growth and maturational activities of the various cartilages examined attained their completion at a very early stage of life (1-3 months postnatally). Thereafter, a phase of steady state prevails, followed by a gradual but continuous decline in the various metabolic activities. The morphological findings illustrate the nature of the age-related structural changes. The present findings indicate that a skeletal tissue such as cartilages of various types, tends to age early during the lifespan of the animal.

Aging↗
PubMed1986

Dexamethasone and 8-bromo-cyclic AMP depress the incorporation of [3H]thymidine into mouse condylar cartilage by different pathways.

We examined whether cyclic AMP (cAMP) affects the incorporation of [3H]thymidine into cartilage cells and, if so, whether this action could be related to the inhibitory effect of glucocorticoid hormones on the growth of ossifying cartilage. Incorporation of [3H]thymidine into trichloroacetic acid-precipitable material by mouse cartilage was measured concomitantly with the concentration of cAMP. Dexamethasone (1 mumol/l) significantly (P less than 0.05) depressed the incorporation of [3H]thymidine. The cAMP analogue 8-bromo-cAMP (0.01-1 mmol/l) also depressed the incorporation of the radionucleotide in a dose-dependent fashion. When various concentrations of 8-bromo-cAMP were added with dexamethasone (1 mumol/l), no apparent changes took place compared with the effect of dexamethasone alone. The phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (0.2-1 mmol/l) elicited an inhibitory effect on [3H]thymidine incorporation and a stimulatory influence on cartilage cAMP concentrations. Dexamethasone, at doses (0.01-1 mumol/l) causing significant inhibition of [3H]thymidine incorporation, failed to increase cartilage levels of cAMP. It seems, therefore, that the depressive effect of dexamethasone on [3H]thymidine incorporation in condylar cartilage is not mediated through an increase of cAMP in the tissue.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1986

Further characterization of spontaneous arthritic changes in murine squamo-mandibular joint: histopathological aspects.

The appearance of age-related ulcerative changes in the mouse mandibular condyle were evaluated by light and electron microscopy examinations. Fibrillations appeared along the articular surface and in deeper tissue regions, as early as at six months of age. Such changes were characterized by a marked loss of the tissue's cellularity and by a marked reduction in matrix metachromasia and safranin-0 staining. These microscopical changes were accompanied by a reduced reactivity for both ruthenium red and colloidal iron binding, as noted ultrastructurally. At the same time, increasing numbers of erythrocytes appeared to be adhered to the surface irregularities and were also found in deeper regions within the articular lesions. Using morphological criteria, it became apparent that the degenerative changes of aging articular cartilage started at the more superficial regions of the tissue and only thereafter proceeded toward the chondro-osseous junction. Also, with the advancement of age, the degenerative changes became more severe.

Aging↗
PubMed1986

Morphology and in vivo growth characteristics of an atypical murine proliferative osseous lesion induced in vitro.

Mandibular condyles of late embryonic NMRI mice were used to study the effect of the FBR murine osteosarcoma virus in an in vitro tissue culture system. Chondroprogenitor cells and chondroblastic cells present in the condylar tissue normally undergo rapid differentiation in vitro which results in an advanced stage of bone formation. The infection of condyles with FBR murine osteosarcoma virus induced the transformation of bone progenitor cells and the formation of an atypical proliferative osseous lesion. In markedly disorganized tissue many spindle-like cells, giant cells, and pleomorphic cells were seen together with the formation of large bone spicules and the heavy mineralization of osteoid-like material and of the remaining cartilage. Fibroblast-like cells were found to penetrate from the perichondrial zone into the condylar mass and also into the underlying collagen sponge. The in vivo growth characteristics of FBR murine osteosarcoma virus-infected condyles after 3 days in culture were studied via s.c. transplantation into syngeneic mice. Control condyles developed normal trabecular bone, whereas the infected condyles induced a strong cellular response with the presence of atypical cells and newly formed connective tissue and bone in situ. These observations raise the possibility of a novel approach for further investigations related to numerous aspects of virus-induced osteosarcomagenesis.

Animals↗
PubMed1986

Effects of systemic glucocorticoids on the degradation of glycosaminoglycans in the mandibular condylar cartilage of newborn mice.

This investigation studied the early in vivo effects of triamcinolone hexacetonide, a potent fluorinated analogue of cortisol, on the degradation of sulfated proteoglycans in condylar cartilage of newborn mice. While determining the rate of [35S]sulfate release in test and control specimens, it became evident that in hormone-treated animals there was a dose-dependent retardation of the isotope clearance. Further, triamcinolone was found to have increased the half-life of condylar glycosaminoglycans (GAGs) from 16 h in control animals to 31.4 h in hormone-treated ones. Dexamethasone, another fluorinated analogue of cortisol, and progesterone evoked a similar effect. On the other hand, hydrocortisone and cortisone induced a much milder effect, whereas the non-glucocorticoid steroid deoxycorticosterone did not affect the turnover of radiosulfate in this tissue. Clearance of the isotope from the serum was faster in the hormone-treated animals. Further, the hormone led to a decrease in the overall content of GAGs, a feature that lasted for 24 h. These data tend to imply that, concomitant with the inhibitory effects of glucocorticoids on proteoglycan synthesis in cartilage, they also induce a transient depressive effect upon the degradation of proteoglycans and thereby interfere with the normal growth and development of this cartilage.

Animals↗
PubMed1986

Studies on hormonal regulation of the growth of the craniofacial skeleton: IV. Specific binding sites for glucocorticoids in condylar cartilage and their involvement in the biological effects of glucocorticoids on cartilage cell growth.

Using a whole-tissue binding assay and cell-free binding measurements indicated the presence of a specific steroid receptor for triamcinolone in cartilage cells of neonatal mouse mandibular condyle. Analysis of receptor levels showed that whole-tissue preparations bound 1360 fmol triamcinolone/mg protein. Affinity measurements revealed a dissociation constant of 7.6 X 10(-9) M. There was a close correlation between triamcinolone inhibition of DNA synthesis and steroid occupancy of whole-tissue receptors. The inhibitory effect of triamcinolone upon DNA synthesis could be significantly reduced by "blocking" the respective receptors with cortexolone. All the cartilage cells in the condyle revealed distinct intracellular labeling using [3H] dexamethasone autoradiography. Hence, neonatal condylar cartilage, an active site of endochondral bone formation in the craniofacial skeleton, can be regarded as a genuine target tissue for the biological effects of glucocorticoids.

Animals↗
PubMed1985

Cartilage surface charge. A possible determinant in aging and osteoarthritic processes.

Polycationic labels such as cationized ferritin and colloidal iron were used to evaluate the surface negative charges over the mandibular condyles of ICR mice. The effects of neuraminidase, hyaluronidase, pronase, and collagenase on the binding of cationized ferritin and colloidal iron particles to the condylar articular surface were also studied. The results of this study clearly indicate that the surface area of the cartilaginous condyle is negatively charged and that its composition consists mainly of a collagenous material embedded within a proteinaceous matrix. With age, a substantial decrease in the density of negative charges took place along the surface area and, in particular, in the context of sialic acid residues. It is, therefore, possible that the reduction in cartilage surface charge might be associated with the onset of osteoarthritic changes commonly seen in aging humans and experimental animals.

Aging↗
PubMed1985

Morphologic and cytochemical changes in maturing and osteoarthritic articular cartilage in the temporomandibular joint of mice.

We studied the light microscopic, ultrastructural, and cytochemical characteristics of the temporomandibular joints of male ICR mice, from early neonatal life until they reached senescence, when spontaneous osteoarthritis is a common phenomenon. Aging of mandibular condylar cartilage was accompanied by decreasing total proteoglycan content and by an unmasking of collagen fibers, with no shift in collagen type. Fibronectin was also commonly present on the articular surface of specimens from old animals. Chondrocytes of aged mice contained an increased number of lysosomes, and their adjacent matrix vesicles reacted positively for acid phosphatase and arylsulfatase, but not for alkaline phosphatase. Such vesicles were also found to be devoid of calcium complexes and, thus, did not appear to be involved in the mineralization process. Similar age-related changes have been described in human mandibular condyles; hence, the male ICR mouse could serve as a useful model for studies of spontaneous osteoarthritis in the human mandibular joint.

Acid Phosphatase↗
PubMed1985

Evidence of Schwann cell degeneration in the aging mouse motor end-plate region.

An ultrastructural evaluation was made of the changes encountered in the motor endplates of the gastrocnemius muscle of old mice in comparison with young-adult mice. In old, 27-month-old mice, a great variety of changes were recorded. The main abnormalities occurred in the axon terminals, the myoneural junctional folds, and the endomysial space. Loss of synaptic vesicles and mitochondria and vacuolization of axon terminals were common. Shrinkage and degenerative changes of axon terminals were noticed. The end-plate abnormalities were closely associated with an increased amount of collagen tissue, degenerated Schwann cells, and cell debris. Degenerated Schwann cells were not encountered in young-adult mice. In the old group, endplates of normal appearance occurred in the proportion of 40%. The young group had normally structured end-plates in the proportion of 85%. The lesions described are not specific for the aging process.

Aging↗
PubMed1985

Structural study on the effect of 24,25-dihydroxyvitamin D3 on epiphyseal growth plate in suckling mice.

The in vivo effects of 24,25(OH)2D3 on cellular structure and organization, matrix metachromasia and mineralization were studied in epiphyseal growth plate of normal neonatal mice. A relatively low dose of the metabolite, 40 ng/kg body weight, significantly increased the overall size of humeral growth plate and the zone of cellular proliferation. By and large, the tissue's response to the metabolite did not change with the increase in dose administered except for a decrease in the number of chondroblasts. 24,25(OH)2D3 led to significant increases in the metachromatic reaction of the cartilaginous matrix, but appeared to depress the mineralization process. Qualitative structural changes were noted in chondroblasts and hypertrophic chondrocytes. 24,25(OH)2D3 affected the osteoblastic and osteocytic populations of cells in the metaphysis and diaphysis of the humerus. High doses of 24,25(OH)2D3 brought about distinct atrophic changes in the above cells. These findings indicate that excessive doses of 24,25(OH)2D3 in an intact animal may lead to retardative effects upon bone growth.

Animals↗
PubMed1985

Structural and histochemical study of the effect of 1,25-dihydroxyvitamin D3 on long-bone growth center in suckling mice.

The in vivo effects of 1 alpha,25-dihydroxyvitamin D3 [1,25(OH)2D3] on cellular structure and response, matrix metachromasia and mineralization have been studied in the epiphysis and growth plate of humeri in normal neonatal mice. A relatively low dose of the metabolite, 50 ng/kg body weight, significantly increased the overall size of humeral growth plate, the zone of proliferating cells and that of hypertrophic chondrocytes. The response of the tissue to the metabolite changed with the increase in dose administered so that it was only the zone of proliferation that still showed increases in size in comparison to untreated controls. 1,25 (OH)2D3 led to an increase in the metachromatic reaction of the cartilage matrix in the chondroblastic zone, and to marked increase in matrix mineralization in the hypertrophic zone. Qualitative changes were also noted in the structure of chondroblasts and hypertrophic chondrocytes. 1,25(OH)2D3 affected the osteoblastic and osteocytic populations of cells along the metaphysis and diaphysis of the humerus. High doses of 1,25(OH)2D3 brought about distinct atrophic changes in the above cells. Chondrocytes in the epiphysis were found to synthesize type I collagen and fibronectin. These findings indicate that excessive doses of 1,25(OH)2D3 in an intact growing animal affect the normal differentiative pathway of prechondroblasts and thereby affect long bone development.

Animals↗
PubMed1985

Receptor-mediated glucocorticoid inhibition of cell proliferation in mouse growth cartilage in vitro.

The growth hormone of neonatal facial cartilage from ICR mice is inhibited by glucocorticoid treatment in vitro. A reduction of the overall tissue weight is accompanied by a substantial decrease in the protein content of the tissue. For the first 48 h in culture, hormone-treated cartilage undergoes a complete standstill in protein gain, and only thereafter the protein content increases, yet is markedly smaller than that of control specimens. Further, a significant reduction in the DNA content is seen already by 24 h, a feature that intensifies by 48 h. A slight recovery takes place thereafter. The reduction in DNA concentration is accompanied by a significant decrease in [3H]thymidine incorporation in acid-insoluble material. The inhibition of DNA synthesis by triamcinolone acetonide is protein- and RNA-synthesis-dependent. Autoradiographic examinations reveal that young cartilage cells are heavily labelled with [3H]dexamethasone and that this labelling is specific. To further substantiate the involvement of glucocorticoid-specific receptors in the latter's inhibitory effects, tissues were treated with cortexolone, this apparently 'masking' the cytosolic receptors for glucocorticoids, and thereby succeeded to eliminate the growth-inhibitory effect of triamcinolone. These results provide evidence for a receptor-mediated set of responses to glucocorticoids in these cartilage cells.

17-Hydroxycorticosteroids↗
PubMed1984

In vitro precocious accumulation of calcium and matrix vesicles formation in young cartilage cells: specific effects of corticosteroids.

The present study examined the effects of various corticosteroid and noncorticosteroid hormones upon the ultrastructure of chondroprogenitor cells and chondroblasts in an organ-culture system of late fetal condylar cartilage. Corticosteroids, (triamcinolone acetonide, dexamethasone, corticosterone) at concentrations of 10(-6) - 10(-8)M stimulated markedly a precocious formation of matrix vesicles by chondroblasts. This stimulation was accompanied by a significant accretion of calcium complexes intra- and extracellularly in both the chondroprogenitor cell population and chondroblasts in vitro, as well as in the newly induced matrix vesicles. Nonglucocorticoid steroids (progesterone, estradiol, testosterone, cortexolone) did not evoke similar effects. Progesterone and testosterone, however, seemed to adversely affect the ultrastructure of the cartilage cells, whereas estradiol appeared to have a favorable effect on the morphology of cultured condylar cartilage.

Adrenal Cortex Hormones↗
PubMed1984

Studies on hormonal regulation of the growth of the craniofacial skeleton: III. Effects of 24,25 (OH)2D3 on cartilage growth in the mandibular condyle of suckling mice.

This study examined the influence of 24,25 (OH)2D3, an active metabolite of vitamin D, on the growth and development of cartilage cells in condylar cartilage of suckling mice. It became evident that when the hormone was administered even at high doses, it did not significantly affect the incorporation of [3H]thymidine, but led to a marked decrease in the number of both chondroblasts and hypertrophic chondrocytes. At the same time, condyle of hormone-treated mice reached an increase in the number of mesenchymelike cells within the chondroprogenitor zone. High values of correlation were noted between the overall dimensions of the condylar cartilage and those of the chondroblastic and hypertrophic zones. The hormone also significantly reduced the degree of matrix metachromasia (indicative of proteoglycan content) and concomitantly altered the mineralization pattern of the cartilaginous matrix. This study indicates that in young animals increased doses of 24,25(OH)2D3 do not affect the proliferative activity of chondroprogenitor cells yet possess an inhibitory effect upon the capacity of the latter cells to differentiate into chondroblasts. The hormone also seems to affect the already differentiated cells--chondroblasts and hypertrophic chondrocytes--both structurally as well as metabolically. In so doing, this metabolite of vitamin D affects the normal process of endochondral bone formation in one of the mandible's main growth sites. Thus, in the developing animal, elevated concentrations of 24,25(OH)2D3 may impair the growing mandible's ability to achieve its normal size and shape.

24,25-Dihydroxyvitamin D 3↗
PubMed1984

Reconstruction of the acetabular wall with bone graft in arthroplasty of the hip.

The fate of autologous bone grafts under a layer of methylmethacrylate cement, previously investigated in dogs, was observed in eight patients. The grafts were incorporated into the acetabular bone within six to ten months. Reinforcement of the inadequate acetabulum with autologous bone graft is necessary in order to withstand the long-term repetitive loading in patients with total hip arthroplasties. Reconstruction of the medial acetabular wall (using autologous bone chips), combined with fixation of the socket with methylmethacrylate, was successful in all eight patients with a follow-up period of two to six years.

Acetabulum↗
PubMed1983

In vitro transformation of chondroprogenitor cells into osteoblasts and the formation of new membrane bone.

Mandibular condyles of fetal mice 19 to 20 days in utero were kept in an organ culture system for up to 10 days. After 2 days in culture the cartilage of the mandibular condyle appeared to have maintained all its inherent structural characteristics, including its various cell layers: chondroprogenitor, chondroblastic, and hypertrophic. After 5 days in culture no chondroblasts could be seen and, instead, the entire cartilage was occupied by hypertrophic chondrocytes. At the same time, the mesenchymal cells at the chondroprogenitor zone differentiated with osteoblasts which produced osteoid. Light microscopic examinations showed that the newly formed osteoid did not stain with acidic toluidine blue or with alcian blue, but stained intensively with the van Gieson stain and with Periodic acid-Schiff (PAS). The osteoid reacted with antibodies against type I collagen but not with antibodies against type II collagen. Electron microscopic examinations showed that the mineralization appeared to be associated with collagen fibers in bone rather than with matrix vesicles in the cartilage. The process of bone formation progressed with time and by the 10th day new bone replaced almost the entire cartilage, thus forming an expanded layer of membrane bone. This in vitro system represents an experimental model whereby undifferentiated precursor cells transform into osteoblasts with the subsequent formation of a typical membrane bone.

Animals↗