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Biomedical subjects

M Silbermann

Publications and source records attributed to M Silbermann.

At least 37 records · Page 2Linked to original sources

Transformation of fetal secondary cartilage into embryonic bone in organ cultures of human mandibular condyles.

Mandibular condyles of human fetuses, 14-21 weeks in utero, were kept in an organ culture system for up to 60 days. After 6 days in culture, the cartilage of the mandibular condyle appeared to have maintained its inherent structural characteristics, including all its various layers: chondroprogenitor, chondroblastic, and hypertrophic. After 12 days in culture, no chondroblasts could be seen; instead, the entire cartilage was occupied by hypertrophic chondrocytes. At the same time, the mesenchymal cells in the vicinity of the chondroprogenitor zone differentiated into osteoblast-like cells that produced type I collagen. The progenitor cells were still actively incorporating 3H-thymidine. The newly formed osteoid-like tissue lacked both metachromatic reactivity and a response to antibodies against chondroitin sulfate. Instead, the tissue reacted positively for osteocalcin (bone gla-protein). The process of new bone formation further progressed and, by the 20th day in culture, the new bone reacted positively for type I collagen, osteonectin, and to a lesser extent for chondroitin sulfate. The osteoid also underwent mineralization as revealed by both the von Kóssa stain and vital staining with tetracycline. The above feature appeared even more intense in 40-day-old cultures. After 60 days, the newly formed bone contained osteoblasts and osteocytes, whereas the extracellular matrix revealed a high degree of matrix polarization. The results of the present study recapitulate findings reported for organ cultures of mice mandibular condyles. However, the in vitro process of de novo bone formation in human specimens requires a 6-fold longer culture time than that needed for mice condyles.

Animals↗

Effect of growth hormone on gastrocnemius muscle of aged rats after immobilization: biochemistry and morphology.

Immobilization of limbs of aged animals is associated with swift muscular damage and atrophy. We investigated the effect of rat growth hormone (rGH) on immobilized hindlimb muscles of 26-mo-old rats. Administration of rGH significantly reduced muscle weight loss and muscle protein oxidation caused by immobilization. Capillary blood volume, measured by photoplethysmography of the hindlimb, showed a 34% reduction in immobilized animals, which was eliminated by rGH. The activity of creatine phosphokinase in immobilized gastrocnemius muscle was significantly reduced by immobilization. This damage was diminished by rGH administration. Similarly, the increase in acid phosphatase activity in immobilized muscle was reduced after rGH treatment. Morphologically, marked muscle atrophy and fiber disorientation were observed in immobilized limbs. Therapy with rGH prevented some of these changes. These results indicate that administration of rGH may provide a useful means to attenuate the degenerative effects of limb immobilization of aged rats, as evident from physiological, biochemical, and morphological parameters.

Acid Phosphatase↗

Insulin-like growth factor I accelerates proliferation and differentiation of cartilage progenitor cells in cultures of neonatal mandibular condyles.

The present study examined the effect of exogenous IGF-I on growth and development of neonatal cartilage of the mandible condyle. Condylar cartilage was cultured as organ culture. The explants were cultured on top of collagen sponges in medium containing 2% fetal calf serum and were treated with IGF-I at doses ranging from 3.25 to 26 nmol/l for up to six days. IGF-I was found to increase significantly the uptake of [3H]-thymidine and [35S]-sulfate in a dose-related manner. The enhanced cellular proliferation, along with the increased synthesis of proteoglycans, resulted in a substantially larger mass of tissue in the organ culture system. The nature of the IGF-I stimulative effect was further studied through the use of a tissue culture system whereby a separated chondroprogenitor zone is cultured under conditions which favor its development at first into cartilage and then into bone. Using this culture system, we could show that IGF-I induces merely the de novo chondrogenesis process. This was reflected in the appearance of relatively large amounts of cartilage specific antigens such as type II collagen, cartilage proteoglycans, chondrocalcin and 100 KDa protein. Yet, no bone specific antigens were significantly increased, as is the case with GH effects. These results indicate that IGF-I is a strong chondrogenetic agent. But, unlike growth hormone, it does not seem to stimulate bone formation.

Analysis of Variance↗

The early postnatal development of the murine mandibular condyle is regulated by endogenous insulin-like growth factor-I.

Skeletal growth during the early postnatal period is thought to be GH-independent, and is probably regulated by intrinsic growth factors. We studied the involvement of locally produced insulin like growth factor-I (IGF-I) in the growth of the neonatal mandibular condyle. Immunofluorescence studies revealed intense staining with antibodies to IGF-I in the mandibular condyle of 2-day-old ICR mice. We have also shown that these mandibular condyles contain specific high-affinity binding sites (Kd = 0.157 nmol/l) for IGF-I (427 fmol/mg). Autoradiographical studies of iodinated IGF-I revealed that the distribution of the receptors for IGF-I was parallel to that of IGF-I production, mainly in the younger zones of the condyle, namely the chondroprogenitor and the chondroblast cell layers. Immunoinhibition of IGF-I resulted in an almost complete inhibition (-91%) of thymidine incorporation into DNA, as well as in marked degenerative changes in the morphological appearance of the condyle. Our studies support the hypothesis that early postnatal growth is dependent on the paracrine activity of endogenous GH-independent IGF-I.

Animals↗

[Autologous transplantation of rib perichondrium in treatment of deep cartilage defects of the knee joint of sheep. Morphologic comparison of two resorbable fixation methods].

The purpose of this study was to examine the fate of autologous perichondrial grafts after transplantation into cartilage lesions in weight-bearing joints, with reference to different fixation methods and different regimens of postoperative functional treatment. Full-thickness osteochondral lesions were drilled in the articular surface of knee joints in 36 sheep. The defects were filled with autologous rib perichondrial grafts, which were secured by either collagen sponges (group A; n = 12 animals) or fibrin glue (group B; n = 12 animals). Defects not filled with perichondrial grafts served as controls (group C; n = 12 animals). Following 1 week of immobilization of the treated leg, the plaster was removed and the animals were allowed to move freely. Animals were sacrificed after 4, 8, 12 and 16 weeks. The grafts were removed and investigated histologically. In contrast to weight-bearing areas and control defects, hyaline-like cartilage formation could be noted in non-weight-bearing areas after 4 weeks. This newly formed cartilage revealed strong metachromasie following staining with acidic toluidine-blue and reacted positively with PAS, indicating de novo synthesis of proteoglycans and glycoproteins. Enzyme-histochemistry for alkaline and acid phosphatase activity showed positive reactivity only at the base of the transplants. Quantitatively, hyalin-like cartilage was achieved in 40/48 transplants from the non-weight-bearing area (20/24 transplants fixated with collagen sponge (group A); and 20/24 fixated with fibrin glue (group B). In control defects only fibrous scar tissue was detectable. In transplants from the weight-bearing area only 7/48 (24 of group A and B each) a differentiation into hyalin-like cartilage was visible.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Chondroclasts and endothelial cells collaborate in the process of cartilage resorption.

The condylar cartilage of the young rat is a major growth center of the craniofacial complex. Differences between the mechanism that results in bone formation from growth centers in the epiphyseal plates of long bones are dictated primarily by the different character of the mineralization of the cartilage. In this ultrastructural study we demonstrate that the terminal hypertrophic chondrocytes undergo apoptosis and disintegration while simultaneously chondroclasts dissolve gaps in the calcified cartilage that engulfs them. The latter are also phagocytizing debris of the chondrocytes. The chondroclasts are intimately followed by tube-forming endothelial cells that most probably coalesce to create extensions of the invading capillaries into the evacuated lacunae. The chondroclasts have ultrastructural features similar to osteoclasts. They are multinucleate, are rich in mitochondria and vacuoles, form clear zones that adhere to the spicules of the calcified cartilage, and also form a sort of ruffled border. The latter is not as elaborate and orderly arranged as is known from osteoclasts. The capillaries that follow orient the stroma cells to the evacuated lacunae and, together with the calcified cartilaginous scaffold, supply the adequate environmental conditions for the stroma cells to differentiate into osteoblasts and to build up trabecular bone.

Animals↗

The threshold of age in exercise and antioxidants action.

Physical activity and exercise are important factors in determining the quality of life in old animals and humans. With age there is a slow but significant reduction in muscle mass and ability to perform certain physical activities. This may be due to changes with the age of muscle composition and protein turnover, as well as decrease of trophic influence in neural control of muscles of old individuals. Exercise in general was shown to improve muscle performance even in old age. However a concept of threshold of age in exercise was advanced forward in the 1970s. Accordingly, the idea was that for a given exercise of a particular duration and intensity there is a certain age beyond which this exercise may not have a positive influence, but can become detrimental to the exercising animal or human. Recent studies on the effect of antioxidants such as Vitamins C and E and selenium have shown that these agents could decrease the free radical associated muscle damage caused by extensive exercise. Thus, administration of these antioxidants especially vitamins C and E may reduce the oxidative damage due to exercise, and may alter the threshold of age by delaying it to an older age.

Adult↗

Structural changes in condylar cartilage following prolonged exposure to the human parathyroid hormone fragment (hPTH) 1-34 in vitro.

This investigation presents the structural changes in condylar cartilage incubated in the presence of human parathyroid hormone (1-34) in an organ culture system for 6 to 12 days. Control cultures maintained their cartilaginous characteristics whereas human parathyroid hormone (1-34)-treated cultures revealed the following modifications: (1) The chondroprogenitor cell zone at the apical region of the explant underwent a substantial enlargement. The cells changed from a mesenchyme-like morphology into polygonal, glycogen-rich cells that were tightly attached to each other by a fibrillar intercellular matrix, but even by 12 days the apical region was comprised of healthy cells. (2) The mineralizing zone in the hypertrophic cartilage revealed a change in its cellular population. Hypertrophic chondrocytes were replaced by cells with amoeboid extensions and large numbers of secretory granules or vesicles. Based upon the above findings it appears that the chondroprogenitor cells that are initially stimulated to proliferate, are being suppressed from subsequent differentiation into chondroblasts; and that hypertrophic chondrocytes apparently undergo a dedifferentiation process followed by development into an as yet unknown cell population.

Animals↗

Interactions between growth hormone and dexamethasone in skeletal growth and bone structure of the young mouse.

The present study investigated the interactions of growth hormone (GH) and glucocorticoid on skeletal growth and bone structure in young mice. The purpose of this study was to examine the possible prevention by GH of the damage inflicted by dexamethasone (Dex) at sites of skeletal growth and ossification. Dex (1 mg/kg) with or without rat GH (rGH) or bovine GH (bGH), 1 mg/kg, was given for 4 weeks, from age 3-7 weeks, to female ICR mice. Tibiae, humerus, and vertebrae were analyzed morphometrically and biochemically. Growth, as determined by the mouse weight, tibial length, and humerus protein content was found to be compromised by dexamethasone. This was prevented by rGH or bGH. The epiphyseal growth plate width, trabecular bone volume, cortical bone width, mineral bone content, and alkaline and acid phosphatase activity were decreased by dexamethasone. These were prevented by rGH or by bGH. The findings of the present study suggest that in the mouse, GH can decrease or even avoid some of the pathological features in growing bones inflicted by high-dose glucocorticoid treatment.

Analysis of Variance↗

Autologous rib perichondrial grafts in experimentally induced osteochondral lesions in the sheep-knee joint: morphological results.

The purpose of the present study was to examine the fate of autologous perichondrial grafts after transplantation into cartilage lesions in weight-bearing joints. Osteochondral lesions were made in the articular surface of knee joints in 36 sheep. The defects were filled with autologous rib perichondrial grafts which were secured by either collagen sponges (12 animals) or fibrin glue (12 animals). Defects without perichondrial grafts served as controls (12 animals). Following 1 week of immobilization of the operated leg, the plaster was removed and the animals were allowed to move freely. Animals were sacrificed after 4, 8, 12 and 16 weeks. The grafts were removed and investigated histologically. In contrast to weight-bearing areas and control defects, hyaline-like cartilage formation was seen in non-weight-bearing areas after 4 weeks. This newly formed cartilage revealed strong metachromasia following staining with acidic toluidine blue and reacted positively with periodic acid-Schiff, indicating de novo synthesis of proteoglycans and glycoproteins. Scanning electron microscopy and examinations with polarized light confirmed a hyaline cartilage-like architecture for the surface area as well as for the fibre orientation of the whole graft. Enzyme histochemistry for alkaline and acid phosphatase activity showed positive reactivity only at the base of the transplants.

Animals↗

Muscle microcirculatory impairment following acute compartment syndrome in the dog.

Visualization of the intramuscular microcirculation during and after compartmental syndrome was studied by microangiograms and histologic cross sections. A marked reduction in the circulation of the endomysial capillary network was found during compartment tamponade, whereas the perimysium arteriolar system was patent. Revascularization took place by formation of distorted blood vessels accompanied by intramuscular hematomas in muscles 7 and 14 days after the compartment insult. The cross sections show massive fibroblastic activity around blood vessels that caused concealed intramuscular pressure-ischemic contracture resulting in the foci of myofibrillar necrosis seen within normal muscle tissue. The muscle located in the tamponaded compartment profusely bleeds when it is touched, even though its viability is in doubt. The explanation for this clinical observation might be the abnormal intramuscular revascularization that was found in this work.

Acute Disease↗

Structural characterization of the mandibular condyle in human fetuses: light and electron microscopy studies.

Mandibular condyles from 18- to 20-week-old human fetuses were examined in the light and electron microscope with particular attention to intratissue organization and extracellular matrix. In the human fetus the condyle has been divided into five layers: (1) the most superficial, articular layer, (2) chondroprogenitor cell layer, (3) condroblast cell layer, (4) nonmineralized hypertrophic cell layer, and (5) mineralized hypertrophic cell layer. The articular layer is rich in collagen fibers (mostly of the type I collagen), but the cells seldom divide. By contrast, in the chondroprogenitor cell layer and upper part of the chondroblastic cell layer mitosis gives rise to new cells. The matrix in the latter layer is composed of thick banded 'lucent' fibrils in a loose feltwork of granules representing cartilage proteoglycans. The daughter cells in the progenitor cell layer undergo differentiation which is apparently completed along the lower border of the mineralized hypertrophic cell layer--the ossification front. The matrix in the hypertrophic cell layer reveals distinct matrix vesicles that undergo mineralization and subsequently coalesce to form larger sheets of mineralized extracellular matrix. Mineralized cartilage serves as a backbone for new bone formation as marrow-derived osteoblasts and osteoclasts attach to remnants of mineralized cartilage, which enables the turning on of the remodeling cycles involved in new bone formation. It can be concluded that the process of endochondral ossification as has been reported in lower animals is recapitulated in the human fetus, thus the dynamics associated with condylar morphogenesis is maintained through phylogeny.

Cartilage↗

The rib perichondrium. An anatomical study in sheep of a tissue used as transplant in the treatment of hyaline-cartilage defects.

Perichondrial tissue is of great importance as a transplant for the repair of hyaline-cartilage defects of synovial joints because of its potential to produce hyaline-like cartilage. The histological and histochemical character and fiber orientation of rib perichondrial grafts prior to a transplantation procedure for the repair of full-thickness hyaline-cartilage defects is analyzed in a sheep model. Compared to other descriptions of the rib perichondrium of other different species and humans, the perichondrial architecture of different origins demonstrates the same histological, histochemical character and fiber orientation. The three different layers of this tissue are: the outer fibrous layer, the central part called proliferation zone and the inner part towards the underlying rib called transition zone. The fibrous layer demonstrates a wave-like extracellular configuration and contains a few fibrocytes. The proliferation zone is only a few cell rows thick and contains single cells with an oval shape and longitudinal fibrocyte-like nucleus. Because of the potential of these cells to produce cartilage this layer is called proliferation or cambium layer. The transition zone contains more oval-shaped cells with rounded nuclei and demonstrates a more chondrocytic character. Some cells lie in groups of 2 and more cells like in chondrons. The extracellular matrix shows an increasing stain towards the rib cartilage indicating new cartilage production. The fiber orientation is longitudinal in the fibrous part and vertical in the transition zone. Between these layers, fibers deviate from the longitudinal to the vertical orientation corresponding to the light-microscopically visible proliferation zone.

Animals↗

Isolation of a cathepsin B-encoding cDNA from murine osteogenic cells.

Cathepsin B-encoding cDNA (CTSB) clones have been isolated from a lambda gt10 library of a murine osteosarcoma by differential screening during a search for genes which are typically expressed during osteogenic differentiation in mouse mandibular condyles in vitro. Sequencing of the CTSB 3' end revealed that the isolated sequence contained an 825-bp 3'-noncoding region, the polyadenylation signal and the poly(A) tail. The enhanced CTSB expression during the early stages of the enchondral ossification-like process in mandibular condyles in vitro suggests that CTSB participates in the degradation of cartilage matrix prior to the synthesis of bone matrix proteins.

Animals↗

Immunohistochemical studies of the extracellular matrix in the condylar cartilage of the human fetal mandible: collagens and noncollagenous proteins.

This study provides data concerning the cells and their extracellular matrix in prenatal human mandibular condylar cartilage. The latter cartilage represents a secondary type of cartilage since it develops late in the morphogenesis of the craniofacial skeleton. The cartilage of the mandibular condyle is actively involved in endochondral ossification, thus showing all the phases of cartilage growth, maturation, and mineralization that precedes de novo bone formation. The present study focused on the localization and distribution of the major macromolecules that are normally encountered in cartilage and bone, including collagens, proteoglycans, fibronectin, osteonectin, osteocalcin, alkaline phosphatase, and anchorin CII. It became clear that the mineralized zone of the cartilage already contained bone-specific antigens; thus the above zone might serve as an essential propagative predecessor in the ossification process.

Alkaline Phosphatase↗

In vitro response of neonatal condylar cartilage to simultaneous exposure to the parathyroid hormone fragments 1-34, 28-48, and 53-84 hPTH.

Mandibular condylar explants of neonatal ICR mice were maintained as serum-free organ culture systems and were used to study the effects of three synthetic fragments of human parathyroid hormone (hPTH) on the morphology of the organ and its ability to incorporate [3H]thymidine. Forty-eight-hour incubation with hPTH (1-34), at a concentration of 0.5 microgram/ml caused an increase of 88% in DNA synthesis and a marked increase in the size of the chondroprogenitor zone. The mitogenic effect of hPTH (1-34) was decreased to 34% over control levels when the fragment hPTH (28-48) was added to the system. However, the addition of the latter fragment brought about a marked enhancement in the mineralization of the cartilaginous extracellular matrix along with the formation of an appreciable amount of new bone. The de novo osseous tissue was attached to the mineralized cartilage. When the carboxyl-terminal fragment hPTH (53-84) was added together with the other two fragments, the mitogenic effect of hPTH (1-34) was completely abolished and the respective cultures incorporated [3H]thymidine even less than untreated control cultures. Moreover, the addition of hPTH (53-84) to the culture system led to distinct structural features throughout the mineralized hypertrophic cartilage. The latter contained a mixture of cells within an unorganized extracellular matrix. Untreated control cultures lacked such structures, but contained the various cell zones as normally seen in neonatal condylar cartilage. Therefore, it seems reasonable to suggest that each of the three fragments tested induces a biological effect on neonatal cartilage and might be involved in the normal process of endochondral ossification.

Animals↗

Structural changes in aging bone: osteopenia in the proximal femurs of female mice.

A computerized image analysis system was used to quantitate age-related changes in the structure of the proximal femur in CW-1 female mice, ranging from 3 to 32 months of age. Morphological findings revealed a progressive thinning of bone trabeculae within the femoral head, accompanied by the development of marrow cavities in the cortical bone of the femoral neck and in the subchondral bone. As a result, the compact bone in senescent mice acquired an appearance similar to trabecular bone. Quantitative image analysis revealed a similarity in the pattern of changes in the three types of bone: cortical, trabecular, and subchondral. Bone density increased from 3 to 12 months of age and subsequently declined. A similar pattern was noted for the changes in the thickness of the cortical and the subchondral bone. Regression analysis revealed that the changes with age fitted a second-order model; thus it was possible to predict the age of maximal values for each parameter. Hence, the age of maximal bone density for cortical, trabecular, and subchondral bone was 12.3, 14.8, and 18.0 months, respectively. The rate of bone loss after 12 months was most prominent for trabecular bone (1.47% per month), so that by 32 months of age its overall mass had declined by 57% in comparison to peak values seen at 12 months of age (p less than 0.001). The density of the subchondral and cortical bones decreased at a slower rate (0.6% to 0.8% per month) and at the age of 32 months their values had decreased by 12% to 18% in comparison to those at 12 months (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗