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J Glowacki

Publications and source records attributed to J Glowacki.

At least 73 records · Page 4Linked to original sources

The effect of deficiencies of manganese and copper on osteoinduction and on resorption of bone particles in rats.

Subcutaneous implantation of devitalized demineralized bone powers (DBP) and mineral-containing bone particles (BP) into rats raised on either a control (C), low manganese and low copper (L), or manganese-deplete (D) diet, allowed the separate evaluation of bone formation and of bone resorption, respectively. DBP failed to induce chondrogenesis or osteogenesis in D rats. Cartilage formation was delayed in the L rats compared to C rats. There was significantly less resorption of BP by L and D rats than C rats. These results show multiple cellular effects of long-term manganese (Mn) and copper (Cu) deficiencies on bone metabolism including decreased osteogenesis and a decrease in osteoclast activity.

Aging↗

Expression of differentiated function by mineralizing cultures of chicken osteoblasts.

This report documents osteoblast differentiation in vitro, as demonstrated by the 50-100X increase of proteins which are known markers of the osteoblast phenotype. Collagen type I and osteocalcin synthesis and accumulation, alkaline phosphatase activity, and matrix calcification show similar temporal relationships that are analogous to those seen during in vivo bone development. Chicken embryonic osteoblast progenitor cells were selected by initial growth at low densities in minimal medium. Upon subcultivation into nutrient-enriched medium at higher cell densities, near homogeneous populations of osteoblasts were obtained as demonstrated by the greater than 80% enrichment of cells positive for alkaline phosphatase activity. A comparison was made between cells grown in the presence or absence of 10 mM beta-glycerolphosphate (beta-GPO4), a chemical stimulant of matrix calcification, as a function of time. Cultures treated with beta-GPO4 showed visible calcification at Day 12 when culture monolayers became confluent. By Day 30, numerous large foci of calcification were visible and a 20-fold increase in calcium (Ca) content was observed. In contrast, untreated cultures had only a 3-fold increase in Ca content with many smaller diffuse areas of calcification. DNA, RNA, and total protein levels were nearly identical between the two cultures, indicating that beta-GPO4 had no marked effect on either cell proliferation or transcriptional activity. The major collagen type produced by either culture was type I, with no detectable type III as determined by CNBr peptide mapping and delayed reduction analysis. Alkaline phosphatase activity showed a rapid approximately 50-fold induction by Day 18 and remained elevated in control cultures. However, cultures treated with beta-GPO4 demonstrated a rapid 80% decline of enzyme activity after 18 days. In contrast, total osteocalcin levels showed a 100-fold induction by Day 18 and remained elevated in both control and beta-GPO4-treated cultures throughout the time period examined. While the overall levels of osteocalcin were the same in beta-GPO4-treated and untreated cultures, 2- to 5-fold more osteocalcin was associated with the more mineralized matrices of the beta-GPO4-treated cultures. In order to confirm the association of osteocalcin with areas of mineralization, co-localization of mineral to osteocalcin and collagen was carried out by combining vital labeling with tetracycline and immunofluorescent staining with anti-osteocalcin and anti-collagen antibodies. Both collagen and osteocalcin showed strong localization with areas of mineralization.(ABSTRACT TRUNCATED AT 400 WORDS)

Alkaline Phosphatase↗

Impaired recruitment and differentiation of osteoclast progenitors by osteocalcin-deplete bone implants.

This is a report of an experimental system to study differentiation of bone-resorbing osteoclasts and demonstrates that osteocalcin, an extracellular bone-specific component, is necessary for the recruitment of osteoclast progenitor cells. The subcutaneous implantation of devitalized bone particles (BPs) elicits the recruitment and differentiation of osteoclasts that resorb the BPs. In a previous study, we showed by histomorphometric analysis that BPs that were deficient in osteocalcin were resorbed only 60% as well as normal BPs. In this study, the mechanism of this difference was investigated by measurements of recruitment, differentiation and activity of bone resorbing cells by normal and osteocalcin-deficient BP. Mononuclear cells were attracted to control BPs soon after implantation. In dramatic contrast, cellularity was depressed around osteocalcin-deficient BPs with very few mononuclear cells within the implant on day 5 (35% of control cellularity). In implants of normal BPs, tartrate-resistant acid phosphatase-positive multinucleated cells were evident by day 5; very few appeared in implants of osteocalcin-deplete BPs even by day 12. The amount of tartrate-resistant acid phosphatase activity in homogenates of the osteocalcin-deficient bone particle specimens not only lagged behind controls but never reached the maximum activity of control BP specimens. These data support the hypothesis that osteocalcin may function as a matrix signal in the recruitment and/or activation of cells for bone resorption.

Acid Phosphatase↗

Engraftment of a clonal bone marrow stromal cell line in vivo stimulates hematopoietic recovery from total body irradiation.

Whether bone marrow stromal cells of donors contribute physiologically to hematopoietic stem cell reconstitution after marrow transplantation is unknown. To determine the transplantability of nonhematopoietic marrow stromal cells, stable clonal stromal cell line (GB1/6) expressing the a isoenzyme of glucose-6-phosphate isomerase (Glu6PI-a, D-glucose-6-phosphate ketol-isomerase; EC 5.3.1.9) was derived from murine long-term bone marrow cultures and made resistant to neomycin analogue G418 by retroviral gene transfer. GB1/6 cells were fibronectin+, laminin+, and collagen-type IV+ and collagen type I-; these GB1/6 cells supported in vitro growth of hematopoietic stem cells forming colony-forming units of spleen cells (CFU-S) and of granulocytes, erythrocytes, and macrophage/megakarocytes (CFU-GEMM) in the absence of detectable growth factors interleukin 3 (multi-colony-stimulating factor), granulocyte/macrophage colony-stimulating factor, granulocyte-stimulating factor, or their poly(A)+ mRNAs. The GB1/6 cells produced macrophage colony-stimulating factor constitutively. Recipient C57BL/6J (glucose-6-phosphate isomerase b) mice that received 3-Gy total-body irradiation and 13 Gy to the right hind limb were injected i.v. with GB1/6 cells. Engrafted mice demonstrated donor-originating Glu6PI-a+ stromal cells in marrow sinuses in situ 2 mo after transplantation and a significantly enhanced hematopoietic recovery compared with control irradiated nontransplanted mice. Continuous (over numerous passages) marrow cultures derived from transplanted mice demonstrated G418-resistant, Glu6PI-a+ stromal colony-forming cells and greater cumulative production of multipotential stem cells of recipient origin compared with cultures established from irradiated, nontransplanted control mice. These data are evidence for physiological function in vivo of a transplanted bone marrow stromal cell line.

Animals↗

Comparison of multinucleated cells elicited in rats by particulate bone, polyethylene, or polymethylmethacrylate.

Osteoclasts, the multinucleated resorbing cells of bone, are identified by their characteristic morphology, unique cell membrane specializations, and more recently by the presence of cell surface antigens recognized by monoclonal antibodies. They are derived from mononuclear precursor cells of hematogenous origin. The precise relationship between osteoclasts and other types of tissue giant cells is unknown. This study was designed to examine factors involved in the recruitment and differentiation of multinucleated cells and to investigate the relationship between so-called foreign body giant cells and bone-resorbing osteoclasts. Particles of various materials were implanted into subcutaneous pockets in rats. Histological, histochemical, and electron microscopic evaluations were made of specimens harvested 12 days after implantation. Large, foamy multinucleated cells were evident around particles of devitalized bone, polyethylene, and polymethylmethacrylate. Bone particles showed scalloped surfaces and were partially resorbed. Those cells adjacent to the bone particles stained positive for tartrate-resistant acid phosphatase, in contrast to the multinucleated cells adjacent to the other materials. All the cell types had extensive rough endoplasmic reticulum, abundant mitochondria, cytoplasmic vacuoles and dense bodies, giant centrospheres, and areas of fusion of plasma membranes. Cells in lacunae on the surface of the bone particles showed a clear zone of attachment to the bone substrate and ruffled borders, a feature characteristic of in osso osteoclasts. Although the giant cells occasionally displayed an extensive clear zone of attachment to the polyethylene and polymethylmethacrylate particles, no ruffled borders were detected. The results of these studies show that the multinucleated cells elicited in response to different materials, although sharing many common features, do demonstrate certain features that are substrate specific.(ABSTRACT TRUNCATED AT 250 WORDS)

Acid Phosphatase↗

Osteoclastic features of cells that resorb bone implants in rats.

The subcutaneous implantation of devitalized, mineral-containing bone particles in rats elicits the recruitment and differentiation of multinucleated cells and the rapid resorption of the bone. Cytochemical and ultrastructural characteristics were studied 12 days after implantation of bone. Tartrate-resistant acid phosphatase activity was demonstrated in these cells, especially at the cell/bone interface. Mast cells were seen vicinal to, but not in contact with the osteoclasts and the bone particles. Electron microscopic evaluation revealed that many multinucleated cells in lacunae on the surfaces of the bone particles displayed features characteristic of in osso osteoclasts, including clear zones of attachment to the bone substrate, and ruffled borders overlying bone with frayed collagen fibers. Many cells contained multiple giant centrospheres; this finding suggests that fusion can occur between multinucleated cells. This model may be useful to characterize the differentiation and regulation of bone-resorbing cells.

Acid Phosphatase↗

Osteoclasts can be induced in fish having an acellular bony skeleton.

Kelp bass (Paralabrax clathratus) and leopard sharks (Triakis semifasciata) are characterized by an acellular (anosteocytic) bony skeleton and a focally calcified cartilaginous endoskeleton, respectively. These skeletal forms are not considered to function as mineral reservoirs. Previous studies showed that implanted bone particles are resorbed in rats by large multinucleated cells with ultrastructural features (ruffled borders) characteristic of osteoclasts. We tested the ability of fish to resorb bone matrix and to adapt to reduced salinity conditions. Bone particles were implanted in sharks and bass maintained in seawater (34 ppt, 40.5 mg of calcium per dl) or in diluted seawater (26 ppt, 28.5 mg of calcium per dl). Sera and elicited tissues were harvested 4 weeks later. In sharks, bone particles were not resorbed, and multinucleated cells were not evident under either normal or hyposalinity conditions. Shark sera were isoosmolar with the seawater or diluted seawater, with serum chemistries of the hyposalinity group reflecting the 23% reduction in environmental minerals and electrolytes, compared to sharks in normal seawater. In marked contrast, bass adapted to diluted seawater resorbed bone particles and maintained normal serum chemistries. Electron microcopy showed that the bone particles were surrounded by large, foamy multinucleated cells, many with membrane specializations typical of osteoclasts from higher vertebrates, i.e., extensive clear zones apposed to intact bone matrix and active ruffled borders overlying areas of matrix undergoing dissolution. Although osteoclasts had not been described in these fish, this study shows that bass have stem cells that can be stimulated to differentiate into bone-resorbing osteoclasts.

Journal Article↗

An ultrastructural study of mast cell interactions in hemangiomas.

Hemangiomas, the most common tumors of infancy, are characterized by a postnatal period of rapid growth, followed by a phase of gradual involution. The proliferative phase is characterized by increased numbers of endothelial and mast cells and thickened basement membrane. Ultrastructural analysis of hemangiomas in the late proliferative phase showed that mast cells had numerous fingerlike processes aligned parallel to the outer lamina of the thickened basement membranes surrounding the vessels and to the surfaces of opposing cell membranes. We found evidence of different types of interactions between mast cells and adjacent connective tissue cells (fibroblasts, macrophages, multinucleated giant cells, and plasma cells) in the perivascular regions of the lesions. Intercellular contacts were observed in areas where these mast cell processes were in close association to the opposing cell membrane. Areas of membrane fusions were seen between cell types. Coated vesicles and pinocytotic vesicles were present along the periphery of cells adjacent to mast cells. Occasionally, cytoplasmic bridges were found between mast cells and fibroblasts. These ultrastructural findings suggest the proliferation and involution of hemangiomas are determined by interactions between the various types of cells found in the lesions.

Cell Communication↗

Calcitonin produces hypercalcemia in leopard sharks.

Calcitonin was detected by RIA in sera from four marine species, leopard sharks (Triakis semifasciata), horn sharks (Heterodontus francisci), thornback rays (Platyrhinoides triseriata), and kelp bass (Paralabrax clathratus). These animals have levels of calcitonin and calcium higher than freshwater and terrestrial species have. The administration of salmon calcitonin to bass (4 micrograms/kg BW) produced hypocalcemia and hypophosphatemia as has been reported for other bony vertebrates. In marked contrast, calcitonin produced a prompt hypercalcemia in sharks; the average was 9.8% increase in serum calcium in nine animals with no attendant change in phosphorus. These findings demonstrate that calcitonin can increase serum calcium in sharks. Because shark skeleton is composed of cartilage, this hypercalcemic effect of calcitonin does not require a bony skeleton.

Animals↗

Demineralized bone implants.

There are three mechanisms of bone formation that underlie the use of the different types of implants. In osteogenesis, viable osteoblasts and preosteoblasts are transplanted from one part of the body to the site where new bone is needed; cancellous marrow grafts are an example of such osteogenic engraftment. In osteoconduction, the implant does not provide many viable cells but rather acts as a scaffolding for the ingrowth of new bone from the margins of the defect with the concurrent resorption of the implant; cortical bone grafts or banked bone segments are examples of this "creeping substitution." In osteoinduction, the implant stimulates the transformation of connective tissue to produce endochondral bone, even in extraskeletal sites; demineralized bone implants promote bone formation by osteoinduction. The physiology, cell biology, biochemistry, and endocrinologic regulation of induced osteogenesis are areas of active investigation. Fresh autogenous cancellous bone grafts are preferred for non-stress-bearing defects, but are often of limited availability for extensive procedures, especially in infants. Demineralized bone implants have been used successfully in certain types of craniomaxillofacial, orthopedic, periodontal, and hand reconstruction. Tissue transformation may become as important to reconstructive surgery as is tissue transplantation.

Animals↗

Induced osteogenesis--the biological principle and clinical applications.

Complex orthopedic and craniofacial surgical procedures have stimulated renewed interest in the physiology of bone grafting. The experimental and clinical evidence that a nonvital implant can stimulate local host cells to become bone-forming cells (osteoinduction) is reviewed. Studies of induced osteogenesis were derived from old clinical observations of heterotopic ossification in scars and muscle. When demineralized bone is placed within soft tissues of a rat, a remarkable sequence of endochondral ossification occurs. In contrast, mineral-containing bone particles stimulate obligatory resorption by host mono- and multinucleated cells. The biochemical, endocrine, and physicochemical aspects of induced osteogenesis are actively being investigated. Present understanding of the mechanism of bone induction is discussed along with descriptions of preliminary clinical trials with allogeneic demineralized implants.

Abnormalities, Multiple↗

The use of demineralized xenogeneic bone implants to correct phalangeal defects: a case report.

Demineralized bovine bone blocks and powder were used to reconstruct phalangeal defects created by excision of bulging endochondromas . Bony reconstruction was documented by roentgenography at 9 weeks and by biopsy at 7 months. Follow-up at 22 months has shown stability of the implants, further ossification, and no recurrence of the phalangeal endochondromas . The use of xenogeneic demineralized bone implants promotes induced osteogenesis and rapid healing, avoids harvesting procedures, and offers a potentially unlimited supply of banked material.

Adolescent↗

Resorption of implanted bone prepared from normal and warfarin-treated rats.

Bone that was virtually depleted of the vitamin K-dependent protein, osteocalcin, and 93% reduced in the concentration of its characteristic amino acid, gamma-carboxyglutamic acid, was obtained from rats treated with warfarin for 6 wk. Osteocalcin-deficient bone particles were resistant to resorption when implanted subcutaneously in normal rats. The relative resorption was 60% of control bone, as measured by histomorphometry as percent of bone particles in the field. Additionally, the number of multinucleated cells around the bone particles was reduced by 54%. These data suggest that osteocalcin is an essential component for bone matrix to elicit progenitor-cell recruitment and differentiation necessary for bone resorption.

Animals↗

Augmentation of rat mandibular ridge with demineralized bone implants.

Demineralized bone implants were placed as onlays in submucosal pockets on the edentulous segment of the rat mandible, between the incisor and posterior teeth. These implants, predictably, induced osteogenesis. The mass of induced bone and implant was united to the ridge by two weeks, and there was little or no resorption over a six-month follow-up period. This study suggests that demineralized bone implants may be useful for mandibular ridge augmentation.

Alveolar Ridge Augmentation↗

The effects of heparin and protamine on resorption of bone particles.

Implantation of bone particles into rats initiates rapid, reproducible resorption that can be quantitated by histomorphometric analysis. Mast cells are found with the mononuclear and multinucleated cells that digest the bone. One of the constituents of mast cell secretory granules, heparin, is known to regulate collagenase activity. This study shows that 1) exogenous heparin stimulated resorption of implanted bone to 141% of control values and that 2) protamine, a heparin antagonist, reduced resorption to rates 50% of controls.

Animals↗

Congenital vascular lesions: clinical application of a new classification.

Two hundred and ninety-seven patients with 375 pediatric vascular lesions were followed from 1967 to 1981. By history and physical examination, 96% of childhood vascular lesions can be classified as hemangiomas or malformations. Hemangiomas are often not present at birth (40%), but make their appearance during the first month. A proliferative phase, lasting an average of 3 months, is followed by a slow, but eventually complete involution. A "perfect" cosmetic result is more likely when involution is complete before age 6. Malformations are always present at birth, their growth is commensurate with the patient's, and they never involute. Analysis of clinical characteristics fails to identify a subgroup of hemangiomas destined for early involution.

Adolescent↗

Cell shape and phenotypic expression in chondrocytes.

The relationship between cell shape, proliferation, and phenotypic expression was studied in human chondrocytes. Shape was controlled independent of serum concentration, anchorage, and cell density by alteration of substratum adhesiveness with poly(2-hydroxyethyl methacrylate) (poly[HEMA]). Cells that were held rounded displayed features of the chondrocyte phenotype; i.e., they were round, proliferated slowly, incorporated low levels of [3H]thymidine into DNA, and incorporated large amounts of 35SO4 into glycosaminoglycans. In contrast, cells that were held flat were fibroblast-like: they exhibited flattened morphology, more rapid growth, greater incorporation of [3H]thymidine, and less incorporation of 35SO4. These studies suggest that cell shape may play an important role in phenotypic expression in chondrocytes.

Cartilage↗