Biomedical subjects

E B Hunziker

Publications and source records attributed to E B Hunziker.

At least 55 records · Page 3Linked to original sources

PubMed1999

Hypertrophy of growth plate chondrocytes in vivo is accompanied by modulations in the activity state and surface area of their cytoplasmic organelles.

The rate of longitudinal bone growth is regulated primarily by modulations in the activity of epiphyseal plate hypertrophic chondrocytes, these being manifested as changes in cell and matrix volume. It was the purpose of this study to ascertain whether the cytoplasmic organelles representing the cellular production apparatus, i.e. rough endoplasmic reticulum, Golgi apparatus and mitochondria, contribute to these changes by modulating their rate of activity or by increasing/decreasing the surface area and/or volume of their membranes. Using rats at different stages of growth, the surface areas and volumes of the three organellar systems were quantified in epiphyseal plate chondrocytes at the onset and termination of hypertrophy by ultrastructural stereology. Matrix synthesis during the same span was assessed by monitoring the production of its principal components, namely, fibrillar collagen (ultrastructural morphometry) and glycosaminoglycans (quantitative (35)S-autoradiography). Each organelle adapts to increases (21- to 35-day-old rats) and decreases (35- to 80-day-old rats) in growth rate by its own individual combination of the two alternative mechanisms, but modulations in the level of activity predominate over alterations in the surface area or volume of their membranes. These findings point to the danger of relying solely on data gleaned from a quantitative ultrastructural analysis of organellar parameters and emphasise the necessity of conducting functional assays in parallel, as performed here.

Animals↗

PubMed1999

Articular cartilage repair: are the intrinsic biological constraints undermining this process insuperable?

This article reviews the experimental and clinical strategies currently in use or under development for the treatment of articular cartilage lesions. The vast majority of protocols under investigation pertain to the treatment of full-thickness defects (i.e., those which penetrate the subchondral bone and trabecular-bone spaces) rather than partial-thickness ones (i.e., those which are confined to the substance of articular cartilage tissue itself). This bias probably reflects the circumstance that partial-thickness defects do not heal spontaneously whereas full-thickness ones below a critical size do, albeit transiently. And it is, of course, a seemingly easier task to manipulate a process which is readily set in train than it is to overcome an induction-problem which Nature herself has not solved. Indeed, the reasons for this inert state of partial-thickness defects have only recently been elucidated, and these are briefly discussed. However, the main body of this review deals with the various transplantation concepts implemented for the repair of full-thickness defects. These fall into two broad categories: tissue-based (entailing the grafting of perichondrial, periosteal, cartilage or bone-cartilage material) and cell-based (utilizing chondroblasts, chondrocytes, periochondrial cells or mesenchymal stem cells). Cell-based systems are further subdivided according to whether cells are transplanted within a matrix (biodegradable, non-biodegradable or synthetic) or free in suspension. Thus far, the application of cell suspensions has always been combined with the grafting of a periosteal flap. The strengths and weaknesses of each concept are discussed.

Cartilage Diseases↗

PubMed1999

Normal skeletal development of mice lacking matrilin 1: redundant function of matrilins in cartilage?

Matrilin 1, or cartilage matrix protein, is a member of a novel family of extracellular matrix proteins. To date, four members of the family have been identified, but their biological role is unknown. Matrilin 1 and matrilin 3 are expressed in cartilage, while matrilin 2 and matrilin 4 are present in many tissues. Here we describe the generation and analysis of mice carrying a null mutation in the Crtm gene encoding matrilin 1. Anatomical and histological studies demonstrated normal development of homozygous mutant mice. Northern blot and biochemical analyses show no compensatory up-regulation of matrilin 2 or 3 in the cartilage of knockout mice. Although matrilin 1 interacts with the collagen II and aggrecan networks of cartilage, suggesting that it may play a role in cartilage tissue organization, studies of collagen extractability indicated that collagen fibril maturation and covalent cross-linking were unaffected by the absence of matrilin 1. Ultrastructural analysis did not reveal any abnormalities of matrix organization. These data suggest that matrilin 1 is not critically required for cartilage structure and function and that matrilin 1 and matrilin 3 may have functionally redundant roles.

Animals↗

PubMed1999

Biologic repair of articular cartilage. Defect models in experimental animals and matrix requirements.

Articular cartilage lesions are a commonly encountered medical problem, for which there exists no satisfactory solution at present. Although many treatment protocols are in current clinical use, few of these have been the subject of rigorous evaluation in animal studies. Such an evaluation requires not only the selection of a suitable animal species, but also careful consideration of the defect parameters, not per se, but in relation to the biologic setting, and of whether this model is of relevance when transposed to the human situation. The current study addresses some of the common pitfalls into which investigators unknowingly stumble and which, unless avoided, can render their studies less meaningful in the human context. Because most treatment protocols entail the deposition of a space filling matrix within the defect void, the important attributes which such a material should possess also are described in this article.

Animals↗

PubMed1998

Endochondral ossification is dependent on the mechanical properties of cartilage tissue and on intracellular signals in chondrocytes.

Skeletal elements are formed either by replacing a performed cartilagenous matrix template in a process called endochondral ossification or directly from mesenchyme by a process known as membranous ossification. Longitudinal growth of bones is achieved by growth plates where calcified cartilage is converted into bone. To investigate the role of extracellular matrix as well as intracellular signaling pathways in the formation and growth of bone, the genes coding for type II collagen and cyclic guanosine 3',5'-monophosphate (cGMP)-dependent protein kinase (cGK) II, were disrupted. It is demonstrated that loss of Col2a1 or cGKII led to abnormal endochondral ossification and skeletal development. In cGKII -/- mice, bones derived by membranous ossification developed normally while bones derived by endochondral ossification were shortened. This growth defect was not associated with a general metabolic disturbance. In Col2a1 knockout mice, endochondral ossification was completely absent, whereas membraneous ossification was not affected. Despite the defects in bone formation, invasion of blood vessels into bone cavities and formation of bone marrow occurred in Col2a1-null mice. Taken together, the phenotypes of these two knockout mice show that chondrocytes need a well-functioning extracellular matrix scaffold and a normal cGMP-signaling system for endochondral ossification to form a normal skeleton.

Animals↗

PubMed1998

Effects of injurious compression on matrix turnover around individual cells in calf articular cartilage explants.

The effects of mechanical injury on the metabolism of cartilage matrix are of interest for understanding the pathogenesis of osteoarthrosis and the development of strategies for cartilage repair. The purpose of the present study was to examine the effects of injury on matrix turnover in a calf articular cartilage explant system for which the effects of mechanical loading on cell activity and the cell-mediated pathways of matrix metabolism are already well characterized. New methods of quantitative autoradiography were used in combination with established biochemical and biomechanical techniques for the analysis of cell and matrix responses to acute mechanical injury, with particular attention to the processes of localized matrix turnover in the cell-associated matrices of individual chondrocytes. Matrix deposition and turnover around cells in control explants was spatially dependent, with the highest rates of proteoglycan deposition and turnover and the lowest rates of collagen deposition (as indicated by [3H]proline autoradiography) occurring in the pericellular matrix. Injurious compression was associated with (a) an abrupt decrease in the tensile load-carrying capacity of the collagen matrix, apparently associated with mechanical failure of the tissue, (b) a considerable but subtotal decrease in cell viability, marked by the emergence of an apparently inactive cell population interspersed within catabolically active but abnormally large cells, and (c) sustained, elevated rates of proteoglycan turnover, particularly in the cell-associated matrices of apparently viable cells, which involved the increased release of aggregating species in addition to a spectrum of degradation fragments that were also in controls. These results may represent an in vitro model for the responses of chondrocytes and the cartilage extracellular matrix to mechanical injury.

Animals↗

PubMed1998

Changes in cell, matrix compartment, and fibrillar collagen volumes between growth-plate zones.

To define the contributions of changes in cell, matrix compartment, and fibrillar collagen volumes to longitudinal bone growth, we measured the differences in cell, pericellular/territorial matrix and interterritorial matrix volumes, and fibrillar collagen concentrations between the upper proliferative and lower hypertrophic zones of the proximal tibial physes of six miniature pigs. The mean numerical density of cells decreased from 110,000 cells/mm3 in the upper proliferative zone to 59,900 cells/mm3 in the lower hypertrophic zone. The mean cell volume increased nearly 5-fold (from 1,174 to 5,530 microm3), and the total matrix volume per cell increased 46% (from 8,040 to 11,760 microm3/cell) between the upper proliferative and lower hypertrophic zones. Both the pericellular/territorial matrix volume per cell and the interterritorial matrix volume per cell increased between the upper proliferative and lower hypertrophic zones; the pericellular/territorial matrix volume per cell increased 61% (from 4,580 to 7,390 microm3/cell), whereas the interterritorial matrix volume per cell increased 26% (from 3,460 to 4,370 microm3/cell). The total increase in mean cell volume of 4,356 microm3 exceeded the total increase in mean matrix volume per cell of 3,720 microm3; the total mean pericellular/territorial matrix volume per cell increased more than the total mean interterritorial matrix volume per cell (2,810 compared with 910 microm3/cell). Fibrillar collagen concentration was greater in the interterritorial matrix than in the pericellular/territorial matrix in both zones and increased in both matrix compartments between the upper proliferative and lower hypertrophic zones. The amount of fibrillar collagen per cell also increased in both matrix compartments between the upper proliferative and lower hypertrophic zones (from 1,720 to 3,100 microm3/cell in the pericellular/territorial matrix and from 1,490 to 2,230 microm3/cell in the interterritorial matrix; thus, the total amount of fibrillar collagen per cell increased from 3,210 to 5,530 microm3/cell). Growth rate was inversely related to the cell numerical density in the upper proliferative and lower hypertrophic zones and was directly related to interterritorial matrix volume per cell in the upper proliferative zone and to pericellular/territorial matrix volume per cell in the lower hypertrophic zone. These results show that cell enlargement contributes more to longitudinal bone growth than does increased matrix volume, that increased pericellular/territorial matrix volume makes a greater contribution to growth than does increased interterritorial matrix volume, and that the total amount of fibrillar collagen per cell increases between the upper proliferative and lower hypertrophic zones. The differences between the two matrix compartments in increase in volume, fibrillar collagen concentration, and amount of fibrillar collagen per cell strongly suggest that they differ not only in matrix organization but in rate of matrix accumulation and assembly and that these differences give the two compartments different roles in skeletal growth.

Animals↗

PubMed1998

Confined compression of articular cartilage: linearity in ramp and sinusoidal tests and the importance of interdigitation and incomplete confinement.

Experimental and theoretical methods were used to investigate the linearity of the stress response of articular cartilage to ramp and sinusoidal tests in confined compression, as well as the role of cartilage-porous platen and lateral confinement boundary conditions in determining material responses. With respect to linearity, we posed the question as to whether the elicited stress responses to ramp compression, ramp release and sinusoidal tests were similar. With respect to boundary conditions we inquired as to the necessity of specifying a detailed interdigitating contact with the porous filter and of specifying the level of confinement present at the lateral edge of the disk. We found that the stress responses to the three types of tests were dissimilar, with ramp compression the only test exhibiting linear behavior. Ramp release from a static compression offset was non-linear in a manner such that the cartilage maintained a compressive stress higher than expected by a linear theory. Sinusoidal compression also displayed a non-linear response consistent with the presence of a release phase in each cycle. The actual boundary conditions present at the cartilage/porous-filter interface were visualized histologically. Areas (tens of microns) of cartilage in contact with the metal of the filter were interspersed with areas expanded into the pores of the filter. Finite-element analysis incorporating this information suggested that precise specification of this interface and of the level of the extent of lateral confinement would be necessary for the estimation of material properties, such as the hydraulic permeability, from these tests. The trends of the linearity studies did not appear to be significantly affected by the problems posed by these difficult to quantitate boundary conditions. The non-linear cartilage response to release and sinusoidal displacements therefore appear to be physiologically interesting. The maintained, that is higher than would be linear, compressive stress observed during release may be a beneficial adaptation to repeated loading where temporal variations in tissue stresses would be minimized.

Animals↗

PubMed1998

Differential effects of IGF-1 and TGF beta-2 on the assembly of proteoglycans in pericellular and territorial matrix by cultured bovine articular chondrocytes.

OBJECTIVES: Knowledge of matrix assembly is necessary to understand the pathogenesis of disease processes and to find solutions for repair of articular cartilage lesions. The influence of growth factors on matrix assembly is largely unknown. We investigated whether, and to what degree, insulin-like growth factor (IGF-1) and transforming growth factor beta-2 (TGF beta-2) influence proteoglycan synthesis and accumulation in the cell-associated matrix compartment (consisting of pericellular and territorial matrix) compared to the further-removed matrix compartment (consisting of the interterritorial matrix). DESIGN: Bovine articular chondrocytes were cultured in alginate beads for day 13. The effect of addition of 25 ng/ml IGF-1 or 25 ng/ml TGF beta-2 during the last 7 days in culture was determined. Cell-associated and further-removed matrix compartments were separated by centrifugation after sodium citrate/EDTA treatment. The amount of DNA, the total amount of proteoglycans and the amount of newly synthesized proteoglycans were analyzed biochemically. Morphometric analysis on electron micrographs was used to calculate the volumes of the cell-associated and further-removed matrix components. RESULTS: It was demonstrated in control beads that 25 +/- 8% of the proteoglycans were laid down in the cell-associated matrix compartment compared with 75 +/- 8% in the further-removed matrix compartment. The cell-associated matrix compartment in intact beads could be recognized in electron microscopy by a delineation of dense amorph material. Morphometric evaluation showed a relative volume of the cell-associated matrix compartment of 5.2 +/- 0.6% compared with 91.3 +/- 0.8% of the further-removed matrix compartment and 3.5 +/- 0.3% of the area occupied by cells. Combination of biochemical and morphometric results showed that the concentration of proteoglycans in the cell-associated matrix compartment was 3.63 +/- 0.32 mg/ml. By adding IGF-1 or TGF beta-2, the amount of both total accumulated proteoglycans and newly synthesized [35S]proteoglycans at day 13 in culture increased. In addition to an overall rise in proteoglycan content, IGF-1 significantly increased (24%) the percentage of proteoglycans laid down in the cell-associated matrix compartment while not changing the relative volume of this compartment (5.2 +/- 0.8%). This leads to a 82% (P < 0.05) increase in the proteoglycan concentration in the cell-associated matrix compartment compared to control beads. In contrast, TGF beta-2 significantly decreased (24%) the relative amount of proteoglycans in the cell-associated matrix compartment which was paralleled by a reduction of the relative volume from 5.2 +/- 0.6 to 3.6 +/- 1.4%. This leads to a significant increase of 87% of the proteoglycan concentration in the cell-associated matrix compartment. CONCLUSIONS: This study demonstrates that both IGF-1 and TGF beta-2 significantly but differently influence proteoglycan synthesis and accumulation in the cell-associated matrix compartment of cultured bovine chondrocytes in alginate. Both growth factors increase the concentration of proteoglycans in the cell-associated matrix compartment. However, addition of TGF beta-2 to bovine articular chondrocytes cultured in alginate beads for 13 days results in a significant reduction of the relative volume of the pericellular matrix compartment compared to controls, indicating differences in assembly of the matrix.

Animals↗

PubMed1998

Mechanical compression alters proteoglycan deposition and matrix deformation around individual cells in cartilage explants.

We have used new techniques of cell-length scale quantitative autoradiography to assess matrix synthesis, deposition, and deformation around individual chondrocytes in mechanically compressed cartilage explants. Our objectives were to: (1) quantify the effects of static and dynamic compression on the deposition of newly synthesized proteoglycans into cell-associated and further-removed matrices; (2) measure cell-length scale matrix strains and morphological changes of the cell and matrix associated with tissue compression; and (3) relate microscopic physical stimuli to changes in proteoglycan synthesis as functions of compression level and position within mechanically compressed explants. Results indicate a high degree of structural organization in the extracellular matrix, with the pericellular matrix associated with the most rapid rates of proteoglycan deposition, and greatest sensitivity to mechanical compression. Static compression could stimulate directional deposition of secreted proteoglycans around chondrocytes, superimposed on an inhibition of proteoglycan synthesis; these events followed trends for compressive strain in the cell-associated matrix. Conversely, proteoglycan synthesis and pericellular deposition was stimulated by dynamic compression. Results suggest that cell-matrix interactions in the cell-associated matrix may be a particularly important aspect of the chondrocyte response to mechanical compression, possibly involving macromolecular transport limitations and morphological changes associated with fluid flow and local compaction of the matrix around cells.

Animals↗

PubMed1998

Removal of proteoglycans from the surface of defects in articular cartilage transiently enhances coverage by repair cells.

Lesions within the articular cartilage layer of synovial joints do not heal spontaneously. Some repair cells may appear, but their failure to become established may be related to problems of adhesion to proteoglycan-rich surfaces. We therefore investigated whether controlled enzymatic degradation of surface proteoglycan molecules to a depth of about 1 microm, using chondroitinase ABC, would improve coverage by repair cells. We created superficial lesions (1.0 x 0.2 x 5 mm) in the articular cartilage of mature rabbit knees and treated the surfaces with 1 U/ml of chondroitinase ABC for four minutes. The defects were studied by histomorphometry and electron microscopy at one, three and six months. At one month, untreated lesions were covered to a mean extent of 28% by repair cells; this was enhanced to a mean of 53% after enzyme treatment. By three months, the mean coverage of both control and chondroitinase-ABC-treated defects had diminished dramatically to 0.2% and 13%, respectively, but at six months both untreated and treated lesions had a similar coverage of about 30%, not significantly different from that achieved in untreated knees at one month. These findings suggest that, with time, chondrocytes near the surface of the defect may compensate for the loss of proteoglycans produced by enzyme treatment, thereby restoring the inhibitory properties of the matrix as regards cell adhesion. This supposition was confirmed by electron microscopy. Our results have an important bearing on attempts made to induce healing responses by transplanting chondrogenic cells or by applying growth factors.

Animals↗

PubMed1997

[Lesions of articular cartilage and their treatment].

Lesions of human articular cartilage most often are the result of a pathogenetically unclear disease process eventually leading to primary osteoarthrosis. The increasing incidence of traumatic injuries during sport activities, leading to lesions of cartilage and bone, is also worth noting. In addition, osteochondrosis dissecans and hereditary dysplasias of the skeletal system may lead to osseous and cartilaginous lesions. Adult articular cartilage has only very limited repair capacity in response to acute and chronic insults. Even now, diagnostic tools and methods of diagnosing cartilaginous lesions are still far from accurate. Lesions seen by conventional imaging technology chiefly represent late and irreversible damage of joint cartilage structures. The present review first summarizes current basic knowledge of joint anatomy and physiology, to enable the reader to understand better the unique nature of cartilaginous lesions and the difficulties and controversies involved in old and novel experimental methodologies in cartilage repair. It then critically evaluates developments in orthopedic therapy for repair of articular cartilage lesions.

Adult↗

PubMed1997

Ultrastructure of adult human articular cartilage matrix after cryotechnical processing.

The ultrastructure of adult human articular cartilage matrix is reexamined in tissue processed according to recently improved cryotechniques [Studer et al. (1995) J. Microsc., 179:321-332]. In truely vitrified tissue, a network of fine cross-banded filaments (10-15 nm in diameter) with a periodicity characteristic of collagen fibrils is seen throughout the extracellular substance, even within the pericellular compartment, which has hitherto been deemed free of such components. Proteoglycans fill the interstices between these entities as a homogeneously distributed granular mass; they do not manifest a morphologically identifiable reticular structure. Longitudinally sectioned collagen fibrils exhibit variations in thickness and kinking; they tend to align with their periodic banding in register and are frequently seen to split or fuse along their longitudinal course. The tendency of fibrils to form bundles is greater in deeper zones than in more superficial ones. A duality in the orientation of fibrils and fibril bundles is observed within the interterritorial matrix compartment: superimposed upon the well-characterized arcade-like structure formed by one subpopulation is another, more randomly arranged one. The classical concepts of matrix organization thus need to be modified and refined to encompass these findings.

Cartilage, Articular↗

PubMed1997

Optical and mechanical determination of Poisson's ratio of adult bovine humeral articular cartilage.

The equilibrium stiffness of articular cartilage is controlled by flow-independent elastic properties (Young's modulus, ES, and Poisson's ratio, v(s)) of the hydrated tissue matrix. In the current study, an optical (microscopic) method has been developed for the visualization of boundaries of cylindrical bovine humeral head articular cartilage disks (n = 9), immersed in physiological solution, and compressed in unconfined geometry. This method allowed a direct, model-independent estimation of Poisson's ratio of the tissue at equilibrium, as well as characterization of the shape changes of the sample during the nonequilibrium dynamic phase. In addition to optical analyses, the equilibrium behavior of cartilage disks in unconfined and confined ramp-stress relaxation tests provided a direct estimation of the aggregate modulus, H(a) and Young's modulus and, indirectly, Poisson's ratio for the articular cartilage. The mean value for Poisson's ratio obtained from the optical analysis was 0.185 +/- 0.065 (mean +/- S.D., n = 9). Values of elastic parameters obtained from the mechanical tests were 0.754 +/- 0.198 MPa, 0.677 +/- 0.223 MPa, and 0.174 +/- 0.106 for H(a), ES, and v(s), respectively (mean +/- S.D., n = 7). The similar v(s)-values obtained with optical and mechanical techniques imply that, at equilibrium for these two tests, the isotropic model is acceptable for mechanical analysis. However, the microscopic technique revealed that the lateral expansion, especially during the initial phase of relaxation, was inhomogeneous through the tissue depth. The superficial cartilage zone expanded less than the radial zone. The zonal differences in expansion were attributed to the known zonal differences in the fibrillar collagen architecture and proteoglycan concentration.

Analysis of Variance↗

PubMed1997

A new biological glue for cartilage-cartilage interfaces: tissue transglutaminase.

In this study, we used an in vitro model to test the capacity of tissue transglutaminase to increase the adhesive strength at a cartilage-cartilage interface. Full-thickness cartilage-bone cylinders were prepared from fresh adult bovine shoulder joints, and the superficial half of the hyaline cartilage was then removed to provide a plane surface. Tissue transglutaminase was applied to the freshly cut surface of one cylinder, and a calcium-chloride solution (to act as an activating agent) was applied to that of the other. The cartilage surfaces were immediately apposed, one on top of the other, and an eighty-gram weight was applied to the upper cylinder for ten minutes at 37 degrees Celsius under defined humidity conditions. A measured force was then applied transversely to the upper cylinder until it was displaced from the lower one (which was clamped in a holding device), and the force recorded at this point was taken as a measure of the adhesive strength achieved at the cartilage-cartilage interface. The adhesive strength increased linearly with an increasing concentration of tissue transglutaminase (0.25 to 2.75 milligrams per milliliter) and was enhanced by increasing the duration of incubation, but it was not influenced by the level of humidity. The adhesive strength was improved by as much as 40 per cent when the cartilage surfaces had been pretreated with chondroitinase AC or hyaluronidase to remove glycosaminoglycan chains of proteoglycans, which are largely responsible for the intrinsic anti-adhesive properties of cartilage.

Adhesiveness↗

PubMed1996

Visualization by magnetic resonance imaging of focal cartilage lesions in the excised mini-pig knee.

This study showed that magnetic resonance imaging can be used to visualize partial thickness cartilage lesions, 0.7 x 10 mm in area and 0.5 mm in depth, surgically induced in the femur (femoropatellar compartment) of a mini-pig knee joint. Formalin-fixed joints, intact as well as disarticulated, were studied by high resolution imaging using a 2.35 T, 31 cm horizontal-bore superconducting magnet. The two-dimensional and three-dimensional spatial resolutions achievable were as follows: 0.12 x 0.23 mm (two-dimensional) and 0.35 x 0.35 x 0.35 mm (three-dimensional) for the intact joint, and 0.08 x 0.08 mm (two-dimensional) and 0.14 x 0.14 x 0.27 mm (three-dimensional) for the disarticulated joint. These results demonstrate that magnetic resonance imaging, together with edge detection and volume rendering, can be used to visualize focal cartilage lesions.

Animals↗

PubMed1996

Vitrified articular cartilage reveals novel ultra-structural features respecting extracellular matrix architecture.

The quality of cryosections prepared from high pressure frozen bovine articular cartilage has been recently evaluated by systematic electron diffraction analysis, and vitrification found to be zone-dependent. The lower radial layer was optimally frozen throughout the entire section thickness (150 microns), whereas in the upper radial, transitional and superficial layers this was achieved down to a depth of only approximately 5-50 microns. These differences were found to correlate proportionally with proteoglycan concentration and inversely with water content. In the current investigation, extracellular matrix ultrastructure was examined in high pressure frozen material (derived from the lower radial zone of young adult bovine articular cartilage), by both cryoelectron microscopy of cryosections and by conventional transmission electron microscopy of freeze-substituted and embedded samples. Several novel features were revealed, in particular, the existence of a fine filamentous network; this consisted of elements 10-15 nm in diameter and with a regular cross-banded structure similar to that characterising collagen fibrils. These filaments were encountered throughout the entire extracellular space, even within the pericellular region, which is generally believed to be free of filamentous or fibrillar components. The proteoglycan-rich interfibrillar/filamentous space manifested a fine granular appearance, there being no evidence of the reticular network previously seen in suboptimally frozen material.

Animals↗

PubMed1996

Growth promoting effects of growth hormone and IGF-I are additive in experimental uremia.

Exogenous growth hormone (GH) stimulates the endogenous production of IGF-I and improves growth in uremia. We investigated whether exogenous IGF-I is also able to improve uremic growth failure in rats and whether the growth promoting effects of GH and IGF-I are additive. In female 150 g uremic (subtotal nephrectomy, NX) Sprague-Dawley rats, both rhGH in doses from 2 X 1.25 to 2 X 10 IU/kg bid s.c. and rhIGF-I in doses from 2 X 0.5 to 2 X 4.0 mg/kg bid s.c. caused a dose-dependent increase in weight gain and length gain. However, endogenous production of GH was suppressed by both agents. Peptide hormone treatment did not affect cumulative food intake, but significantly increased food efficiency ratio (weight gain/food intake). Concomitant s.c. treatment with maximally effective doses of rhGH (12 X 5 IU/kg bid) and of rhIGF-I (2 X 2 mg/kg bid) resulted in additive growth promoting effects in NX and pair-fed control (CO) animals during the observation period of 12 days. Cumulative length gain was 3.2 +/- 0.5 cm in solvent-treated NX-animals, 4.1 +/- 0.5 cm with rhGH (+ 28% above solvent), 4.2 +/- 0.6 cm with rhIGF-I (+ 31%) and 4.9 +/- 0.5 cm with both peptides (+ 53%). The food efficiency ratio was 0.16 +/- 0.05 in solvent NX, 0.33 +/- 0.04 with rhGH (+ 106% above solvent), 0.23 +/- 0.02 with rhIGF-I (+ 44%), and 0.38 +/- 0.02 with both peptides (+ 138%). Histomorphometric analysis and measurements of length gain by fluorescence microscopy in the upper tibial metaphysis confirmed the growth promoting effects of both peptide hormones. The serum concentrations of IGF binding protein (BP)-4 (Western ligand blotting analysis) and of IGFBP-2 (immunoblot) were increased in uremic animals whereas IGFBP-3 was unchanged. Treatment with IGF-I and/or rhGH increased serum concentration of IGF-I but did not change the IGFBP pattern. rhIGF-I lowered blood glucose levels within one to two hours after injection. The effect was most pronounced during the first treatment day and declined thereafter. Concomitant treatment with rhGH attenuated the glucose lowering effect of rhIGF-I (glucose serum concentration at day one: 120 +/- 11 mg% in solvent NX, 50 +/- 21 mg% with rhIGF-I, 80 +/- 24 mg% with both peptides). It is concluded that: (i) IGF-I is able to stimulate growth in NX animals but suppresses endogenous GH production in the long run; (ii) the concomitant treatment with IGF-I and GH has additive effects on growth; and (iii) concomitant treatment with rhGH prevents hypoglycemia that is noted with rhIGF-I alone.

Animals↗