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Biomedical subjects

E van der Zee

Publications and source records attributed to E van der Zee.

12 recordsLinked to original sources

[Guided bone regeneration for single tooth replacements by oral implants].

Single tooth replacement is a good indication for an oral implant. However, bone regeneration, is often required to improve the bone quantity and the contour of the overlying soft tissues. Guided Bone Regeneration (GBR) using membranes is a good technique to restore local bone defects within a six months period. With this technique it is crucial that a membrane properly separates the bony and soft tissues and creates a space around a bony defect for bone ingrowth. This article describes the indications and contra-indications for GBR, the technique and the possible complications.

Bone Regeneration↗

EGF and IL-1 alpha modulate the release of collagenase, gelatinase and TIMP-1 as well as the release of calcium by rabbit calvarial bone explants.

Matrix metalloproteinases (MMPs), among which is collagenase (MMP-1), are likely to be involved in various steps of the bone resorption process. As both production of these enzymes and bone resorption appear to be mediated by cytokines, we investigated the effects of two cytokines, IL-1 alpha and EGF, on the release of collagenase, gelatinase A (MMP-2), gelatinase B (MMP-9), TIMP-1 and calcium by rabbit calvariae. It was found that all these parameters increased under the influence of these cytokines. The release of calcium--used as a parameter of bone resorption--was highest in the combined presence of the cytokines. Although the absolute and relative enhancement by a combination of IL-1 alpha and EGF was most pronounced for collagenase (7-fold), both gelatinase A (5-fold) and gelatinase B (1.5-fold) had increased simultaneously. Calvariae produced a high level of MMP inhibitor (TIMP-1), especially under the influence of the cytokines; periosteum released little inhibitor. It is concluded that IL-1 alpha and EGF are likely to play a modulating role in the process of bone resorption.

Analysis of Variance↗

Cytokines modulate routes of collagen breakdown. Review with special emphasis on mechanisms of collagen degradation in the periodontium and the burst hypothesis of periodontal disease progression.

In this paper, we review recent work on collagen degradation, 2 main routes of breakdown are described and their relevance during healthy and inflammatory conditions of the periodontium is discussed. Special attention is paid to the possible role of cytokines, in particular interleukin 1 (IL-1) and transforming growth factor beta (TGF-beta), on the modulation of collagen phagocytosis and metalloproteinase production. IL-1 has been shown to have a dual function in collagen digestion. It inhibits the intracellular phagocytic pathway, but at the same time, it strongly promotes extracellular digestion by inducing the release of collagenolytic enzymes like collagenase. TGF-beta has an opposite effect on both pathways and antagonizes IL-1. Collagenase is released in an inactive form, and a considerable fraction of the proenzyme may become incorporated in the extracellular matrix. This reservoir of latent enzyme can be activated (for instance by plasmin), leading to a sudden and extensive breakdown of the collagenous fibre meshwork. It is suggested that this phenomenon may also take place during progressive periodontitis and could explain an episodic nature of collagenolysis, clinically resulting in bursts of attachment loss (burst hypothesis).

Animals↗

Net-shaped hydroxyapatite implants for release of agents modulating periodontal-like tissues.

Periodontal-like tissues and, in particular, alveolar bone- and root cementum-like material can theoretically be modulated by release of biochemical agents such as bisphosphonate (PCP), growth hormone (GH) and alkaline phosphatase (ALP) from the implant surface. The present research focused on porous ceramic hydroxyapatite (PCHA) implants. In the past the PCHA implants were machined on a lathe out of simple blocks of PCHA ceramic. This was a tedious and cumbersome method, often resulting in implants with undesirable characteristics: different porosities, cracks and fractures. Therefore a moulding technique was developed to sinter near-net-shaped PCHA implants at 2 different sintering temperatures: 1170 degrees C and 1280 degrees C, resulting in PCHA implants with porosities of 62.06% (PCHA type 1) and 40.74% (PCHA type 2), respectively. After 1 h incubation in a 10(-2) M solution of PCP, the total amounts adsorbed onto PCHA type 1 and type 2 were 114.9 +/- 2.1 micrograms and 46.1 +/- 1.5 micrograms, respectively. This was approximately 5 times higher than after incubation for 1 wk in a 10(-4) M solution of PCP. The total amounts of PCP released after the observation period of 75 d from PCHA type 1 and type 2 after incubation in the 10(-2) M solution were 103.1 +/- 1.8 micrograms and 42.8 +/- 1.5 micrograms, respectively. The total amounts released from type 1 and 2 after incubation in the 10(-4) M solution were 7.4 +/- 0.4 micrograms and 4.1 +/- 0.1 micrograms, respectively. After 2 wk of incubation in a liver/bone/kidney ALP solution the total amount of ALP adsorbed onto PCHA type 1 implants was 5039 +/- 412 mU/ml. The total amounts of ALP released were 4674 +/- 438 mU/ml and 53 +/- 20 mU/ml after 1 and 2 wk, respectively. The release of ALP was high at the beginning but slowed down thereafter. It was evident that despite the well-known high bonding affinity of PCP to HA the release of PCP occurred steadily, over a long period of time in vitro.

Adsorption↗

Unbiased stereological estimation of the total number of synapses in a brain region.

Modern stereological methods have been used to make unbiased estimates of the total number of synapses in the striatum radiatum of the hippocampal CA1 region of five rabbits. The approach used involved a two stage analysis and is generally applicable to all parts of the nervous system. During the first stage of the analysis, the reference volume was estimated by point counting, at the light microscope level, according to the Cavalieri principle. During the second stage, the numerical density of synapses was estimated with dissectors at the electron microscopic level. The total number of synapses was calculated as the product of the numerical density and the volume of the region. The sampling with points and dissectors was carried out in all three dimensions of the entire CA1 region in a manner that ensured that all parts of the region and all synapses within it had equal probabilities of being sampled. An analysis of the precision of the estimate of total synapse number has been performed in terms of the variances of volume and synaptic numerical density at different levels of sampling, i.e. at the level of points, sections, individual animals and group of animals. Detailed descriptions of the procedures used to estimate the total number of synapses, evaluate the precision of the estimates, and optimize the sampling scheme are provided.

Analysis of Variance↗

Phagocytosis and intracellular digestion of collagen, its role in turnover and remodelling.

Collagens of most connective tissues are subject to continuous remodelling and turnover, a phenomenon which occurs under both physiological and pathological conditions. Degradation of these proteins involves participation of a variety of proteolytic enzymes including members of the following proteinase classes: matrix metalloproteinases (e.g. collagenase, gelatinase and stromelysin), cysteine proteinases (e.g. cathepsin B and L) and serine proteinases (e.g. plasmin and plasminogen activator). Convincing evidence is available indicating a pivotal role for matrix metalloproteinases, in particular collagenase, in the degradation of collagen under conditions of rapid remodelling, e.g. inflammation and involution of the uterus. Under steady state conditions, such as during turnover of soft connective tissues, involvement of collagenase has yet to be demonstrated. Under these circumstances collagen degradation is likely to take place particularly within the lysosomal apparatus after phagocytosis of the fibrils. We propose that this process involves the following steps: (i) recognition of the fibril by membrane-bound receptors (integrins?), (ii) segregation of the fibril, (iii) partial digestion of the fibril and/or its surrounding non-collagenous proteins by matrix metalloproteinases (possibly gelatinase), and finally (iv) lysosomal digestion by cysteine proteinases, such as cathepsin B and/or L. Modulation of this pathway is carried out under the influence of growth factors and cytokines, including transforming growth factor beta and interleukin 1 alpha.

Animals↗

Cytokine-induced endogenous procollagenase stored in the extracellular matrix of soft connective tissue results in a burst of collagen breakdown following its activation.

Numerous data strongly suggest the involvement of cytokines and the matrix metalloproteinase collagenase (MMP-1) in the pathogenesis of periodontitis. Recently, we have demonstrated that, upon culturing under the influence of IL-1 alpha + EGF, a large amount of inactive procollagenase (MMP-1) is stored in the extracellular matrix of periosteal tissue. We now show that this endogenous reservoir of proenzyme can be operative after activation with plasmin and is able to induce a rapid and almost complete breakdown of the collagenous extracellular matrix. The level of collagen degradation following activation showed a strong correlation with the amount of proenzyme that was incorporated in the tissue. The highest level of degradation (70% of the total amount of collagenous proteins) was found with the IL-1 alpha + EGF-treated explants, followed by those treated with IL-1 alpha alone (35%). Explants cultured with EGF or in the absence of cytokines, containing only small amounts of procollagenase, showed little collagen breakdown following plasmin activation (7%). Inhibition of metalloproteinases by EDTA, or blockage of plasmin by PMSF, prevented the degradation in all explants irrespective of the amount of proenzyme present in the tissue. Our findings demonstrate that endogenous proenzyme stored in a native connective tissue matrix can be activated at a later time interval which results in a massive breakdown of the tissue. This study shows a possible pathway of collagenase-induced breakdown without recent de novo synthesis of the enzyme. Such a sequence may be operative in chronic inflammatory diseases, such as periodontitis, where production of procollagenase under the influence of cytokines spans a longer time period, whereas breakdown is often characterized by a cyclic behaviour.

Analysis of Variance↗

Cytokines modulate phagocytosis and intracellular digestion of collagen fibrils by fibroblasts in rabbit periosteal explants. Inverse effects on procollagenase production and collagen phagocytosis.

Degradation of fibrillar collagen may occur in the extracellular space by enzymes, such as the metalloproteinase collagenase, or in the lysosomal apparatus of fibroblasts following phagocytosis. As the mechanisms involved in the regulation of the latter process are unknown, we investigated possible modulating effects of the cytokines epidermal growth factor (EGF), platelet-derived growth factor (PDGF), interleukin-1 alpha (IL-1 alpha) and transforming growth factor-beta (TGF-beta) on both collagen phagocytosis and the release of collagenase in an in vitro model employing periosteal tissue explants. The data demonstrated that the level of intracellular collagen digestion could be influenced by cytokines: IL-1 alpha inhibited and TGF-beta enhanced phagocytosis of fibrillar collagen by periosteal fibroblasts, whereas the cytokines had an opposite effect on the release of procollagenase. In combination, IL-1 alpha and TGF-beta proved to have an antagonizing effect on either parameter. PDGF and EGF had no effect on phagocytosis or collagenase release. The level of phagocytosed collagen correlated positively with the actual breakdown of collagen as assessed by the release of hydroxyproline but negatively with the level of released procollagenase. Our findings demonstrated that cytokines are able to modulate both the phagocytosis of collagen fibrils by fibroblasts and their subsequent intracellular breakdown, as well as the release of procollagenase, an enzyme considered crucial for extracellular collagenolysis. Moreover, our data show a negative correlation between these two parameters. It is concluded that IL-1 alpha, EGF and TGF-beta may be important in modulating the contribution of the intracellular and extracellular route of collagen breakdown.

Animals↗

Cytokines modulate contraction of periosteal explants from rabbit calvariae.

Periosteal explants obtained from rabbit calvariae are introduced as a model system to study contraction of soft connective tissue. Culturing of these explants resulted in a substantial time-dependent decrease of the surface area which coincided with a similar decrease in volume. A 75% reduction of the initial explant surface area was observed after a culture period of 72h in the presence of 10% serum. Also in the absence of serum contraction of explants was found, indicating that serum components were not essential. Following devitalization of the explants no contraction did occur. A strong inhibition of contraction was found following interference with the formation of microfilaments, microtubules or intracellular cyclic-AMP levels by using cytochalasin B, colchicine or dibutyryl-cAMP, respectively. These data indicated that viable cells and an intact cytoskeleton were a prerequisite for contraction to occur. A number of cytokines (EGF, aFGF, bFGF, IGF-1, PDGF, TGF-beta and IL-1alpha) was tested for their ability to influence contraction. IL-1alpha was shown to inhibit contraction from the 48h culture period on. Anti-IL-1alpha-serum completely abolished this effect. The IL-1alpha-inducible inhibition of contraction was also partially blocked by indomethacin. TGF-beta enhanced contraction dose-dependently during the 24-48h culture period, whereas TGF-1 and IL-1alpha, added to the cultures in combination, proved to antagonize each other. The other growth factors did neither influence contraction not the IL-1alpha-induced inhibition of contraction.

Actin Cytoskeleton↗

Immunolocalisation of collagenase in rabbit periosteal tissue explants and extraction of the enzyme. The effect of the cytokines IL-1 alpha and EGF.

The effect of interleukin-1 alpha (IL-1 alpha) and murine epidermal growth factor (EGF) on incorporation of endogenously produced collagenase in the extracellular matrix of soft connective tissue was studied in an in vitro model system using periosteal explants obtained from rabbit calvariae. Immunohistochemical analysis indicated the highest level of collagenase in explants cultured for 72 hours with IL-1 alpha in combination with EGF. Most enzyme appeared to be associated with the extracellular matrix, but labeling was also found in numerous fibroblast-like cells. Explants cultured in the presence of IL-1 alpha alone contained less enzyme and in periostea treated without cytokines, or with EGF alone, only a faint label, if any, was seen. Freshly isolated, non-cultured periostea contained no detectable enzyme. Extraction of collagenase from periostea revealed that: (1) non-cultured periosteum did not contain detectable levels of enzyme. (2) The amount of total activatable enzyme synergistically increased (10-fold) under the influence of IL-1 alpha and EGF, whereas IL-1 alpha alone showed a 4-fold enhancement compared to control or EGF-incubated explants. (3) The latent fraction of the enzyme was synergistically increased (up to 100-fold or more) in periostea cultured in the presence of IL-1 alpha + EGF (21.17 mU/explant versus 0.05 mU/explant in controls). (4) Active collagenase, on the other hand, appeared to be present in a relatively high concentration in explants cultured without cytokines (2.45 mU/explant versus 0.36 mU/explant in IL-1 alpha + EGF-treated explants).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Interleukin-1 alpha and epidermal growth factor synergistically enhance the release of collagenase by periosteal connective tissue in vitro.

The effects of recombinant human interleukin-1 alpha (IL-1 alpha) and murine epidermal growth factor (EGF) on the release of collagenase were studied in an in vitro model system using periosteal explants from rabbit calvariae. Following an incubation period of 72 h it was shown that IL-1 alpha in combination with EGF (IL-1 alpha + EGF) induced a synergistic increase in the amount of collagenase released by periosteal explants. This increase appeared to be at least 10-fold. Most of the enzyme was present in a latent form since the increase in enzyme activity was only detectable after activation by APMA and the molecular weight as determined in immunoblots corresponded to the latent form of this enzyme. Incubations carried out with IL-1 alpha alone resulted in a 2- to 4-fold increase of total enzyme activity, whereas the amount of collagenase in media of EGF-treated periosteal did not surpass control values. A neutralizing anti-IL-1 alpha antibody completely blocked the enhanced release of collagenase as induced both by IL-1 alpha and by IL-1 alpha + EGF. Indomethacin partially prevented the IL-1 alpha + EGF-induced increase in enzyme release, suggesting the involvement of prostaglandins. The amount of tissue inhibitor of metalloproteinases (TIMP) as determined by ELISA was slightly elevated in culture media obtained from all cytokine-treated explants. Comparable results were obtained by Western blot analysis as well as by a functional bioassay. It is suggested that the concomitant presence of the cytokines IL-1 alpha and EGF may play an important role in collagenase-mediated degradation of collagen.

Animals↗

The effects of intraventricular carbachol injections on the free-running activity rhythm of the hamster.

The effects of light on the circadian pacemaker in the suprachiasmatic nucleus (SCN) are mediated by the retinohypothalamic tract (RHT) and by the retinogeniculosuprachiasmatic tract (RGST). The neurotransmitter of the RGST is neuropeptide Y. The RHT may contain glutamate and aspartate. Recent evidence indicates that acetylcholine could also be involved in phase shifting by light. We determined that intraventricular injections with an acetylcholine agonist, carbachol, induces phase advances during the subjective day and phase delays during the early subjective night. No differences were observed between phase shifts induced in constant darkness and those induced in continuous light. A dose-response curve for carbachol was described at circadian time 6 (CT6). Injections at CT14 with various dosages of carbachol indicated the same dose dependency for this circadian time. Finally, carbachol injections in split animals resulted in similar responses of the two components of the split activity rhythm.

Animals↗