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1.
Acta Biomater ; 115: 197-209, 2020 10 01.
Article in English | MEDLINE | ID: mdl-32814142

ABSTRACT

Hydrogels used in regenerative medicine are often designed to allow cellular infiltration, degradation, and neovascularization. Low molecular weight hydrogels (LMWHs), formed by self-assembly via non-covalent interactions, are gaining significant interest because they are soft, easy to use and injectable. We propose LMWHs as suitable body implant materials that can stimulate tissue regeneration. We produced four new LMWHs with molecular entities containing nucleic acid and lipid building blocks and analyzed the foreign body response upon subcutaneous implantation into mice. Despite being infiltrated with macrophages, none of the hydrogels triggered detrimental inflammatory responses. Most macrophages present in the hydrogel-surrounding tissue acquired an immuno-modulatory rather than inflammatory phenotype. Concomitantly, no fibrotic capsule was formed after three weeks. Our glyconucleolipid LMWHs exhibited different degradation kinetics in vivo and in vitro. LMWHs with high angiogenic properties in vivo, were found to release glyconucleoside (glucose covalently linked to thymidine via a triazole moiety) as a common by-product of in vitro LMWH degradation. Chemically synthesized glyconucleoside exhibited angiogenic properties in vitro in scratch assays with monolayers of human endothelial cells and in vivo using the chick chorioallantoic membrane assay. Collectively, LMWHs hold promise as efficient scaffolds for various regenerative applications by displaying good biointegration without causing fibrosis, and by promoting angiogenesis through the release of a pro-angiogenic degradation product. STATEMENT OF SIGNIFICANCE: The main limitations of biomaterials developed in the field of tissue engineering remains their biocompatibility and vascularisation properties. In this context, we developed injectable Low Molecular Weight Hydrogels (LMWH) exhibiting thixotropic (reversible gelation) and thermal reversible properties. LMWH having injectability is of great advantage since it allows for their delivery without wounding the surrounding tissues. The resulting gels aim at forming scaffolds that the host cells colonize without major inflammation, and that won't be insulated by a strong fibrosis reaction. Importantly, their molecular degradation releases a product (a glycosyl-nucleoside conjugate) promoting angiogenesis. In this sense, these LMWH represent an important advance in the development of biomaterials promoting tissue regeneration.


Subject(s)
Endothelial Cells , Hydrogels , Animals , Biocompatible Materials , Heparin, Low-Molecular-Weight , Hydrogels/pharmacology , Mice , Tissue Engineering
2.
Biomaterials ; 229: 119577, 2020 01.
Article in English | MEDLINE | ID: mdl-31704466

ABSTRACT

The durability of prosthetic arteriovenous (AV) grafts for hemodialysis access is low, predominantly due to stenotic lesions in the venous outflow tract and infectious complications. Tissue engineered blood vessels (TEBVs) might offer a tailor-made autologous alternative for prosthetic grafts. We have designed a method in which TEBVs are grown in vivo, by utilizing the foreign body response to subcutaneously implanted polymeric rods in goats, resulting in the formation of an autologous fibrocellular tissue capsule (TC). One month after implantation, the polymeric rod is extracted, whereupon TCs (length 6 cm, diameter 6.8 mm) were grafted as arteriovenous conduit between the carotid artery and jugular vein of the same goats. At time of grafting, the TCs were shown to have sufficient mechanical strength in terms of bursting pressure (2382 ±â€¯129 mmHg), and suture retention strength (SRS: 1.97 ±â€¯0.49 N). The AV grafts were harvested at 1 or 2 months after grafting. In an ex vivo whole blood perfusion system, the lumen of the vascular grafts was shown to be less thrombogenic compared to the initial TCs and ePTFE grafts. At 8 weeks after grafting, the entire graft was covered with an endothelial layer and abundant elastin expression was present throughout the graft. Patency at 1 and 2 months was comparable with ePTFE AV-grafts. In conclusion, we demonstrate the remodeling capacity of cellularized in vivo engineered TEBVs, and their potential as autologous alternative for prosthetic vascular grafts.


Subject(s)
Blood Vessel Prosthesis Implantation , Blood Vessel Prosthesis , Carotid Arteries/surgery , Jugular Veins/surgery , Renal Dialysis , Tissue Engineering , Vascular Patency
3.
Curr Res Transl Med ; 64(4): 171-177, 2016.
Article in English | MEDLINE | ID: mdl-27939455

ABSTRACT

The importance of proper skin wound healing becomes evident when our body's repair mechanisms fail, leading to either non-healing (chronic) wounds or excessive repair (fibrosis). Chronic wounds are a tremendous burden for patients and global healthcare systems and are on the rise due to their increasing incidence with age and diabetes. Curiously, these same risk factors also sign responsible for the development of hypertrophic scarring and organ fibrosis. Activated repair cells - myofibroblasts - are the main producers and organizers of extracellular matrix which is needed to restore tissue integrity after injury. Too many myofibroblasts working for too long cause tissue contractures that ultimately obstruct organ function. Insufficient myofibroblast activation and activities, in turn, prevents normal wound healing. This short review puts a spotlight on the myofibroblast for those who seek therapeutic targets in the context of dysregulated tissue repair. "Keep your myofibroblasts in balance" is the message.


Subject(s)
Myofibroblasts/physiology , Wound Healing/physiology , Animals , Apoptosis , Cicatrix/metabolism , Cicatrix/pathology , Contracture/etiology , Contracture/pathology , Contracture/prevention & control , Extracellular Matrix/metabolism , Fibrosis/pathology , Fibrosis/prevention & control , Granulation Tissue/pathology , Humans , Intercellular Signaling Peptides and Proteins/physiology
4.
Br J Dermatol ; 164(1): 125-34, 2011 Jan.
Article in English | MEDLINE | ID: mdl-20804491

ABSTRACT

BACKGROUND: Epidermolytic hyperkeratosis (bullous congenital ichthyosiform erythroderma), characterized by ichthyotic, rippled hyperkeratosis, erythroderma and skin blistering, is a rare autosomal dominant disease caused by mutations in keratin 1 or keratin 10 (K10) genes. A severe phenotype is caused by a missense mutation in a highly conserved arginine residue at position 156 (R156) in K10. OBJECTIVES: To analyse molecular pathomechanisms of hyperproliferation and hyperkeratosis, we investigated the defects in mechanosensation and mechanotransduction in keratinocytes carrying the K10(R156H) mutation. METHODS: Differentiated primary human keratinocytes infected with lentiviral vectors carrying wild-type K10 (K10(wt)) or mutated K10(R156H) were subjected to 20% isoaxial stretch. Cellular fragility and mechanosensation were studied by analysis of mitogen-activated protein kinase activation and cytokine release. RESULTS: Cultured keratinocytes expressing K10(R156H) showed keratin aggregate formation at the cell periphery, whereas the filament network in K10(wt) cells was normal. Under stretching conditions K10(R156H) keratinocytes exhibited about a twofold higher level of filament collapse compared with steady state. In stretched K10(R156H) cells, higher p38 activation, higher release of tumour necrosis factor-α and RANTES but reduced interleukin-1ß secretion compared with K10(wt) cells was observed. CONCLUSIONS: These results demonstrate that the R156H mutation in K10 destabilizes the keratin intermediate filament network and affects stress signalling and inflammatory responses to mechanical stretch in differentiated cultured keratinocytes.


Subject(s)
Chemokine CCL5/metabolism , Hyperkeratosis, Epidermolytic , Keratin-10/metabolism , Keratinocytes/metabolism , Tumor Necrosis Factor-alpha/metabolism , p38 Mitogen-Activated Protein Kinases/metabolism , Blotting, Western , Cells, Cultured , Fluorescent Antibody Technique , Humans , Hyperkeratosis, Epidermolytic/genetics , Hyperkeratosis, Epidermolytic/metabolism , Interleukin-1beta/metabolism , Keratin-10/genetics , Keratin-10/ultrastructure , Keratinocytes/physiology , Keratinocytes/ultrastructure , Mutation , Polymerase Chain Reaction/methods , RNA, Messenger/metabolism , Stress, Mechanical
5.
Diabet Med ; 26(12): 1296-300, 2009 Dec.
Article in English | MEDLINE | ID: mdl-20002485

ABSTRACT

AIMS: Individuals with diabetic retinopathy (DR) represent a high-risk group who would benefit from intensive metabolic control and risk factor management. This brief report examines quality of care among diabetic patients attending a tertiary retinal clinic. METHODS: A cross-sectional survey, notes review, and slit-lamp examination was conducted in 139 diabetic patients attending a specialist retinal clinic to assess the quality of comprehensive diabetes care. DR was graded according to the Early Treatment Diabetic Retinopathy Study scale. RESULTS: The prevalence of non-proliferative DR (NPDR) and proliferative DR (PDR) was 39.6 and 35.2%, respectively. The prevalence of microalbuminuria in patients with no DR, NPDR and PDR was 32, 54.1 and 68.8%, respectively. Glycaemic control was suboptimal (mean HbA(1c) 8.0 +/- 1.8%) and 15.8% were current smokers. Drugs affecting the renin-angiotensin system were used by only 61.9% of patients with both DR and microalbuminuria, and aspirin by only 35.3%. CONCLUSIONS: These data suggest that diabetes care in this high-risk population with established microvascular complications was suboptimal. Specialist clinics dealing with diabetic complications may be a setting where quality improvement strategies to reduce morbidity and mortality should be focused.


Subject(s)
Delivery of Health Care/standards , Diabetes Mellitus, Type 1/therapy , Diabetes Mellitus, Type 2/therapy , Diabetic Retinopathy/epidemiology , Adult , Aged , Albuminuria/epidemiology , Cross-Sectional Studies , Diabetes Mellitus, Type 1/blood , Diabetes Mellitus, Type 1/complications , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/complications , Diabetic Retinopathy/diagnosis , Female , Glycated Hemoglobin/analysis , Health Knowledge, Attitudes, Practice , Humans , Male , Middle Aged , Outcome and Process Assessment, Health Care , Prevalence
6.
Int J Clin Pharmacol Ther ; 47(10): 643-8, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19825328

ABSTRACT

BACKGROUND AND AIM: Diclofenac is a nonsteroidal antiinflammatory drug with potent analgesic and anti-inflammatory properties. An immediate-release formulation containing a low dose of 12.5 mg diclofenac-potassium (-K) is marketed as over the counter (OTC) medication in most European countries. An immediate-release formulation containing 25 mg diclofenac-K has now also been approved for OTC use. This study assessed the bioequivalence of two immediate-release diclofenac formulations when administered at the same dose. SUBJECTS AND METHODS: A randomized, crossover, open-label study was conducted in 29 healthy volunteers to assess the bioequivalence of single 25 mg dose of diclofenac-K in two formulations: 2 x 12.5 mg as film-coated tablets and 1 x 25 mg as sugar-coated tablet. Blood samples for pharmacokinetic analyses were obtained over 10 hours post administration. RESULTS: Plasma time courses of diclofenac were similar for the tested formulations. Mean AUC yen was 798 +/- 281 ng x h/ml (mean +/- SD) for the film-coated and 776 +/- 249 ng x h/ml for the sugar-coated formulation, respectively. The 2-sided 90% confidence interval for the mean test/reference ratio of AUCinfinity (95.5 - 107.5) fell within the predetermined equivalence range of 80 - 125%. Both formulations were rapidly absorbed; median time to maximal plasma concentration was 35 min in each group. Adverse events (peripheral erythema on the hand, headache, hypoesthesia) reported during the study were of mild severity and were considered as unlikely to be drug-related. CONCLUSION: The two diclofenac-K immediate release formulations were pharmacokinetically similar. It can be concluded that the new sugar-coated tablet formulation is equivalent to the available film-coated tablet formulation with respect to the extent of diclofenac absorption.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacokinetics , Diclofenac/pharmacokinetics , Nonprescription Drugs/pharmacokinetics , Adult , Anti-Inflammatory Agents, Non-Steroidal/administration & dosage , Anti-Inflammatory Agents, Non-Steroidal/adverse effects , Area Under Curve , Cross-Over Studies , Diclofenac/administration & dosage , Diclofenac/adverse effects , Excipients/chemistry , Female , Humans , Male , Nonprescription Drugs/administration & dosage , Nonprescription Drugs/adverse effects , Tablets , Therapeutic Equivalency , Young Adult
8.
Physiol Res ; 58(2): 193-202, 2009.
Article in English | MEDLINE | ID: mdl-18380533

ABSTRACT

Microvessels respond to metabolic stimuli (e.g. pO(2)) and hemodynamic forces (e.g. shear stress and wall stress) with structural adaptations including angiogenesis, remodeling and pruning. These responses could be mediated by differential gene expression in endothelial and smooth muscle cells. Therefore, rat mesenteric arteries and veins were excised by microsurgery, and mRNA expression of four angioadaptation-related genes was quantified by real time duplex RT-PCR in equal amounts of total RNA, correlated to two different house keeping genes (beta-actin, GAPDH). The results show higher expression of VEGFA, TIE2, and ANG2 in arteries than in veins, but equal expression of ADAMTS1. Higher availability of VEGFA mRNA in endothelial cells of arteries shown here could contribute to the maintenance of mechanically stressed blood vessels and counteract pressure-induced vasoconstriction.


Subject(s)
ADAM Proteins/genetics , Angiopoietin-2/genetics , Mesenteric Arteries/physiology , Mesenteric Veins/physiology , Receptor, TIE-2/genetics , Vascular Endothelial Growth Factor A/genetics , ADAMTS1 Protein , Adaptation, Physiological/physiology , Animals , Endothelium, Vascular/physiology , Gene Expression/physiology , Male , Microcirculation/physiology , Muscle, Smooth, Vascular/physiology , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Vasoconstriction/physiology
9.
Int J Clin Pharmacol Ther ; 46(4): 180-6, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18397691

ABSTRACT

OBJECTIVE: Over the last few years, there has been concern regarding the cardiovascular safety of selective cyclooxygenase (COX)-2 inhibitors and high-dose regimens of nonsteroidal anti-inflammatory drugs (NSAIDs). On the other hand, those compounds which elicit an almost complete (> 95%) and continuous suppression of platelet COX-1 may represent an exception. Apart from aspirin, which irreversibly inactivates COX-1, high-dose naproxen causes sustained COX-1 inhibition throughout the dose interval in some individuals. The present study examines whether naproxen sodium after a single dose administration and at steady state using "over-the-counter (OTC) doses" produces sufficient COX-1 inhibition. COX-2 inhibition was assessed concomitantly. METHODS: Ex vivo inhibition of COX enzymes and the pharmacokinetics of naproxen were assessed in four volunteers receiving 220 mg naproxen sodium b.i.d. for 7 days. Blood samples were obtained pre-dose, at specified time points after the first dose on Day 1, and 12 hours after the previous evening dose on Days 2, 3, 4, 5 and 8. Recovery was assessed up to 36 hours after the last dose. Coagulation-induced thromboxane B2 formation and lipopolysaccharide-induced prostaglandin E2 synthesis were measured ex vivo in human whole blood as indices of COX-1 and COX-2 activity. RESULTS: Maximal inhibition after a single dose and at steady state were as follows: 94% and 93% (COX-1), and 79% and 85% (COX-2). A greater than 95% COX-1 inhibition was observed transiently in 2 of 4 volunteers at the time of maximal plasma concentration after a single-dose administration and in 1 of 4 volunteers throughout the 12-hour dose interval at steady state. For both isoenzymes, COX inhibition correlated with naproxen plasma levels (ex vivo IC50 values of 35.48 micromol/l (COX-1) and 64.62 micromol/l (COX-2)). CONCLUSIONS: Administration of naproxen sodium at OTC doses was associated with a profound inhibition of both COX enzymes. Although low-dose naproxen may elicit a virtually complete COX-1 inhibition in some individuals, it does not mimic the reliable, sustained and complete COX-1 blockade produced by aspirin. In conclusion, prolonged treatment with 220 mg naproxen sodium b.i.d. is not expected to provide sufficient cardioprotection in all patients, but may influence platelet function in some.


Subject(s)
Cyclooxygenase Inhibitors/administration & dosage , Cyclooxygenase Inhibitors/pharmacology , Naproxen/administration & dosage , Naproxen/pharmacology , Adult , Aged , Chromatography, High Pressure Liquid , Cyclooxygenase 1/blood , Cyclooxygenase 2/blood , Dinoprostone/blood , Dose-Response Relationship, Drug , Escherichia coli , Female , Humans , In Vitro Techniques , Isoenzymes/antagonists & inhibitors , Isoenzymes/blood , Lipopolysaccharides/pharmacology , Male , Middle Aged , Platelet Aggregation/drug effects , Thromboxane B2/blood
10.
Oncogene ; 27(21): 3032-44, 2008 May 08.
Article in English | MEDLINE | ID: mdl-18071320

ABSTRACT

The role of cyclooxygenase-2 (COX-2) in cancer remains controversial. Using cervical carcinoma cells (HeLa), the present study investigates the involvement of COX-2 in apoptosis elicited by the chemotherapeutics paclitaxel, cisplatin and 5-fluorouracil. Each compound led to a profound induction of COX-2 expression and prostaglandin (PG) synthesis, accompanied by a substantial decrease of viability and enhanced apoptosis. Cells were significantly less sensitive to apoptotic death when either COX-2 expression or its activity was suppressed by small-interfering RNA (siRNA) and by the selective COX-2 inhibitor NS-398, respectively. Experiments performed to clarify how COX-2 leads to apoptosis revealed a profound proapoptotic action of PGD2 and its dehydration product, 15-deoxy-Delta(12,14) PGJ2 (15d-PGJ2). In line with these findings, chemotherapeutics-induced apoptosis was prevented by siRNA targeting lipocalin-type PGD synthase (L-PGDS), which catalyses the isomerization of PGH(2) to PGD2. Moreover, apoptosis by chemotherapeutics, PGD2 and 15d-PGJ2 was suppressed by the peroxisome proliferator-activated receptor gamma (PPARgamma) antagonist, GW-9662 or PPARgamma siRNA. Finally, a COX-2-dependent apoptotic mechanism of all investigated chemotherapeutics was confirmed in human lung cancer cells (A549) as well as in another cervical carcinoma cell line (C33A). Collectively, this study suggests COX-2 induction and synthesis of L-PGDS-derived, PPARgamma-activating PGs as a decisive target by which several chemotherapeutics induce apoptosis. COX-2 is therefore suspected to sensitize cancer cells to apoptotic death under certain circumstances, suggesting that COX-2 inhibition during cancer therapy could diminish its efficacy.


Subject(s)
Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Cyclooxygenase 2/metabolism , Intramolecular Oxidoreductases/metabolism , Lipocalins/metabolism , Uterine Cervical Neoplasms/drug therapy , Antineoplastic Agents/therapeutic use , Base Sequence , Female , HeLa Cells , Humans , PPAR gamma/physiology , RNA, Small Interfering , Uterine Cervical Neoplasms/enzymology , Uterine Cervical Neoplasms/pathology
11.
Oncogene ; 26(39): 5822-7, 2007 Aug 23.
Article in English | MEDLINE | ID: mdl-17369856

ABSTRACT

Cancer cell invasion is one of the crucial events in local spreading, growth and metastasis of tumors. The present study investigates the mechanism underlying the anti-invasive action of the chemotherapeutic cisplatin. In human cervical carcinoma cells (HeLa), cisplatin caused a time- and concentration-dependent suppression of cell invasion through Matrigel. Inhibition of invasion was accompanied by upregulation of tissue inhibitor of matrix metalloproteinases-1 (TIMP-1), whereas levels of matrix metalloproteinase-2 (MMP-2), MMP-9 and TIMP-2 remained unchanged. Cisplatin's effects on TIMP-1 expression and invasion were associated with phosphorylations of p38 and p42/44 mitogen-activated protein kinases and were abrogated by specific inhibitors of both pathways. The impact of TIMP-1 in the anti-invasive action of cisplatin was proven by transfecting cells with small interfering RNA targeting TIMP-1, which completely reversed suppression of invasion by cisplatin. A functional relevance of TIMP-1 upregulation was substantiated by findings showing a concentration-dependent inhibition of Matrigel invasion by recombinant TIMP-1. The essential role of TIMP-1 in the anti-invasive action of cisplatin was confirmed using another human cervical carcinoma cell line (C33A) and human lung carcinoma cells (A549). Altogether, our data demonstrate a hitherto unknown mechanism by which cisplatin exerts its antimetastatic properties on highly invasive cancer cells.


Subject(s)
Antineoplastic Agents/pharmacology , Cisplatin/pharmacology , Gene Expression Regulation, Enzymologic , Lung Neoplasms/drug therapy , Neoplasm Invasiveness/prevention & control , Tissue Inhibitor of Metalloproteinase-1/metabolism , Collagen/metabolism , Drug Combinations , Enzyme Activation , HeLa Cells/drug effects , Humans , Laminin/metabolism , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Mitogen-Activated Protein Kinases/metabolism , Phosphorylation , Proteoglycans/metabolism , Tissue Inhibitor of Metalloproteinase-2/metabolism , Up-Regulation
12.
Handb Exp Pharmacol ; (177): 65-93, 2007.
Article in English | MEDLINE | ID: mdl-17087120

ABSTRACT

Antipyretic analgesics are a group of heterogeneous substances including acidic (nonsteroidal antiinflammatory drugs, NSAIDs) and nonacidic (paracetamol, pyrazolinones) drugs. Moreover, various selective cyclooxygenase-2 (COX-2) inhibitors with improved gastrointestinal tolerability as compared with conventional NSAIDs have been established for symptomatic pain treatment in recent years. The present review summarizes the pharmacology of all of these drugs with particular emphasis on their rational use based on the diverse pharmacokinetic characteristics and adverse drug reaction profiles. Referring to the current debate, potential mechanisms underlying cardiovascular side effects associated with long-term use of COX inhibitors are discussed.


Subject(s)
Acetaminophen/pharmacology , Analgesics, Non-Narcotic/pharmacology , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Cyclooxygenase 2 Inhibitors/pharmacology , Pyrazoles/pharmacology , Acetaminophen/pharmacokinetics , Acetaminophen/therapeutic use , Analgesics, Non-Narcotic/pharmacokinetics , Analgesics, Non-Narcotic/therapeutic use , Animals , Anti-Inflammatory Agents, Non-Steroidal/pharmacokinetics , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Cyclooxygenase 2 Inhibitors/pharmacokinetics , Cyclooxygenase 2 Inhibitors/therapeutic use , Humans , Pain/drug therapy , Pyrazoles/pharmacokinetics , Pyrazoles/therapeutic use
13.
Orthopade ; 36(1): 23-31, 2007 Jan.
Article in German | MEDLINE | ID: mdl-17171384

ABSTRACT

The pharmacotherapy of musculoskeletal pain remains of high importance in Western countries. The present review concentrates on the use of acidic (nonsteroidal anti-inflammatory drugs) and nonacidic (paracetamol, selective cyclooxygenase-2 inhibitors) antipyretic analgesics in the therapy of musculoskeletal pain disorders with particular emphasis on the diverse pharmacokinetic properties and unwanted side effects of these substances.


Subject(s)
Analgesics, Non-Narcotic/classification , Analgesics, Non-Narcotic/therapeutic use , Musculoskeletal Diseases/complications , Musculoskeletal Diseases/drug therapy , Pain/drug therapy , Pain/etiology , Analgesics, Non-Narcotic/adverse effects , Humans , Practice Guidelines as Topic , Practice Patterns, Physicians'
14.
Int J Clin Pharmacol Ther ; 44(9): 397-400, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16995327

ABSTRACT

OBJECTIVE: We reported recently that celecoxib inhibits the metabolism of the cytochrome P450 (CYP)2D6 substrate metoprolol in volunteers. Valdecoxib, the active metabolite of parecoxib, has also been claimed to interfere with the metabolism of CYP2D6 substrates. However, little support for this contention is available despite the intensive use of parecoxib in the perioperative setting. Therefore, the objective of this study was to examine the effect of valdecoxib on the pharmacokinetics of the clinically relevant CYP2D6 substrate metoprolol. METHODS: An open, randomized, 3-period crossover study was performed in 15 healthy male volunteers. Metoprolol (50 mg) was given in all 3 periods without, or following a 7-day pre-treatment with valdecoxib (20 mg, o.d.) or rofecoxib (25 mg, o.d.), to achieve steady state conditions of COX-2 inhibitors in Periods 2 and 3. In a small group of extensive metabolizers (EM/EM), short-term application of twice the dose was investigated. RESULTS: No effect of valdecoxib (20 mg/d) or rofecoxib (25 mg/d) were detected on the area under the plasma concentration-time curve of metoprolol (323 +/- 333 to 324 +/- 296 or 309 +/- 256 microg x h/l) or at a higher dose. No significant changes of pharmacokinetic or pharmacodynamic parameters of metoprolol were apparent. CONCLUSION: We conclude that, at therapeutic doses, valdecoxib and rofecoxib do not influence the CYP2D6 substrate metoprolol.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Cyclooxygenase 2 Inhibitors/pharmacology , Cytochrome P-450 CYP2D6/metabolism , Isoxazoles/pharmacology , Metoprolol/pharmacokinetics , Sulfonamides/pharmacology , Adult , Cross-Over Studies , Cytochrome P-450 CYP2D6/genetics , Humans , Lactones/pharmacology , Male , Metoprolol/blood , Prodrugs/pharmacology , Sulfones/pharmacology
15.
Int J Immunopathol Pharmacol ; 19(1): 35-48, 2006.
Article in English | MEDLINE | ID: mdl-16569344

ABSTRACT

Binding of thrombospondin-1 (TSP-1) to the CD36 receptor inhibits angiogenesis and induces apoptosis in endothelial cells (EC). Conversely, matrix-bound TSP-1 supports vessel formation. In this study we analyzed the shear stress-dependent expression of TSP-1 and CD36 in endothelial cells in vitro and in vivo to reveal its putative role in the blood flow-induced remodelling of vascular networks. Shear stress was applied to EC using a cone-and-plate apparatus and gene expression was analyzed by RT-PCR, Northern and Western blot. Angiogenesis in skeletal muscles of prazosin-fed (50 mg/l drinking water; 4 d) mice was assessed by measuring capillary-to-fiber (C/F) ratios. Protein expression in whole muscle homogenates (WMH) or BS-1 lectin-enriched EC fractions (ECF) was analyzed by Western blot. Shear stress downregulated TSP-1 and CD36 expression in vitro in a force- and time-dependent manner sustained for at least 72 h and reversible by restoration of no-flow conditions. In vivo, shear stress-driven increase of C/F in prazosin-fed mice was associated with reduced expression of TSP-1 and CD36 in ECF, while TSP-1 expression in WMH was increased. Down-regulation of endothelial TSP-1/CD36 by shear stress suggests a mechanism for inhibition of apoptosis in perfused vessels and pruning in the absence of flow. The increase of extra-endothelial (e.g. matrix-bound) TSP-1 could support a splitting type of vessel growth.


Subject(s)
CD36 Antigens/biosynthesis , Endothelial Cells/metabolism , Endothelial Cells/physiology , Neovascularization, Pathologic/physiopathology , Stress, Mechanical , Thrombospondin 1/biosynthesis , Adrenergic alpha-Antagonists/pharmacology , Alkaline Phosphatase/metabolism , Animals , Blotting, Northern , Blotting, Western , Cells, Cultured , Hemodynamics/physiology , Humans , Mice , Mice, Inbred C57BL , Neovascularization, Pathologic/metabolism , Prazosin/pharmacology , RNA/biosynthesis , RNA/isolation & purification , Reverse Transcriptase Polymerase Chain Reaction , Transcription, Genetic
16.
Mol Biol Cell ; 15(9): 4310-20, 2004 Sep.
Article in English | MEDLINE | ID: mdl-15240821

ABSTRACT

Myofibroblasts of wound granulation tissue, in contrast to dermal fibroblasts, join stress fibers at sites of cadherin-type intercellular adherens junctions (AJs). However, the function of myofibroblast AJs, their molecular composition, and the mechanisms of their formation are largely unknown. We demonstrate that fibroblasts change cadherin expression from N-cadherin in early wounds to OB-cadherin in contractile wounds, populated with alpha-smooth muscle actin (alpha-SMA)-positive myofibroblasts. A similar shift occurs during myofibroblast differentiation in culture and seems to be responsible for the homotypic segregation of alpha-SMA-positive and -negative fibroblasts in suspension. AJs of plated myofibroblasts are reinforced by alpha-SMA-mediated contractile activity, resulting in high mechanical resistance as demonstrated by subjecting cell pairs to hydrodynamic forces in a flow chamber. A peptide that inhibits alpha-SMA-mediated contractile force causes the reorganization of large stripe-like AJs to belt-like contacts as shown for enhanced green fluorescent protein-alpha-catenin-transfected cells and is associated with a reduced mechanical resistance. Anti-OB-cadherin but not anti-N-cadherin peptides reduce the contraction of myofibroblast-populated collagen gels, suggesting that AJs are instrumental for myofibroblast contractile activity.


Subject(s)
Adherens Junctions/metabolism , Fibroblasts/cytology , Fibroblasts/metabolism , Myoblasts/cytology , Myoblasts/metabolism , Actins/metabolism , Animals , Biomechanical Phenomena , Cadherins/metabolism , Cell Differentiation , Cells, Cultured , Collagen , Female , Gels , Models, Biological , Rats , Rats, Wistar , Skin/cytology , Skin/injuries , Skin/metabolism , Wound Healing/physiology
17.
Scand J Rheumatol ; 33(1): 1-6, 2004.
Article in English | MEDLINE | ID: mdl-15124935

ABSTRACT

The enzyme cyclooxygenase (COX) was shown to exist as two distinct isoforms about a decade ago. COX-1 is constitutively expressed as a 'housekeeping' enzyme in nearly all tissues, and mediates physiological responses (e.g. cytoprotection of the stomach, and platelet aggregation). On the other hand, COX-2, expressed by cells involved in inflammation (e.g. macrophages, monocytes, synoviocytes), has emerged as the isoform that is primarily responsible for the synthesis of prostanoids involved in acute and chronic inflammatory states. Consequently, the hypothesis that selective inhibition of COX-2 might have therapeutic actions similar to those of non-steroidal anti-inflammatory drugs, but without causing gastrointestinal side effects, was the rationale for the development of selective inhibitors of the COX-2 isoenzyme. Selective COX-2 inhibitors currently used in the clinic are the sulphonamides celecoxib and valdecoxib (parecoxib is a prodrug of valdecoxib), as well as the methylsulphones rofecoxib and etoricoxib. Furthermore, the phenylacetic acid derivative lumiracoxib has gained permission recently to be marketed in Europe. This review discusses the clinically relevant similarities and differences of these substances, with particular emphasis on their diverse pharmacokinetic characteristics.


Subject(s)
Cyclooxygenase Inhibitors/pharmacokinetics , Cyclooxygenase Inhibitors/therapeutic use , Rheumatic Diseases/drug therapy , Anti-Inflammatory Agents, Non-Steroidal/pharmacokinetics , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Biological Availability , Clinical Trials as Topic , Cyclooxygenase Inhibitors/adverse effects , Drug Interactions , Humans , Rheumatic Diseases/diagnosis , Risk Factors , Sensitivity and Specificity
18.
Am J Ophthalmol ; 132(5): 792-4, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11704049

ABSTRACT

PURPOSE: The purpose of this study was to investigate the role of leptin in choroidal neovascularization. METHODS: We examined the localization of leptin by immunohistochemistry in nine choroidal neovascular membranes surgically excised from patients with age-related macular degeneration, idiopathic choroidal neovascularization, and ocular histoplasmosis. Controls included omission of primary antibody, use of an irrelevant primary antibody and leptin staining of posterior segment of four normal donor eyes. RESULTS: Leptin was present in eight membranes and appeared vesicular, within the cytoplasm. The more vascular membranes and those consisting of a larger number of retinal pigment epithelium cells were associated with greater leptin staining. Leptin was not seen in the posterior segment of the four normal eyes. CONCLUSION: We suggest that leptin plays an active role in choroidal neovascularization, although further experiments are necessary to establish a causal relationship.


Subject(s)
Choroidal Neovascularization/metabolism , Leptin/metabolism , Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Immunoenzyme Techniques , Macular Degeneration/metabolism , Male , Middle Aged , Platelet Endothelial Cell Adhesion Molecule-1/metabolism
19.
Eur J Cell Biol ; 80(10): 615-9, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11713865

ABSTRACT

The vesicular protein synaptobrevin contributes to two mutually exclusive complexes in mature synapses. Synaptobrevin tightly interacts with the plasma membrane proteins syntaxin and SNAP 25 forming the SNARE complex as a prerequisite for exocytotic membrane fusion. Alternatively, synaptobrevin binds to the vesicular protein synaptophysin. It is unclear whether SNARE complex formation is diminished or facilitated when synaptobrevin is bound to synaptophysin. Here we show that the synaptophysin-synaptobrevin complex is increased in adult rat brain after repeated synaptic hyperactivity in the kindling model of epilepsy. Two days after the last kindling-induced stage V seizure the relative amount of synaptophysin-synaptobrevin complex obtained by co-immunoprecipitation from cortical and hippocampal membranes was increased twofold compared to controls. By contrast the relative amounts of various synaptic proteins as well as that of the SNARE complex did not change in membrane preparations from kindled rats compared to controls. The increased amount of synaptophysin-synaptobrevin complex in kindled rats supports the idea that this complex represents a reserve pool for synaptobrevin enabling synaptic vesicles to adjust to an increased demand for synaptic efficiency. We conclude that the synaptophysin-synaptobrevin interaction is involved in activity-dependent plastic changes in adult rat brain.


Subject(s)
Amygdala/metabolism , Epilepsy/metabolism , Membrane Proteins/metabolism , Presynaptic Terminals/metabolism , Synaptophysin/metabolism , Vesicular Transport Proteins , Age Factors , Amygdala/physiopathology , Animals , Epilepsy/physiopathology , Kindling, Neurologic/physiology , Nerve Tissue Proteins/metabolism , Neuronal Plasticity/physiology , Qa-SNARE Proteins , R-SNARE Proteins , Rats , SNARE Proteins , Synaptosomal-Associated Protein 25
20.
Am J Pathol ; 159(3): 1009-20, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11549593

ABSTRACT

We have examined the role of mechanical tension in myofibroblast differentiation using two in vivo rat models. In the first model, granulation tissue was subjected to an increase in mechanical tension by splinting a full-thickness wound with a plastic frame. Myofibroblast features, such as stress fiber formation, expression of ED-A fibronectin and alpha-smooth muscle actin (alpha-SMA) appeared earlier in splinted than in unsplinted wounds. Myofibroblast marker expression decreased in control wounds starting at 10 days after wounding as expected, but persisted in splinted wounds. In the second model, granuloma pouches were induced by subcutaneous croton oil injection; pouches were either left intact or released from tension by evacuation of the exudate at 14 days. The expression of myofibroblast markers was reduced after tension release in the following sequence: F-actin (2 days), alpha-SMA (3 days), and ED-A fibronectin (5 days); cell density was not affected. In both models, isometric contraction of tissue strips was measured after stimulation with smooth muscle agonists. Contractility correlated always with the level of alpha-SMA expression, being high when granulation tissue had been subjected to tension and low when it had been relaxed. Our results support the assumption that mechanical tension is crucial for myofibroblast modulation and for the maintenance of their contractile activity.


Subject(s)
Fibroblasts/cytology , Granulation Tissue/physiology , Muscle, Smooth/cytology , Actins/metabolism , Animals , Biomarkers , Cell Differentiation/physiology , Female , Isometric Contraction/physiology , Muscle, Smooth/physiology , Rats , Rats, Wistar , Stress, Mechanical , Wound Healing/physiology
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