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Grenegård, Magnus
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Publications (10 of 53) Show all publications
Södergren, A., Svensson Holm, A.-C., Ramström, S., Lindström, E., Grenegård, M. & Öllinger, K. (2016). Thrombin-induced lysosomal exocytosis in human platelets is dependent on secondary activation by ADP and regulated by endothelial-derived substances. Platelets, 27(1), 86-92
Open this publication in new window or tab >>Thrombin-induced lysosomal exocytosis in human platelets is dependent on secondary activation by ADP and regulated by endothelial-derived substances
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2016 (English)In: Platelets, ISSN 0953-7104, E-ISSN 1369-1635, Vol. 27, no 1, p. 86-92Article in journal (Refereed) Published
Abstract [en]

Exocytosis of lysosomal contents from platelets has been speculated to participate in clearance of thrombi and vessel wall remodelling. The mechanisms that regulate lysosomal exocytosis in platelets are, however, still unclear. The aim of this study was to identify the pathways underlying platelet lysosomal secretion and elucidate how this process is controlled by platelet inhibitors. We found that high concentrations of thrombin induced partial lysosomal exocytosis as assessed by analysis of the activity of released N-acetyl--glucosaminidase (NAG) and by identifying the fraction of platelets exposing the lysosomal-associated membrane protein (LAMP)-1 on the cell surface by flow cytometry. Stimulation of thrombin receptors PAR1 or PAR4 with specific peptides was equally effective in inducing LAMP-1 surface expression. Notably, lysosomal exocytosis in response to thrombin was significantly reduced if the secondary activation by ADP was inhibited by the P2Y(12) antagonist cangrelor, while inhibition of thromboxane A(2) formation by treatment with acetylsalicylic acid was of minor importance in this regard. Moreover, the NO-releasing drug S-nitroso-N-acetyl penicillamine (SNAP) or the cyclic AMP-elevating eicosanoid prostaglandin I-2 (PGI(2)) significantly suppressed lysosomal exocytosis. We conclude that platelet inhibitors that mimic functional endothelium such as PGI(2) or NO efficiently counteract lysosomal exocytosis. Furthermore, we suggest that secondary release of ADP and concomitant signaling via PAR1/4- and P2Y(12) receptors is important for efficient platelet lysosomal exocytosis by thrombin.

Place, publisher, year, edition, pages
TAYLOR & FRANCIS INC, 2016
Keywords
ADP receptors; endothelium; exocytosis; lysosome; platelet physiology; protease activated receptors (PAR); thrombin
National Category
Clinical Medicine Biological Sciences
Identifiers
urn:nbn:se:liu:diva-125318 (URN)10.3109/09537104.2015.1042446 (DOI)000368717700011 ()25970449 (PubMedID)
Note

Funding Agencies|County Council of Ostergotland; Swedish Research Council

Available from: 2016-02-24 Created: 2016-02-19 Last updated: 2018-10-25
Svensson Holm, A.-C., Grenegård, M., Ollinger, K. & Lindström, E. (2014). Inhibition of 12-lipoxygenase reduces platelet activation and prevents their mitogenic function. Platelets, 25(2), 111-117
Open this publication in new window or tab >>Inhibition of 12-lipoxygenase reduces platelet activation and prevents their mitogenic function
2014 (English)In: Platelets, ISSN 0953-7104, E-ISSN 1369-1635, Vol. 25, no 2, p. 111-117Article in journal (Refereed) Published
Abstract [en]

The aim of the present study was to investigate the role of 12-lipoxygenase (12-LOX) on platelet-induced airway smooth muscle cell (ASMC) proliferation. Co-incubation of platelets and ASMC caused platelet activation as determined by morphological changes. Simultaneously, reactive oxygen species (ROS)-generation was detected and ASMC proliferation (measured by using the MTS assay) increased significantly. Furthermore, we found that the 12-LOX inhibitors cinnamyl-3,4-dihydroxy-α-cyanocinnamate (CDC) and Baicalein prevented platelet activation in a co-cultures of platelets and ASMC. The inhibitory effect of CDC and Baicalein on platelets was also registered in a pure platelet preparation. Specifically, the 12-LOX inhibitors reduced collagen-induced platelet aggregation both in the presence and absence of external added fibrinogen. Importantly, platelet-induced ASMC proliferation and ROS production generated during the platelet/ASMC interaction was significantly inhibited in the presence of 12-LOX inhibitors. In conclusion, our findings reveal that 12-LOX is crucial for the observed enhancement of ASMC proliferation in co-cultures of platelets and ASMC. The present result suggests that 12-LOX activity is important in the initial step of platelet/ASMC interaction and platelet activation. Such action of 12-LOX represents a potential important mechanism that may contribute to platelet-induced airway remodelling.

Place, publisher, year, edition, pages
Informa Healthcare, 2014
Keywords
12-lipoxygenase, airway remodelling, airway smooth muscle, platelet-induced proliferation
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-99371 (URN)10.3109/09537104.2013.783688 (DOI)000331905100006 ()23534390 (PubMedID)
Available from: 2013-10-16 Created: 2013-10-16 Last updated: 2017-12-06Bibliographically approved
Tengdelius, M., Lee, C.-J., Grenegård, M., Griffith, M., Påhlsson, P. & Konradsson, P. (2014). Synthesis and biological evaluation of fucoidan-mimetic glycopolymers through cyanoxyl-mediated free-radical polymerization. Biomacromolecules, 15(7), 2359-2368
Open this publication in new window or tab >>Synthesis and biological evaluation of fucoidan-mimetic glycopolymers through cyanoxyl-mediated free-radical polymerization
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2014 (English)In: Biomacromolecules, ISSN 1525-7797, E-ISSN 1526-4602, Vol. 15, no 7, p. 2359-2368Article in journal (Refereed) Published
Abstract [en]

The sulfated marine polysaccharide fucoidan has been reported to have health benefits ranging from antivirus and anticancer properties to modulation of high blood pressure. Hence, they could enhance the biological function of materials for biomedical applications. However, the incorporation of fucoidan into biomaterials has been difficult, possibly due to its complex structure and lack of suitable functional groups for covalent anchoring to biomaterials. We have developed an approach for a rapid synthesis of fucoidanmimetic glycopolymer chains through cyanoxyl-mediated free-radical polymerization, a method suitable for chain-end functionalizing and subsequent linkage to biomaterials. The resulting sulfated and nonsulfated methacrylamido alpha-L-fucoside glycopolymers fucoidan-mimetic properties were studied in HSV-1 infection and platelet activation assays. The sulfated glycopolymer showed similar properties to natural fucoidan in inducing platelet activation and inhibiting HSV-1 binding and entry to cells, thus indicating successful syntheses of fucoidan-mimetic glycopolymers.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2014
National Category
Chemical Sciences Clinical Medicine Physical Sciences
Identifiers
urn:nbn:se:liu:diva-109382 (URN)10.1021/bm5002312 (DOI)000339090500003 ()24813544 (PubMedID)
Available from: 2014-08-15 Created: 2014-08-15 Last updated: 2017-12-05Bibliographically approved
Peng, X., Ramström, S., Kurz, T., Grenegård, M. & Segelmark, M. (2014). The neutrophil serine protease PR3 induces shape change of platelets via the Rho/Rho kinase and Ca2+ signaling pathways. Thrombosis Research, 134(2), 418-425
Open this publication in new window or tab >>The neutrophil serine protease PR3 induces shape change of platelets via the Rho/Rho kinase and Ca2+ signaling pathways
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2014 (English)In: Thrombosis Research, ISSN 0049-3848, E-ISSN 1879-2472, Vol. 134, no 2, p. 418-425Article in journal (Refereed) Published
Abstract [en]

Introduction: Proteinase 3 (PR3) is released from neutrophil azurophilic granules and exerts complex effects on the inflammatory process. PR3 catalyzes the degradation of a number of macromolecules, but the consequences on blood cells are less well defined. In the present study, the effect of PR3 on human platelets was thoroughly investigated. Methods: The experiments were performed on washed platelets freshly isolated from blood donated by healthy human volunteers. Platelets shape change and aggregation was measured on a Chrono-Log aggregometer. The phosphorylated form of MYPT1 was visualized by immunostaining. Platelet activation was further evaluated by flow cytometry. Results: PR3 induced platelet shape change but not aggregation. Flow cytometry analysis showed that PR3 induced no P-selectin expression or binding of fibrinogen to the platelets, and it did not change the activation in response to PAR1- or PAR4-activating peptides or to thrombin. Furthermore, Fura-2 measurement and immuno-blotting analysis, respectively, revealed that PR3 stimulated small intracellular Ca2+ mobilization and Thr696-specific phosphorylation of the myosin phosphatase target subunit 1 (MYPT1). Separate treatment of platelets with the Rho/Rho kinase inhibitor Y-27632 and the intracellular Ca2+ chelator BAPTA/AM reduced the shape change induced by PR3 whereas concurrent treatment completely inhibited it. Conclusion: The data shows that the neutrophil protease PR3 is a direct modulator of human platelets and causes shape change through activation of the Rho/Rho kinase and Ca2+ signaling pathways. This finding highlights an additional mechanism in the complex interplay between neutrophils and platelets.

Place, publisher, year, edition, pages
Elsevier, 2014
Keywords
ANCA-associated vasculitis; Proteinase PR3; Platelet shape change; Rho/Rho kinase signaling pathway; Ca2+ signaling pathway
National Category
Clinical Medicine
Identifiers
urn:nbn:se:liu:diva-111292 (URN)10.1016/j.thromres.2014.06.001 (DOI)000341309200035 ()24993595 (PubMedID)
Note

Funding Agencies|Swedish Renal Foundation; Asp Foundation

Available from: 2014-10-14 Created: 2014-10-14 Last updated: 2017-12-05
Boknäs, N., Faxälv, L., Sanchez Centellas, D., Wallstedt, M., Ramström, S., Grenegård, M. & Lindahl, T. (2014). Thrombin-induced platelet activation via PAR4: pivotal role for exosite II. Thrombosis and Haemostasis, 112(3), 558-565
Open this publication in new window or tab >>Thrombin-induced platelet activation via PAR4: pivotal role for exosite II
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2014 (English)In: Thrombosis and Haemostasis, ISSN 0340-6245, Vol. 112, no 3, p. 558-565Article in journal (Refereed) Published
Abstract [en]

Thrombin-induced platelet activation via PAR1 and PAR4 is an important event in haemostasis. Although the underlying mechanisms responsible for ensuring efficient PAR1 activation by thrombin have been extensively studied, the potential involvement of recognitions sites outside the active site of the protease in thrombin-induced PAR4 activation is largely unknown. In this study, we developed a new assay to assess the importance of exosite I and II for PAR4 activation with alpha- and gamma-thrombin. Surprisingly, we found that exosite II is critical for activation of PAR4. We also show that this dependency on exosite II likely represents a new mechanism, as it is unaffected by blockage of the previously known interaction between thrombin and glycoprotein Ib alpha.

Place, publisher, year, edition, pages
Schattauer Gmbh, 2014
National Category
Clinical Medicine
Identifiers
urn:nbn:se:liu:diva-111269 (URN)10.1160/TH13-12-1013 (DOI)000341547000015 ()24990072 (PubMedID)
Note

Funding Agencies|Swedish Research Council [K2010-65X-15060-07-3, K2013-65X-15060-10-3]; Swedish Heart and Lung Foundation [20100219, 20120263]

Available from: 2014-10-15 Created: 2014-10-14 Last updated: 2017-12-05Bibliographically approved
Junker, J., Lönnqvist, S., Rakar, J., Karlsson, L. K., Grenegård, M. & Kratz, G. (2013). Differentiation of human dermal fibroblasts towards endothelial cells. Differentiation, 85(3), 67-77
Open this publication in new window or tab >>Differentiation of human dermal fibroblasts towards endothelial cells
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2013 (English)In: Differentiation, ISSN 0301-4681, E-ISSN 1432-0436, Vol. 85, no 3, p. 67-77Article in journal (Refereed) Published
Abstract [en]

The ultimate goal of vascular tissue engineering is the production of functional grafts for clinical use. Difficulties acquiring autologous endothelial cells have motivated the search for alternative cell sources. Differentiation of dermal fibroblasts towards several mesenchymal lineages as well as endothelial cells has been proposed. The aim of the present study was to investigate the endothelial differentiation capacity of human dermal fibroblasts on a gene expression, protein expression and functional physiological level. Endothelial differentiation of fibroblasts was induced by culturing cells in 30% human serum, but not in fetal calf serum. Expression of proteins and genes relevant for endothelial function and differentiation was increased after induction. Furthermore, fibroblasts exposed to 30% human serum displayed increased uptake of low-density lipoprotein and formation of capillary-like networks. The results of this study may have an impact on cell sourcing for vascular tissue engineering, and the development of methods for vascularization of autologous tissue engineered constructs.

Place, publisher, year, edition, pages
Wiley-Blackwell / Elsevier, 2013
Keywords
Differentiation; Endothelial cell; Fibroblast; Tissue engineering; Human serum
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-96492 (URN)10.1016/j.diff.2013.01.005 (DOI)000320766300001 ()
Available from: 2013-08-23 Created: 2013-08-20 Last updated: 2017-12-06
Elvers, M., Grenegård, M., Khoshjabinzadeh, H., Muenzer, P., Borst, O., Tian, H., . . . Fälker, K. (2012). A novel role for phospholipase D as an endogenous negative regulator of platelet sensitivity. Cellular Signalling, 24(9), 1743-1752
Open this publication in new window or tab >>A novel role for phospholipase D as an endogenous negative regulator of platelet sensitivity
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2012 (English)In: Cellular Signalling, ISSN 0898-6568, E-ISSN 1873-3913, Vol. 24, no 9, p. 1743-1752Article in journal (Refereed) Published
Abstract [en]

Platelet aggregation, secretion and thrombus formation play a critical role in primary hemostasis to prevent excessive blood loss. On the other hand, uncontrolled platelet activation leads to pathological thrombus formation resulting in myocardial infarction or stroke. Stimulation of heterotrimeric G-proteins by soluble agonists or immunoreceptor tyrosine based activation motif-coupled receptors that interact with immobilized ligands such as the collagen receptor glycoprotein (GP) VI lead to the activation of phospholipases that cleave membrane phospholipids to generate soluble second messengers. Platelets contain the phospholipases (PL) D1 and D2 which catalyze the hydrolysis of phosphatidylcholine to generate the second messenger phosphatidic acid (PA). The production of PA is abrogated by primary alcohols that have been widely used for the analysis of PLD-mediated processes. However, it is not clear if primary alcohols effectively reduce PA generation or if they induce PLD-independent cellular effects. In the present study we made use of the specific PLD inhibitor 5-fluoro-2-indolyl des-chlorohalopemide (FIPI) and show for the first time, that FIPI enhances platelet dense granule secretion and aggregation of human platelets. Further. FIPI has no effect on cytosolic Ca2+ activity but needs proper Rho kinase signaling to mediate FIPI-induced effects on platelet activation. Upon FIPI treatment the phosphorylation of the PKC substrate pleckstrin was prominently enhanced suggesting that FIPI affects PKC-mediated secretion and aggregation in platelets. Similar effects of FIPI were observed in platelets from mouse wild-type and Pld1(-/-) mice pointing to a new role for PLD2 as a negative regulator of platelet sensitivity.

Place, publisher, year, edition, pages
Elsevier, 2012
Keywords
PLD; Platelets; Secretion; Aggregation; Regulation
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-80685 (URN)10.1016/j.cellsig.2012.04.018 (DOI)000306621000003 ()
Available from: 2012-08-29 Created: 2012-08-29 Last updated: 2017-12-07
Junker, J. P., Lonnqvist, S., Karlsson, L. K., Grenegard, M. & Kratz, G. (2012). Endothelial differentiation of human dermal fibroblasts in JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, vol 6, issue SI, pp 149-149. In: JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE (pp. 149-149). John Wiley and Sons, 6(SI)
Open this publication in new window or tab >>Endothelial differentiation of human dermal fibroblasts in JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, vol 6, issue SI, pp 149-149
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2012 (English)In: JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, John Wiley and Sons , 2012, Vol. 6, no SI, p. 149-149Conference paper, Published paper (Refereed)
Abstract [en]

n/a

Place, publisher, year, edition, pages
John Wiley and Sons, 2012
Series
JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, ISSN 1932-6254
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-84337 (URN)000308313001150 ()
Available from: 2012-10-05 Created: 2012-10-05 Last updated: 2012-10-05
Junker, J. P., Lönnqvist, S., Karlsson, L. K., Grenegård, M. & Kratz, G. (2012). ENDOTHELIAL DIFFERENTIATION OF HUMAN DERMAL FIBROBLASTS in WOUND REPAIR AND REGENERATION, vol 20, issue 2, pp A27-A27. In: WOUND REPAIR AND REGENERATION (pp. A27-A27). Wiley-Blackwell, 20(2)
Open this publication in new window or tab >>ENDOTHELIAL DIFFERENTIATION OF HUMAN DERMAL FIBROBLASTS in WOUND REPAIR AND REGENERATION, vol 20, issue 2, pp A27-A27
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2012 (English)In: WOUND REPAIR AND REGENERATION, Wiley-Blackwell , 2012, Vol. 20, no 2, p. A27-A27Conference paper, Published paper (Refereed)
Abstract [en]

n/a

Place, publisher, year, edition, pages
Wiley-Blackwell, 2012
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-76620 (URN)000300943100060 ()
Available from: 2012-04-13 Created: 2012-04-13 Last updated: 2012-04-13
Svensson Holm, A.-C., Bengtsson, T., Grenegård, M. & Lindström, E. G. (2012). Hyaluronic acid influence on platelet-induced airway smooth muscle cell proliferation. Experimental Cell Research, 318(5), 632-640
Open this publication in new window or tab >>Hyaluronic acid influence on platelet-induced airway smooth muscle cell proliferation
2012 (English)In: Experimental Cell Research, ISSN 0014-4827, E-ISSN 1090-2422, Vol. 318, no 5, p. 632-640Article in journal (Refereed) Published
Abstract [en]

Hyaluronic acid (HA) is one of the main components of the extracellular matrix (ECM) and is expressed throughout the body including the lung and mostly in areas surrounding proliferating and migrating cells. Furthermore, platelets have been implicated as important players in the airway remodelling process, e.g. due to their ability to induce airway smooth muscle cell (ASMC) proliferation. The aim of the present study was to investigate the role of HA, the HA-binding surface receptor CD44 and focal adhesion kinase (FAK) in platelet-induced ASMC proliferation. Proliferation of ASMC was measured using the MTS-assay, and we found that the CD44 blocking antibody and the HA synthase inhibitor 4-Methylumbelliferone (4-MU) significantly inhibited platelet-induced ASMC proliferation. The interaction between ASMC and platelets was studied by fluorescent staining of F-actin. In addition, the ability of ASMC to synthesise HA was investigated by fluorescent staining using biotinylated HA-binding protein and a streptavidin conjugate. We observed that ASMC produced HA and that a CD44 blocking antibody and 4-MU significantly inhibited platelet binding to the area surrounding the ASMC. Furthermore, the FAK-inhibitor PF 573228 inhibited platelet-induced ASMC proliferation. Co-culture of ASMC and platelets also resulted in increased phosphorylation of FAK as detected by Western blot analysis. In addition, 4-MU significantly inhibited the increased FAK-phosphorylation. In conclusion, our findings demonstrate that ECM has the ability to influence platelet-induced ASMC proliferation. Specifically, we propose that HA produced by ASMC is recognised by platelet CD44. The platelet/HA interaction is followed by FAK activation and increased proliferation of co-cultured ASMC. We also suggest that the mitogenic effect of platelets represents a potential important and novel mechanism that may contribute to airway remodelling.

Place, publisher, year, edition, pages
Elsevier, 2012
Keywords
Airway smooth muscle; Airway remodelling; Extracellular matrix; Hyaluronic acid; CD44; Focal adhesion kinase; Platelets
National Category
Basic Medicine
Identifiers
urn:nbn:se:liu:diva-75382 (URN)10.1016/j.yexcr.2011.12.011 (DOI)000300966300021 ()22227408 (PubMedID)
Note
funding agencies|strategic areas Cardiovascular Inflammation Research Centre (CIRC)||Material in Medicine, the Heart- and Lung foundation||County of Ostergotlands Lan||STIFUD||Olle E||Swedish Research Council||Available from: 2012-02-28 Created: 2012-02-28 Last updated: 2018-01-12
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