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Direct activation of platelets by addition of CaCl(2) leads coagulation of platelet-rich plasma

BACKGROUND: Based on the notion that full activation of platelets is required for a growth factor release, in regenerative dentistry, platelet-rich plasma (PRP) in liquid form is usually clotted by addition of CaCl(2) in glassware before topical implantation. However, there has been no evidence as t...

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Autores principales: Toyoda, Toshihisa, Isobe, Kazushige, Tsujino, Tetsuhiro, Koyata, Yasuo, Ohyagi, Fumitaka, Watanabe, Taisuke, Nakamura, Masayuki, Kitamura, Yutaka, Okudera, Hajime, Nakata, Koh, Kawase, Tomoyuki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6068057/
https://www.ncbi.nlm.nih.gov/pubmed/30066050
http://dx.doi.org/10.1186/s40729-018-0134-6
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author Toyoda, Toshihisa
Isobe, Kazushige
Tsujino, Tetsuhiro
Koyata, Yasuo
Ohyagi, Fumitaka
Watanabe, Taisuke
Nakamura, Masayuki
Kitamura, Yutaka
Okudera, Hajime
Nakata, Koh
Kawase, Tomoyuki
author_facet Toyoda, Toshihisa
Isobe, Kazushige
Tsujino, Tetsuhiro
Koyata, Yasuo
Ohyagi, Fumitaka
Watanabe, Taisuke
Nakamura, Masayuki
Kitamura, Yutaka
Okudera, Hajime
Nakata, Koh
Kawase, Tomoyuki
author_sort Toyoda, Toshihisa
collection PubMed
description BACKGROUND: Based on the notion that full activation of platelets is required for a growth factor release, in regenerative dentistry, platelet-rich plasma (PRP) in liquid form is usually clotted by addition of CaCl(2) in glassware before topical implantation. However, there has been no evidence as to which is better, full or partial activation of platelets, for minimizing the loss of growth factors and improving the controlled release of growth factors from coagulated PRP. To address this matter, here, we primarily examined direct effects of CaCl(2) on platelets in PBS and on coagulation in citrated PRP. METHODS: PRP was prepared from healthy volunteers’ blood. Platelets’ actions were monitored by scanning electron microscopy, flow cytometry, digital holographic microscopy, and immunofluorescent staining. Clot formation was examined in plasma. RESULTS: In plasma-free PBS, 0.1% CaCl(2) immediately upregulated CD62P and CD63, causing a release of microparticles and fibrinogen/fibrin; consequently, platelets aggregated and adhered to polystyrene culture dishes with enlargement of their attachment area. In a clot formation assay in plasma, CaCl(2) initially induced platelet aggregation, which triggered loop-like matrix formation and subsequently induced coagulation on a watch glass. Such changes were not clearly observed either with PRP in a plastic dish or in platelet-poor plasma on a watch glass: coagulation was delayed in both conditions. CONCLUSIONS: These findings indicate that besides the well-known coagulation pathway, which activates platelets via thrombin conversion in a coagulation cascade, CaCl(2) directly activates platelets, which then facilitate clot formation independently and in cooperation with the coagulation pathway.
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spelling pubmed-60680572018-08-13 Direct activation of platelets by addition of CaCl(2) leads coagulation of platelet-rich plasma Toyoda, Toshihisa Isobe, Kazushige Tsujino, Tetsuhiro Koyata, Yasuo Ohyagi, Fumitaka Watanabe, Taisuke Nakamura, Masayuki Kitamura, Yutaka Okudera, Hajime Nakata, Koh Kawase, Tomoyuki Int J Implant Dent Research BACKGROUND: Based on the notion that full activation of platelets is required for a growth factor release, in regenerative dentistry, platelet-rich plasma (PRP) in liquid form is usually clotted by addition of CaCl(2) in glassware before topical implantation. However, there has been no evidence as to which is better, full or partial activation of platelets, for minimizing the loss of growth factors and improving the controlled release of growth factors from coagulated PRP. To address this matter, here, we primarily examined direct effects of CaCl(2) on platelets in PBS and on coagulation in citrated PRP. METHODS: PRP was prepared from healthy volunteers’ blood. Platelets’ actions were monitored by scanning electron microscopy, flow cytometry, digital holographic microscopy, and immunofluorescent staining. Clot formation was examined in plasma. RESULTS: In plasma-free PBS, 0.1% CaCl(2) immediately upregulated CD62P and CD63, causing a release of microparticles and fibrinogen/fibrin; consequently, platelets aggregated and adhered to polystyrene culture dishes with enlargement of their attachment area. In a clot formation assay in plasma, CaCl(2) initially induced platelet aggregation, which triggered loop-like matrix formation and subsequently induced coagulation on a watch glass. Such changes were not clearly observed either with PRP in a plastic dish or in platelet-poor plasma on a watch glass: coagulation was delayed in both conditions. CONCLUSIONS: These findings indicate that besides the well-known coagulation pathway, which activates platelets via thrombin conversion in a coagulation cascade, CaCl(2) directly activates platelets, which then facilitate clot formation independently and in cooperation with the coagulation pathway. Springer Berlin Heidelberg 2018-08-01 /pmc/articles/PMC6068057/ /pubmed/30066050 http://dx.doi.org/10.1186/s40729-018-0134-6 Text en © The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Research
Toyoda, Toshihisa
Isobe, Kazushige
Tsujino, Tetsuhiro
Koyata, Yasuo
Ohyagi, Fumitaka
Watanabe, Taisuke
Nakamura, Masayuki
Kitamura, Yutaka
Okudera, Hajime
Nakata, Koh
Kawase, Tomoyuki
Direct activation of platelets by addition of CaCl(2) leads coagulation of platelet-rich plasma
title Direct activation of platelets by addition of CaCl(2) leads coagulation of platelet-rich plasma
title_full Direct activation of platelets by addition of CaCl(2) leads coagulation of platelet-rich plasma
title_fullStr Direct activation of platelets by addition of CaCl(2) leads coagulation of platelet-rich plasma
title_full_unstemmed Direct activation of platelets by addition of CaCl(2) leads coagulation of platelet-rich plasma
title_short Direct activation of platelets by addition of CaCl(2) leads coagulation of platelet-rich plasma
title_sort direct activation of platelets by addition of cacl(2) leads coagulation of platelet-rich plasma
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6068057/
https://www.ncbi.nlm.nih.gov/pubmed/30066050
http://dx.doi.org/10.1186/s40729-018-0134-6
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