Toyoda, T., Isobe, K., Tsujino, T. et al. Direct activation of platelets by addition of CaCl2 leads coagulation of platelet-rich plasma.
Int J Implant Dent 4, 23 (2018). https://doi.org/10.1186/s40729-018-0134-6
Download citation
Received: 13 March 2018
Accepted: 11 May 2018
Published: 01 August 2018
DOI: https://doi.org/10.1186/s40729-018-0134-6
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 ...
The study design and consent forms of all the procedures performed were approved by the ethics committee for human participants of the Niigata University School of Medicine (Niigata, Japan) in accordance with the Helsinki Declaration of 1964 as revised in 2013. Written informed consents to participate in the study were obtained from all the participants.
Written informed consent for the publicati...
You can also search for this author in
PubMed Google Scholar
You can also search for this author in
PubMed Google Scholar
You can also search for this author in
PubMed Google Scholar
TT, KI, TT, and TK conceived and designed the study, performed the experiments and data analysis, and wrote the manuscript. YK, FO, TW, ...
Tokyo Plastic Dental Society, Kita-ku, Tokyo, Japan
Toshihisa Toyoda, Kazushige Isobe, Tetsuhiro Tsujino, Yasuo Koyata, Fumitaka Ohyagi, Taisuke Watanabe, Masayuki Nakamura, Yutaka Kitamura & Hajime Okudera
Bioscience Medical Research Center, Niigata University Medical and Dental Hospital, Niigata, Japan
Koh Nakata
Division of Oral Bioengineering, Institute of Medicine and Dentistry, ...
The authors appreciate Shoshin EM (Okazaki, Japan) for giving an opportunity to use DHM.
Because an article performed and prepared in parallel is now submitted elsewhere for publication, the authors do not wish to share their data at present time.
Blitz A. Pump thrombosis—a riddle wrapped in a mystery inside an enigma. Ann Cardiothor Surg. 2014;3:450–71.
Kawase T, Watanabe T, Okuda K. Platelet-rich plasma and its derived platelet concentrates: what dentists involved in cell-based regenerative therapy should know. Nihon Shishubyou Gakkai Kaishi. 2017;59:68–76. (in Japanese)
Download references
Kobayashi M, Kawase T, Horimizu M, Okuda K, Wolff LF, Yoshie H. A proposed protocol for the standardized preparation of PRF membranes for clinical use. Biologicals. 2012;40:323–9.
Kawase T, Okuda K, Nagata M, Tsuchimochi M, Yoshie H, Nakata K. Non-invasive, quantitative assessment of the morphology of gamma-irradiated human mesenchymal stem cells and periosteal cells using digital holographic m...
Marx RE, Carlson ER, Eichstaedt RM, Schimmele SR, Strauss JE, Georgeff KR. Platelet-rich plasma: growth factor enhancement for bone grafts. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1998;85:638–46.
Anitua E. Plasma rich in growth factors: preliminary results of use in the preparation of future sites for implants. Int J Oral Maxillofac Implants. 1999;14:529–35.
Masuki H, Okudera T, Wa...
Acid citrate dextrose solution
Adenosine diphosphate
Calcium-sensing receptor
Digital holographic microscopy
Phosphate-buffered saline
Platelet-poor plasma
Platelet-rich fibrin
Platelet-rich plasma
Scanning electron microscope
Tris-buffered saline
Thromboxane A2
In addition to the well-known intrinsic coagulation pathway, which activates platelets via thrombin conversion, added Ca2+ may directly activate washed platelets and promote clot formation alone and in cooperation with the coagulation pathway as illustrated in Fig. 8. Therefore, it is possible to control platelet activation levels and the consequent growth factor release by modulating not only in...
On the other hand, in PRF prepared from fresh whole-blood without anticoagulants, it is thought that the centrifugal force increases the contact of factor XII with a glass surface, thereby primarily forming fibrin clots. Although platelets can also be activated by a glass surface, the centrifugal force accumulates platelets at and just below the interface between the red blood cell fraction and pl...
An alternative possibility may be related to the status of platelets stored in a low-Ca2+ environment: platelets constantly increase thromboxane A2 (TXA2) production and tend to easily aggregate under the influence of the TXA2 autocrine loop [19]. Therefore, it is thought that platelets prepared from citrated whole-blood samples continuously repeat Ca2+ discharge from intracellular Ca2+ stores and...
It is well known that platelets are activated by adenosine diphosphate (ADP), thrombin, epinephrine, thromboxane A2, collagen, and many other compounds and thus aggregate through binding of fibrinogen and glycoprotein IIb/IIIa receptors and upregulate surface antigens known as “platelet activation markers”: CD62P and CD63 [13, 14]. During activation, platelet morphology generally changes from ...
The surface microstructures of the formed fibrin clots are depicted in Fig. 6. The loop-like substances that initially formed in PRP on watch glasses were composed of abundant aggregated platelets, and relatively smaller amounts of fibrin fibers were deposited around platelet aggregates (Fig. 6a). By contrast, in the final version of fibrin clots, platelet aggregates were hardly detectable and m...
Morphological changes of (and microparticle release and fibrin formation by) Ca2+-stimulated platelets are shown in Fig. 1. When washed platelets were treated with 0.1% CaCl2 in sample tubes and plated in culture dishes for the last 5 min of treatment (control, 2 min), platelets’ ability to adhere to the dish bottom surface increased with the duration of Ca2+ treatment (Fig. 1a). On the othe...
Imaging by DHM (HoloMonitor M4; Phase Holographic Imaging AB, Lund, Sweden) was performed as described elsewhere [12]. The data were analyzed using specialized software, HoloStudio M4 (Phase Holographic Imaging AB). For surface roughness and area analysis, after a series of images were captured, the grayscale images were converted to the black-and-white format by the Otsu method, and the cell iden...
As described in the subsection above, the platelet fractions were prepared and treated with CaCl2 in polypropylene sample tubes. At the end of incubation, the platelets were fixed with an equal volume of a commercial fixative, ThromboFix (Beckman-Coulter, Brea, CA, USA), for 30 min, washed twice with PBS, and probed simultaneously with phycoerythrin (PE)-conjugated mouse monoclonal anti-CD41 and ...
Blood samples were collected from eight nonsmoking healthy male volunteers at ages from 32 to 68 years. The study design and consent forms of all the procedures performed were approved by the ethics committee for human participants of the Niigata University School of Medicine (Niigata, Japan) in accordance with the Helsinki Declaration of 1964 as revised in 2013.
Peripheral blood (~ 9 mL) was...
In this study, we attempted to dissociate platelets from the coagulation pathway and to evaluate possible direct action of Ca2+ on platelet functions in citrated whole-blood samples. In addition, in response to recent increasingly frequent requests for scheduled or outsourced, but not immediate on-site, preparation of various platelet concentrate types [4, 5], we examined time course changes in pl...
Since Marx’s report [1], platelet-rich plasma (PRP) and subsequently modified PRP derivatives have been widely applied in regenerative dentistry. Unlike self-clotted platelet-rich fibrin (PRF), for better handling efficiency and minimizing the loss of growth factors to diffusion, PRP and some other derivatives in liquid form are usually clotted by addition of exogenous coagulation factors, such ...
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 CaCl2 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 improvin...