Possible anti-inflammatory effects of transcription factors contained in platelet microparticles and periodontal regeneration

• Project/Area Number: 18K09595
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 57030:Conservative dentistry-related
• Research Institution: Niigata University
• Principal Investigator: Kawase Tomoyuki 新潟大学, 医歯学系, 准教授 (90191999)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 血小板 / 多血小板血漿 / microparticle / 抗炎症 / PPARγ / 増殖因子 / α顆粒 / マイクロパーティクル / 抗炎症作用 / 歯周再生

Outline of Final Research Achievements

Scanning electron microscopy demonstrated that when stimulated with 0.1% CaCl2, washed platelets released microparticles (MPs), of which diameter was 1 μm or smaller, into extracellular spaces. To support this morphological data, we used flowcytometry (FCM) to detect specific surface antigens of MPs. However, our FCM failed to clearly distinguish such MPs from contaminated debris.
Immunofluorescent staining demonstrated that PPARγ was distributed in the cytoplasm of platelet in resting state and that the stimulation with CaCl2 facilitated its release and diffusion to the extracellular spaces. This kind of change in distribution was very similar to representative growth factors, such as PDGF and TGFβ1, contained in α granules of platelets. Thus, it is suggested that PPARγ is released from platelets in a form of MP upon activation.

Academic Significance and Societal Importance of the Research Achievements

多血小板血漿(PRP)の組織再生能力は広く知られているが，抗炎症作用については，その実態が解明されていない．本研究によって，PPARγが血小板中に含まれ，活性化刺激によって血小板外に放出される所見が得られたことは，組織再生機序を包括的に理解するうえで貴重な糸口となりうると確信する．
また，同じく研究途上にあるPRPの抗菌作用とともに，今後のPRP臨床応用に組織再生以外のあたらしい意義(治療効果)をもたらし，停滞気味のPRP研究の発展を加速させることが期待される．

Research Products

• [Int'l Joint Research] University of Bern(スイス)
• [Int'l Joint Research] Federal Fluminense University(ブラジル)
• [Journal Article] Fluorescent cytochemical detection of polyphosphates associated with human platelets (2021)
  • Author(s): Sato A, Aizawa H, Tsujino T, Isobe K, Watanabe T, Kitamura Y, Kawase T
  • Journal Title: Int J Mol Sci Volume: 22 Issue: 3 Pages: 1040-1040
  • DOI: 10.3390/ijms22031040
  • Peer Reviewed / Open Access
• [Journal Article] Distribution and quantification of activated platelets in platelet-rich fibrin matrices (2020)
  • Author(s): Sato A, Kawabata H, Aizawa H, Tsujino T, Isobe K, Watanabe T, Kitamura Y, Miron RJ, Kawase T
  • Journal Title: Platelets Volume: in press Issue: 1 Pages: 110-115
  • DOI: 10.1080/09537104.2020.1856359
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Platelet adhesion on commercially pure titanium plates in vitro III: effects of calcium phosphate-blasting on titanium plate biocompatibility (2020)
  • Author(s): Nakamura M, Aizawa H, Kawabata H, Sato A, Watanabe T, Isobe K, Kitamura Y, Tanaka T, Kawase T
  • Journal Title: Int J Implant Dent Volume: 6 Issue: 1 Pages: 74-74
  • DOI: 10.1186/s40729-020-00270-2
  • Peer Reviewed / Open Access
• [Journal Article] Quantitative Near-Infrared Imaging of Platelets in Platelet-Rich Fibrin (PRF) Matrices: Comparative Analysis of Bio-PRF, Leukocyte-Rich PRF, Advanced-PRF and Concentrated Growth Factors (2020)
  • Author(s): Aizawa H, Tsujino T, Watanabe T, Isobe K, Kitamura Y, Sato A, Yamaguchi S, Okudera H, Kawase T
  • Journal Title: Int J Mol Sci Volume: 21 Issue: 12 Pages: 4426-4426
  • DOI: 10.3390/ijms21124426
  • Peer Reviewed / Open Access
• [Journal Article] Concentrated growth factor matrices prepared using silica-coated plastic tubes are distinguishable from those prepared using glass tubes in platelet distribution: Application of a novel near-infrared imaging-based, quantitative technique (2020)
  • Author(s): Yamaguchi S, Aizawa H, Sato A, Tsujino T, Isobe K, Kitamura Y, Watanabe T, Okudera H, Mourao CF, Kawase T
  • Journal Title: Front Bioeng Biotechnol Volume: 8 Pages: 600-600
  • DOI: 10.3389/fbioe.2020.00600
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A comparative study of the effects of anticoagulants on pure platelet-rich plasma quality and potency. (2020)
  • Author(s): Aizawa H, Kawabata H, Sato A, Masuki H, Watanabe T, Tsujino T, Isobe K, Nakamura M, Nakata K, Kawase T.
  • Journal Title: Biomedicines Volume: 8 Issue: 3 Pages: 42-42
  • DOI: 10.3390/biomedicines8030042
  • Peer Reviewed / Open Access
• [Journal Article] Acute cytotoxic effects of silica microparticles used for coating of plastic blood-collection tubes on human periosteal cells. (2020)
  • Author(s): Masuki H, Isobe K, Kawabata H, Tsujino T, Yamaguchi S, Watanabe T, Sato A, Aizawa H, Mourao CF, Kawase T.
  • Journal Title: Odontology Volume: in press Issue: 4 Pages: 545-552
  • DOI: 10.1007/s10266-020-00486-z
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Distribution of platelets, TGFβ1, PDGF-BB, VEGF, MMP9 and fibronectin in advanced platelet-rich fibrin (A-PRF) and concentrated growth factors (CGF) matrices. (2020)
  • Author(s): Takahashi A, Tsujino T, Yamaguchi S, Isobe K, Watanabe T, Kitamura Y, Okuda K, Nakata K, Kawase T.
  • Journal Title: Journal of Investigative Clinical Dentistry Volume: in press Issue: 4
  • DOI: 10.1111/jicd.12458
  • Peer Reviewed
• [Journal Article] Platelet adhesion on commercially pure titanium plates in vitro II. Immunofluorescence visualization of PDGF-B, TGFβ1, and PPARγ released from activated, adherent platelets. (2019)
  • Author(s): Tsujino T, Takahashi A, Isobe, K, Kitamura Y, Okuda K, Nakata K, Kawase T.
  • Journal Title: Dentistry Journal Volume: 7 Issue: 4 Pages: 109-109
  • DOI: 10.3390/dj7040109
  • Peer Reviewed / Open Access
• [Journal Article] Striking differences in platelet distribution between advanced-platelet-rich fibrin and concentrated growth factors: effects of silica-containing plastic tubes. (2019)
  • Author(s): Tsujino T, Masuki H, Nakamura M, Isobe K, Kawabata H, Aizawa H, Watanabe T, Kitamura Y, Okudera H, Okuda K, Nakata K, Kawase T.
  • Journal Title: Journal of Functional Biomaterial Volume: 10 Issue: 3 Pages: 43-43
  • DOI: 10.3390/jfb10030043
  • Peer Reviewed / Open Access
• [Journal Article] Platelet adhesion on commercially pure titanium plates in vitro I. Effects of plasma components and involvement of the von Willebrand factor and fibronectin. (2019)
  • Author(s): Takahashi A, Takahashi S, Tsujino T, Isobe K, Watanabe T, Kitamura Y, Watanabe Tk, Nakata K, Kawase T.
  • Journal Title: International Journal of Implant Dentistry Volume: 5 Issue: 1 Pages: 5-5
  • DOI: 10.1186/s40729-019-0160-z
  • Peer Reviewed / Open Access
• [Presentation] 多血小板フィブリン基質中の血小板分布に関する免疫組織化学的研究: 遠心条件と採血管の影響 (2020)
  • Author(s): 辻野哲弘， 川端秀男， 北村 豊，佐藤 篤 ，礒邉和重，山口貞博，奥寺 元，川瀬知之
  • Organizer: 第50回日本口腔インプラント学会学術大会
• [Presentation] 多血小板フィブリン調製に使用するシリカコート採血管に含まれるシリカ微粒子はヒト骨膜細胞に急性細胞毒性を示す (2020)
  • Author(s): 増木英郎，辻野哲弘，渡辺泰典，北村 豊，佐藤 篤，中村雅之，奥寺 元，川瀬知之
  • Organizer: 第50回日本口腔インプラント学会学術大会
• [Presentation] Platelet adhesion on commercially pure titanium plates in vitro: effects of plasma components and involvement of the von Willebrand factor and fibronectin. (2020)
  • Author(s): Kitamura Y, Takahashi A, Isobe K, Tsujino T, Watanabe T, Kawase T.
  • Organizer: Academy of Osseointegration 2020 Annual meeting
  • Int'l Joint Research
• [Presentation] Evidence for contamination of silica microparticles in advanced platelet-rich fibrin matrix prepared using silica-coated plastic tubes. (2020)
  • Author(s): Tsujino T, Watanabe T, Kitamura Y, Isobe K, Tanaka T, Kawase T.
  • Organizer: Academy of Osseointegration 2020 Annual meeting
  • Int'l Joint Research
• [Presentation] Spectrophotometric determination of platelet aggregation activity. (2020)
  • Author(s): Watanabe T, Tsujino T, Kitamura Y, Watanabe T, Isobe K, Kawase T.
  • Organizer: Academy of Osseointegration 2020 Annual meeting
  • Int'l Joint Research
• [Presentation] Platelet-rich fibrin extract: a promising fetal bovine serum alternative in explant cultures of human periosteal sheets for alveolar bone augmentation. (2020)
  • Author(s): Kawase T, Tsujino T, Watanabe T, Isobe K, Kitamura Y.
  • Organizer: Academy of Osseointegration 2020 Annual meeting
  • Int'l Joint Research
• [Presentation] In vitro における血小板の平坦純チタン表面への接着: 血漿成分の影響およびvonWillebrand factorとfibronectin の関与． (2019)
  • Author(s): 高橋 璋, 川瀬知之, 髙橋章太郎．
  • Organizer: 第49回日本口腔インプラント学会学術大会
• [Book] Understanding Platelet-Rich Fibrin (2021)
  • Author(s): Miron, RJ, et al.
  • Total Pages: 384
  • Publisher: QUINTESSENCE PUBLISHING CO., INC.
  • ISBN: 9781647240493