Advertisement
Basic Research| Volume 40, ISSUE 11, P1810-1814, November 2014

Download started.

Ok

Effects of Platelet-rich Plasma and Cell Coculture on Angiogenesis in Human Dental Pulp Stem Cells and Endothelial Progenitor Cells

Published:August 28, 2014DOI:https://doi.org/10.1016/j.joen.2014.07.022

      Highlights

      • We culture 2 kinds of cells (human dental pulp stem cells and endothelial progenitor cells) and prepare platelet-rich plasma successfully.
      • We coculture human dental pulp stem cells and endothelial progenitor cells to investigate the potential effects on vasculogenic differentiation.
      • We confirm that platelet-rich plasma not only maintains the growth and proliferation of cells but also promotes vasculogenesis.
      • We confirm that coculture can promote vasculogenesis.
      • Thus, platelet-rich plasma can be used in tissue engineering and might even be applied in tooth regeneration in the future.

      Abstract

      Introduction

      Platelet-rich plasma (PRP) has been described as platelet concentrate. Growth factors released by activated platelets can improve wound vasculogenesis and enhance wound healing. In this study, we used PRP instead of serum to culture human dental pulp stem cells (hDPSCs) and endothelial progenitor cells (EPCs) and investigated revascularization ability. The effect of hDPSC and EPC coculture on vasculogenesis was also studied.

      Methods

      PRP was prepared by secondary centrifugation. Real-time polymerase chain reaction and Western blotting were used to determine the expression of vasculogenesis-related factors vascular endothelial growth factor, platelet-derived growth factor, fetal liver kinase 1 (Flk-1), and stromal cell-derived factor 1 (SDF-1) in cultured hDPSCs and EPCs. The cells were divided into 4 groups: EPCs + 10% fetal bovine serum (FBS), EPCs + 10% PRP, EPCs + hDPSCs + 10% FBS, and EPCs + hDPSCs + 10% PRP. Then, the formation of vessel-like structures was tested by the tube formation assay.

      Results

      On day 3, the expression levels of all the markers in the coculture groups were much higher than in the single-culture groups and were also higher in the PRP groups compared with the FBS groups (P < .05), except for SDF-1. Expression levels were significantly higher in the experimental groups (EPCs + 10% PRP, EPCs + hDPSCs + 10% FBS, and EPCs + hDPSCs + 10% PRP) than in the control group (EPCs + 10% FBS) and in the PRP groups/coculture groups compared with the FBS groups/single-culture groups (P < .01). The tube formation assay showed the area of vessel-like structures formed by the PRP group to be larger than in the FBS group (P < .05).

      Conclusions

      PRP and coculture can both promote vasculogenesis, and PRP can promote EPCs to form vessel-like structures.

      Key Words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Journal of Endodontics
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Eppley B.L.
        • Woodell J.E.
        • Higgins J.
        Platelet quantification and growth factor analysis from platelet-rich plasma: implications for wound healing.
        Plast Reconstr Surg. 2004; 114: 1502-1508
        • Pietramaggiori G.
        • Kaipainen A.
        • Czeczuga J.M.
        • et al.
        Freeze-dried platelet-rich plasma shows beneficial healing properties in chronic wounds.
        Wound Repair Regen. 2006; 14: 573-580
        • Robson M.C.
        • Phillips L.G.
        • Thomason A.
        • et al.
        Recombinant human platelet-derived growth factor-BB for the treatment of chronic pressure ulcers.
        Ann Plast Surg. 1992; 29: 193-201
        • Heldin C.-H.
        • Westermark B.
        Mechanism of action and in vivo role of platelet-derived growth factor.
        Physiol Rev. 1999; 79: 1283-1316
        • Beck J.
        • Evans D.
        • Tonino P.M.
        • et al.
        The biomechanical and histologic effects of platelet-rich plasma on rat rotator cuff repairs.
        Am J Sports Med. 2012; 40: 2037-2044
        • Roy S.
        • Driggs J.
        • Elgharably H.
        • et al.
        Platelet-rich fibrin matrix improves wound angiogenesis via inducing endothelial cell proliferation.
        Wound Repair Regen. 2011; 19: 753-766
        • Yang H.S.
        • Shin J.
        • Bhang S.H.
        • et al.
        Enhanced skin wound healing by a sustained release of growth factors contained in platelet-rich plasma.
        Exp Mol Med. 2011; 43: 622-629
        • Ravari H.
        • Hamidi-Almadari D.
        • Salimifar M.
        • et al.
        Treatment of non-healing wounds with autologous bone marrow cells, platelets, fibrin glue and collagen matrix.
        Cytotherapy. 2011; 13: 705-711
        • Tashnizi M.A.
        • Alamdari D.H.
        • Khayami M.E.
        • et al.
        Treatment of non-healing sternum wound after open-heart surgery with allogenic platelet-rich plasma and fibrin glue-preliminary outcomes.
        Indian J Plast Surg. 2013; 46: 538-542
        • Albanese A.
        • Licata M.E.
        • Polizzi B.
        • et al.
        Platelet-rich plasma (PRP) in dental and oral surgery: from the wound healing to bone regeneration.
        Immun Ageing. 2013; 10: 23
        • Kaul R.P.
        • Godhi S.S.
        • Singh A.
        Autologous platelet rich plasma after third molar surgery: a comparative study.
        J Maxillofac Oral Surg. 2012; 11: 200-205
        • Boswell S.G.
        • Cole B.J.
        • Sundman E.A.
        • et al.
        Platelet-rich plasma: a milieu of bioactive factors.
        Arthroscopy. 2012; 28: 429-439
        • Ma D.
        • Gao J.
        • Wu B.
        • et al.
        Changes in proliferation and osteogenic differentiation of stem cells from deep caries in vitro.
        J Endod. 2012; 38: 796-802
        • Reinhold J.
        • Christina L.
        • Mervyn W.
        • et al.
        Outgrowth endothelial cells: characterization and their potential for reversing ischemic retinopathy.
        Invest Ophthalmol Vis Sci. 2010; 51: 5906-5913
        • Noriko O.
        • Qin L.
        • Yuko H.
        • et al.
        Optimized method for culturing outgrowth endothelial progenitor cells.
        Inflamm Regen. 2011; 31: 219-227
        • Mooren R.E.
        • Hendriks E.J.
        • van den Beucken J.J.
        • et al.
        The effect of platelet-rich plasma in vitro on primary cells: rat osteoblast-like cells and human endothelial cells.
        Tissue Eng Part A. 2010; 16: 3159-3172
        • Dohan Ehrenfest D.M.
        • Rasmusson L.
        • Albrektsson T.
        Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF).
        Trends Biotechnol. 2009; 27: 158-167
        • Ogino Y.
        • Ayukawa Y.
        • Kukita T.
        • et al.
        Platelet-rich plasma suppresses osteoclastogenesis by promoting the secretion of osteoprotegerin.
        J Periodontal Res. 2009; 44: 217-224
        • Zaky S.H.
        • Ottonello A.
        • Strada P.
        • et al.
        Platelet lysate favours in vitro expansion of human bone marrow stromal cells for bone and cartilage engineering.
        J Tissue Eng Regen Med. 2008; 2: 472-481
        • Dissanayaka W.L.
        • Zhan X.
        • Zhang C.
        • et al.
        Coculture of dental pulp stem cells with endothelial cells enhances osteo-/odontogenic and angiogenic potential in vitro.
        J Endod. 2012; 38: 454-463
        • Martin G.
        • Ricucci D.
        • Gibbs J.L.
        • et al.
        Histological findings of revascularized/revitalized immature permanent molar with apical periodontitis using platelet-rich plasma.
        J Endod. 2013; 39: 138-144
        • Torabinejad M.
        • Faras H.
        A clinical and histological report of a tooth with an open apex treated with regenerative endodontics using platelet-rich plasma.
        J Endod. 2012; 38: 864-868
        • Jadhav G.
        • Shah N.
        • Logani A.
        Revascularization with and without platelet-rich plasma in nonvital, immature, anterior teeth: a pilot clinical study.
        J Endod. 2012; 38: 1581-1587
        • Foster T.E.
        • Puskas B.L.
        • Mandelbaum B.R.
        • et al.
        Platelet-rich plasma: from basic science to clinical applications.
        Am J Sports Med. 2009; 37: 2259-2272
        • Dong Z.
        • Li B.
        • Liu B.
        • et al.
        Platelet-rich plasma promotes angiogenesis of prefabricated vascularized bone graft.
        J Oral Maxillofac Surg. 2012; 70: 2191-2197
        • Medina R.J.
        • O'Neill C.L.
        • Humphreys M.W.
        • et al.
        Outgrowth endothelial cells: characterization and their potential for reversing ischemic retinopathy.
        Invest Ophthalmol Vis Sci. 2010; 51: 5906-5913