Skip to main content
SpringerLink
Log in

Platelet-rich plasma as supplement and scaffold for the culture of Vero cell line

  • Original Article
  • Published:
Research on Biomedical Engineering Aims and scope Submit manuscript

Abstract

Purpose

Tissue engineering performs the culture of cells on scaffolds, aiming at the restoration of damaged tissue. For the cell culture into the scaffold, it is necessary to establish certain conditions, such as the type of supplementation of the culture medium. Therefore, the aim of this research was to evaluate the platelet-rich plasma (PRP) as a supplement and scaffold for Vero cell culture, in a one case report study.

Methods

Calcium chloride was added to the PRP, obtained by apheresis. After PRP retraction, serum from PRP (SPRP) and the clot from PRP (CPRP) were obtained. SPRP was used as a supplement to the culture medium, and CPRP was utilized as a scaffold.

Results

The evaluation of SPRP as a supplement showed that there was no statistical difference in cell viability compared to cultures supplemented in the standard way, with fetal bovine serum (FBS), after 24 h of culture. The morphological analysis, carried out on the third and fifth days, did not verify changes in the typical morphology of Vero cells cultured with SPRP. For the use of CPRP, its structure was observed. The CPRP was formed by an irregular network of fibers with different diameters, having a mean value of 0.210 ± 0.097 μ. In the analysis of CPRP as a scaffold, it was verified that the cells adhered and spread over the biomaterial.

Conclusion

Therefore, in Vero cell cultures, the results suggest that SPRP and CPRP can be used as a supplement and scaffold, respectively.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Anitua E, Andia I, Ardanza B, Nurden P, Nurden AT. Autologous platelets as a source of proteins for healing and tissue regeneration. Thromb Haemost. 2004;91(1):4–15.

    Article  Google Scholar 

  • Anitua E, Andía I, Sanchez M, Azofra J, del Mar Zalduendo M, de la Fuente M, et al. Autologous preparations rich in growth factors promote proliferation and induce VEGF and HGF production by human tendon cells in culture. J Orthop Res. 2005;23(2):281–6.

    Article  Google Scholar 

  • Anitua E, Zalduendo MM, Alkhraisat MH, Orive G. Release kinetics of platelet-derived and plasma-derived growth factors from autologous plasma rich in growth factors. Ann Anat. 2013;195(5):461–6.

    Article  Google Scholar 

  • Badami AS, Kreke MR, Thompson MS, Riffle JS, Goldstein AS. Effect of fiber diameter on spreading, proliferation, and differentiation of osteoblastic cells on electrospun poly (lactic acid) substrates. Biomaterials. 2006;27(4):596–606.

    Article  Google Scholar 

  • Barbanti SH, Zavaglia CAC, Duek EAR. Polímeros bioreabsorvíveis na engenharia de tecidos. Polímeros. 2005;15(1):13–21.

    Article  Google Scholar 

  • Bashur CA, Dahlgren LA, Goldstein AS. Effect of fiber diameter and orientation on fibroblast morphology and proliferation on electrospun poly(d,l-lactic-co-glycolic acid) meshes. Biomaterials. 2006;27(33):5681–8.

    Article  Google Scholar 

  • Brunner D, Frank J, Appl H, Schöffl H, Pfaller W, Gstraunthaler G. Serum-free cell culture: the serum-free media interactive online database. ALTEX. 2010;27(1):53–62.

    Article  Google Scholar 

  • Carlos MML, Freitas PDF d S. Estudo da cascata de coagulação sangüínea e seus valores de referência. Acta Vet Bras. 2007;1(2):49–55.

    Google Scholar 

  • Carneiro MO, Barbieri CH, Neto JB. O gel de plasma rico em plaquetas propicia a regeneração da cartilagem articular do joelho de ovelhas. Acta Ortop Bras. 2013;21(2):80–6.

    Article  Google Scholar 

  • Cavallo C, Roffi A, Grigolo B, Mariani E, Pratelli L, Merli G, et al. Platelet-rich plasma: the choice of activation method affects the release of bioactive molecules. Biomed Res Int. 2016;2016:1–7.

    Article  Google Scholar 

  • Chachques JC, Herreros J, Trainini J, Juffe A, Rendal E, Prosper F, et al. Autologous human serum for cell culture avoids the implantation of cardioverter-defibrillators in cellular ardiomyoplasty. Int J Cardiol. 2004;95(Suppl 1):S29–33.

    Article  Google Scholar 

  • Christopherson GT, Song H, Mao HQ. The influence of fiber diameter of electrospun substrates on neural stem cell differentiation and proliferation. Biomaterials. 2009;30(4):556–64.

    Article  Google Scholar 

  • Cunha C, Panseri S, Antonini S. Emerging nanotechnology approaches in tissue engineering for peripheral nerve regeneration. Nanomedicine. 2011;7(1):50–9.

    Article  Google Scholar 

  • Ehrenfest DMD, Del Corso M, Diss A, Mouhyi J, Charrier J-B. Three-dimensional architecture and cell composition of a Choukroun’s platelet-rich fibrin clot and membrane. J Periodontol. 2010;81(4):546–55.

    Article  Google Scholar 

  • El-Sharkawy H, Kantarci A, Deady J, Hasturk H, Liu H, Alshahat M, et al. Platelet-rich plasma: growth factors and pro- and anti-inflammatory properties. J Periodontol. 2007;78(4):661–9.

    Article  Google Scholar 

  • Esposito AR, Lucchesi C, Ferreira BMP, Duek EAR. Estudo da interação células vero/plga após a modificação da superfície por plasma de oxigênio. Matéria (Rio J). 2007;12(1):164–72.

    Article  Google Scholar 

  • Fallouh L, Nakagawa K, Sasho T, Arai M, Kitahara S, Wada Y, et al. Effects of autologous platelet-rich plasma on cell viability and collagen synthesis in injured human anterior cruciate ligament. J Bone Joint Surg Am. 2010;92(18):2909–16.

    Article  Google Scholar 

  • Gawai KT, Sobhana CR. Clinical evaluation of use of platelet rich plasma in bone healing. J Maxillofac Oral Surg. 2015;14(1):67–80.

    Article  Google Scholar 

  • Gstraunthaler G. Alternatives to the use of fetal bovine serum: serum-free cell culture. ALTEX. 2003;20(4):275–81.

    Google Scholar 

  • Johnson MC, Meyer AA, deSerres S, Herzog S, Peterson HD. Persistence of fetal bovine serum proteins in human keratinocytes. J Burn Care Rehabil. 1990;11(6):504–9.

    Article  Google Scholar 

  • 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(5):323–9.

    Article  Google Scholar 

  • Kocaoemer A, Kern S, Klüter H, Bieback K. Human AB serum and thrombin-activated platelet-rich plasma are suitable alternatives to fetal calf serum for the expansion of mesenchymal stem cells from adipose tissue. Stem Cells. 2007;25(5):1270–8.

    Article  Google Scholar 

  • Kurita M, Aiba-Kojima E, Shigeura T, Matsumoto D, Suga H, Inoue K, et al. Differential effects of three preparations of human serum on expansion of various types of human cells. Plast Reconstr Surg. 2008;122(2):438–48.

    Article  Google Scholar 

  • Lombello CB, Malmonge SM, Wada ML. Morphology of fibroblastic cells cultured on poly (HEMA-co-AA) substrates. Cytobios. 2000;101(397):115–22.

    Google Scholar 

  • Malavolta EA, Gracitelli MEC, Sunada EE, Benegas E, Prada F de S, Bolliger Neto R, et al. Plasma rico em plaquetas no reparo artroscópico das roturas completas do manguito rotador. Rev Bras Ortop. 2012;47(6):741–7.

    Article  Google Scholar 

  • Marx RE. Platelet-rich plasma (PRP): what is PRP and what is not PRP? Implant Dent. 2001;10(4):225–8.

    Article  Google Scholar 

  • Marx RE. Platelet-rich plasma: evidence to support its use. J Oral Maxillofac Surg. 2004;62(4):489–96.

    Article  Google Scholar 

  • Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assay. J Immunol Methods. 1983;65(1–2):55–63.

    Article  Google Scholar 

  • O’Brien FJ. Biomaterials & scaffolds for tissue engineering. Mater Today. 2011;14(3):88–95.

    Article  Google Scholar 

  • Pinto JMN, Pizani NS, Kang HC, Silva LAK. Application of platelet-rich plasma in the treatment of chronic skin ulcer - case report. An Bras Dermatol. 2014;89(4):638–40.

    Article  Google Scholar 

  • Pretorius E, Vieira WA, Oberholzer HM, Auer REJ. Comparative scanning electron microscopy of platelets and fibrin networks of human and differents animals. J Morphol. 2009;27(1):69–76.

    Google Scholar 

  • Ryan EA, Mockros LF, Weisel JW, Lorand L. Structural origins of fibrin clot rheology. Biophys J. 1999;77(5):2813–26.

    Article  Google Scholar 

  • Santos AR Jr, Wada MLF. Polímeros biorreabsorvíveis como substrato para cultura de células e engenharia tecidual. Polímeros. 2007;17(4):308–17.

    Article  Google Scholar 

  • Wang H-L, Avila G. Platelet rich plasma: myth or reality? Eur J Dent. 2007;1(4):192–4.

    Google Scholar 

  • Weibrich G, Hansen T, Kleis W, Buch R, Hitzler WE. Effect of platelet concentration in platelet-rich plasma on peri-implant bone regeneration. Bone. 2004;34(4):665–71.

    Article  Google Scholar 

  • Wirth C, Comte V, Lagneau C, Exbrayat P, Lissac M, Jaffrezic-Renault N, et al. Nitinol surface roughness modulates in vitro cell response: a comparison between fibroblasts and osteoblasts. Mater Sci Eng C. 2005;25(1):51–60.

    Article  Google Scholar 

Download references

Acknowledgments

The authors are grateful to the company Criogênesis for kindly providing the PRP. The authors also acknowledge the Multiuser Central Facilities (UFABC) for the experimental support in the electron microscopy observation.

Funding

The authors thank the Federal University of ABC and the Coordination for the Improvement of Higher Education Personnel (CAPES) for the financial support.

Author information

Authors and Affiliations

  1. Center for Engineering, Modeling and Applied Social Sciences, Federal University of ABC, São Bernardo do Campo, São Paulo, Brazil

    Débora Carajiliascov Ferraraz, Leonardo Ribeiro Rodrigues & Christiane Bertachini Lombello

  2. Hematology and Hemotherapy of Hospital das Clínicas, Faculty of Medicine, University of São Paulo, São Paulo, São Paulo, Brazil

    Nelson Hidekazu Tatsui

  3. Criogênesis Biotecnologia Ltda., São Paulo, São Paulo, Brazil

    Nelson Hidekazu Tatsui

Authors
  1. Débora Carajiliascov Ferraraz
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nelson Hidekazu Tatsui
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Leonardo Ribeiro Rodrigues
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christiane Bertachini Lombello
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Débora Carajiliascov Ferraraz.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ferraraz, D.C., Tatsui, N.H., Rodrigues, L.R. et al. Platelet-rich plasma as supplement and scaffold for the culture of Vero cell line. Res. Biomed. Eng. 35, 1–9 (2019). https://doi.org/10.1007/s42600-019-00001-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s42600-019-00001-6

Keywords

Search

Navigation