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Comparative effect of platelet-rich plasma, platelet-poor plasma, and fetal bovine serum on the proliferative response of periodontal ligament cell subpopulations

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Abstract

Objectives

Cell-based therapies involve the need to expand cell cultures ex vivo for their subsequent implantation in an autologous manner. An important limitation regarding this technology is the use of fetal bovine serum (FBS) that has critical safety limitations. Platelet-derived fractions represent an autologous source of growth factors that may be used for the expansion of these cell cultures. Periodontal ligament (PDL) cells comprise a heterogeneous cell population that may not necessarily respond in a uniform manner to proliferative stimuli. The aim of this study was to evaluate the ability of two platelet-derived fractions, platelet-rich plasma (PRP) and platelet-poor plasma (PPP) and FBS on the proliferative response of different subpopulations of PDL cell cultures.

Materials and methods

PDL cells were characterized and then exposed to PRP, PPP, or FBS during 2, 5, or 14 days to analyze cell proliferation and clonogenic capability. Cell proliferation was evaluated through immunofluorescence for Ki67 and by tracing carboxyfluorescein diacetate succinimidyl ester (CFSE) dye in combination with mesenchymal stem cell markers using flow cytometry.

Results

Both PRP and PPP stimulated PDL cell proliferation and their clonogenic ability. We found a significant increase of CD73- and CD90-positive cells after PRP or PPP treatment, compared to FBS. Otherwise, no differences were found regarding the response of CD146-or CD105-positive cells when stimulated with PRP, PPP, or FBS.

Conclusion

PRP and PPP can stimulate the proliferation and clonogenicity of PDL cell populations including cells positive for CD90 and CD73 markers.

Clinical relevance

These findings may have implications for future therapies aiming to stimulate periodontal regeneration using autologous growth factors.

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Acknowledgements

This work was supported by the projects FONDECYT 11121294, PUENTE P1704/2017 (CM), and FONDECYT 1130618 (PS) Millennium Institute on Immunology and Immunotherapy (P09/016-F, to AK). The flow cytometry experiments were performed at the Flow Cytometry Core Facility, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile. We acknowledge the contribution of tissue and blood donors.

The selected hybridoma monoclonal antibodies were PD31, developed by Elizabeth A. Wayner and Gregory Vercellotti and P8B1 by Elizabeth A. Wayner/Tucker LeBien, from the Fred Hutchinson Cancer Research Center, Seattle, WA. These antibodies were obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA, 52242.

Funding

The work was supported by the Fondo Nacional de Desarrollo Científico y Tecnológico (FONDECYT) from the Chilean Government and Puente research grant by Pontificia Universidad Católica de Chile.

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Authors and Affiliations

  1. School of Dentistry, Facultad de Medicina, Pontificia Universidad Católica de Chile, Santiago, Chile

    Constanza E. Martínez & Patricio C. Smith

  2. Millennium Institute of Immunology and Immunotherapy, Departamento de Genética Molecular y Microbiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile

    Roberto Gómez & Alexis M. Kalergis

  3. Departamento de Endocrinología, Facultad de Medicina, Pontificia Universidad Católica de Chile, Santiago, Chile

    Alexis M. Kalergis

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  1. Constanza E. Martínez
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Correspondence to Constanza E. Martínez.

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All procedures performed in studies involving samples from human participants were in accordance with the ethical standards of the Faculty of Medicine Ethical Committee at Pontificia Universidad Católica de Chile and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Informed consent was obtained from all individual participants included in the study.

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Martínez, C.E., Gómez, R., Kalergis, A.M. et al. Comparative effect of platelet-rich plasma, platelet-poor plasma, and fetal bovine serum on the proliferative response of periodontal ligament cell subpopulations. Clin Oral Invest 23, 2455–2463 (2019). https://doi.org/10.1007/s00784-018-2637-1

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