Clinical Oral Investigations

, Volume 23, Issue 2, pp 813–820 | Cite as

Influence of pH on osteoclasts treated with zoledronate and alendronate

  • Francisco Javier Manzano-Moreno
  • Javier Ramos-Torrecillas
  • Elvira de Luna-Bertos
  • Rebeca Illescas-Montes
  • Timothy R. Arnett
  • Concepción RuizEmail author
  • Olga García-Martínez
Original Article



The objectives of this study were to analyze the effect of pH on the growth and activity of osteoclasts treated with different doses of two nitrogen-containing BPs, zoledronate and alendronate.

Materials and Methods

Murine osteoclasts cultured on dentine disks were treated with zoledronate (50 or 500 nM) or alendronate (500 or 5 μM) at two different pH values (7.4 or 7.0). Osteoclasts were counted with transmitted light microscopy, apoptosis/necrosis was studied with flow cytometry and confocal microscopy, and resorption pit number and depth were calculated using reflected light and scanning electron microscopy.


The osteoclast count on dentine disks was significantly (p < 0.001) reduced by zoledronate or alendronate treatment at pH 7.0 in comparison to treatment with the same doses at pH 7.4 and untreated disks (controls). The percentage of apoptotic cells was significantly increased by treatment with 500 nM zoledronate or 5 μM alendronate at pH 7.0 in comparison to the same doses at pH 7.4. The number and depth of resorption pits were significantly lower in disks treated at each BP dose studied than in untreated controls at pH 7.0.


Zoledronate and alendronate at therapeutic doses have an adverse effect on the viability and resorptive activity of osteoclasts when the local medium pH is reduced.

Clinical relevance

These findings suggest that periodontal or peri-implant oral cavity infection may be a key trigger of the cascade of events that lead to BRONJ.


Bisphosphonates Osteonecrosis Jaw Activity Growth Apoptosis 



We thank Mark Turmaine for assistance with scanning electron micrographs.


This study was supported by research group BIO277 (Junta de Andalucía).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in this study involving animals were in accordance with the ethical standards of the ethical committee of the University of Granada.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Francisco Javier Manzano-Moreno
    • 1
  • Javier Ramos-Torrecillas
    • 2
    • 3
  • Elvira de Luna-Bertos
    • 2
    • 3
  • Rebeca Illescas-Montes
    • 2
    • 4
  • Timothy R. Arnett
    • 5
  • Concepción Ruiz
    • 2
    • 3
    • 6
    Email author
  • Olga García-Martínez
    • 2
    • 3
  1. 1.Biomedical Group (BIO277), Department of Stomatology, School of DentistryUniversity of GranadaGranadaSpain
  2. 2.Instituto Investigación Biosanitaria, ibs.GranadaGranadaSpain
  3. 3.Biomedical Group (BIO277), Department of Nursing, Faculty of Health SciencesUniversity of GranadaGranadaSpain
  4. 4.Biomedical Group (BIO277), Department of Nursing, Faculty of Health SciencesUniversity of GranadaMelillaSpain
  5. 5.Department of Cell and Developmental BiologyUniversity College LondonLondonUK
  6. 6.Institute of Neuroscience, Parque Tecnológico Ciencias de la Salud, Armilla (Granada)University of GranadaGranadaSpain

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