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Zoledronic acid at subtoxic dose extends osteoblastic stage span of primary human osteoblasts

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Abstract

Objective

This study aimed to check the effect of zoledronic acid (ZA) at subtoxic dose on human osteoblasts (HOs) in terms of cell viability, apoptosis occurrence, and differentiation induction. ZA belongs to the family of bisphosphonates (BPs), largely used in the clinical practice for the treatment of bone diseases, often associated with jaw osteonecrosis onset. Their pharmacological action consists in the direct block of the osteoclast-mediated bone resorption along with indirect action on osteoblasts.

Materials and methods

HOs were treated choosing the highest limit concentration (10−5 M) which does not induce toxic effects. Live/dead staining, flow cytometry, mitochondrial membrane potential assay, osteocalcin western blotting, gp38 RT-PCR, collagen type I, PGE2, and IL-6 ELISA assays were performed.

Results

Similar viability level between control and ZA-treated samples is found along with no significant increase of apoptotic and necrotic cells in ZA-treated sample. To establish if an early apoptotic pathway was triggered, Bax expression and mitochondrial membrane potential were evaluated finding a higher protein expression in control sample and a good integrity of mitochondrial membrane in both experimental points. Type I collagen secretion and alkaline phosphatase (ALP) activity appear increased in ZA-treated sample, osteocalcin expression level is reduced in ZA-treated cells, whereas no modifications of gp38 mRNA level are evidenced. No statistical differences are identified in PGE2 secretion level whereas IL-6 secretion is lower in ZA-treated HOs with respect to control ones.

Conclusions

These results highlight that ZA, delaying the osteoblastic differentiation process versus the osteocytic lineage, strengthens its pharmacological activity enhancing bone density.

Clinical relevance

The knowledge of ZA effects on osteoblasts at subtoxic dose allows to improve therapeutic protocols in order to strengthen drug pharmacological activity through a combined action on both osteoclastic and osteoblastic cells.

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Acknowledgments

The authors would like to thank Dott.ssa Piera Sozio for drug purity determinations, Dott.ssa Alessandra di Paolo for the kind gift of Zometa, and MIUR FIRB-Accordi di Programma 2010 Project “Processi degenerativi dei tessuti mineralizzati del cavo orale, impiego di biomateriali e controllo delle interazioni con i microrganismi dell’ambiente” for the fellowships attributed to Dr. V.L Zizzari and Dr. M. De Colli.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Section of Human Anatomy, Department of Pharmacy, University “G. d’Annunzio” Chieti-Pescara, Via dei Vestini 31, 66100, Chieti, Italy

    Susi Zara, Marianna De Colli, Viviana di Giacomo, Vincenzo Luca Zizzari, Chiara Di Nisio, Marialucia Gallorini & Amelia Cataldi

  2. Department of Medical, Oral and Biotechnological Sciences, University “G. d’Annunzio” Chieti-Pescara, Via dei Vestini 31, 66100, Chieti, Italy

    Umberto Di Tore & Stefano Tetè

  3. Department of Medicine and Sciences of Aging, University “G. d’Annunzio” Chieti-Pescara, Via dei Vestini 31, 66100, Chieti, Italy

    Vincenzo Salini

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  1. Susi Zara
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Correspondence to Susi Zara.

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Zara, S., De Colli, M., di Giacomo, V. et al. Zoledronic acid at subtoxic dose extends osteoblastic stage span of primary human osteoblasts. Clin Oral Invest 19, 601–611 (2015). https://doi.org/10.1007/s00784-014-1280-8

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  • DOI: https://doi.org/10.1007/s00784-014-1280-8

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