Translational hurdles for tissue engineering

An in vitro analysis of commonly used local anaesthetics on skeletal stem cell survival

E. R. Tayton, J. O. Smith, A. Aarvold, S. Kalra, D. G. Dunlop, R. O. C. Oreffo

Abstract

When transferring tissue regenerative strategies involving skeletal stem cells to human application, consideration needs to be given to factors that may affect the function of the cells that are transferred. Local anaesthetics are frequently used during surgical procedures, either administered directly into the operative site or infiltrated subcutaneously around the wound. The aim of this study was to investigate the effects of commonly used local anaesthetics on the morphology, function and survival of human adult skeletal stem cells.

Cells from three patients who were undergoing elective hip replacement were harvested and incubated for two hours with 1% lidocaine, 0.5% levobupivacaine or 0.5% bupivacaine hydrochloride solutions. Viability was quantified using WST-1 and DNA assays. Viability and morphology were further characterised using CellTracker Green/Ethidium Homodimer-1 immunocytochemistry and function was assessed by an alkaline phosphatase assay. An additional group was cultured for a further seven days to allow potential recovery of the cells after removal of the local anaesthetic.

A statistically significant and dose dependent reduction in cell viability and number was observed in the cell cultures exposed to all three local anaesthetics at concentrations of 25% and 50%, and this was maintained even following culture for a further seven days.

This study indicates that certain local anaesthetic agents in widespread clinical use are deleterious to skeletal progenitor cells when studied in vitro; this might have relevance in clinical applications.

Footnotes

  • E. Tayton and J. Smith were joint first authors for this study. The authors would like to thank the orthopaedic surgeons at Southampton General Hospital for provision of bone marrow samples. They also acknowledge the excellent technical support of Ms E. Ralph.

    No benefits in any form have been received or will be received from a commercial party related directly or indirectly to the subject of this article.

  • Supplementary material. A further opinion by Professor D. Hukins is available with the electronic version of this article on our website at www.boneandjoint.org.uk/site/education/further_op

  • Received October 11, 2011.
  • Accepted February 17, 2012.
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