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Bupivacaine, ropivacaine, and morphine: comparison of toxicity on human hamstring-derived stem/progenitor cells

  • Experimental Study
  • Published:
Knee Surgery, Sports Traumatology, Arthroscopy Aims and scope

Abstract

Purpose

Bupivacaine, ropivacaine, and morphine are commonly administered intraarticularly after anterior cruciate ligament (ACL) reconstruction. However, their effects on human tendon stem/progenitor cells (TSPC) have not been studied. Therefore, this study investigates the cytotoxicity of these analgetics on TSPC.

Methods

Cells were isolated from human hamstring grafts of 3 female (age 15, 16 and 59) and 2 male patients (age 16 and 47). Cells were incubated using 0.5% bupivacaine, 0.5/0.75% ropivacaine, and 0.025% morphine. Cell viability was assessed after 0.5, 2, and 6 h using live/dead assay. Metabolic activity and apoptosis were measured by WST- and Annexin-V-FACS-assay after 2 h.

Results

Cell viability remained unchanged after 0.5 h in all groups, while treatment with bupivacaine and 0.5/0.75% ropivacaine resulted in a complete cell loss after 6 h. Contrarily, morphine showed no cytotoxic effect. Cell viability and metabolism were significantly reduced after treatment with bupivacaine (22.1; 8.3%) and 0.75% ropivacaine (56.5; 23.8%), while 0.5% ropivacaine and morphine showed no significant difference compared with controls. Apoptosis was significantly induced after incubation with bupivacaine (58.1%) and 0.75% ropivacaine (26.2%), whereas 0.5% ropivacaine only led to a slight induction compared with morphine and controls.

Conclusions

Clinically administered concentrations of bupivacaine (0.5%) and ropivacaine (0.75%) have a significant cytotoxic effect on human TSPC in vitro, while ropivacaine in a concentration of 0.5% has a mild but not significant effect on apoptosis and cell metabolism. In contrast, morphine does not affect cell survival, metabolism, or apoptosis. Knowing that morphine provides comparable to even prolonged pain reduction after ACL reconstruction, the presented in vitro study suggests morphine as a potentially less toxic analgetic drug for intraarticular application in clinical practice.

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Acknowledgments

We thank Dr. David Anz, Division of Clinical Pharmacology, University of Munich (LMU), for the collaboration and support regarding the flow cytometry analysis. Florian Haasters acknowledges the support of the Friedrich-Baur-Foundation, LMU Munich (project 0019/2008) and by the Faculty of Medicine, LMU Munich (FöFoLe, project 660). Hans Polzer was supported by the Friedrich-Baur-Foundation, LMU Munich (project 0018/2008), the scientific publisher council of MMW, Munich and by the Faculty of Medicine, LMU Munich (FöFoLe, project 704). Wolf Christian Prall was supported by the Faculty of Medicine, LMU Munich (FöFoLe, project 565). Denitsa Docheva and Matthias Schieker acknowledge the support of the German Research Foundation (DFG) (project DO1414/1-1) and the Bavarian Research Foundation (ForZebRA).

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Authors and Affiliations

  1. Experimental Surgery and Regenerative Medicine, Department of Surgery, Ludwig-Maximilians-University Munich, Nussbaumstr. 20, 80336, Munich, Germany

    Florian Haasters, Hans Polzer, Wolf Christian Prall, Maximilian Michael Saller, Julia Kohler, Stefan Grote, Wolf Mutschler, Denitsa Docheva & Matthias Schieker

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  1. Florian Haasters
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  2. Hans Polzer
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Correspondence to Matthias Schieker.

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F. Haasters and H. Polzer have contributed equally to this work.

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Haasters, F., Polzer, H., Prall, W.C. et al. Bupivacaine, ropivacaine, and morphine: comparison of toxicity on human hamstring-derived stem/progenitor cells. Knee Surg Sports Traumatol Arthrosc 19, 2138–2144 (2011). https://doi.org/10.1007/s00167-011-1564-3

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