Lipophilicity but not stereospecificity is a major determinant of local anaesthetic-induced cytotoxicity in human T-lymphoma cells

Background and objectives Local neurotoxicity of local anaesthetics is a well known phenomenon which is determined by lipophilicity. Recent reports have indicated the relevance of local anaesthetic-induced cytotoxicity also in nonneuronal tissues. This study re-evaluates the role of lipophilicity in local anaesthetic cytotoxicity in nonneuronal cells. In addition, the toxicities of pipecoloxylidine S(−) enantiomers were investigated. Methods Local anaesthetic-induced cytotoxicity was investigated in vitro in T-lymphoma cells (Jurkat). Cells were incubated with each of eight different local anaesthetics, two esters and six amides. Annexin V-fluorescein isothiocyanate and 7-aminoactinomycin D double staining followed by flow cytometry were used to investigate the fraction of early apoptotic cells as well as the overall cell death. The concentrations leading to 50% cell death (LC50) were calculated and compared. In a second step, we compared the toxicities of S(−) bupivacaine and the racemate as well as R(+) and S(−) ropivacaine. Results Concentration-dependent cytotoxicity was observed for all investigated local anaesthetics. Apoptosis was seen at low concentrations, whereas necrosis was observed at higher concentrations. LC50 values of the different local anaesthetics yielded the following decreasing order of toxicity: tetracaine, bupivacaine, ropivacaine, prilocaine, procaine, lidocaine, articaine and mepivacaine. Toxicity correlated with octanol/buffer partition coefficients, but was independent of the ester or amide linkage. There was no effect of stereoisomerism on apoptosis and necrosis. Conclusion Moderate correlations for cytotoxicity with lipophilicity and clinical potency of local anaesthetics can be found in nonneuronal cells that are less than those reported previously with neuronal cells. Structural factors such as ester or amide linkage or stereospecificity do not have any influence on cytotoxicity. Although S(−) enantiomers may be advantageous with regard to systemic toxicity, they have no advantage in respect of local cytotoxicity in vitro.

[1]  M. Hollmann,et al.  Benzethonium Increases the Cytotoxicity of S(+)-Ketamine in Lymphoma, Neuronal, and Glial Cells , 2010, Anesthesia and analgesia.

[2]  M. Hollmann,et al.  Ketamine induces apoptosis via the mitochondrial pathway in human lympocytes and neuronal cells , 2012 .

[3]  A. Pearsall,et al.  Apoptosis and mitochondrial dysfunction in human chondrocytes following exposure to lidocaine, bupivacaine, and ropivacaine. , 2010, The Journal of bone and joint surgery. American volume.

[4]  M. Szczodry,et al.  In vivo effects of single intra-articular injection of 0.5% bupivacaine on articular cartilage. , 2010, The Journal of bone and joint surgery. American volume.

[5]  F. Essmann,et al.  Apoptosis induction by different local anaesthetics in a neuroblastoma cell line. , 2009, British journal of anaesthesia.

[6]  T. Shin,et al.  Only tetracaine and not other local anaesthetics induce apoptosis in rat cortical astrocytes. , 2009, British journal of anaesthesia.

[7]  M. Seeberger,et al.  Bupivacaine concentrations in the lumbar cerebrospinal fluid of patients during spinal anaesthesia. , 2009, British journal of anaesthesia.

[8]  T. Blanck,et al.  Cytotoxicity of Local Anesthetics in Human Neuronal Cells , 2009, Anesthesia and analgesia.

[9]  X. Capdevila,et al.  Infectious Risk of Continuous Peripheral Nerve Blocks , 2009, Anesthesiology.

[10]  B. Graf,et al.  The toxicity of local anesthetics: the place of ropivacaine and levobupivacaine , 2008, Current opinion in anaesthesiology.

[11]  P. Lirk,et al.  In Vitro, Inhibition of Mitogen-Activated Protein Kinase Pathways Protects Against Bupivacaine- and Ropivacaine-Induced Neurotoxicity , 2008, Anesthesia and analgesia.

[12]  Q. Hogan.  Pathophysiology of Peripheral Nerve Injury During Regional Anesthesia , 2008, Regional Anesthesia & Pain Medicine.

[13]  P. Lirk,et al.  In Vitro, Lidocaine-Induced Axonal Injury Is Prevented by Peripheral Inhibition of the p38 Mitogen-Activated Protein Kinase, but Not by Inhibiting Caspase Activity , 2007, Anesthesia and analgesia.

[14]  C. Chu,et al.  Lidocaine Exhibits Dose- and Time-Dependent Cytotoxic Effects on Bovine Articular Chondrocytes in Vitro , 2007, The American journal of sports medicine.

[15]  K. Schulze-Osthoff,et al.  Lidocaine induces apoptosis via the mitochondrial pathway independently of death receptor signaling , 2012 .

[16]  P. Sirvent,et al.  Effects of Intermittent Femoral Nerve Injections of Bupivacaine, Levobupivacaine, and Ropivacaine on Mitochondrial Energy Metabolism and Intracellular Calcium Homeostasis in Rat Psoas Muscle , 2007, Anesthesiology.

[17]  S. Sakura,et al.  Epidural Lidocaine Induces Dose-Dependent Neurologic Injury in Rats , 2006, Anesthesia and analgesia.

[18]  S. Sakura,et al.  Comparative Somatic and Visceral Antinociception and Neurotoxicity of Intrathecal Bupivacaine, Levobupivacaine, and Dextrobupivacaine in Rats , 2006, Anesthesiology.

[19]  Y. Oyama,et al.  Cytometric analysis of lidocaine-induced cytotoxicity: a model experiment using rat thymocytes. , 2006, Toxicology.

[20]  K. Ohta,et al.  Lidocaine-induced apoptosis and necrosis in U937 cells depending on its dosage. , 2005, Biomedical research.

[21]  X. Capdevila,et al.  Continuous Peripheral Nerve Blocks in Hospital Wards after Orthopedic Surgery: A Multicenter Prospective Analysis of the Quality of Postoperative Analgesia and Complications in 1,416 Patients , 2005, Anesthesiology.

[22]  Sung Joong Lee,et al.  Bupivacaine Induces Apoptosis via ROS in the Schwann Cell Line , 2005, Journal of dental research.

[23]  F. Nicoletti,et al.  Comparative effects of levobupivacaine and racemic bupivacaine on excitotoxic neuronal death in culture and N-methyl-D-aspartate-induced seizures in mice. , 2005, European journal of pharmacology.

[24]  B. Graf,et al.  The Long Term Myotoxic Effects of Bupivacaine and Ropivacaine After Continuous Peripheral Nerve Blocks , 2005, Anesthesia and analgesia.

[25]  R. Fink,et al.  Differential Effects of Bupivacaine and Ropivacaine Enantiomers on Intracellular Ca2+ Regulation in Murine Skeletal Muscle Fibers , 2005, Anesthesiology.

[26]  G. Gores,et al.  Mitochondrial Injury and Caspase Activation by the Local Anesthetic Lidocaine , 2004, Anesthesiology.

[27]  R. Fink,et al.  The Acute Myotoxic Effects of Bupivacaine and Ropivacaine After Continuous Peripheral Nerve Blockades , 2003, Anesthesia and analgesia.

[28]  B. Allaouchiche,et al.  The Induction of Apoptosis by Local Anesthetics: A Comparison Between Lidocaine and Ropivacaine , 2003, Anesthesia and analgesia.

[29]  P. Friederich,et al.  Lidocaine-induced cell death in a human model of neuronal apoptosis. , 2002, European journal of anaesthesiology.

[30]  B. Graf,et al.  Differences in Cardiotoxicity of Bupivacaine and Ropivacaine Are the Result of Physicochemical and Stereoselective Properties , 2002, Anesthesiology.

[31]  Y. Nozawa,et al.  Involvement of the Mitogen-activated Protein Kinase Family in Tetracaine-induced PC12 Cell Death , 2002, Anesthesiology.

[32]  J. Mazat,et al.  Absence of Stereospecific Effects of Bupivacaine Isomers on Heart Mitochondrial Bioenergetics , 2000, Anesthesiology.

[33]  G. Lyons,et al.  Epidural pain relief in labour: potencies of levobupivacaine and racemic bupivacaine. , 1998, British journal of anaesthesia.

[34]  J. Levine,et al.  Lidocaine toxicity in primary afferent neurons from the rat. , 1998, The Journal of pharmacology and experimental therapeutics.

[35]  G. Novotny,et al.  Inactivation of baroafferents leads to loss of barosensitivity without changes in nerve morphology. , 1998, Journal of the autonomic nervous system.

[36]  W. Kirch,et al.  Clinical Pharmacokinetics of Articaine , 1997, Clinical pharmacokinetics.

[37]  B. Graf,et al.  Stereospecific Effect of Bupivacaine Isomers on Atrioventricular Conduction in the Isolated Perfused Guinea Pig Heart , 1997, Anesthesiology.

[38]  A. Bollen,et al.  Local Anesthetic Neurotoxicity Does Not Result from Blockade of Voltage-Gated Sodium Channels , 1995, Anesthesia and analgesia.

[39]  C Haanen,et al.  A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. , 1995, Journal of immunological methods.

[40]  I Schmid,et al.  A rapid method for measuring apoptosis and dual-color immunofluorescence by single laser flow cytometry. , 1994, Journal of immunological methods.

[41]  Michael J. Cousins,et al.  Neural Blockade in Clinical Anesthesia and Management of Pain , 1988 .

[42]  B. R. Fink,et al.  A Comparative in Vivo Study of Local Neurotoxicity of Lidocaine, Bupivacaine, 2‐Chloroprocaine, and a Mixture of 2‐Chloroprocaine and Bupivacaine , 1982, Anesthesia and analgesia.