caa a22 4500 605459134 CHVBK 20210128100232.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s00167-014-3312-y doi (NATIONALLICENCE)springer-10.1007/s00167-014-3312-y Local anesthetic cytotoxicity on human mesenchymal stem cells during chondrogenic differentiation [Elektronische Daten] [Anita Breu, Ingrid Scheidhammer, Richard Kujat, Bernhard Graf, Peter Angele] Purpose: This study was to investigate the cytotoxic potency of local anesthetics on human mesenchymal stem cells during chondrogenesis. Methods: Aggregates were created from density-gradient centrifugation-separated bone marrow-derived mesenchymal stem cells. After 7, 14, and 21days, aggregates were analyzed histologically and immunohistochemically and exposed to equipotent concentrations of bupivacaine, ropivacaine, and mepivacaine for 1h. Cell viability, apoptosis, and necrosis were determined using live-dead and caspase staining. Additionally, following a 1-h exposure on day 7, aggregates were cultured under chondrogenic conditions until day 21 to assess the effects of local anesthetics on differentiation potency of mesenchymal stem cells. Results: In the course of chondrogenesis, mesenchymal stem cells were embedded in varying amount and structure of cartilage-specific extracellular matrix. Contents of sulfated glycosaminoglycan, type I and II collagen increased from day 7 to day 21. Compared to control, death rates of mesenchymal stem cells were significantly elevated 1day after treatment at 7 and 14days. Four days after exposure, death rates were 13-15% at 7 and 11-17% at 14days. Mesenchymal stem cell viability in aggregates at 21days was unchanged to controls. The width of the superficial aggregate zone containing stem cell necrosis decreased with elongated differentiation time. Apoptosis rates were elevated in the edge regions of aggregates, reaching maximum values 4days after treatment. Local anesthetic exposure on day 7 reduced Collagen II, but not DNA contents in aggregates at 21days. Bupivacaine, ropivacaine, and mepivacaine did not differ in mesenchymal stem cell cytotoxicity in aggregates. Conclusion: Local anesthetic exposure results in cytotoxicity of mesenchymal stem cells undergoing chondrogenesis, especially in superficial layers. Therefore, induced cell damage should be avoided during chondrogenesis of mesenchymal stem cells, particularly early after cartilage repair. European Society of Sports Traumatology, Knee Surgery, Arthroscopy (ESSKA), 2014 Local anesthetics nationallicence Mesenchymal stem cells nationallicence Chondrogenesis nationallicence Chondrotoxicity nationallicence Breu Anita Department of Anesthesiology, University Medical Center Regensburg, Franz-Josef-Strauss-Allee 11, 93042, Regensburg, Germany aut Scheidhammer Ingrid Department of Anesthesiology, University Medical Center Regensburg, Franz-Josef-Strauss-Allee 11, 93042, Regensburg, Germany aut Kujat Richard Department of and Trauma Surgery, University Medical Center Regensburg, 93041, Regensburg, Germany aut Graf Bernhard Department of Anesthesiology, University Medical Center Regensburg, Franz-Josef-Strauss-Allee 11, 93042, Regensburg, Germany aut Angele Peter Department of and Trauma Surgery, University Medical Center Regensburg, 93041, Regensburg, Germany aut Knee Surgery, Sports Traumatology, Arthroscopy Springer Berlin Heidelberg 23/4(2015-04-01), 937-945 0942-2056 23:4<937 2015 23 167 https://doi.org/10.1007/s00167-014-3312-y text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00167-014-3312-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Breu Anita Department of Anesthesiology, University Medical Center Regensburg, Franz-Josef-Strauss-Allee 11, 93042, Regensburg, Germany aut NATIONALLICENCE 700 1- Scheidhammer Ingrid Department of Anesthesiology, University Medical Center Regensburg, Franz-Josef-Strauss-Allee 11, 93042, Regensburg, Germany aut NATIONALLICENCE 700 1- Kujat Richard Department of and Trauma Surgery, University Medical Center Regensburg, 93041, Regensburg, Germany aut NATIONALLICENCE 700 1- Graf Bernhard Department of Anesthesiology, University Medical Center Regensburg, Franz-Josef-Strauss-Allee 11, 93042, Regensburg, Germany aut NATIONALLICENCE 700 1- Angele Peter Department of and Trauma Surgery, University Medical Center Regensburg, 93041, Regensburg, Germany aut NATIONALLICENCE 773 0- Knee Surgery, Sports Traumatology, Arthroscopy Springer Berlin Heidelberg 23/4(2015-04-01), 937-945 0942-2056 23:4<937 2015 23 167