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Knee Surgery, Sports Traumatology, Arthroscopy
Published in: Knee Surgery, Sports Traumatology, Arthroscopy 4/2015

01-04-2015 | Experimental Study

Local anesthetic cytotoxicity on human mesenchymal stem cells during chondrogenic differentiation

Authors: Anita Breu, Ingrid Scheidhammer, Richard Kujat, Bernhard Graf, Peter Angele

Published in: Knee Surgery, Sports Traumatology, Arthroscopy | Issue 4/2015

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Abstract

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 21 days, aggregates were analyzed histologically and immunohistochemically and exposed to equipotent concentrations of bupivacaine, ropivacaine, and mepivacaine for 1 h. 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 1 day after treatment at 7 and 14 days. Four days after exposure, death rates were 13–15 % at 7 and 11–17 % at 14 days. Mesenchymal stem cell viability in aggregates at 21 days 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 4 days after treatment. Local anesthetic exposure on day 7 reduced Collagen II, but not DNA contents in aggregates at 21 days. 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.
Literature
1.
Angele P, Yoo JU, Smith C, Mansour J, Jepsen KJ, Nerlich M, Johnstone B (2003) Cyclic hydrostatic pressure enhances the chondrogenic phenotype of human mesenchymal progenitor cells differentiated in vitro. J Orthop Res 21:451–457CrossRefPubMed
2.
Asik M, Ciftci F, Sen C, Erdil M, Atalar A (2008) The microfracture technique for the treatment of full-thickness articular cartilage lesions of the knee: midterm results. Arthroscopy 24:1214–1220CrossRefPubMed
3.
Berde CB, Strichartz GR (2010) Local anesthetics. In: Miller RD, Eriksson LI, Fleisher LA, Wiener-Kronish JP, Young WL (eds) Miller’s anesthesia. Curchill Livingstone Elsevier, Philadelphia, pp 913–940
4.
Breu A, Rosenmeier K, Kujat R, Angele P, Zink W (2013) The cytotoxicity of bupivacaine, ropivacaine, and mepivacaine on human chondrocytes and cartilage. Anesth Analg 117:514–522CrossRefPubMed
5.
Breu A, Eckl S, Zink W, Kujat R, Angele P (2013) Cytotoxicity of local anesthetics on human mesenchymal stem cells in vitro. Arthroscopy 29:1676–1684CrossRefPubMed
6.
Chadwick RB, Conrad MP, McGinnis MD, Johnston-Dow L, Spurgeon SL, Kronick MN (1996) Heterozygote and mutation detection by direct automated fluorescent DNA sequencing using a mutant Taq DNA polymerase. Biotechniques 20:676–683PubMed
7.
Chu CR, Izzo NJ, Coyle CH, Papas NE, Logar A (2008) The in vitro effects of bupivacaine on articular chondrocytes. J Bone Joint Surg Br 90:814–820CrossRefPubMed
8.
Chu CR, Coyle CH, Chu CT, Szczodry M, Seshadri V, Karpie JC, Cieslak KM, Pringle EK (2010) In vivo effects of single intra-articular injection of 0.5 % bupivacaine on articular cartilage. J Bone Joint Surg Am 92:599–608CrossRefPubMed
9.
Farkas B, Kvell K, Czompoly T, Illes T, Bardos T (2010) Increased chondrocyte death after steroid and local anesthetic combination. Clin Orthop Relat Res 468:3112–3120CrossRefPubMedCentralPubMed
10.
Graf BM, Zausig Y, Zink W (2005) Current status and clinical relevance of studies of minimum local-anaesthetic concentration (MLAC). Curr Opin Anaesthesiol 18:241–245CrossRefPubMed
11.
Grishko V, Xu M, Wilson G, Pearsall AW (2010) Apoptosis and mitochondrial dysfunction in human chondrocytes following exposure to lidocaine, bupivacaine, and ropivacaine. J Bone Joint Surg Am 92:609–618CrossRefPubMed
12.
Hsu SM, Raine L, Fanger H (1981) Use of avidin–biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29:577–580CrossRefPubMed
13.
Jenei-Lanzl Z, Straub RH, Dienstknecht T, Huber M, Hager M, Grassel S, Kujat R, Angele MK, Nerlich M, Angele P (2010) Estradiol inhibits chondrogenic differentiation of mesenchymal stem cells via nonclassic signaling. Arthritis Rheum 62:1088–1096CrossRefPubMed
14.
Johnstone B, Hering TM, Caplan AI, Goldberg VM, Yoo JU (1998) In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp Cell Res 238:265–272CrossRefPubMed
15.
Karpie JC, Chu CR (2007) Lidocaine exhibits dose- and time-dependent cytotoxic effects on bovine articular chondrocytes in vitro. Am J Sports Med 35:1621–1627CrossRefPubMed
16.
Khan WS, Johnson DS, Hardingham TE (2010) The potential of stem cells in the treatment of knee cartilage defects. Knee 17:369–374CrossRefPubMed
17.
Koyonos L, Yanke AB, McNickle AG, Kirk SS, Kang RW, Lewis PB, Cole BJ (2009) A randomized, prospective, double-blind study to investigate the effectiveness of adding DepoMedrol to a local anesthetic injection in postmeniscectomy patients with osteoarthritis of the knee. Am J Sports Med 37:1077–1082CrossRefPubMed
18.
Kujat R, Rose C, Wrobel KH (1993) The innervation of the bovine ductus deferens: comparison of a modified acetylcholinesterase-reaction with immunoreactivities of cholinacetyltransferase and panneuronal markers. Histochemistry 99:231–239CrossRefPubMed
19.
Kuroda R, Ishida K, Matsumoto T, Akisue T, Fujioka H, Mizuno K, Ohgushi H, Wakitani S, Kurosaka M (2007) Treatment of a full-thickness articular cartilage defect in the femoral condyle of an athlete with autologous bone-marrow stromal cells. Osteoarth Cartil 15:226–231CrossRef
20.
Lee JI, Sato M, Ushida K, Mochida J (2011) Measurement of diffusion in articular cartilage using fluorescence correlation spectroscopy. BMC Biotechnol 11:19CrossRefPubMedCentralPubMed
21.
Lucchinetti E, Awad AE, Rahman M, Feng J, Lou PH, Zhang L, Ionescu L, Lemieux H, Thebaud B, Zaugg M (2012) Antiproliferative effects of local anesthetics on mesenchymal stem cells: potential implications for tumor spreading and wound healing. Anesthesiology 116:841–856CrossRefPubMed
22.
Marret E, Gentili M, Bonnet MP, Bonnet F (2005) Intra-articular ropivacaine 0.75 % and bupivacaine 0.50 % for analgesia after arthroscopic knee surgery: a randomized prospective study. Arthroscopy 21:313–316CrossRefPubMed
23.
Miskulin M, Maldini B (2006) Outpatient arthroscopic knee surgery under multimodal analgesic regimens. Arthroscopy 22:978–983CrossRefPubMed
24.
Piper SL, Kim HT (2008) Comparison of ropivacaine and bupivacaine toxicity in human articular chondrocytes. J Bone Joint Surg Am 90:986–991CrossRefPubMed
25.
Punwar S, Khan WS (2011) Mesenchymal stem cells and articular cartilage repair: clinical studies and future direction. Open Orthop J 5(Suppl 2):296–301CrossRefPubMedCentralPubMed
26.
Rahnama R, Wang M, Dang AC, Kim HT, Kuo AC (2013) Cytotoxicity of local anesthetics on human mesenchymal stem cells. J Bone Joint Surg Am 95:132–137CrossRefPubMed
27.
Rao AJ, Johnston TR, Harris AH, Smith RL, Costouros JG (2014) Inhibition of chondrocyte and synovial cell death after exposure to commonly used anesthetics: chondrocyte apoptosis after anesthetics. Am J Sports Med 42:50–58CrossRefPubMed
28.
Slabaugh MA, Friel NA, Cole BJ (2010) Rapid chondrolysis of the knee after anterior cruciate ligament reconstruction: a case report. J Bone Joint Surg Am 92:186–189CrossRefPubMed
29.
Sulc K, Neuwirt J, Travnicek T, Kobylka P, Radikovska E (1977) Bone marrow cell separation on Ficoll gradient. Haematologia (Budap) 11:41–46
30.
Takahashi T, Tanaka M, Ikeuchi M, Sadahiro T, Tani T (2004) Pain in arthroscopic knee surgery under local anesthesia. Acta Orthop Scand 75:580–583CrossRefPubMed
31.
Taylor KB, Jeffree GM (1969) A new basic metachromatic dye, I:9-dimethyl methylene blue. Histochem J 1:199–204CrossRefPubMed
32.
Wakitani S, Nawata M, Tensho K, Okabe T, Machida H, Ohgushi H (2007) Repair of articular cartilage defects in the patello-femoral joint with autologous bone marrow mesenchymal cell transplantation: three case reports involving nine defects in five knees. J Tissue Eng Regen Med 1:74–79CrossRefPubMed
33.
Zink W, Seif C, Bohl JR, Hacke N, Braun PM, Sinner B, Martin E, Fink RH, Graf BM (2003) The acute myotoxic effects of bupivacaine and ropivacaine after continuous peripheral nerve blockades. Anesth Analg 97:1173–1179CrossRefPubMed
34.
Zink W, Graf BM (2003) Toxicology of local anesthetics. Clinical, therapeutic and pathological mechanisms. Anaesthesist 52:1102–1123CrossRefPubMed
35.
Zink W, Graf BM (2004) Benefit-risk assessment of ropivacaine in the management of postoperative pain. Drug Saf 27:1093–1114CrossRefPubMed
Metadata
Title
Local anesthetic cytotoxicity on human mesenchymal stem cells during chondrogenic differentiation
Authors
Anita Breu
Ingrid Scheidhammer
Richard Kujat
Bernhard Graf
Peter Angele
Publication date
01-04-2015
Publisher
Springer Berlin Heidelberg
Published in
Knee Surgery, Sports Traumatology, Arthroscopy / Issue 4/2015
Print ISSN: 0942-2056
Electronic ISSN: 1433-7347
DOI
https://doi.org/10.1007/s00167-014-3312-y

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