<i>In vitro</i> chondrotoxicity of bupivacaine, levobupivacaine and ropivacaine and their effects on caspase activity in cultured canine articular chondrocytes

Abstract

Bupivacaine, levobupivacaine and ropivacaine are potent, long acting, amide-type local anesthetics that have several clinical applications including intra-articular administration. The objectives of this study were to evaluate their in vitro effects on cell viability and caspase activity to elucidate whether they activate the extrinsic or intrinsic pathways of apoptosis in canine articular chondrocytes.Chondrocytes in monolayer culture were treated with culture medium as the control, or with 0.062% (0.62 mg/mL) bupivacaine, 0.062% levobupivacaine, and 0.062% ropivacaine for 24 hr. Cell viability was evaluated using the live/dead, 3-(4,5-dimehylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT), and Cell Counting Kit-8 (CCK-8) assays. Evaluation of caspase-3, caspase-8, and caspase-9 activity was performed using colorimetric assays. The MTT and CCK-8 assays were used to evaluate the effect of caspase inhibitors on local anesthetic chondrotoxicity. All three local anesthetics decreased chondrocyte viability after 24 hr (P ˂ 0.001). Apoptosis was induced through both the extrinsic and intrinsic pathways.Bupivacaine increased caspase-3, caspase-8, and caspase-9 activity (P ˂ 0.001). Levobupivacaine increased caspase-3 (P = 0.03) while ropivacaine did not significantly upregulate activity for all three caspases. Caspase inhibition did not suppress bupivacaine chondrotoxicity whereas inhibition of caspase-8 and caspase-9 decreased ropivacaine chondrotoxicity and mildly attenuated levobupivacaine chondrotoxicity. In summary, the level of chondrotoxicity, the type of caspase activated, the level of caspase activation, and the response to caspase inhibitors was dependent on the type of local anesthetic.Therefore, ropivacaine may be a safer choice for intra-articular administration compared to levobupivacaine and bupivacaine.

Rapid Osteoarthritis and Femoral Head Collapse: A Case Series

“…Research suggests that their application could induce cell death and structural changes in the articular cartilage in a matter of weeks [ 8 ]. Recent in vitro research by Mwale et al in 2023 found that bupivacaine, levobupivacaine, and ropivacaine triggered apoptosis, as indicated by increased caspase-3 activity, within 24 h of induction of the drug [ 9 ]. Properly evaluating the balance between short-term pain relief benefits and potential long-term cartilage risks is essential when considering what combination of IA injections to utilize.…”

Comment on &quot;Comparison of the pericapsular nerve group block with the intra-articular and quadratus lumborum blocks in primary total hip arthroplasty: a randomized controlled trial&quot;

“…We acknowledge that the sentence in the introduction highlighted by Dr. Raghuraman could be confusing. It would be more accurate to state that numerous meta-analyses have examined different facets of laparoscopic cholecystectomy [3], and a subset of them have placed a specific emphasis on pain management and analgesic needs.…”

“…Therefore, ropivacaine would have been a better choice for this study, especially at such high doses. Additionally, the authors could have followed up with their patients for chondrotoxicity, which would have added more evidence regarding this LA associated side effect [ 3 ].…”

“…Pain scores were higher in the intraarticular group than in the other groups at 3 and 6 h postoperatively. With an intraarticular injection of LA, the joint capsule has a greater surface area for drug absorption, which may explain the poor analgesia provided by this block in the initial hours after surgery [ 3 ]. In addition, comparing the effect of an intraarticular block on quadriceps muscle function is futile, as drug deposition is limited to the articular surface, and thus does not affect the nerve supply to any muscle.…”