Lidocaine induces ROCK-dependent membrane blebbing and subsequent cell death in rabbit articular chondrocytes

Abstract

Local anesthetics are administered intraarticularly for pain control in orthopedic clinics and surgeries. Although previous studies have shown that local anesthetics can be toxic to chondrocytes, the underlying cellular mechanisms remain unclear. The present study investigates acute cellular responses associated with lidocaine-induced toxicity to articular chondrocytes. Rabbit articular chondrocytes were exposed to lidocaine and their morphological changes were monitored with live cell microscopy. The viability of chondrocytes was evaluated using a fluorescence based LIVE/DEAD assay. Acute treatment of chondrocytes with lidocaine (3-30 mM) induced spherical protrusions on the cell surface (so called "membrane blebbing") in a time-and concentration-dependent manner. The concentration-response relationship for the lidocaine effect was shifted leftward by elevating extracellular pH, as expected for the nonionized lidocaine being involved in the bleb formation. ROCK (Rho-kinase) inhibitors Y-27632 and fasudil completely prevented the lidocaine-induced membrane blebbing, suggesting that ROCK activation is required for bleb formation. Caspase-3 levels were unchanged by 10 mM lidocaine (p ¼ 0.325) and a caspase inhibitor z-VAD-fmk did not affect the lidocaine-induced blebbing (p ¼ 0.964). GTP-RhoA levels were significantly increased (p < 0.001), but Rho inhibitor-1 failed to suppress the membrane blebbing (p ¼ 0.875). Lidocaine (30 mM) reduced the cell viability of isolated chondrocytes (p < 0.001) and in situ chondrocytes (p < 0.001). The chondrotoxicity was attenuated by pretreatment of cells with ROCK inhibitors or a myosin-II inhibitor blebbistatin (p < 0.001). These findings suggest that lidocaine induces ROCK-dependent membrane blebbing and thereby produces a cytotoxic effect on chondrocytes.

Revealing oxidative damage to enzymes of carbohydrate metabolism in yeast: An integration of 2D DIGE, quantitative proteomics, and bioinformatics

“…Similarly, S. cerevisiae grown aerobically are more resistant to oxidants than those grown anaerobically [38]. In addition, examination of lidocaine toxicity in articular chondrocytes from rabbits demonstrated an LC 50 of 30 mM over a 4-h time course [24]. These studies support the high concentrations of stressors necessary to cause 50% cell death in the current experiments examining aerobically grown, stationary phase S. cerevisiae.…”

“…Prior investigation on lidocaine toxicity in multiple models has yielded extensive information regarding its physiological effects of ROS and RNS generation, mitochondrial depolarization, calcium dysregulation, and apoptotic‐driven cell death . Targeted proteomics has granted some insight into lidocaine‐induced cell death through protein kinase C (PKC) activation in human gingival cells and Rho‐kinase activation in rabbit chondrocytes . However, despite the abundance of knowledge in lidocaine‐induced physiological changes, the mechanisms of lidocaine toxicity at the whole‐proteome level have not been adequately explored.…”

Ultrasound‐guided lower forearm median nerve block in open surgery for trigger thumb in 1‐ to 3‐year‐old children: A randomized trial

“…Therefore, they do not recommend it as a high priority. Furthermore, several studies have suggested that local anesthetics can be toxic to chondrocytes and cartilage, which cause cellular death and cartilage structure collapse . In the present study, the puncture point was far from the wrist and thumb joint and could thus avoid the potential toxicity of local anesthetics to chondrocytes and cartilage.…”

P2X7 ionotropic receptor is functionally expressed in rabbit articular chondrocytes and mediates extracellular ATP cytotoxicity

“…During the acclimation period, the health status of animals was checked every day and no adverse events were observed. Articular chondrocytes were isolated using the enzymatic dissociation procedure described previously [14]. In brief, rabbits were deeply anesthetized with an i.m.…”

“…Two distinct families of purinergic receptors, P1 receptors for adenosine and P2 receptors for ATP, are widely distributed. P2 receptors are further divided into ligand-gated ion channel receptors, P2X (P2X1-7), and G protein-coupled P2Y (P2Y 1,2,4,6,[11][12][13][14] subfamilies. The P2X7 receptor belongs to the P2X ionotropic receptor family, but it was originally identified as a unique ATP receptor termed the P2Z receptor due to its peculiar functional features which are distinct from P2X and P2Y receptors [11].…”