In vitro exposure of human fibroblasts to local anaesthetics impairs cell growth

Abstract

SummaryLidocaine, bupivacaine or ropivacaine are used routinely to manage perioperative pain. Sparse data exist evaluating the effects of local anaesthetics (LA) on fibroblasts, which are involved actively in wound healing. Therefore, we investigated the effects of the three LA to assess the survival, viability and proliferation rate of fibroblasts. Human fibroblasts were exposed to 0·3 mg/ml and 0·6 mg/ml of each LA for 2 days, followed by incubation with normal medium for another 1, 4 or 7 days (group 1). Alternatively, cells were incubated permanently with LA for 3, 6 or 9 days (group 2). Live cell count was assessed using trypan blue staining. Viability was measured by the tetrazolium bromide assay. Proliferation tests were performed with the help of the colorimetric bromodeoxyuridine assay. Production of reactive oxygen species (ROS) was determined, measuring the oxidation of non-fluorescent-2,7Ј-dichlorofluorescin. Treatment of cells with the three LA showed a concentration-dependent decrease of live cells, mitochondrial activity and proliferation rate. Group arrangement played a significant role for cell count and proliferation, while exposure time influenced viability. Among the analysed LA, bupivacaine showed the most severe cytotoxic effects. Increased production of ROS correlated with decreased viability of fibroblasts in lidocaine-and bupivacaine-exposed cells, but not upon stimulation with ropivacaine. This study shows a concentration-dependent cytotoxic effect of lidocaine, bupivacaine and ropivacaine on fibroblasts in vitro, with more pronounced effects after continuous incubation. A possible mechanism of cell impairment could be triggered by production of ROS upon stimulation with lidocaine and bupivacaine.

Comparação dos efeitos da infiltração com bupivacaína, lidocaína e tramadol na cicatrização de feridas em ratos

“…While our study found results similar to previous studies on the effects of bupivacaine and lidocaine on wound healing [30][31][32][33][34][35][36][37][38][39][40][41] , no significant difference was found between tramadol and the control group. The antibacterial properties of local anesthetics and other agents used in wound infiltration are important.…”

“…Other research found that lidocaine and bupivacaine inhibited collagen synthesis in fibroblast tissue cultures, and had cytotoxic effects on different cell lines [30][31][32][33][34][35] .…”

“…Another study on the effects of local anesthetic on human fibroblast found that lidocaine, bupivacaine and ropivacaine produced dose-dependent cytotoxic effects on human fibroblast 31 . Research on lidocaine in wound infiltration of rats found collagenization and effects on mast cell numbers in the wound 36 .…”

“…Research on the effect of these and other local anesthetic agents used for wound infiltration on wound healing is limited and results are controversial 3,[28][29][30][31] .…”

“…For this reason, it is important to know not just the effects of the wound infiltration agents on postoperative pain but their effect in detail on wound healing and whether they are a cause of morbidity in clinical use 3,[28][29][30][31] . Previous research using experimental models and fibroblast tissue cultures from surgical wounds had looked at the effect of local anesthetic agents such as bupivacaine, prilocaine and lidocaine on wound healing 3,28-31 .…”