Cytotoxicity of local anesthetics on rabbit adipose‑derived mesenchymal stem cells during early chondrogenic differentiation

Abstract

Local anesthetics (LAs) are commonly used to provide peri-operative pain control in the peripheral joints. In the field of regenerative medicine, adipose-derived mesenchymal stem cells (ADMSCs) are gaining attention as a cellular source for repair and regeneration in degenerative diseases. However, previous studies have demonstrated that the commonly used drugs lidocaine, ropivacaine, bupivacaine and mepivacaine may be toxic to human chondrocytes, which has raised concerns over whether they exert similar negative effects on ADMSCs during early chondrogenic differentiation. In the present in vitro study, the cytotoxicity of different LAs to ADMSCs was determined during early chondrogenic differentiation. At concentrations similar to those after physiological dilution once injected into the degenerative tissues, LAs (1% lidocaine, 0.5% bupivacaine, 0.5% ropivacaine or 2% mepivacaine) and PBS (control group) were incubated with rabbit ADMSCs (rADMSCs) for 60 min. Following further culture for 3 or 7 days, the cell viability, apoptosis and morphological alterations of chondrogenic differentiation were measured by determining the mitochondrial activity, by flow cytometric analysis, Safranine Fast Green double staining and reverse transcription-quantitative polymerase chain reaction of chondrogenesis-associated genes. The results indicated that the mitochondrial activity in rADMSC was decreased and the apoptotic rate was increased, following treatment with LAs (P<0.05). Lidocaine (1%) was less cytotoxic to rADMSCs during early chondrogenesis compared with other LAs. The expression levels of chondrogenesis-associated markers, including collagen I, collagen III and sex-determining region Y box 9 were all decreased at day 3 following exposure to LAs compared with the control group (P<0.05). The expression levels of these chondrogenesis-associated genes began to increase on day 7 following exposure but remained lower compared with the control group (P<0.05). Of note, 2% mepivacaine and 1% lidocaine exhibited a less pronounced negative effect on chondrogenesis-associated gene expression compared with other LAs. Therefore, the present study concluded that LAs are cytotoxic to rADMSCs during early chondrogenesis. Attention should be paid to the different types of LA selected in conjunction with ADMSC injection therapy.

Effect of five dental pulp capping agents on cell proliferation, viability, apoptosis and mineralization of human dental pulp cells

“…Although the concentrations of different materials were consistent, it is still necessary to compare the influence of different concentrations of materials. Furthermore, the maximum observation time was 7 days, which referred to certain previous studies (17,23). Inevitable passaging of cells in vitro may limit the evaluation time.…”

Investigating the Cytotoxic and Anti-Bacterial Activity of Commercially Available Local Anesthetics: An In-Vitro Analysis

“…These results were in consensus to the findings of Wu et al and colleagues, who investigated the effects of anesthetics on MSCs derived from rabbit adipose cells and concluded that 1% lidocaine showed little toxic effects when compared with other anesthetic agents at different concentrations. 10 Similarly, Shoshani et al and Berger et al studied the effects of lidocaine along with epinephrine and demonstrated that there was no effect on the viability of adipocytes after being exposed to this combination. 23 In line with the ideas of Celeste et al and colleagues, it was concluded that toxicity of lidocaine was dose and time-dependent, their study shed light on the fact that at low doses, lidocaine did not effect mitochondrial functions of fibroblast of the oral mucosa.…”

“…Toxicity of anesthetics is linked to their lipophilicity, thus lidocaine being less lipophilic makes it less toxic when compared to other amide anesthetic agents, validating their secure use in dental practice. 10 In case of clinical implications, a toxic anesthetic would augment the possibility of oral paresthesia, especially when administrating an inferior alveolar nerve block as studies report 89% chances of lingual nerve being involved. 12,24 Secondly, muscular stiffness is also linked to the myotoxic effects of anesthetics resulting in degenerative changes in masseter and medial pterygoid.…”

“…Numerous in-vitro cytotoxic studies have been published on local anesthesia to assess their effects on cell viability using mammalian cell lines. 7, 10 Breu et al and Eckl et al studied the relationship of the viability of MSCs to the time of exposure at different concentrations of local anesthetic agents and deduced that MSC's viability depends on the local anesthetic agent used, its concentration and the time of exposure. Cell viability was evaluated using Annexin-V stain and results were analyzed quantitatively using a flow cytometer.…”

Effect of local anesthetics on viability and differentiation of various adult stem/progenitor cells

“…LAs are also administered into the shoulder joint to treat rotator cuff injuries and diseases [5]. In addition, fat aspiration for a cosmetic purpose or isolation of autologous adipose-derived stem/progenitor cells (ADSCs) routinely involves an administration of LAs [6][7][8][9][10].…”