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

Abstract

Background Local anesthetics (LAs) are widely used to control pain during various clinical treatments. One of the side effects of LAs, cytotoxicity, has been investigated in various cells including stem/progenitor cells. However, our understanding of the effects of LAs on the differentiation capacity of stem/progenitor cells still remains limited. Therefore, a comparative study was conducted to investigate the effects of multiple LAs on viability and multi-lineage differentiation of stem/progenitor cells that originated from various adult tissues. Method Multiple types of stem/progenitor cells, including bone marrow mesenchymal stem/progenitor cells (MSCs), dental pulp stem/progenitor cells (DPSCs), periodontal ligament stem/progenitor cells (PDLSCs), and tendon-derived stem/progenitor cells, were either obtained from a commercial provider or isolated from adult human donors. Lidocaine (LD) and bupivacaine (BP) at various doses (1×, 0.75×, 0.5×, and 0.25× of each physiological dose) were applied to the different stem/progenitor cells for an hour, followed by induction of fibrogenic, chondrogenic, osteogenic, and adipogenic differentiation. Live/dead and MTT assays were performed at 24 h after the LD or BP treatment. At 2 weeks, qRT-PCR was conducted to evaluate the gene expressions associated with differentiation. After 4 weeks, multiple biochemical staining was performed to evaluate matrix deposition. Results At 24 h after LD or BP treatment, 1× and 0.75× physiological doses of LD and BP showed significant cytotoxicity in all the tested adult stem/progenitor cells. At 0.5×, BP resulted in higher viability than the same dose LD, with variance between cell types. Overall, the gene expressions associated with fibrogenic, chondrogenic, osteogenic, and adipogenic differentiation were attenuated in LD or BP pre-treated stem/progenitor cells, with notable dose-effect and dependence on types. In contrast, certain doses of LD and/or BP were found to increase specific gene expression, depending on the cell types. Conclusion Our data suggest that LAs such as LD and BP affect not only the viability but also the differentiation capacity of adult stem/progenitor cells from various anatomical sites. This study sheds light on stem cell applications for tissue regeneration in which isolation and transplantation of stem cells frequently involve LA administration.

Comprehensive analysis of local anesthetics affecting adipose stem cells, in vitro

“…Historically, lidocaine is considered to be one of the most cytotoxic local anesthetics. 9 Interestingly, ropivacaine did not reveal cytotoxic effects in concentrations up to 10 for brief and 1 mM for extended durations of exposure to ASCs. Mepivacaine may be used in concentrations up to 10 mM, but cytotoxic effects could occur.…”

“…3,8 However, they are considered cytotoxic in concentrations commonly found in ready-to-use medical products, which would render them impracticable when handling ASCs. [9][10][11][12][13] In addition, they are known to inhibit the differentiation capacity of mesenchymal stem cells (ie, in ASCs). 12,13 Thus, it was historically recommended to avoid using LAs when harvesting or transplanting ASCs.…”

“…3,8 Numerous in vitro studies propagate cytotoxic effects of LAs on a variety of different cell types, including ASCs, which would diminish the number of vital cells in fat grafts and cause graft atrophy. [9][10][11][12][13] Therefore, it was historically recommended to avoid LAs when handling ASCs. 13 In contrast, a recent clinical study investigating the effect of lidocaine on cell numbers and viability of various cell populations of the stromal vascular fraction (ie, ASCs during liposuction) found no evidence of objecting the use of lidocaine for human lipograft harvest.…”

Metabolomic Impact of Lidocaine on a Triple Negative Breast Cancer Cell Line

“…Indeed, if the cell culture allows for a simple experimental approach, both the nutritional conditions (excess glucose in the culture medium) and the oxygen concentrations (ranging from hyperoxia to hypoxia in some parts of the flask if it is not agitated) often do not allow extrapolation of experimental results to clinical use. Moreover, our in vitro study doesn’t assess the impact of lidocaine on the microenvironment while it has been shown that local anesthetics could affect viability and differentiation capacity of adult stem/progenitor cells ( Kim et al, 2020 ; Kubrova et al, 2021 ). Those effects on mesenchymal stem cells could influence wound healing or tumor spreading after surgery ( Lucchinetti et al, 2012 ).…”

Quantitative Analysis of the Membrane Affinity of Local Anesthetics Using a Model Cell Membrane

“…Due to the amphiphilic nature of LA compounds, they partition into cell membranes and perturb the cell membrane matrix by modifying its physiochemical properties, such as permeability, fluidity, and electrostatic potential, which eventually affect membrane protein functions [8][9][10]. This indirect interaction model explains the mechanism of some local anesthetics and the associated toxic effects caused by LAs such as lidocaine, tetracaine, bupivacaine, and procaine [11,12]. Recently, several approaches were reported to analyze the effects of LAs on model-cell membranes.…”