Effect of Lidocaine on Viability and Gene Expression of Human Adipose–derived Mesenchymal Stem Cells: An in vitro Study

Abstract

Objective To assess the biologic effects of lidocaine on the viability, proliferation, and function of human adipose tissue–derived mesenchymal stromal/stem cells (MSCs) in vitro. Methods Adipose‐derived MSCs from three donors were exposed to lidocaine at various dilutions (2 mg/mL to 8 mg/mL) and exposure times (0.5 to 4 hours). Cell number and viability, mitochondrial activity, and real‐time reverse‐transcriptase quantitative polymerase chain reaction (RT‐qPCR) were analyzed at 0 (immediate effects) or 24 and 48 hours (recovery effects) after treatment with lidocaine. Results Trypan blue staining showed that increasing concentrations of lidocaine decreased the number of observable viable cells. 3‐[4,5,dimethylthiazol‐2‐yl]‐5‐[3‐carboxymethoxy‐phenyl]‐2‐[4‐sulfophenyl]‐2H‐tetrazolium (MTS) assays revealed a concentration‐ and time‐ dependent decline of mitochondrial activity and proliferative ability. Gene expression analysis by RT‐qPCR revealed that adipose‐derived MSCs exposed to lidocaine express robust levels of stress response/cytoprotective genes. However, higher concentrations of lidocaine caused a significant downregulation of these genes. No significant differences were observed in expression of extracellular matrix (ECM) markers COL1A1 and DCN except for COL3A1 (P < .05). Levels of messenger RNA (mRNA) for proliferation markers (CCNB2, HIST2H4A, P < .001) and MKI67 (P < .001) increased at 24 and 48 hours. Expression levels of several transcription factors— including SP1, PRRX1, and ATF1—were modulated in the same manner. MSC surface markers CD44 and CD105 demonstrated decreased expression immediately after treatment, but at 24 and 48 hours postexposure, the MSC markers showed no significant difference among groups. Conclusion Lidocaine is toxic to MSCs in a dose‐ and time‐ dependent manner. MSC exposure to high (4‐8 mg/mL) concentrations of lidocaine for prolonged periods can affect their biologic functions. Although the exposure time in vivo is short, it is essential to choose safe concentrations when applying lidocaine along with MSCs to avoid compromising the viability and potency of the stem cell therapy.

Lidocaine intensifies the anti-osteogenic effect on inflammation-induced human dental pulp stem cells via mitogen-activated protein kinase inhibition

“…It has been shown that lidocaine has a cytotoxic effect on adipose-derived MSCs during early chondrogenic differentiation and attenuates the fibrogenic, chondrogenic, osteogenic, and adipogenic differentiation of bone marrow–derived MSCs, DPSCs, periodontal ligament stem/progenitor cells, and tendon-derived stem/progenitor cells. 45 , 46 , 47 However, there have been no studies on the effect of lidocaine on the osteogenic differentiation of inflammation-induced MSCs. Therefore, this is the first study to demonstrate that lidocaine attenuates osteogenic differentiation of hDPSCs induced with LPS/TNF-α through the regulation of MAPK pathways.…”

“… 23 Lidocaine is the most widely used amide-type LAs, is known to have anti-inflammatory effects in many experimental studies, and has a cytotoxic effect on MSCs. 24 , 25 These findings raise the question of how lidocaine influences osteogenic differentiation of hDPSCs induced by an inflammatory response, and there is no research yet to clarify this question.…”

Histone and Histone Acetylation-Related Alterations of Gene Expression in Uninvolved Psoriatic Skin and Their Effects on Cell Proliferation, Differentiation, and Immune Responses

“…In non-dividing cells, HIST2H2AA3 participates in the terminal differentiation program [31] (Figure 2). HIST2H4A is commonly used as a marker for proliferation [27,28] (Figure 2). Therefore, differential expression of these histones may contribute to proliferation and differentiation-related alterations in psoriasis.…”

“…On the other hand, they differ in the number and position of post-transcription modification sites at their globular core and N-terminal tail, allowing them to carry out distinct and specialized roles, including the regulation of tissue-and cell-type-specific functions. These functions include the regulation of proliferation [27,28], cell fate commitment [29], hematopoiesis [30], differentiation [31], macrophage [32] and T cell immune responses, and mutagenesis of immunoglobins [33,34].…”

Mesenchymal Stem/Progenitor Cells: The Prospect of Human Clinical Translation

“…MSCs subjected to lidocaine at various dilutions (2 mg/ml to 8 mg/ml) and exposure times (0.5 to 4 hours) showed upregulation of genes normally associated with responses to stress and cytoprotective mechanisms, while higher concentration of lidocaine (8 mg/ml and more) resulted in a significant drop in gene expression. Exposure of MSCs to high concentrations of lidocaine for prolonged periods was shown to negatively affect MSC viability, proliferation, and/or functions [ 531 ]. A recent study investigated the effect of lidocaine applied during tumescent local anesthesia prior to liposuction.…”