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Raminta Vaichiulevichiena Raminta Vaichiulevichiena Scilit Preprints.org Google Scholar 1 , Ilona Uzieliena Ne , Vitaly Novitskiy Sorg , Google , Echo Alaburda Aidas Alaburda Scilit Preprints.org Google Scholar 4 and Eiva Bernotienė Eiva Bernotiensė 1, Google Scholar 4 and Eiva Bernotienė Eiva Bernotienlars 1, Google Scholar Preprints.
Received: 2023 March 7 / Revised: 2023 March 31 / Adopted: 2023 April 6 / Published: 2023 April 7
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Electrical stimulation (ES) is often used in various biomedical fields in vitro and in vivo. Many studies have shown the beneficial effects of ES on cellular functions, including metabolism, proliferation, and differentiation. The application of ES to cartilage tissue to increase extracellular matrix formation is interesting because cartilage cannot repair its own damage due to its vascular nature and lack of cells. Various ES methods have been used to induce chondrogenic differentiation in chondrocytes and stem cells; However, there is a large gap in the systematization of ES protocols used for chondrogenic cell differentiation. This review focuses on the application of ES in chondrogenesis of chondrocytes and mesenchymal stem cells for cartilage tissue regeneration. The effects of different types of ES on cell function and chondrogenic differentiation are reviewed, with a systematic presentation of ES protocols and their beneficial effects. In addition, 3D models of cartilage using cells in ES-based scaffolds/hydrogels were noted and recommendations were made to report the use of ES in different studies to ensure the appropriate consolidation of ES knowledge. This review provides new insights into further applications of in vitro ES studies, which hold promise for further cartilage repair techniques.
Electrical stimulation (ES) has received much attention as a physical stimulus used for tissue engineering and treatment of various diseases such as movement, psychiatry and seizures to reduce pain and improve quality of life [1, 2, 3]. . ES are often used to stimulate cells in vitro and in vivo, triggering several intracellular pathways involved in the regulation of cell metabolism, proliferation, migration, and differentiation [ 4 , 5 ]. A meta-analysis of clinical trials showed that neuromuscular electrical stimulation or current disruption can improve pain management and physical function in patients with knee osteoarthritis [6, 7].
ES has proven to be a useful tool for cartilage tissue engineering. Cartilage consists only of insensitive cells – chondrocytes [8, 9]. Because of the voltage-gated deficit of Na
Channels, these nonexcitable cells cannot generate action potentials in response to membrane depolarization [10]. Because the number of chondrocytes in cartilage is very low and their ability to repair damage to the extracellular matrix (ECM) is poor, cartilage is prone to degenerative diseases such as osteoarthritis (OA) [11]. Various procedures for arthroscopic cartilage intervention are currently being developed, such as chondroplasty, microfracture or mosaicplasty [12, 13, 14], as well as more modern technologies such as autologous chondrocyte implantation [15]; However, the method has not yet been approved for clinical use, as its efficacy remains to be confirmed. Tissue engineering technology based on stem cells, especially those using mesenchymal stem cells (MSCs) derived from mature tissue that can differentiate into chondrocytes, appears to be a promising therapeutic approach to repair cartilage damage. ES has been shown to be an important part of cartilage engineering with stem cells because it promotes the chondrogenic differentiation of MSCs even in the absence of growth factors [ 16 , 17 ].
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The lack of a standard EU protocol in network engineering leads to the challenge of determining the exact mechanism of the effect, because the electrical parameters (voltage, pulse or stimulation duration, frequency and field strength) can vary by several orders of magnitude.
This review summarizes ES applied in chondrogenic differentiation experiments. ES modes such as continuous, static, cyclic or pulsed stimulation are described, highlighting their beneficial effects and limitations. ES parameters and effects in the context of chondrogenesis are presented to increase the consolidation of knowledge in this field and direct research towards the development of optimized and more standardized protocols.
ES induces electrical stress in the cell and changes the membrane potential, which, depending on the protocol used, can lead to the activation of ion channels and other voltage-sensitive proteins or to the permeabilization of the plasma membrane. This process determines the flow of various ions across the plasma membrane [18]. Altered ion concentration leads to activation of different gene expression [19], production and secretion of growth and transcription factors [16, 20], cell adhesion [21] and cell-cell interaction molecules [22]. Typically, ES application uses a low electric field that causes a change in the membrane potential by an average of one-tenth of mV and initiates the movement of the voltage-sensing domain, which leads to a conformational change and opens the voltage-gated ion channel (Fig. 1). The most important voltage-gated channel for chondrogenic differentiation is the L-type voltage-gated calcium channel (VGCC), which regulates the expression of chondrogenesis markers (SOX9, COL2A1, Ihh) in vitro and limb development in vivo [23]. However, when cells are exposed to a high-intensity pulsed electric field (PEF), the cell plasma membrane is polarized and a significant transmembrane potential (TMP) occurs [ 24 , 25 ]. When a critical TMP threshold is reached, often called 1 V [26], hydrophilic pores are formed in the membrane, increasing the permeability of the membrane to exogenous molecules [27]. This phenomenon, called electroporation or electropermeabilisation [28], has been used for gene and drug delivery, tissue ablation, protein extraction, and food processing. Depending on the PEF parameters, electroporation can be reversible or irreversible [ 29 , 30 ]. In mammalian cells, electroporation can be triggered at 400-800 V/cm PEF [31], while a lower field (<400 V/cm) can cause a phenomenon known as electroendocytosis, which means increased absorption of macromolecules after cell exposure. to low temperature. electric field [32, 33]. In recent years, scientific articles focusing on the effects of high-intensity PEF stimulation have also begun to appear.
ES has been widely applied in various fields of cell research. A Web of Science Clarivate Analytics analysis using the keywords “electrical stimulation” and “cell” filtered 11,180 articles published since 2008. The most dominant keywords were related to electrical aspects (electrodes, devices, currents and amplitudes), while the most frequently used for biological aspects are expression, receptors, differentiation and inhibition (Figure 2). The dominant keyword “mouse” refers to the application of ES in in vivo studies with mice as model organisms.
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Cell membranes are the first to respond to ES and are critical for maintaining membrane potential, cellular homeostasis, and controlling the exchange of nutrients, waste products, and chemical molecules important for signaling [ 34 ]. ES activates several independent signaling pathways; therefore, it is difficult to establish a direct relationship between ES and specific cellular responses. ES is known to activate JNK/CREB-STAT3, ERK/JNK/STAT3 and wnt/β-catenin signaling pathways, leading to increased phosphorylation of JNK, CREB and STAT3 [35] and β-catenin protein expression [36]. .
Electric fields and fluid shear stress are hypothesized to activate signaling pathways similar to integrin receptors [ 37 ]. One of the main cellular responses to ES is the opening of voltage-gated calcium channels (VGCCs) and an increase in calcium (Ca).
) inside the cell [38]. Both chondrocytes and mesenchymal stem cells have VGCCs, which can be regulated by external chemical or physical stimuli [ 39 , 40 ]. Such an increase in intracellular Ca
It is also observed in vitro after mechanical stimulation [41]. In addition, remodeling of cytoskeletal structures, including a denser f-actin texture and aligned actin filament orientation, has been observed in response to ES [42], as well as vice versa when mechanical stimulation induces intracellular electrical signals through mechanotransduction [37]. ]. The interaction between ES and mechanical loading can be used in cartilage tissue engineering as a scaffold that cannot withstand mechanical stress and can be replaced.
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