Abstract
Previously, we reported that millimeter wave promoted the chondrocyte proliferation by pushing cell cycle progression. Activation of K+ channels plays an essential role in the stimulating of extracellular matrix (ECM) synthesis and the cell proliferation in chondrocytes. While it is unclear if millimeter wave enhances ECM synthesis and proliferation of chondrocytes by regulating K+ channel activity, we here investigated the effects of millimeter waves on ECM synthesis, chondrocyte proliferation and ion channels in the primary chondrocyte culture. We found that millimeter waves led to the increase of chondrocyte viability, the morphological changes of chondrocyte, and the F-actin distortion and remodeling. Ultrastructural analysis showed that treated chondrocytes contained an expansion of mitochondria and granular endoplasmic reticulum, and a high number of cytoplasmic vesicles in the cytoplasm compared to untreated cells, suggesting millimeter waves increased the energy metabolism and protein synthesis of chondrocytes. The analysis of differential ion channels’ genes expression further showed an obvious increase of Kcne1, Kcnj3 and Kcnq2. To determine the role of voltage-gated K+ channel in chondrocyte, we blocked the voltage-gated K+ channel with 10 mM tetraethylammonium (TEA) and treated chondrocytes with millimeter waves. The results indicated that TEA significantly negated the promotion of millimeter waves for the ECM synthesis and chondrocyte proliferation. Our results support the hypothesis that millimeter waves promote the synthesis of ECM and the proliferation of chondrocyte by regulating the voltage-gated K+ channel.
Similar content being viewed by others
References
Perkins GL, Derfoul A, Ast A, Hall DJ (2005) An inhibitor of the stretch-activated cation receptor exerts a potent effect on chondrocyte phenotype. Differentiation 73:199–211
Zhang L, Hu J, Athanasiou KA (2009) The role of tissue engineering in articular cartilage repair and regeneration. Crit Rev Biomed Eng 37:1–57
Wohlrab D, Lebek S, Krüger T, Reichel H (2002) Influence of ion channels on the proliferation of human chondrocytes. Biorheology 39:55–61
Wohlrab D, Wohlrab J, Reichel H, Hein W (2001) Is the proliferation of human chondrocytes regulated by ionic channels? J Orthop Sci 6:155–159
Wu QQ, Chen Q (2000) Mechanoregulation of chondrocyte proliferation, maturation, and hypertrophy: ion-channel dependent transduction of matrix deformation signals. Exp Cell Res 256:383–391
Mobasheri A, Trujillo E, Arteaga MF, Martín-Vasallo P (2012) Na(+), K(+)-ATPase Subunit Composition in a Human Chondrocyte Cell Line; Evidence for the Presence of α1, α3, β1, β2 and β3 Isoforms. Int J Mol Sci 13:5019–5034
Hoffmann EK, Pedersen SF (2011) Cell volume homeostatic mechanisms: effectors and signalling pathways. Acta Physiol (Oxf) 202:465–485
Brini M, Carafoli E (2000) Calcium signalling: a historical account, recent developments and future perspectives. Cell Mol Life Sci 57:354–370
Archer CW, Francis-West P (2003) The chondrocyte. Int J Biochem Cell Biol 35:401–404
Mobasheri A (1999) Regulation of Na+, K+-ATPase density by the extracellular ionic and osmotic environment in bovine articular chondrocytes. Physiol Res 48:509–512
Mobasheri A, Trujillo E, Bell S, Carter SD, Clegg PD, Martín-Vasallo P, Marples D (2004) Aquaporin water channels AQP1 and AQP3, are expressed in equine articular chondrocytes. Vet J 168:143–150
Guilak F (2000) The deformation behavior and viscoelastic properties of chondrocytes in articular cartilage. Biorheology 37:27–44
Mobasheri A, Lewis R, Ferreira-Mendes A, Rufino A, Dart C, Barrett-Jolley R (2012) Potassium channels in articular chondrocytes. Channels (Austin) 6:416–425
Clark RB, Hatano N, Kondo C, Belke DD, Brown BS, Kumar S, Votta BJ, Giles WR (2010) Voltage-gated K+ currents in mouse articular chondrocytes regulate membrane potential. Channels (Austin) 4:179–191
Ponce A (2006) Expression of voltage dependent potassium currents in freshly dissociated rat articular chondrocytes. Cell Physiol Biochem 18:35–46
Funabashi K, Ohya S, Yamamura H, Hatano N, Muraki K, Giles W, Imaizumi Y (2010) Accelerated Ca2+ entry by membrane hyperpolarization due to Ca2+-activated K+ channel activation in response to histamine in chondrocytes. Am J Physiol Cell Physiol 298:C786–C797
Lewis R, Feetham CH, Barrett-Jolley R (2011) Cell volume regulation in chondrocytes. Cell Physiol Biochem 28:1111–1122
Wohlrab D, Vocke M, Klapperstück T, Hein W (2004) Effects of potassium and anion channel blockers on the cellular response of human osteoarthritic chondrocytes. J Orthop Sci 9:364–3671
Yan Q, Feng Q, Beier F (2010) Endothelial nitric oxide synthase deficiency in mice results in reduced chondrocyte proliferation and endochondral bone growth. Arthritis Rheum 62:2013–2022
Lewis R, Asplin KE, Bruce G, Dart C, Mobasheri A, Barrett-Jolley R (2011) The role of the membrane potential in chondrocyte volume regulation. J Cell Physiol 226:2979–2986
Qusous A, Geewan CS, Greenwell P, Kerrigan MJ (2011) siRNA-mediated inhibition of Na(+)-K(+)-2Cl− cotransporter (NKCC1) and regulatory volume increase in the chondrocyte cell line C-20/A4. J Membr Biol 243:25–34
Xia L, Luo QL, Lin HD, Zhang JL, Guo H, He CQ (2012) The effect of different treatment time of millimeter wave on chondrocyte apoptosis, caspase-3, caspase-8, and MMP-13 expression in rabbit surgically induced model of knee osteoarthritis. Rheumatol Int 32:2847–2856
Wu G, Sferra T, Chen X, Chen Y, Wu M, Xu H, Peng J, Liu X (2011) Millimeter wave treatment inhibits the mitochondrion-dependent apoptosis pathway in chondrocytes. Mol Med Report 4:1001–1006
Miryutova NF, Levitskii EF, Kozhemyakin AM, Mavlyautdinova IM (2001) Millimeter waves in the treatment of neurological manifestations of vertebral osteochondrosis. Crit Rev Biomed Eng 29:613–621
Li X, Ye H, Yu F, Cai L, Li H, Chen J, Wu M, Chen W, Lin R, Li Z, Zheng C, Xu H, Wu G, Liu X (2012) Millimeter wave treatment promotes chondrocyte proliferation via G1/S cell cycle transition. Int J Mol Med 29:823–831
Li X, Du M, Liu X, Chen W, Wu M, Lin J, Wu G (2010) Millimeter wave treatment promotes chondrocyte proliferation by upregulating the expression of cyclin-dependent kinase 2 and cyclin A. Int J Mol Med 26:77–84
Nishimura I, Chano T, Kita H, Matsusue Y, Okabe H (2011) RB1CC1 protein suppresses type II collagen synthesis in chondrocytes and causes dwarfism. J Biol Chem 286:43925–43932
Mobasheri A, Gent TC, Nash AI, Womack MD, Moskaluk CA, Barrett-Jolley R (2007) Evidence for functional ATP-sensitive (K(ATP)) potassium channels in human and equine articular chondrocytes. Osteoarthritis Cartilage 15:1–8
Clark RB, Kondo C, Belke DD, Giles WR (2011) Two-pore domain K+ channels regulate membrane potential of isolated human articular chondrocytes. J Physiol 589:5071–5089
Clarke OB, Gulbis JM (2012) Oligomerization at the membrane: potassium channel structure and function. Adv Exp Med Biol 747:122–136
Mizuno S, Ogawa R (2011) Using changes in hydrostatic and osmotic pressure to manipulate metabolic function in chondrocytes. Am J Physiol Cell Physiol 300:C1234–C1245
Okazaki R, Sakai A, Uezono Y, Ootsuyama A, Kunugita N, Nakamura T, Norimura T (2001) Sequential changes in transforming growth factor (TGF)-beta1 concentration in synovial fluid and mRNA expression of TGF-beta1 receptors in chondrocytes after immobilization of rabbit knees. J Bone Miner Metab 19:228–235
Bader DL, Ohashi T, Knight MM, Lee DA, Sato M (2002) Deformation properties of articular chondrocytes: a critique of three separate techniques. Biorheology 39:69–78
Sánchez JC, Danks TA, Wilkins RJ (2003) Mechanisms involved in the increase in intracellular calcium following hypotonic shock in bovine articular chondrocytes. Gen Physiol Biophys 22:487–500
Browning JA, Saunders K, Urban JP, Wilkins RJ (2004) The influence and interactions of hydrostatic and osmotic pressures on the intracellular milieu of chondrocytes. Biorheology 41:299–308
Pingguan-Murphy B, El-Azzeh M, Bader DL, Knight MM (2006) Cyclic compression of chondrocytes modulates a purinergic calcium signalling pathway in a strain rate- and frequency-dependent manner. J Cell Physiol 209:389–397
Mobasheri A, Gent TC, Womack MD, Carter SD, Clegg PD, Barrett-Jolley R (2005) Quantitative analysis of voltage-gated potassium currents from primary equine (Equus caballus) and elephant (Loxodonta africana) articular chondrocytes. Am J Physiol Regul Integr Comp Physiol 289:R172–R180
Wu QQ, Chen Q (2000) Mechanoregulation of chondrocyte proliferation, maturation, and hypertrophy: ion-channel dependent transduction of matrix deformation signals. Exp Cell Res 256:383–391
Ouadid-Ahidouch H, Roudbaraki M, Delcourt P, Ahidouch A, Joury N, Prevarskaya N (2004) Functional and molecular identification of intermediate-conductance Ca(2 +)-activated K(+) channels in breast cancer cells: association with cell cycle progression. Am J Physiol Cell Physiol 287:C125–C134
MacFarlane SN, Sontheimer H (2000) Changes in ion channel expression accompany cell cycle progression of spinal cord astrocytes. Glia 30:39–48
Braun GS, Veh RW, Segerer S, Horster MF, Huber SM (2002) Developmental expression and functional significance of Kir channel subunits in ureteric bud and nephron epithelia. Pflugers Arch 445:321–330
Wang Z (2004) Roles of K+ channels in regulating tumour cell proliferation and apoptosis. Pflugers Arch 448:274–286
Matta C, Fodor J, Szíjgyártó Z, Juhász T, Gergely P, Csernoch L, Zákány R (2008) Cytosolic free Ca2+ concentration exhibits a characteristic temporal pattern during in vitro cartilage differentiation: a possible regulatory role of calcineurin in Ca-signalling of chondrogenic cells. Cell Calcium 44:310–323
Lang F, Shumilina E, Ritter M, Gulbins E, Vereninov A, Huber SM (2006) Ion channels and cell volume in regulation of cell proliferation and apoptotic cell death. Contrib Nephrol 152:142–160
Hoffmann EK (2011) Ion channels involved in cell volume regulation: effects on migration, proliferation, and programmed cell death in non adherent EAT cells and adherent ELA cells. Cell Physiol Biochem 28:1061–1078
Acknowledgments
This work was supported by the Developmental Fund of Chen Keji Integrative Medicine (Grant No. CKJ2010013&CKJ2010035), Youth Foundation of Fujian Provincial Health Bureau (Grant No. 2011-2-31) and Natural Science Foundation of Fujian Province (Grant No. 2011J05076).
Conflict of interest
All authors have no conflicts of interest.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
About this article
Cite this article
Li, X., Liu, C., Liang, W. et al. Millimeter wave promotes the synthesis of extracellular matrix and the proliferation of chondrocyte by regulating the voltage-gated K+ channel. J Bone Miner Metab 32, 367–377 (2014). https://doi.org/10.1007/s00774-013-0513-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00774-013-0513-2