Abstract
Cell membranes are susceptible to biophysical damages. These biophysical damages often present themselves in challenging oxidative environments, such as in chronic inflammation. Here we report the damage evolution after single myoblasts were individually subjected to femtosecond (fs) laser photoporation on their plasma membranes under normal and oxidative conditions. A well-characterized tunable fs laser was coupled with a laser scanning confocal microscope. The post-damage wound evolution was documented by real-time imaging. The fs laser could generate a highly focused hole at a targeted site of the myoblast plasma membrane. The initial hole size depended on the laser dosage in terms of power and exposure duration. With the same laser power and irradiation duration, photoporation invoked bigger holes in the oxidative groups than in the control. Myoblasts showed difficulty in repairing holes with initial size beyond certain threshold. Within the threshold, holes could apparently be resealed within 100 s under the normal condition; while in oxidative condition, the resealing process could take 100–300 s. The hole-resealing capacity of myoblasts was compromised under oxidative stress particularly when the oxidative exposure was chronic. It is interesting to note that brief exposure to oxidative stress apparently could promote resealing in myoblasts after photoporation.
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Abreu-Blanco, M. T., J. M. Verboon, and S. M. Parkhurst. Cell wound repair in Drosophila occurs through three distinct phases of membrane and cytoskeletal remodeling. J. Cell Biol. 193(3):455–464, 2011.
Abreu-Blanco, M. T., J. M. Verboon, and S. M. Parkhurst. Single cell wound repair: dealing with life’s little traumas. Bioarchitecture 1(3):114–121, 2011.
Antkowiak, M., M. L. Torres-Mapa, D. J. Stevenson, K. Dholakia, and F. J. Gunn-Moore. Femtosecond optical transfection of individual mammalian cells. Nat. Protoc. 8(6):1216–1233, 2013.
Behl, C. Alzheimer’s disease and oxidative stress: implications for novel therapeutic approaches. Prog. Neurobiol. 57(3):301–323, 1999.
Benink, H. A., and W. M. Bement. Concentric zones of active RhoA and Cdc42 around single cell wounds. J. Cell Biol. 168(3):429–439, 2005.
Cachofeiro, V., M. Goicochea, S. G. de Vinuesa, P. Oubiña, V. Lahera, and J. Luño. Oxidative stress and inflammation, a link between chronic kidney disease and cardiovascular disease. Kidney Int. 74:S4–S9, 2008.
Cai, C., H. Masumiya, N. Weisleder, N. Matsuda, M. Nishi, M. Hwang, J. Ko, P. Lin, A. Thornton, X. Zhao, Z. Pan, S. Komazaki, M. Brotto, H. Takeshima, and J. Ma. MG53 nucleates assembly of cell membrane repair machinery. Nat. Cell Biol. 11(1):56–64, 2008.
Celis, J. E. Microinjection of somatic cells with micropipettes: comparison with other transfer techniques. Biochem. J. 223(2):281, 1984.
Davies, K. J. Oxidative stress, antioxidant defenses, and damage removal, repair, and replacement systems. IUBMB Life 50(4–5):279–289, 2000.
Dean, D. A., D. Machado-Aranda, K. Blair-Parks, A. V. Yeldandi, and J. L. Young. Electroporation as a method for high-level nonviral gene transfer to the lung. Gene Ther. 10(18):1608–1615, 2003.
Gattass, R. R., and E. Mazur. Femtosecond laser micromachining in transparent materials. Nat. Photon. 2(4):219–225, 2008.
Halliwell, B. Oxidative stress and cancer: have we moved forward? Biochem. J. 401:1–11, 2007.
Harding, H. P., Y. Zhang, H. Zeng, I. Novoa, P. D. Lu, M. Calfon, N. Sadri, C. Yun, B. Popko, R. Paules, D. F. Stojdl, J. C. Bell, T. Hettmann, J. M. Leiden, and D. Ron. An integrated stress response regulates amino acid metabolism and resistance to oxidative stress. Mol. Cell 11(3):619–633, 2003.
Hu, Yaxin, Jennifer M. F. Wan, and Alfred C. H. Yu. Membrane perforation and recovery dynamics in microbubble-mediated sonoporation. Ultrasound Med. Biol. 39(12):2393–2405, 2013.
Liu, J., T. N. Lewis, and M. R. Prausnitz. Non-invasive assessment and control of ultrasound-mediated membrane permeabilization. Pharm. Res. 15(6):918–924, 1998.
Mandato, C. A., and W. M. Bement. Contraction and polymerization cooperate to assemble and close actomyosin rings around Xenopus oocyte wounds. J. Cell Biol. 154(4):785–798, 2001.
McNeil, A. K., U. Rescher, V. Gerke, and P. L. McNeil. Requirement for annexin A1 in plasma membrane repair. J. Biol. Chem. 281(46):35202–35207, 2006.
McNeil, P. L., and R. A. Steinhardt. Loss, restoration, and maintenance of plasma membrane integrity. J. Cell Biol. 137(1):1–4, 1997.
McNeil, P. L., and R. A. Steinhardt. Plasma membrane disruption: repair, prevention, adaptation. Annu. Rev. Cell Dev. Boil. 19(1):697–731, 2003.
Pawley, J. Handbook of Biological Confocal Microscopy. Springer Science & Business Media, 2006.
Powers, S. K., A. N. Kavazis, and J. M. McClung. Oxidative stress and disuse muscle atrophy. J. Appl. Physiol. 102(6):2389–2397, 2007.
Serpersu, E. H., K. Kinosita, and T. Y. Tsong. Reversible and irreversible modification of erythrocyte membrane permeability by electric field. Biochim. Biophys. Acta. (BBA)-Biomembr. 812(3):779–785, 1985.
Siu, P. M., Y. Wang, and S. E. Alway. Apoptotic signaling induced by H2O2-mediated oxidative stress in differentiated C2C12 myotubes. Life Sci. 84(13):468–481, 2009.
Sonnemann, K. J., and W. M. Bement. Wound repair: toward understanding and integration of single-cell and multicellular wound responses. Annu. Rev. Cell Dev. Boil. 27:237–263, 2011.
Sun, S., S. Wong, A. Mak, and M. Cho. Impact of oxidative stress on cellular biomechanics and rho signaling in C2C12 myoblasts. J. Biomech. 47(15):3650–3656, 2014.
Tatham, P. E., and B. D. Gomperts. Rat mast cells degranulate in response to microinjection of guanine nucleotide. J. Cell Sci. 98(2):217–224, 1991.
Terasaki, M., K. Miyake, and P. L. McNeil. Large plasma membrane disruptions are rapidly resealed by Ca2+-dependent vesicle–vesicle fusion events. J. Cell Biol. 139(1):63–74, 1997.
Wong, S. W., S. Sun, M. Cho, K. K. Lee, and A. F. T. Mak. H2O2 exposure affects myotube stiffness and actin filament polymerization. Ann. Biomed. Eng. 1–11, 2014.
Yao, Y., Z. Xiao, S. W. Wong, Y. Hsu, T. Cheng, C. Chang, L. Bian, and A. Mak. The effects of oxidative stress on the compressive damage thresholds of C2C12 mouse myoblasts—implications for deep tissue injury. Cells. Ann. Biomed. Eng. 43(2):287–296, 2015.
Zhou, Y., J. Shi, J. Cui, and C. X. Deng. Effects of extracellular calcium on cell membrane resealing in sonoporation. J Control Release 126(1):34–43, 2008.
Acknowledgments
The authors thank the Hong Kong Research Grants Council for its General Research Funding support for this work through the project Grant CUHK415413.
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The authors declare that they have no conflict of interests.
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Associate Editor James Tunnell oversaw the review of this article.
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Duan, X., Chan, K.T., Lee, K.K.H. et al. Oxidative Stress and Plasma Membrane Repair in Single Myoblasts After Femtosecond Laser Photoporation. Ann Biomed Eng 43, 2735–2744 (2015). https://doi.org/10.1007/s10439-015-1341-4
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DOI: https://doi.org/10.1007/s10439-015-1341-4