Abstract
An important issue in tissue biomechanics is to decipher the relationship between the mechanical behavior at macroscopic scale and the organization of the collagen fiber network at microscopic scale. Here, we present a protocol to combine traction assays with multiphoton microscopy in ex vivo murine skin. This multiscale approach provides simultaneously the stress/stretch response of a skin biopsy and the collagen reorganization in the dermis by use of second harmonic generation (SHG) signals and appropriate image processing.
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References
Goulam Houssen Y, Gusachenko I, Schanne-Klein M-C, Allain J-M (2011) Monitoring micrometer-scale collagen organization in rat-tail tendon upon mechanical strain using second harmonic generation microscopy. J Biomech 44:2047–2052
Bancelin S, Lynch B, Bonod-Bidaud C, Ducourthial G, Psilodimitrakopoulos S, Dokladal P, Allain J-M, Schanne-Klein M-C, Ruggiero F (2015) Ex vivo multiscale quantitation of skin biomechanics in wild-type and genetically-modified mice using multiphoton microscopy. Sci Rep 5:17635
Zipfel WR, Williams RM, Christie R, Nikitin AY, Hyman BT, Webb WW (2003) Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation. Proc Natl Acad Sci U S A 100:7075–7080
Strupler M, Pena A-M, Hernest M, Tharaux P-L, Martin J-L, Beaurepaire E, Schanne-Klein M-C (2007) Second harmonic imaging and scoring of collagen in fibrotic tissues. Opt Express 15(7):4054–4065
Chen XY, Nadiarynkh O, Plotnikov S, Campagnola PJ (2012) Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure. Nat Protoc 7(4):654–669
Lynch B, Bancelin S, Bonod-Bidaud C, Guesquin J-B, Ruggiero F, Schanne-Klein M-C, Allain J-M (2017) A novel microstructural interpretation for the biomechanics of mouse skin derived from multiscale characterization. Acta Biomater 50:302–311
Jayyosi C, Affagard J-S, Ducourthial G, Bonod-Bidaud C, Lynch B, Bancelin S, Ruggiero F, Schanne-Klein M-C, Allain J-M, Bruyère-Garnier K, Coret M (2017) Affine kinematics in planar fibrous connective tissues: an experimental investigation. Biomech Model Mechanobiol 16(4):1459–1473
Lynch B, Bonod-Bidaud C, Ducourthial G, Affagard J-S, Bancelin S, Psilodimitrakopoulos S, Ruggiero F, Allain J-M, Schanne-Klein M-C (2017) How aging impacts skin biomechanics: a multiscale study in mice. Sci Rep 7:13750
Sinclair EB, Andarawis-Puri N, Ros SJ, Laudier DM, Jepsen KJ, Hausman MR (2012) Relating applied strain to the type and severity of structural damage in the rat median nerve using second harmonic generation microscopy. Muscle Nerve 46(6):899–907
Wentzell S, Nesbitt RS, Macione J, Kotha S (2013) Measuring strain using digital image correlation of second harmonic generation images. J Biomech 46(12):2032–2038
Chow MJ, Turcotte R, Lin CP, Zhang YH (2014) Arterial extracellular matrix: a Mechanobiological study of the contributions and interactions of elastin and collagen. Biophys J 106(12):2684–2692
Sigal IA, Grimm JL, Jan NJ, Reid K, Minckler DS, Brown DJ (2014) Eye-specific IOP-induced displacements and deformations of human Lamina Cribrosa. Invest Ophth Vis Sci 55(1):1–15
Mauri A, Ehret AE, Perrini M, Maake C, Ochsenbein-Kolble N, Ehrbar M, Oyen ML, Mazza E (2015) Deformation mechanisms of human amnion: quantitative studies based on second harmonic generation microscopy. J Biomech 48(9):1606–1613
Alavi SH, Sinha A, Steward E, Milliken JC, Kheradvar A (2015) Load-dependent extracellular matrix organization in atrioventricular heart valves: differences and similarities. Am J Physiol Heart Circ Physiol 309(2):H276–H284
Nesbitt S, Scott W, Macione J, Kotha S (2015) Collagen fibrils in skin orient in the direction of applied uniaxial load in proportion to stress while exhibiting differential strains around hair follicles. Materials 8(4):1841–1857
Caulk AW, Nepiyushchikh ZV, Shaw R, Dixon JB, Gleason RL (2015) Quantification of the passive and active biaxial mechanical behaviour and microstructural organization of rat thoracic ducts. J R Soc Interface 12(108). ARTN 20150280
Benoit A, Latour G, Schanne-Klein M-C, Allain J-M (2016) Simultaneous microstructural and mechanical characterization of human corneas at increasing pressure. J Mech Behav Biomed Mater 60:93–105
Krasny W, Morin C, Magoariec H, Avril S (2017) A comprehensive study of layer-specific morphological changes in the microstructure of carotid arteries under uniaxial load. Acta Biomater 57:342–351
Serra J (1982) Analysis and mathematical morphology. Academic Press
Bayan C, Levitt JM, Miller E, Kaplan D, Georgakoudi I (2009) Fully automated, quantitative, noninvasive assessment of collagen fiber content and organization in thick collagen gels. J Appl Phys 105(10):102042
Zipfel WR, Williams RM, Webb WW (2003) Nonlinear magic: multiphoton microscopy in the biosciences. Nat Biotech 21(11):1369–1377
Acknowledgments
This work was supported by grants from Ecole Polytechnique (interdisciplinary project) and from Agence Nationale de la Recherche (contracts ANR-10-INBS-04 France BioImaging, ANR-11-EQPX-0029 Morphoscope2, and ANR-13-BS09-0004-02 Metis). The authors thank Vincent de Greef for his help in the technical implantation of the mechanical setup.
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Bancelin, S. et al. (2019). Combination of Traction Assays and Multiphoton Imaging to Quantify Skin Biomechanics. In: Sagi, I., Afratis, N. (eds) Collagen. Methods in Molecular Biology, vol 1944. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9095-5_11
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DOI: https://doi.org/10.1007/978-1-4939-9095-5_11
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