Abstract
The mechanical behavior of soft connective tissue is governed by a dense network of fibrillar proteins in the extracellular matrix. Characterization of this fibrous network requires the accurate extraction of descriptive structural parameters from imaging data, including fiber dispersion and mean fiber orientation. Common methods to quantify fiber parameters include fast Fourier transforms (FFT) and structure tensors; however, information is limited on the accuracy of these methods. In this study, we compared these two methods using test images of fiber networks with varying topology. The FFT method with a band-pass filter was the most accurate, with an error of \(0.7^{\circ } \pm 0.4^{\circ }\) in measuring mean fiber orientation and an error of \(7.4 \pm 3.0\,\%\) in measuring fiber dispersion in the test images. The accuracy of the structure tensor method was approximately five times worse than the FFT band-pass method when measuring fiber dispersion. A free software application, FiberFit, was then developed that utilizes an FFT band-pass filter to fit fiber orientations to a semicircular von Mises distribution. FiberFit was used to measure collagen fibril organization in confocal images of bovine ligament at magnifications of \(63{\times }\) and \(20{\times }\). Grayscale conversion prior to FFT analysis gave the most accurate results, with errors of \(3.3^{\circ } \pm 3.1^{\circ }\) for mean fiber orientation and \(13.3 \pm 8.2\,\%\) for fiber dispersion when measuring confocal images at \(63{\times }\). By developing and validating a software application that facilitates the automated analysis of fiber organization, this study can help advance a mechanistic understanding of collagen networks and help clarify the mechanobiology of soft tissue remodeling and repair.
Similar content being viewed by others
References
Ayres C, Bowlin GL, Henderson SC, Taylor L, Shultz J, Alexander J, Telemeco T, Simpson DG (2006) Modulation of anisotropy in electrospun tissue-engineering scaffolds: analysis of fiber alignment by the fast Fourier transform. Biomaterials 27(32):5524–5534
Ayres CE, Jha BS, Meredith H, Bowman JR, Bowlin GL, Henderson SC, Simpson DG (2008) Measuring fiber alignment in electrospun scaffolds: a user s guide to the 2D fast Fourier transform approach. J Biomater Sci Polym Ed 19(5):603–621
Berens P (2009) CircStat: a MATLAB toolbox for circular statistics. J Stat Softw 31(10):1–21
Bigun J, Bigun T, Nilsson K (2004) Recognition by symmetry derivatives and the generalized structure tensor. IEEE Trans Pattern Anal Mach Intell 26(12):1590–1605
Chamberlain CS, Crowley EM, Kobayashi H, Eliceiri KW, Vanderby R (2011) Quantification of collagen organization and extracellular matrix factors within the healing ligament. Microsc Microanal 17(05):779–787
D’Amore A, Stella JA, Wagner WR, Sacks MS (2010) Characterization of the complete fiber network topology of planar fibrous tissue and scaffolds. Biomaterials 31(20):5345–5354
Driessen NJB, MaJ C, Bouten CVC, Baaijens FPT (2008) Remodelling of the angular collagen fiber distribution in cardiovascular tissues. Biomech Model Mechanobiol 7(2):93–103
Fitzgibbon A, Pilu M, Fisher RB (1999) Direct least square fitting of ellipses. IEEE Trans Pattern Anal Mach Intell 21(5):476–480
Frisch K, Duenwald-Kuehl S, Lakes R, Vanderby R (2012) Quantification of collagen organization using fractal dimensions and Fourier transforms. Acta Histochem 114(2):140–144
Gasser TC, Ogden RW, Ga H (2006) Hyperelastic modelling of arterial layers with distributed collagen fibre orientations. J R Soc Interface 3(6):15–35 (0312002v1)
Girard MJA, Downs JC, Burgoyne CF, Suh JKF (2009) Peripapillary and posterior scleral mechanics—part I: development of an anisotropic hyperelastic constitutive model. J Biomech Eng 131(5):051,011
Gonzalez R, Woods R (2008) Digital image processing, 3rd edn. Pearson Prentice Hall, New Jersey
Gouget CLM, Girard MJ, Ethier CR (2012) A constrained von Mises distribution to describe fiber organization in thin soft tissues. Biomech Model Mechanobiol 11(3–4):475–482
Grytz R, Meschke G, Jonas JB (2011) The collagen fibril architecture in the lamina cribrosa and peripapillary sclera predicted by a computational remodeling approach. Biomech Model Mechanobiol 10(3):371–382
Holzapfel GA, Gasser TC, Ogden RW (2000) A new constitutive framework for arterial wall mechanics and a comparative study of material models. J Elast 61:1–48
Hurschler C, Provenzano PP, Vanderby R (2003) Scanning electron microscopic characterization of healing and normal rat ligament microstructure under slack and loaded conditions. Connect Tissue Res 44(2):59–68
Jahne B (1993) Spatio-temporal image processing: theory and scientific applications. Springer, Berlin
Jones E, Oliphant T, Peterson P, Others (2001) SciPy: open source scientific tools for Python. http://www.scipy.org/
Lanir Y (1981) The fibrous structure of the skin and its relation to mechanical behaviour. In: Marks R, Payne P (eds) Bioengineering and the skin. Springer, Netherlands, pp 93–95
Loerakker S, Obbink-Huizer C, Baaijens FPT (2014) A physically motivated constitutive model for cell-mediated compaction and collagen remodeling in soft tissues. Biomech Model Mechanobiol 13(5):985–1001
Maas SA, Ellis BJ, Ateshian GA, Weiss JA (2012) FEBio: finite elements for biomechanics. J Biomech Eng 134(1):011,005
Machyshyn IM, Bovendeerd PHM, van de Ven AAF, Rongen PMJ, van de Vosse FN (2010) A model for arterial adaptation combining microstructural collagen remodeling and 3D tissue growth. Biomech Model Mechanobiol 9(6):671–687
Makareeva E, Mertz EL, Kuznetsova NV, Sutter MB, DeRidder AM, Wa C, Barnes AM, McBride DJ, Marini JC, Leikin S (2008) Structural heterogeneity of type I collagen triple helix and its role in osteogenesis imperfecta. J Biol Chem 283(8):4787–4798
Mardia KV, Jupp PE (2000) Directional statistics. Wiley, New York
Marquez JP (2006) Fourier analysis and automated measurement of cell and fiber angular orientation distributions. Int J Solids Struct 43(21):6413–6423
Monici M (2005) Cell and tissue autofluorescence research and diagnostic applications. Biotechnol Annu Rev 11(11):227–256
O’Connell B (2012) Oval profile plot plugin for ImageJ. http://rsb.info.nih.gov/ij/plugins/oval-profile.html
Petroll MW, Cavanagh DH, Barry P, Andrews P, Jester JV (1993) Quantitative analysis of stress fiber orientation during corneal wound contraction. J Cell Sci 104:353–363
Polzer S, Gasser TC, Forsell C, Druckmüllerova H, Tichy M, Staffa R, Vlachovsky R, Bursa J (2013) Automatic identification and validation of planar collagen organization in the aorta wall with application to abdominal aortic aneurysm. Microsc Microanal 19:1395–1404
Rezakhaniha R, Agianniotis A, Schrauwen JTC, Griffa A, Sage D, Bouten CVC, Van De Vosse FN, Unser M, Stergiopulos N (2012) Experimental investigation of collagen waviness and orientation in the arterial adventitia using confocal laser scanning microscopy. Biomech Model Mechanobiol 11(3–4):461–473
Sampo J, Takalo J, Siltanen S, Miettinen A, Lassas M, Timonen J (2014) Curvelet-based method for orientation estimation of particles from optical images. Opt Eng 53(3):033,109
Sander EA, Barocas VH (2009) Comparison of 2D fiber network orientation measurement methods. J Biomed Mater Res Part A 88(2):322–331
Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9(7):676–682
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9(7):671–675
Schriefl AJ, Reinisch AJ, Sankaran S, Pierce DM, Holzapfel GA (2012) Quantitative assessment of collagen fibre orientations from two-dimensional images of soft biological tissues. J R Soc Interface 9(76):3081–3093
Tienevez JY (2010) Directionality plugin for ImageJ. http://fiji.sc/Directionality
Tower TT, Neidert MR, Tranquillo RT (2002) Fiber alignment imaging during mechanical testing of soft tissues. Ann Biomed Eng 30(10):1221–1233
Vogel A, Holbrook KA, Steinmann B, Gitzelmann R, Byres PH (1979) Abnormal collagen fibril structure in the Gravis Form (Type I) of Ehlers–Danlos syndrome. Lab Invest 40(2):201–206
Weichsel J, Urban E, Small JV, Schwarz US (2012) Reconstructing the orientation distribution of actin filaments in the lamellipodium of migrating keratocytes from electron microscopy tomography data. Cytom Part A 81A(6):496–507
Woo SLY, Buckwalter JA, Fung YC (1989) Injury and repair of the musculoskeletal soft tissues. J Biomech Eng 111(1):95
Acknowledgments
This project was supported by Institutional Development Awards (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under Grants Nos. P20GM103408 and P20GM109095. We also acknowledge support from The Biomolecular Research Center at Boise State with funding from the National Science Foundation, Grants Nos. 0619793 and 0923535; the MJ Murdock Charitable Trust; and the Idaho State Board of Education.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Morrill, E.E., Tulepbergenov, A.N., Stender, C.J. et al. A validated software application to measure fiber organization in soft tissue. Biomech Model Mechanobiol 15, 1467–1478 (2016). https://doi.org/10.1007/s10237-016-0776-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10237-016-0776-3