Composition and structure of porcine digital flexor tendon-bone insertion tissues
Sandhya Chandrasekaran
Department of Mechanical and Aerospace Engineering, North Carolina State University, R3158 Engineering Building 3, Campus Box 7910, 911 Oval Drive, Raleigh, North Carolina, 27695 USA
Search for more papers by this authorMark Pankow
Department of Mechanical and Aerospace Engineering, North Carolina State University, R3158 Engineering Building 3, Campus Box 7910, 911 Oval Drive, Raleigh, North Carolina, 27695 USA
Search for more papers by this authorKara Peters
Department of Mechanical and Aerospace Engineering, North Carolina State University, R3158 Engineering Building 3, Campus Box 7910, 911 Oval Drive, Raleigh, North Carolina, 27695 USA
Search for more papers by this authorCorresponding Author
Hsiao-Ying Shadow Huang
Department of Mechanical and Aerospace Engineering, North Carolina State University, R3158 Engineering Building 3, Campus Box 7910, 911 Oval Drive, Raleigh, North Carolina, 27695 USA
Correspondence to: H.-Y. S. Huang; e-mail: [email protected]Search for more papers by this authorSandhya Chandrasekaran
Department of Mechanical and Aerospace Engineering, North Carolina State University, R3158 Engineering Building 3, Campus Box 7910, 911 Oval Drive, Raleigh, North Carolina, 27695 USA
Search for more papers by this authorMark Pankow
Department of Mechanical and Aerospace Engineering, North Carolina State University, R3158 Engineering Building 3, Campus Box 7910, 911 Oval Drive, Raleigh, North Carolina, 27695 USA
Search for more papers by this authorKara Peters
Department of Mechanical and Aerospace Engineering, North Carolina State University, R3158 Engineering Building 3, Campus Box 7910, 911 Oval Drive, Raleigh, North Carolina, 27695 USA
Search for more papers by this authorCorresponding Author
Hsiao-Ying Shadow Huang
Department of Mechanical and Aerospace Engineering, North Carolina State University, R3158 Engineering Building 3, Campus Box 7910, 911 Oval Drive, Raleigh, North Carolina, 27695 USA
Correspondence to: H.-Y. S. Huang; e-mail: [email protected]Search for more papers by this authorAbstract
Tendon-bone insertion is a functionally graded tissue, transitioning from 200 MPa tensile modulus at the tendon end to 20 GPa tensile modulus at the bone, across just a few hundred micrometers. In this study, we examine the porcine digital flexor tendon insertion tissue to provide a quantitative description of its collagen orientation and mineral concentration by using Fast Fourier Transform (FFT) based image analysis and mass spectrometry, respectively. Histological results revealed uniformity in global collagen orientation at all depths, indicative of mechanical anisotropy, although at mid-depth, the highest fiber density, least amount of dispersion, and least cellular circularity were evident. Collagen orientation distribution obtained through 2D FFT of histological imaging data from fluorescent microscopy agreed with past measurements based on polarized light microscopy. Results revealed global fiber orientation across the tendon-bone insertion to be preserved along direction of physiologic tension. Gradation in the fiber distribution orientation index across the insertion was reflective of a decrease in anisotropy from the tendon to the bone. We provided elemental maps across the fibrocartilage for its organic and inorganic constituents through time-of-flight secondary ion mass spectrometry (TOF-SIMS). The apatite intensity distribution from the tendon to bone was shown to follow a linear trend, supporting past results based on Raman microprobe analysis. The merit of this study lies in the image-based simplified approach to fiber distribution quantification and in the high spatial resolution of the compositional analysis. In conjunction with the mechanical properties of the insertion tissue, fiber, and mineral distribution results for the insertion from this may potentially be incorporated into the development of a structural constitutive approach toward computational modeling. Characterizing the properties of the native insertion tissue would provide the microstructural basis for developing biomimetic scaffolds to recreate the graded morphology of a fibrocartilaginous insertion. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3050–3058, 2017.
REFERENCES
- 1 Schwartz AG, Lipner JH, Pasteris JD, Genin GM, Thomopoulos S. Muscle loading is necessary for the formation of a functional tendon enthesis. Bone 2013; 55: 44–51.
- 2 Thomopoulos S, Williams GR, Gimbel JA, Favata M, Soslowsky LJ. Variation of biomechanical, structural, and compositional properties along the tendon to bone insertion site. J Orthop Res. 2003; 21: 413–419.
- 3 Thomopoulos S, Williams GR, Soslowsky LJ. Tendon to bone healing: Differences in biomechanical, structural, and compositional properties due to a range of activity levels. J Biomech Eng Trans ASME 2003; 125: 106–113.
- 4 Thomopoulos S, Zampiakis E, Das R, Silva MJ, Gelberman RH. The effect of muscle loading on flexor tendon-to-bone healing in a canine model. J Orthop Res 2008; 26: 1611–1617.
- 5 Benjamin M, Kumai T, Milz S, Boszczyk BM, Boszczyk AA, Ralphs JR. The skeletal attachment of tendons - tendon “entheses”. Comp Biochem Physiol Mol Integr Physiol 2002; 133: 931–945.
- 6 Benjamin M, Evans EJ, Copp L. The histology of tendon attachments to bone in man. J Anat 1986; 149: 89–100.
- 7 Genin GM, Kent A, Birman V, Wopenka B, Pasteris JD, Marquez PJ, Thomopoulos S. Functional grading of mineral and collagen in the attachment of tendon to bone. Biophys J 2009; 97: 976–985.
- 8 Lu HH, Thomopoulos S. Functional attachment of soft tissues to bone: development, healing, and tissue engineering. Annu Rev Biomed Eng 2013; 15: 201–226.
- 9 Thomopoulos S, Genin GM, Galatz LM. The development and morphogenesis of the tendon-to-bone insertion - What development can teach us about healing. J Musculoskelet Neuronal Interact 2010; 10: 35–45.
- 10 Thomopoulos S, Marquez JP, Weinberger B, Birman V, Genin GM. Collagen fiber orientation at the tendon to bone insertion and its influence on stress concentrations. J Biomech 2006; 39: 1842–1851.
- 11 Lu HH, Subramony SD, Boushell MK, Zhang XZ. Tissue engineering strategies for the regeneration of orthopedic interfaces. Ann Biomed Eng 2010; 38: 2142–2154.
- 12 Robertson DB, Daniel DM, Biden E. Soft-tissue fixation to bone. Am J Sports Med 1986; 14: 398–403.
- 13 Schwartz AG, Pasteris JD, Genin GM, Daulton TL, Thomopoulos S. Mineral distributions at the developing tendon enthesis. Plos One 2012; 7: 1–11.
- 14 Wopenka B, Kent A, Pasteris JD, Yoon Y, Thomopoulos S. The tendon-to-bone transition of the rotator cuff: A preliminary raman spectroscopic study documenting the gradual mineralization across the insertion in rat tissue samples. Appl Spectrosc 2008; 62: 1285–1294.
- 15 Genin GM, Hutchinson JW. Composite laminates in plane stress: Constitutive modeling and stress redistribution due to matrix cracking. J Am Ceram Soc 1997; 80: 1245–1255.
- 16 Williams ML. Stress singularities resulting from various boundary conditions in angular corners of plates in extension. J Appl Mech Trans ASME 1952; 19: 526–528.
- 17 Rieppo J, Hallikainen J, Jurvelin JS, Kiviranta I, Helminen HJ, Hyttinen MM. Practical considerations in the use of polarized light microscopy in the analysis of the collagen network in articular cartilage. Microsc Res Tech 2008; 71: 279–287.
- 18 Whittaker P, Canham PB. Demonstration of quantitative fabric analysis of tendon collagen using 2-dimensional polarized-light microscopy. Matrix 1991; 11: 56–62.
- 19 Billiar KL, Sacks MSA. Method to quantify the fiber kinematics of planar tissues under biaxial stretch. J Biomech 1997; 30: 1997.
- 20 Sacks MS, Billiar KB. Chapter 3: Biaxial mechanical behavior of bioprosthetic heart cusps subjected to accelerated testing. In: S Gabbay, R Frater, editors. Advances in Anticalcific and Antidegenerative Treatment of Heart Valve Bioprostheses. Austin: Silent Partners; 1997.
- 21 Sacks MS. Focus on materials with scattered light. Res Dev 1988; 30: 73–78.
- 22
Sacks MS,
Gloeckner DC. Quantification of the fiber architecture and biaxial mechanical behavior of porcine intestinal submucosa. J Biomed Mater Res 1999; 46: 1–10.
10.1002/(SICI)1097-4636(199907)46:1<1::AID-JBM1>3.0.CO;2-7 CAS PubMed Web of Science® Google Scholar
- 23 Sacks MS, Smith DB, Hiester ED. A small angle light scattering device for planar connective tissue microstructural analysis. Ann Biomed Eng 1997; 25: 678–689.
- 24 Axer H, von Keyserlingk DG, Prescher A. Collagen fibers in linea alba and rectus sheaths - II. Variability and biomechanical aspects. J Surg Res 2001; 96: 239–245.
- 25 Hansen KA, Weiss JA, Barton JK. Recruitment of tendon crimp with applied tensile strain. J Biomech Eng Trans ASME 2002; 124: 72–77.
- 26 Franchi M, Fini M, Quaranta M, De Pasquale V, Raspanti M, Giavaresi G, Ottani V, Ruggeri A. Crimp morphology in relaxed and stretched rat Achilles tendon. J Anat 2007; 210: 1–7.
- 27 Tan HY, Chang YL, Lo W, Hsueh CM, Chen WL, Ghazaryan AA, Hu PS, Young TH, Chen SJ, Dong CY. Characterizing the morphologic changes in collagen crosslinked-treated corneas by Fourier transform-second harmonic generation imaging. J Cataract Refract Surg 2013; 39: 779–788.
- 28 Sander EA, Barocas VH. Comparison of 2D fiber network orientation measurement methods. J Biomed Mater Res A 2009; 88a: 322–331.
- 29 Sander EA, Stylianopoulos T, Tranquillo RT, Barocas VH. Image-based biomechanics of collagen-based tissue equivalents multiscale models compared to fiber alignment predicted by polarimetric imaging. IEEE Eng Med Biol Magazine 2009; 28: 10–18.
- 30 Ayres C, Bowlin GL, Henderson SC, Taylor L, Shultz J, Alexander J, Telemeco TA, Simpson DG. Modulation of anisotropy in electrospun tissue-engineering scaffolds: Analysis of fiber alignment by the fast Fourier transform. Biomaterials 2006; 27: 5524–5534.
- 31 Engelmayr GC, Cheng MY, Bettinger CJ, Borenstein JT, Langer R, Freed LE. Accordion-like honeycombs for tissue engineering of cardiac anisotropy. Nat Mater 2008; 7: 1003–1010.
- 32 Polzer S, Gasser TC, Forsell C, Druckmullerova H, Tichy M, Staffa R, Vlachovsky R, Bursa J. Automatic Identification and validation of planar collagen organization in the aorta wall with application to abdominal aortic aneurysm. Microsc Microanal 2013; 19: 1395–1404.
- 33 Chow MJ, Turcotte R, Lin CP, Zhang YH. Arterial extracellular matrix: A mechanobiological study of the contributions and interactions of elastin and collagen. Biophys J 2014; 106: 2684–2692.
- 34 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. Fiji: an open-source platform for biological-image analysis. Nat Methods 2012; 9: 676–682.
- 35 Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods 2012; 9: 671–675.
- 36 Rezakhaniha R, Agianniotis A, Schrauwen JTC, Griffa A, Sage D, Bouten CVC, van de Vosse FN, Unser M, Stergiopulos N. Experimental investigation of collagen waviness and orientation in the arterial adventitia using confocal laser scanning microscopy. Biomech Model Mechanobiol 2012; 11: 461–473.
- 37 Weichsel J, Urban E, Small JV, Schwarz US. Reconstructing the orientation distribution of actin filaments in the lamellipodium of migrating keratocytes from electron microscopy tomography data. Cytometry Part A 2012; 81a: 496–507.
- 38 ChiuHuang C-K, Zhou C, Huang H-YS. In-situ imaging of lithium-ion batteries via the secondary ion mass spectrometry. ASME J Nanotechnol Eng Med 2014; 5, pp. 021002-1-5.
- 39 Barofsky DF, Brinkmalm G, Hakansson P, Sundqvist BUR. Quantitative-determination of kinetic-energy releases from metastable decompositions of sputtered organic ions using a time-of-flight mass-spectrometer with a single-stage ion mirror. Int J Mass Spectrom Ion Process 1994; 131: 283–294.
- 40 Belu AM, Graham DJ, Castner DG. Time-of-flight secondary ion mass spectrometry: Techniques and applications for the characterization of biomaterial surfaces. Biomaterials 2003; 24: 3635–3653.
- 41 Brown BN, Barnes CA, Kasick RT, Michel R, Gilbert TW, Beer-Stolz D, Castner DG, Ratner B, Badylak SF. Surface characterization of extracellular matrix scaffolds. Biomaterials 2010; 31: 428–437.
- 42 Cersoy S, Richardin P, Walter P, Brunelle A. Cluster TOF-SIMS imaging of human skin remains: analysis of a South-Andean mummy sample. J Mass Spectrom 2012; 47: 338–346.
- 43 Layrolle P, Lebugle A. Characterization and reactivity of nanosized calcium phosphates prepared in anhydrous ethanol. Chem Mater 1994; 6: 1996–2004.
- 44 Lu HB, Campbell CT, Graham DJ, Ratner BD. Surface characterization of hydroxyapatite and related calcium phosphates by XPS and TOF-SIMS. Anal Chem 2000; 72: 2886–2894.
- 45 McDonnell LA, Piersma SR, Altelaar AFM, Mize TH, Luxembourg SL, Verhaert PDEM, van Minnen J, Heeren RMA. Subcellular imaging mass spectrometry of brain tissue. J Mass Spectrom 2005; 40: 160–168.
- 46 Nygren H, Johansson BR, Malmberg P. Bioimaging TOF-SIMS of tissues by gold ion bombardment of a silver-coated thin section. Microsc Res Tech 2004; 65: 282–286.
- 47 Ward AJ, Short RD. A Tofsims and Xps investigation of the structure of plasma polymers prepared from the methacrylate series of monomers. 2. The influence of the W/F parameter on structural and functional-group retention. Polymer 1995; 36: 3439–3450.
- 48 Galatz L, Rothermich S, VanderPloeg K, Petersen B, Sandell L, Thomopoulos S. Development of the supraspinatus tendon-to-bone insertion: Localized expression of extracellular matrix and growth factor genes. J Orthop Res 2007; 25: 1621–1628.
- 49 Thomopoulos S, Hattersley G, Rosen V, Mertens M, Galatz L, Williams GR, Soslowsky LJ. The localized expression of extracellular matrix components in healing tendon insertion sites: an in situ hybridization study. J Orthop Res 2002; 20: 454–463.
- 50 Galatz LM, Sandell LJ, Rothermich SY, Das R, Mastny A, Havlioglu N, Silva MJ, Thomopoulos S. Characteristics of the rat supraspinatus tendon during tendon-to-bone healing after acute injury. J Orthop Res 2006; 24: 541–550.
- 51 Wong MWN, Qin L, Lee KM, Leung KS. Articular cartilage increases transition zone regeneration in bone-tendon junction healing. Clin Orthop Relat Res 2009; 467: 1092–1100.
- 52 Wong MWN, Tai KO, Lee KM, Qin L, Leung KS. Bone formation and fibrocartilage regeneration at bone tendon junction with allogeneic cultured chondrocyte pellet interposition. J Bone Miner Res 2007; 22: 1128–1129.
- 53 Rodeo SA, Arnoczky SP, Torzilli PA, Hidaka C, Warren RF. Tendon-healing in a bone tunnel. A biomechanical and histological study in the dog. J Bone Joint Surg Am 1993; 75: 1795–1803.