Abstract

The collagen meshwork plays a central role in the functioning of a range of tissues including cartilage, tendon, arteries, skin, bone and ligament. Because of its importance in function, it is of considerable interest for studying development, disease and regeneration processes. Here, we have used second harmonic generation (SHG) to image human tissues on the hundreds of micron scale, and developed a numerical model to quantitatively interpret the images in terms of the underlying collagen structure on the tens to hundreds of nanometer scale. Focusing on osteoarthritic changes in cartilage, we have demonstrated that this combination of polarized SHG imaging and numerical modeling can estimate fibril diameter, filling fraction, orientation and bundling. This extends SHG microscopy from a qualitative to quantitative imaging technique, providing a label-free and non-destructive platform for characterizing the extracellular matrix that can expand our understanding of the structural mechanisms in disease.

© 2013 Optical Society of America

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  7. C. P. Brown, M.-A. Houle, M. Chen, A. J. Price, F. Légaré, and H. S. Gill, “Damage initiation and progression in the cartilage surface probed by nonlinear optical microscopy,” J. Mech. Behav. Biomed. Mater.5(1), 62–70 (2012).
    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
  23. D. A. D. Parry and A. S. Craig, “Quantitative electron microscope observations of the collagen fibrils in rat-tail tendon,” Biopolymers16(5), 1015–1031 (1977).
    [CrossRef] [PubMed]
  24. M. Rivard, M. Laliberté, A. Bertrand-Grenier, C. Harnagea, C. P. Pfeffer, M. Vallières, Y. St-Pierre, A. Pignolet, M. A. El Khakani, and F. Légaré, “The structural origin of second harmonic generation in fascia,” Biomed. Opt. Express2(1), 26–36 (2011).
    [CrossRef] [PubMed]
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    [CrossRef]
  28. W. S. Hwang, B. Li, L. H. Jin, K. Ngo, N. S. Schachar, and G. N. F. Hughes, “Collagen fibril structure of normal, aging, and osteoarthritic cartilage,” J. Pathol.167(4), 425–433 (1992).
    [CrossRef] [PubMed]
  29. N. D. Broom, M.-H. Chen, and A. Hardy, “A degeneration-based hypothesis for interpreting fibrillar changes in the osteoarthritic cartilage matrix,” J. Anat.199(6), 683–698 (2001).
    [CrossRef] [PubMed]
  30. J. Mertz and L. Moreaux, “Second-harmonic generation by focused excitation of inhomogeneously distributed scatterers,” Opt. Commun.196(1-6), 325–330 (2001).
    [CrossRef]

2013

2012

C. P. Brown, M.-A. Houle, M. Chen, A. J. Price, F. Légaré, and H. S. Gill, “Damage initiation and progression in the cartilage surface probed by nonlinear optical microscopy,” J. Mech. Behav. Biomed. Mater.5(1), 62–70 (2012).
[CrossRef] [PubMed]

X. Chen, O. Nadiarynkh, S. Plotnikov, and P. J. Campagnola, “Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure,” Nat. Protoc.7(4), 654–669 (2012).
[PubMed]

2011

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

M. Rivard, M. Laliberté, A. Bertrand-Grenier, C. Harnagea, C. P. Pfeffer, M. Vallières, Y. St-Pierre, A. Pignolet, M. A. El Khakani, and F. Légaré, “The structural origin of second harmonic generation in fascia,” Biomed. Opt. Express2(1), 26–36 (2011).
[CrossRef] [PubMed]

2010

C. Harnagea, M. Vallières, C. P. Pfeffer, D. Wu, B. R. Olsen, A. Pignolet, F. Légaré, and A. Gruverman, “Two-dimensional nanoscale structural and functional imaging in individual collagen type I fibrils,” Biophys. J.98(12), 3070–3077 (2010).
[CrossRef] [PubMed]

2008

J. C. Mansfield, C. P. Winlove, J. Moger, and S. J. Matcher, “Collagen fiber arrangement in normal and diseased cartilage studied by polarization sensitive nonlinear microscopy,” J. Biomed. Opt.13(4), 044020 (2008).
[CrossRef] [PubMed]

2007

F. Légaré, C. P. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J.93(4), 1312–1320 (2007).
[CrossRef] [PubMed]

2004

A. Zoumi, X. Lu, G. S. Kassab, and B. J. Tromberg, “Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy,” Biophys. J.87(4), 2778–2786 (2004).
[CrossRef] [PubMed]

2003

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nat. Biotechnol.21(11), 1356–1360 (2003).
[CrossRef] [PubMed]

P. Stoller, P. M. Celliers, K. M. Reiser, and A. M. Rubenchik, “Quantitative second-harmonic generation microscopy in collagen,” Appl. Opt.42(25), 5209–5219 (2003).
[CrossRef] [PubMed]

2002

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, “Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological Tissues,” Biophys. J.82(1), 493–508 (2002).
[CrossRef] [PubMed]

E. B. Hunziker, “Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects,” Osteoarthritis Cartilage10(6), 432–463 (2002).
[CrossRef] [PubMed]

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. U.S.A.99(17), 11014–11019 (2002).
[CrossRef] [PubMed]

2001

N. D. Broom, M.-H. Chen, and A. Hardy, “A degeneration-based hypothesis for interpreting fibrillar changes in the osteoarthritic cartilage matrix,” J. Anat.199(6), 683–698 (2001).
[CrossRef] [PubMed]

J. Mertz and L. Moreaux, “Second-harmonic generation by focused excitation of inhomogeneously distributed scatterers,” Opt. Commun.196(1-6), 325–330 (2001).
[CrossRef]

1997

L. Novotny, “Allowed and forbidden light in near-field optics. I. A single dipolar light source,” J. Opt. Soc. Am.14(1), 91–104 (1997).
[CrossRef]

J. A. Buckwalter and H. J. Mankin, “Articular cartilage, part II: degeneration and osteoarthritis, repair, regeneration, and transplantation,” J. Bone Jt. Surg.79-A, 612–633 (1997).

1994

S. Armstrong, R. Read, and P. Ghosh, “The effects of intraarticular hyaluronan on cartilage and subchondral bone changes in an ovine model of early osteoarthritis,” J. Rheumatol.21(4), 680–688 (1994).
[PubMed]

1992

W. S. Hwang, B. Li, L. H. Jin, K. Ngo, N. S. Schachar, and G. N. F. Hughes, “Collagen fibril structure of normal, aging, and osteoarthritic cartilage,” J. Pathol.167(4), 425–433 (1992).
[CrossRef] [PubMed]

1990

N. D. Broom and H. Silyn-Roberts, “Collagen-collagen versus collagen-proteoglycan interactions in the determination of cartilage strength,” Arthritis Rheum.33, 1512–1517 (1990).
[CrossRef] [PubMed]

1986

I. Freund, M. Deutsch, and A. Sprecher, “Connective tissue polarity. optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon,” Biophys. J.50(4), 693–712 (1986).
[CrossRef] [PubMed]

1979

S. Roth and I. Freund, “Second harmonic generation in collagen,” J. Chem. Phys.70(4), 1637–1643 (1979).
[CrossRef]

1977

D. A. D. Parry and A. S. Craig, “Quantitative electron microscope observations of the collagen fibrils in rat-tail tendon,” Biopolymers16(5), 1015–1031 (1977).
[CrossRef] [PubMed]

1976

A. I. Maroudas, “Balance between swelling pressure and collagen tension in normal and degenerate cartilage,” Nature260(5554), 808–809 (1976).
[CrossRef] [PubMed]

1925

A. Benninghoff, “Form und bau der gelenknorpel in ihren bezeihungen zur funktion,” Z. Mikrosk. Anat. Forsch.2(5), 783–862 (1925).
[CrossRef]

Armstrong, S.

S. Armstrong, R. Read, and P. Ghosh, “The effects of intraarticular hyaluronan on cartilage and subchondral bone changes in an ovine model of early osteoarthritis,” J. Rheumatol.21(4), 680–688 (1994).
[PubMed]

Athanasou, N. A.

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

Barzda, V.

Benninghoff, A.

A. Benninghoff, “Form und bau der gelenknorpel in ihren bezeihungen zur funktion,” Z. Mikrosk. Anat. Forsch.2(5), 783–862 (1925).
[CrossRef]

Benson, R.

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

Bertrand-Grenier, A.

Boucher, Y.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Broom, N. D.

N. D. Broom, M.-H. Chen, and A. Hardy, “A degeneration-based hypothesis for interpreting fibrillar changes in the osteoarthritic cartilage matrix,” J. Anat.199(6), 683–698 (2001).
[CrossRef] [PubMed]

N. D. Broom and H. Silyn-Roberts, “Collagen-collagen versus collagen-proteoglycan interactions in the determination of cartilage strength,” Arthritis Rheum.33, 1512–1517 (1990).
[CrossRef] [PubMed]

Brown, C. P.

C. P. Brown, M.-A. Houle, M. Chen, A. J. Price, F. Légaré, and H. S. Gill, “Damage initiation and progression in the cartilage surface probed by nonlinear optical microscopy,” J. Mech. Behav. Biomed. Mater.5(1), 62–70 (2012).
[CrossRef] [PubMed]

Brown, E.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Buckwalter, J. A.

J. A. Buckwalter and H. J. Mankin, “Articular cartilage, part II: degeneration and osteoarthritis, repair, regeneration, and transplantation,” J. Bone Jt. Surg.79-A, 612–633 (1997).

Campagnola, P. J.

X. Chen, O. Nadiarynkh, S. Plotnikov, and P. J. Campagnola, “Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure,” Nat. Protoc.7(4), 654–669 (2012).
[PubMed]

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nat. Biotechnol.21(11), 1356–1360 (2003).
[CrossRef] [PubMed]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, “Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological Tissues,” Biophys. J.82(1), 493–508 (2002).
[CrossRef] [PubMed]

Carr, A.

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

Celliers, P. M.

Chen, M.

C. P. Brown, M.-A. Houle, M. Chen, A. J. Price, F. Légaré, and H. S. Gill, “Damage initiation and progression in the cartilage surface probed by nonlinear optical microscopy,” J. Mech. Behav. Biomed. Mater.5(1), 62–70 (2012).
[CrossRef] [PubMed]

Chen, M.-H.

N. D. Broom, M.-H. Chen, and A. Hardy, “A degeneration-based hypothesis for interpreting fibrillar changes in the osteoarthritic cartilage matrix,” J. Anat.199(6), 683–698 (2001).
[CrossRef] [PubMed]

Chen, X.

X. Chen, O. Nadiarynkh, S. Plotnikov, and P. J. Campagnola, “Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure,” Nat. Protoc.7(4), 654–669 (2012).
[PubMed]

Craig, A. S.

D. A. D. Parry and A. S. Craig, “Quantitative electron microscope observations of the collagen fibrils in rat-tail tendon,” Biopolymers16(5), 1015–1031 (1977).
[CrossRef] [PubMed]

Deutsch, M.

I. Freund, M. Deutsch, and A. Sprecher, “Connective tissue polarity. optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon,” Biophys. J.50(4), 693–712 (1986).
[CrossRef] [PubMed]

diTomaso, E.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Doll, H.

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

El Khakani, M. A.

Freund, I.

I. Freund, M. Deutsch, and A. Sprecher, “Connective tissue polarity. optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon,” Biophys. J.50(4), 693–712 (1986).
[CrossRef] [PubMed]

S. Roth and I. Freund, “Second harmonic generation in collagen,” J. Chem. Phys.70(4), 1637–1643 (1979).
[CrossRef]

Ghosh, P.

S. Armstrong, R. Read, and P. Ghosh, “The effects of intraarticular hyaluronan on cartilage and subchondral bone changes in an ovine model of early osteoarthritis,” J. Rheumatol.21(4), 680–688 (1994).
[PubMed]

Gill, H. S.

C. P. Brown, M.-A. Houle, M. Chen, A. J. Price, F. Légaré, and H. S. Gill, “Damage initiation and progression in the cartilage surface probed by nonlinear optical microscopy,” J. Mech. Behav. Biomed. Mater.5(1), 62–70 (2012).
[CrossRef] [PubMed]

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

Gruverman, A.

C. Harnagea, M. Vallières, C. P. Pfeffer, D. Wu, B. R. Olsen, A. Pignolet, F. Légaré, and A. Gruverman, “Two-dimensional nanoscale structural and functional imaging in individual collagen type I fibrils,” Biophys. J.98(12), 3070–3077 (2010).
[CrossRef] [PubMed]

Hardy, A.

N. D. Broom, M.-H. Chen, and A. Hardy, “A degeneration-based hypothesis for interpreting fibrillar changes in the osteoarthritic cartilage matrix,” J. Anat.199(6), 683–698 (2001).
[CrossRef] [PubMed]

Harnagea, C.

M. Rivard, M. Laliberté, A. Bertrand-Grenier, C. Harnagea, C. P. Pfeffer, M. Vallières, Y. St-Pierre, A. Pignolet, M. A. El Khakani, and F. Légaré, “The structural origin of second harmonic generation in fascia,” Biomed. Opt. Express2(1), 26–36 (2011).
[CrossRef] [PubMed]

C. Harnagea, M. Vallières, C. P. Pfeffer, D. Wu, B. R. Olsen, A. Pignolet, F. Légaré, and A. Gruverman, “Two-dimensional nanoscale structural and functional imaging in individual collagen type I fibrils,” Biophys. J.98(12), 3070–3077 (2010).
[CrossRef] [PubMed]

Hoppe, P. E.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, “Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological Tissues,” Biophys. J.82(1), 493–508 (2002).
[CrossRef] [PubMed]

Houle, M.-A.

C. P. Brown, M.-A. Houle, M. Chen, A. J. Price, F. Légaré, and H. S. Gill, “Damage initiation and progression in the cartilage surface probed by nonlinear optical microscopy,” J. Mech. Behav. Biomed. Mater.5(1), 62–70 (2012).
[CrossRef] [PubMed]

Hughes, G. N. F.

W. S. Hwang, B. Li, L. H. Jin, K. Ngo, N. S. Schachar, and G. N. F. Hughes, “Collagen fibril structure of normal, aging, and osteoarthritic cartilage,” J. Pathol.167(4), 425–433 (1992).
[CrossRef] [PubMed]

Hulley, P. A.

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

Hunziker, E. B.

E. B. Hunziker, “Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects,” Osteoarthritis Cartilage10(6), 432–463 (2002).
[CrossRef] [PubMed]

Hwang, W. S.

W. S. Hwang, B. Li, L. H. Jin, K. Ngo, N. S. Schachar, and G. N. F. Hughes, “Collagen fibril structure of normal, aging, and osteoarthritic cartilage,” J. Pathol.167(4), 425–433 (1992).
[CrossRef] [PubMed]

Jain, R. K.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Jin, L. H.

W. S. Hwang, B. Li, L. H. Jin, K. Ngo, N. S. Schachar, and G. N. F. Hughes, “Collagen fibril structure of normal, aging, and osteoarthritic cartilage,” J. Pathol.167(4), 425–433 (1992).
[CrossRef] [PubMed]

Kassab, G. S.

A. Zoumi, X. Lu, G. S. Kassab, and B. J. Tromberg, “Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy,” Biophys. J.87(4), 2778–2786 (2004).
[CrossRef] [PubMed]

Laliberté, M.

Légaré, F.

C. P. Brown, M.-A. Houle, M. Chen, A. J. Price, F. Légaré, and H. S. Gill, “Damage initiation and progression in the cartilage surface probed by nonlinear optical microscopy,” J. Mech. Behav. Biomed. Mater.5(1), 62–70 (2012).
[CrossRef] [PubMed]

M. Rivard, M. Laliberté, A. Bertrand-Grenier, C. Harnagea, C. P. Pfeffer, M. Vallières, Y. St-Pierre, A. Pignolet, M. A. El Khakani, and F. Légaré, “The structural origin of second harmonic generation in fascia,” Biomed. Opt. Express2(1), 26–36 (2011).
[CrossRef] [PubMed]

C. Harnagea, M. Vallières, C. P. Pfeffer, D. Wu, B. R. Olsen, A. Pignolet, F. Légaré, and A. Gruverman, “Two-dimensional nanoscale structural and functional imaging in individual collagen type I fibrils,” Biophys. J.98(12), 3070–3077 (2010).
[CrossRef] [PubMed]

F. Légaré, C. P. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J.93(4), 1312–1320 (2007).
[CrossRef] [PubMed]

Li, B.

W. S. Hwang, B. Li, L. H. Jin, K. Ngo, N. S. Schachar, and G. N. F. Hughes, “Collagen fibril structure of normal, aging, and osteoarthritic cartilage,” J. Pathol.167(4), 425–433 (1992).
[CrossRef] [PubMed]

Loew, L. M.

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nat. Biotechnol.21(11), 1356–1360 (2003).
[CrossRef] [PubMed]

Lu, X.

A. Zoumi, X. Lu, G. S. Kassab, and B. J. Tromberg, “Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy,” Biophys. J.87(4), 2778–2786 (2004).
[CrossRef] [PubMed]

Malone, C. J.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, “Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological Tissues,” Biophys. J.82(1), 493–508 (2002).
[CrossRef] [PubMed]

Mankin, H. J.

J. A. Buckwalter and H. J. Mankin, “Articular cartilage, part II: degeneration and osteoarthritis, repair, regeneration, and transplantation,” J. Bone Jt. Surg.79-A, 612–633 (1997).

Mansfield, J. C.

J. C. Mansfield, C. P. Winlove, J. Moger, and S. J. Matcher, “Collagen fiber arrangement in normal and diseased cartilage studied by polarization sensitive nonlinear microscopy,” J. Biomed. Opt.13(4), 044020 (2008).
[CrossRef] [PubMed]

Maroudas, A. I.

A. I. Maroudas, “Balance between swelling pressure and collagen tension in normal and degenerate cartilage,” Nature260(5554), 808–809 (1976).
[CrossRef] [PubMed]

Matcher, S. J.

J. C. Mansfield, C. P. Winlove, J. Moger, and S. J. Matcher, “Collagen fiber arrangement in normal and diseased cartilage studied by polarization sensitive nonlinear microscopy,” J. Biomed. Opt.13(4), 044020 (2008).
[CrossRef] [PubMed]

McDonnell, S.

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

McKee, T.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Mertz, J.

J. Mertz and L. Moreaux, “Second-harmonic generation by focused excitation of inhomogeneously distributed scatterers,” Opt. Commun.196(1-6), 325–330 (2001).
[CrossRef]

Millard, A. C.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, “Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological Tissues,” Biophys. J.82(1), 493–508 (2002).
[CrossRef] [PubMed]

Moger, J.

J. C. Mansfield, C. P. Winlove, J. Moger, and S. J. Matcher, “Collagen fiber arrangement in normal and diseased cartilage studied by polarization sensitive nonlinear microscopy,” J. Biomed. Opt.13(4), 044020 (2008).
[CrossRef] [PubMed]

Mohler, W. A.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, “Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological Tissues,” Biophys. J.82(1), 493–508 (2002).
[CrossRef] [PubMed]

Moreaux, L.

J. Mertz and L. Moreaux, “Second-harmonic generation by focused excitation of inhomogeneously distributed scatterers,” Opt. Commun.196(1-6), 325–330 (2001).
[CrossRef]

Murray, D. W.

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

Nadiarynkh, O.

X. Chen, O. Nadiarynkh, S. Plotnikov, and P. J. Campagnola, “Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure,” Nat. Protoc.7(4), 654–669 (2012).
[PubMed]

Ngo, K.

W. S. Hwang, B. Li, L. H. Jin, K. Ngo, N. S. Schachar, and G. N. F. Hughes, “Collagen fibril structure of normal, aging, and osteoarthritic cartilage,” J. Pathol.167(4), 425–433 (1992).
[CrossRef] [PubMed]

Novotny, L.

L. Novotny, “Allowed and forbidden light in near-field optics. I. A single dipolar light source,” J. Opt. Soc. Am.14(1), 91–104 (1997).
[CrossRef]

Olsen, B. R.

C. Harnagea, M. Vallières, C. P. Pfeffer, D. Wu, B. R. Olsen, A. Pignolet, F. Légaré, and A. Gruverman, “Two-dimensional nanoscale structural and functional imaging in individual collagen type I fibrils,” Biophys. J.98(12), 3070–3077 (2010).
[CrossRef] [PubMed]

F. Légaré, C. P. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J.93(4), 1312–1320 (2007).
[CrossRef] [PubMed]

Parry, D. A. D.

D. A. D. Parry and A. S. Craig, “Quantitative electron microscope observations of the collagen fibrils in rat-tail tendon,” Biopolymers16(5), 1015–1031 (1977).
[CrossRef] [PubMed]

Pfeffer, C. P.

M. Rivard, M. Laliberté, A. Bertrand-Grenier, C. Harnagea, C. P. Pfeffer, M. Vallières, Y. St-Pierre, A. Pignolet, M. A. El Khakani, and F. Légaré, “The structural origin of second harmonic generation in fascia,” Biomed. Opt. Express2(1), 26–36 (2011).
[CrossRef] [PubMed]

C. Harnagea, M. Vallières, C. P. Pfeffer, D. Wu, B. R. Olsen, A. Pignolet, F. Légaré, and A. Gruverman, “Two-dimensional nanoscale structural and functional imaging in individual collagen type I fibrils,” Biophys. J.98(12), 3070–3077 (2010).
[CrossRef] [PubMed]

F. Légaré, C. P. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J.93(4), 1312–1320 (2007).
[CrossRef] [PubMed]

Pignolet, A.

M. Rivard, M. Laliberté, A. Bertrand-Grenier, C. Harnagea, C. P. Pfeffer, M. Vallières, Y. St-Pierre, A. Pignolet, M. A. El Khakani, and F. Légaré, “The structural origin of second harmonic generation in fascia,” Biomed. Opt. Express2(1), 26–36 (2011).
[CrossRef] [PubMed]

C. Harnagea, M. Vallières, C. P. Pfeffer, D. Wu, B. R. Olsen, A. Pignolet, F. Légaré, and A. Gruverman, “Two-dimensional nanoscale structural and functional imaging in individual collagen type I fibrils,” Biophys. J.98(12), 3070–3077 (2010).
[CrossRef] [PubMed]

Plotnikov, S.

X. Chen, O. Nadiarynkh, S. Plotnikov, and P. J. Campagnola, “Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure,” Nat. Protoc.7(4), 654–669 (2012).
[PubMed]

Pluen, A.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Price, A. J.

C. P. Brown, M.-A. Houle, M. Chen, A. J. Price, F. Légaré, and H. S. Gill, “Damage initiation and progression in the cartilage surface probed by nonlinear optical microscopy,” J. Mech. Behav. Biomed. Mater.5(1), 62–70 (2012).
[CrossRef] [PubMed]

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

Read, R.

S. Armstrong, R. Read, and P. Ghosh, “The effects of intraarticular hyaluronan on cartilage and subchondral bone changes in an ovine model of early osteoarthritis,” J. Rheumatol.21(4), 680–688 (1994).
[PubMed]

Reiser, K. M.

Rivard, M.

Roth, S.

S. Roth and I. Freund, “Second harmonic generation in collagen,” J. Chem. Phys.70(4), 1637–1643 (1979).
[CrossRef]

Rout, R.

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

Rubenchik, A. M.

Sandkuijl, D.

Schachar, N. S.

W. S. Hwang, B. Li, L. H. Jin, K. Ngo, N. S. Schachar, and G. N. F. Hughes, “Collagen fibril structure of normal, aging, and osteoarthritic cartilage,” J. Pathol.167(4), 425–433 (1992).
[CrossRef] [PubMed]

Seed, B.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Silyn-Roberts, H.

N. D. Broom and H. Silyn-Roberts, “Collagen-collagen versus collagen-proteoglycan interactions in the determination of cartilage strength,” Arthritis Rheum.33, 1512–1517 (1990).
[CrossRef] [PubMed]

Sipe, J. E.

Sprecher, A.

I. Freund, M. Deutsch, and A. Sprecher, “Connective tissue polarity. optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon,” Biophys. J.50(4), 693–712 (1986).
[CrossRef] [PubMed]

Stoller, P.

St-Pierre, Y.

Terasaki, M.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, “Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological Tissues,” Biophys. J.82(1), 493–508 (2002).
[CrossRef] [PubMed]

Tokarz, D.

Tromberg, B. J.

A. Zoumi, X. Lu, G. S. Kassab, and B. J. Tromberg, “Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy,” Biophys. J.87(4), 2778–2786 (2004).
[CrossRef] [PubMed]

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. U.S.A.99(17), 11014–11019 (2002).
[CrossRef] [PubMed]

Tuer, A. E.

Vallières, M.

M. Rivard, M. Laliberté, A. Bertrand-Grenier, C. Harnagea, C. P. Pfeffer, M. Vallières, Y. St-Pierre, A. Pignolet, M. A. El Khakani, and F. Légaré, “The structural origin of second harmonic generation in fascia,” Biomed. Opt. Express2(1), 26–36 (2011).
[CrossRef] [PubMed]

C. Harnagea, M. Vallières, C. P. Pfeffer, D. Wu, B. R. Olsen, A. Pignolet, F. Légaré, and A. Gruverman, “Two-dimensional nanoscale structural and functional imaging in individual collagen type I fibrils,” Biophys. J.98(12), 3070–3077 (2010).
[CrossRef] [PubMed]

Winlove, C. P.

J. C. Mansfield, C. P. Winlove, J. Moger, and S. J. Matcher, “Collagen fiber arrangement in normal and diseased cartilage studied by polarization sensitive nonlinear microscopy,” J. Biomed. Opt.13(4), 044020 (2008).
[CrossRef] [PubMed]

Wu, D.

C. Harnagea, M. Vallières, C. P. Pfeffer, D. Wu, B. R. Olsen, A. Pignolet, F. Légaré, and A. Gruverman, “Two-dimensional nanoscale structural and functional imaging in individual collagen type I fibrils,” Biophys. J.98(12), 3070–3077 (2010).
[CrossRef] [PubMed]

Yeh, A.

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. U.S.A.99(17), 11014–11019 (2002).
[CrossRef] [PubMed]

Zoumi, A.

A. Zoumi, X. Lu, G. S. Kassab, and B. J. Tromberg, “Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy,” Biophys. J.87(4), 2778–2786 (2004).
[CrossRef] [PubMed]

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. U.S.A.99(17), 11014–11019 (2002).
[CrossRef] [PubMed]

Appl. Opt.

Arthritis Rheum.

N. D. Broom and H. Silyn-Roberts, “Collagen-collagen versus collagen-proteoglycan interactions in the determination of cartilage strength,” Arthritis Rheum.33, 1512–1517 (1990).
[CrossRef] [PubMed]

Biomed. Opt. Express

Biophys. J.

C. Harnagea, M. Vallières, C. P. Pfeffer, D. Wu, B. R. Olsen, A. Pignolet, F. Légaré, and A. Gruverman, “Two-dimensional nanoscale structural and functional imaging in individual collagen type I fibrils,” Biophys. J.98(12), 3070–3077 (2010).
[CrossRef] [PubMed]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, “Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological Tissues,” Biophys. J.82(1), 493–508 (2002).
[CrossRef] [PubMed]

I. Freund, M. Deutsch, and A. Sprecher, “Connective tissue polarity. optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon,” Biophys. J.50(4), 693–712 (1986).
[CrossRef] [PubMed]

F. Légaré, C. P. Pfeffer, and B. R. Olsen, “The role of backscattering in SHG tissue imaging,” Biophys. J.93(4), 1312–1320 (2007).
[CrossRef] [PubMed]

A. Zoumi, X. Lu, G. S. Kassab, and B. J. Tromberg, “Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy,” Biophys. J.87(4), 2778–2786 (2004).
[CrossRef] [PubMed]

Biopolymers

D. A. D. Parry and A. S. Craig, “Quantitative electron microscope observations of the collagen fibrils in rat-tail tendon,” Biopolymers16(5), 1015–1031 (1977).
[CrossRef] [PubMed]

J. Anat.

N. D. Broom, M.-H. Chen, and A. Hardy, “A degeneration-based hypothesis for interpreting fibrillar changes in the osteoarthritic cartilage matrix,” J. Anat.199(6), 683–698 (2001).
[CrossRef] [PubMed]

J. Biomed. Opt.

J. C. Mansfield, C. P. Winlove, J. Moger, and S. J. Matcher, “Collagen fiber arrangement in normal and diseased cartilage studied by polarization sensitive nonlinear microscopy,” J. Biomed. Opt.13(4), 044020 (2008).
[CrossRef] [PubMed]

J. Bone Jt. Surg.

J. A. Buckwalter and H. J. Mankin, “Articular cartilage, part II: degeneration and osteoarthritis, repair, regeneration, and transplantation,” J. Bone Jt. Surg.79-A, 612–633 (1997).

J. Chem. Phys.

S. Roth and I. Freund, “Second harmonic generation in collagen,” J. Chem. Phys.70(4), 1637–1643 (1979).
[CrossRef]

J. Mech. Behav. Biomed. Mater.

C. P. Brown, M.-A. Houle, M. Chen, A. J. Price, F. Légaré, and H. S. Gill, “Damage initiation and progression in the cartilage surface probed by nonlinear optical microscopy,” J. Mech. Behav. Biomed. Mater.5(1), 62–70 (2012).
[CrossRef] [PubMed]

J. Opt. Soc. Am.

L. Novotny, “Allowed and forbidden light in near-field optics. I. A single dipolar light source,” J. Opt. Soc. Am.14(1), 91–104 (1997).
[CrossRef]

J. Opt. Soc. Am. B

J. Pathol.

W. S. Hwang, B. Li, L. H. Jin, K. Ngo, N. S. Schachar, and G. N. F. Hughes, “Collagen fibril structure of normal, aging, and osteoarthritic cartilage,” J. Pathol.167(4), 425–433 (1992).
[CrossRef] [PubMed]

J. Rheumatol.

S. Armstrong, R. Read, and P. Ghosh, “The effects of intraarticular hyaluronan on cartilage and subchondral bone changes in an ovine model of early osteoarthritis,” J. Rheumatol.21(4), 680–688 (1994).
[PubMed]

Knee

R. Rout, S. McDonnell, R. Benson, N. A. Athanasou, A. Carr, H. Doll, H. S. Gill, D. W. Murray, P. A. Hulley, and A. J. Price, “The histological features of anteromedial gonarthrosis--the comparison of two grading systems in a human phenotype of osteoarthritis,” Knee18(3), 172–176 (2011).
[CrossRef] [PubMed]

Nat. Biotechnol.

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nat. Biotechnol.21(11), 1356–1360 (2003).
[CrossRef] [PubMed]

Nat. Med.

E. Brown, T. McKee, E. diTomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med.9(6), 796–801 (2003).
[CrossRef] [PubMed]

Nat. Protoc.

X. Chen, O. Nadiarynkh, S. Plotnikov, and P. J. Campagnola, “Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure,” Nat. Protoc.7(4), 654–669 (2012).
[PubMed]

Nature

A. I. Maroudas, “Balance between swelling pressure and collagen tension in normal and degenerate cartilage,” Nature260(5554), 808–809 (1976).
[CrossRef] [PubMed]

Opt. Commun.

J. Mertz and L. Moreaux, “Second-harmonic generation by focused excitation of inhomogeneously distributed scatterers,” Opt. Commun.196(1-6), 325–330 (2001).
[CrossRef]

Osteoarthritis Cartilage

E. B. Hunziker, “Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects,” Osteoarthritis Cartilage10(6), 432–463 (2002).
[CrossRef] [PubMed]

Proc. Natl. Acad. Sci. U.S.A.

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. U.S.A.99(17), 11014–11019 (2002).
[CrossRef] [PubMed]

Z. Mikrosk. Anat. Forsch.

A. Benninghoff, “Form und bau der gelenknorpel in ihren bezeihungen zur funktion,” Z. Mikrosk. Anat. Forsch.2(5), 783–862 (1925).
[CrossRef]

Other

M. Born and E. Wolf, Principles of Optics (Cambridge University Press, 1999).

K. D. Brandt, M. Doherty, and L. S. Lohmander, “Introduction: the concept of osteoarthritis as failure of the diarthrodal joint,” in Osteoarthritis, K. D. Brandt, M. Doherty, and L. S. Lohmander, eds. (Oxford University Press, Oxford, 2003), pp. 69–72.

N. D. Broom, “The collagen framework of articular cartilage: its profound influence on normal and abnormal load-bearing function,” in Collagen, M. E. Nimni, ed. (CRC Press, Boca Raton, 1988), pp. 244–264.

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Figures (6)

Fig. 1
Fig. 1

Graphical depiction of cross-section of the (a) normal and (b) diseased model tissues. The shape of the laser beam propagating in the x direction is shown by the solid lines. The fibrils with the positive χ(2) are indicated by the red/grey points and those with the negative χ(2) are indicated by the blue/black points.

Fig. 2
Fig. 2

Simulated average forward-to-backward ratio of SHG signal from a random distribution representing healthy tissue (a) and bundled fibrils of the same polarization representing diseased tissue (b). Bundles of mixed polarization had no effect on the ratio (not shown). Dependence of forwards signal (arbitrary units) on filling fraction for normal tissue (c). Dependence of backwards signal (arbitrary units) on filling fraction for normal tissue (d).

Fig. 3
Fig. 3

Representative forward and backward SHG images from normal ((a) and (b) respectively) and osteoarthritic ((c) and (d) respectively) cartilage with the pump beam polarized in the direction from the bone to the cartilage surface. Scale bar 50 μm. Image intensity is scaled for presentation.

Fig. 4
Fig. 4

(a) Ratio of forward to backward signal from experiments in normal and diseased joints, with R1 and R2 corresponding to bright and non-bright regions respectively. (b) Representative polarization dependence of forward (black) and backward (red) signal from normal and non-bright regions of diseased samples. (c) Representative polarization dependence of forward and backward signal from bright regions in diseased samples. 0 degrees is the direction from the bone to the cartilage surface.

Fig. 5
Fig. 5

Representative AFM height images from (a) early-stage osteoarthritic tissues, and (b) normal tissues. Osteoarthritic tissues showed regions of characteristically normal (c’, random alignment) and degraded (d’, kinked bundles and increased alignment) collagen meshwork structures. Zoomed-in images of normal (c) and diseased (d) regions are shown.

Fig. 6
Fig. 6

Representative image (2 x 4 mm) indicating damage progression in the deep zone of a diseased sample. Forward SHG, polarized in the direction from the bone to the cartilage surface (see arrow), shows the progression of alignment and bundling from left to right in the early stages of osteoarthritic degradation. It should be noted that forward SHG provides qualitative information showing disease progression, yet does not provide a quantitative description of collagen organization.

Equations (3)

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E =4 k 0 2 Z +( Z ),
2 Z + k 2 Z = 4π n 0 2 P NL ,
P NL ( r ,2 ω 0 )= y ^ χ (2) ( r ) E 0 2 ( r )( e 2i k 0 x δ( ω2 ω 0 )+ e 2i k 0 x δ( ω+2 ω 0 ) )

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