No Access

CHARACTERIZING THREE-DIMENSIONAL MICROSTRUCTURE OF COLLAGEN/CHITOSAN SCAFFOLDS USING MULTIPHOTON MICROSCOPE

Hsin-Yuan Tan

Institute of Biomedical Engineering, College of Engineering and Medicine, National Taiwan University, Taipei, Taiwan

Department of Ophthalmology, Chang Gung Memorial Hospital, College of Medicine, Chang Gung University, Linko, Taiwan

Search for more papers by this author

Wen Lo

Department of Physics, National Taiwan University, Taipei, Taiwan

Search for more papers by this author

Chiu-Mei Hsueh

Department of Physics, National Taiwan University, Taipei, Taiwan

Search for more papers by this author

Chia-Yi Wang

Department of Ophthalmology, Chang Gung Memorial Hospital, College of Medicine, Chang Gung University, Linko, Taiwan

Search for more papers by this author

Sung-Jan Lin

Institute of Biomedical Engineering, College of Engineering and Medicine, National Taiwan University, Taipei, Taiwan

Department of Dermatology, National Taiwan University Hospital, Taipei, Taiwan

Search for more papers by this author

Chen-Yuan Dong

Department of Physics, National Taiwan University, Taipei, Taiwan

Corresponding author: Chen-Yuan Dong, Department of Physics, National Taiwan University, Taipei, Taiwan.

Search for more papers by this author

, and

Tai-Horng Young

Institute of Biomedical Engineering, College of Engineering and Medicine, National Taiwan University, Taipei, Taiwan

Corresponding author: Tai-Horng Young, Institute of Biomedical Engineering, College of Engineering and Medicine, National Taiwan University, Taipei, Taiwan.

Search for more papers by this author

https://doi.org/10.4015/S1016237213500385Cited by:0 (Source: Crossref)

Abstract

In this work, we used multiphoton microscopic system for characterizing three-dimensional microstructure of collagen/chitosan polymeric scaffolds in a noninvasive fashion. Nonlinear optical signals including multiphoton autofluorescence (MAF) and second harmonic generation (SHG) derived from collagen/chitosan scaffolds were collected and analyzed. The three-dimensional porous microstructures of collagen/chitosan scaffolds were visualized by co-localized and evenly distributed MAF and SHG signals. The distribution of collagen and chitosan compositions within miscible collagen/chitosan blends cannot be either localized or differentiated simply using these nonlinear optical signals. However, the intensity of MAF signals in scaffolds was found to be markedly decreased in correlation to the supplementation of chitosan within blends, regardless of collagen/chitosan weight ratios. It therefore implied that the MAF-generating molecules within collagen being altered in miscible collagen/chitosan blends. And the SHG signals also decreased significantly in collagen/chitosan scaffolds with the supplementation of chitosan, regardless of different weight ratios. This finding supported the hypothesis regarding the miscibility of collagen/chitosan blends that triple helix structure of collagen, a proven SHG-generating microstructure, was altered in miscible collagen/chitosan blends. In conclusion, our work demonstrated that multiphoton imaging modality can be versatile for investigating three-dimensional microstructure of miscible polymeric scaffolds in a minimal invasive fashion, and may potentially be applicable in the field of tissue engineering.

Keywords:

References

R. L. Reis et al. , J. R. Soc. Interface 4 , 999 ( 2007 ) . Crossref, Web of Science, Google Scholar
Kudva RA, Keskkula H, Paul DR, Compatibilization of nylon 6/ABS blends using glycidyl methacrylate methyl methacrylate copolymers, Abstr Pap Am Chem S 215:U280, 1998 . Google Scholar
T. H. Young et al. , J. Biomed. Mater. Res. A 64A , 606 ( 2003 ) . Crossref, Web of Science, Google Scholar
C. Y. Gao et al. , Biomaterials 24 , 4833 ( 2003 ) . Crossref, Web of Science, Google Scholar
P. B. Malafaya and R. L. Reis, Key Eng. Mat. 240(2), 39 (2003). Crossref, Google Scholar
M. G. Dunn , S. L. Bourke and J. Kohn , Tissue Eng. 10 , 43 ( 2004 ) . Crossref, Google Scholar
P. V. Moghe et al. , J. Biomed. Mater. Res. B 82B , 284 ( 2007 ) . Crossref, Web of Science, Google Scholar
T. H. Young et al. , Biomaterials 27 , 5079 ( 2006 ) . Crossref, Web of Science, Google Scholar
D. Raghavan et al. , J. Polym. Sci. Pol. Phys. 44 , 492 ( 2006 ) . Crossref, Web of Science, Google Scholar
B. Serrano et al. , J. Fluoresc. 10 , 135 ( 2000 ) . Crossref, Web of Science, Google Scholar
A. E. Ribbe , Trends Polym. Sci. 5 , 333 ( 1997 ) . Web of Science, Google Scholar
R. E. Guldberg et al. , Biomaterials 24 , 481 ( 2003 ) . Crossref, Web of Science, Google Scholar
J. P. Dunkers , M. T. Cicerone and N. R. Washburn , Opt. Express 11 , 3074 ( 2003 ) . Crossref, Web of Science, Google Scholar
K. J. Halbhuber and K. Konig , Ann. Anat. 185 , 1 ( 2003 ) . Crossref, Web of Science, Google Scholar
C. W. Jones et al. , Prog. Histochem. Cyto. 40 , 1 ( 2005 ) . Crossref, Web of Science, Google Scholar
H. Shuman , J. M. Murray and C. Dilullo , Biotechniques 7 , 154 ( 1989 ) . Crossref, Web of Science, Google Scholar
R. Richards-Kortum et al. , Ieee T. Bio.-Med. Eng. 49 , 1168 ( 2002 ) . Crossref, Web of Science, Google Scholar
B. R. Masters and P. T. C. So , Microsc. Microanal. 5 , 282 ( 1999 ) . Crossref, Web of Science, Google Scholar
J. A. Izatt et al. , Ieee J. Sel. Top. Quant. 2 , 1017 ( 1996 ) . Crossref, Web of Science, Google Scholar
B. J. Tromberg , A. Zoumi and A. Yeh , P. Natl. Acad. Sci. USA 99 , 11014 ( 2002 ) . Crossref, Web of Science, Google Scholar
I. Constantididis et al. , Ann. Ny. Acad. Sci. 961 , 298 ( 2002 ) . Crossref, Web of Science, Google Scholar
W. Denk , J. H. Strickler and W. W. Webb , Science 248 , 73 ( 1990 ) . Crossref, Web of Science, Google Scholar
P. T. C. So et al. , Annu. Rev. Biomed. Eng. 2 , 399 ( 2000 ) . Crossref, Web of Science, Google Scholar
W. W. Webb , W. R. Zipfel and R. M. Williams , Nat. Biotechnol. 21 , 1368 ( 2003 ) . Web of Science, Google Scholar
S. Roth and I. Freund , J. Chem. Phys. 70 , 1637 ( 1979 ) . Crossref, Web of Science, Google Scholar
Q. Wu et al. , Biomaterials 26 , 899 ( 2005 ) . Crossref, Web of Science, Google Scholar
D. M. Burland , R. D. Miller and C. A. Walsh , Chem. Rev. 94 , 31 ( 1994 ) . Crossref, Web of Science, Google Scholar
J. Xu et al. , Polymer 48 , 348 ( 2007 ) . Crossref, Web of Science, Google Scholar
W. A. Mohler et al. , Biophys. J. 90 , 693 ( 2006 ) . Crossref, Web of Science, Google Scholar
W. W. Webb et al. , P. Natl. Acad. Sci. USA 100 , 7075 ( 2003 ) . Crossref, Web of Science, Google Scholar
R. M. Williams , W. R. Zipfel and W. W. Webb , Biophys. J. 88 , 1377 ( 2005 ) . Crossref, Web of Science, Google Scholar
P. J. Campagnola et al. , Biophys. J. 82 , 493 ( 2002 ) . Crossref, Web of Science, Google Scholar
L. M. Loew and P. J. Campagnola , Nat. Biotechnol. 21 , 1356 ( 2003 ) . Crossref, Web of Science, Google Scholar
R. D. Bowles et al. , Tissue Eng. A 16 , 1339 ( 2010 ) . Crossref, Web of Science, Google Scholar
W.-L. Chen et al. , Tissue Eng. Pt. C-Meth. 16 , 913 ( 2010 ) . Crossref, Web of Science, Google Scholar
E. Filova et al. , J. Biomed. Opt. 15 , 066011 ( 2010 ) . Crossref, Web of Science, Google Scholar
A.-M. Pena et al. , J. Biomed. Opt. 15 , 056018 ( 2010 ) . Crossref, Web of Science, Google Scholar
M. P. Rubbens et al. , An. Biomed. Eng. 37 , 1263 ( 2009 ) . Crossref, Web of Science, Google Scholar
S. L. Sanos et al. , J. Immunol. 186 , 2835 ( 2011 ) . Crossref, Web of Science, Google Scholar
R. Schade et al. , Int. J. Artificial Organs. 33 , 219 ( 2010 ) . Crossref, Web of Science, Google Scholar
C. Schizas and D. Karalekas , J. Mater. Process. Technol. 209 , 2349 ( 2009 ) . Crossref, Web of Science, Google Scholar
C. Y. Dong et al. , Microsc. Res. Techniq. 71 , 140 ( 2008 ) . Crossref, Web of Science, Google Scholar
H. S. Lee et al. , Tissue Eng. 12 , 2835 ( 2006 ) . Crossref, Google Scholar
C. Y. Dong et al. , Opt. Express 16 , 3818 ( 2008 ) . Crossref, Web of Science, Google Scholar
A. Sionkowska et al. , J. Photochem. Photobiol. A 162 , 545 ( 2004 ) . Crossref, Web of Science, Google Scholar
Q. L. Feng et al. , Biomaterials 24 , 3213 ( 2003 ) . Crossref, Web of Science, Google Scholar
F. Berthod , M. Gingras and I. Paradis , Biomaterials 24 , 1653 ( 2003 ) . Crossref, Web of Science, Google Scholar
C. Y. Gao , D. Y. Wang and J. C. Shen , Polym. Advan. Technol. 14 , 373 ( 2003 ) . Crossref, Web of Science, Google Scholar
D. Thacharodi and K. P. Rao , Int. J. Pharm. 120 , 115 ( 1995 ) . Crossref, Web of Science, Google Scholar
J. S. Chen et al. , Artif. Organs 29 , 104 ( 2005 ) . Crossref, Web of Science, Google Scholar
T. J. Wess et al. , Biomaterials 25 , 795 ( 2004 ) . Crossref, Web of Science, Google Scholar
W. H. Dan et al. , Eur. Polym. J. 43 , 2066 ( 2007 ) . Crossref, Web of Science, Google Scholar
P. Arpornmaeklong et al. , Int. J. Oral Max. Surg. 36 , 328 ( 2007 ) . Crossref, Web of Science, Google Scholar
Y. Sun et al. , Microsc. Res. Techniq. 71 , 140 ( 2008 ) . Crossref, Web of Science, Google Scholar
N. Shanmugasundaram et al. , Biomaterials 22 , 1943 ( 2001 ) . Crossref, Web of Science, Google Scholar
D. Fujimoto , Biochem. Biophys. Res. Commun. 76 , 1124 ( 1977 ) . Crossref, Web of Science, Google Scholar
D. R. Eyre , M. A. Paz and P. M. Gallop , Annu. Rev. Biochem. 53 , 717 ( 1984 ) . Crossref, Web of Science, Google Scholar
W. L. Chen et al. , Tissue Eng. Part C Methods 16 , 913 ( 2010 ) . Crossref, Web of Science, Google Scholar
C. B. Raub et al. , Biophys. J. 92 , 2212 ( 2007 ) . Crossref, Web of Science, Google Scholar