Abstract

Tissue engineered medical products (TEMPs) are often three-dimensional (3D) hybrid materials consisting of a porous scaffold upon which the tissue is grown. However, monitoring of the developing tissue deep within the scaffold is hampered by its turbidity. We have sought new ways to probe the interior of the scaffold with the same resolution as conventional laser scanning confocal microscopy but with greater sensitivity. We present a novel application of optical coherence microscopy (OCM) by combining it with confocal fluorescence microscopy (CFM) to gather simultaneous structural and functional information on TEMPs in a registered fashion. In this work, we describe the collinear OCM and CFM instrument. We demonstrate the utility of this dual-mode technique for TEMPs by imaging fluorescently stained osteoblasts cultured in a polymeric TEMP.

© 2003 Optical Society of America

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References

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  10. Identification of a commercial product is made only to facilitate experimental reproducibility and to adequately describe experimental procedure. In no case does it imply endorsement by NIST or imply that it is necessarily the best product for the experimental procedure.
  11. All uncertainties expressed in accordance to: B. Taylor and C. Kuyatt, NIST Technical Note 1297, 1994 and are based on one standard deviation.
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  15. <a href= "http://www.olympusmicro.com/primer/techniques/fluorescence/fluorotable2.html">http://www.olympusmicro.com/primer/techniques/fluorescence/fluorotable2.html</a>

Cell Transplantation

F. Thorsen, T.-A. Read, M. Lund-Johansen, B. B. Tysnes, and R. Bjerkvig, �??Alginate-encapsulated producer cells: a potential new approach for the treatment of malignant brain tumors,�?? Cell Transplantation. 9, 773-783 (2000).

Electron. Lett.

S. Chinn and E. Swanson, �??Blindness limitations in optical coherence domain reflectometry,�?? Electron. Lett. 29, 2025-2027 (1993).
[CrossRef]

IEEE J. Sel. Top. Quantum Electron.

J. Izatt, M. Kulkarni, H.-S. Wang, K. Kobayashi, and M. Sivak, �??Optical coherence tomography and microscopy in gastrointestinal tissues,�?? IEEE J. Sel. Top. Quantum Electron. 2, 1017-1028 (1996).
[CrossRef]

J. Biomed. Mat. Res.

M. Attawia, J. Devin, and C. Laurencin, �??Immunofluorescence and confocal laser-scanning microscopy studies of osteoblast growth and phenotypic-expression in 3-dimensional degradable synthetic matrices,�??J. Biomed. Mat. Res. 29, 843-848 (1995).
[CrossRef]

N. Washburn, C. Simon, A. Tona, H. Elgendy, A. Karim, and E. Amis, �??Co-extrusion of biocompatible polymers for scaffolds with co-continuous morphology,�?? J. Biomed. Mat. Res. 60, 20-29 (2002).
[CrossRef]

J. Biomed. Mater. Res.

J. M. Anderson, L. G. Cima, S. G. Eskin, et al., �??Tissue engineering in cardiovascular disease - a report,�?? J. Biomed. Mater. Res. 29, 1473-1476 (1995).
[CrossRef]

NIST Technical Note

All uncertainties expressed in accordance to: B. Taylor and C. Kuyatt, NIST Technical Note 1297, 1994 and are based on one standard deviation.

Opt. Express

Opt. Lett.

Polymer Data Handbook

J. O. Iroh, �??Poly(ε-caprolactone)�?? in Polymer Data Handbook, J. E. Mark, ed. (Oxford University Press, New York. 1999).

Proc. SPIE Int. Soc. Opt. Eng.

F. Guzman and J. K. Barton, �??Dual OCT/spectroscopy system for identification of vascular pathology,�??Proc. SPIE Int. Soc. Opt. Eng. 4244, 413 �??414 (2001).

Tissue Eng.

M. Afting, U. Stock, B. Nasseri, I. Pomerantseva, B. Seed, and J. Vacanti, �??Efficient and stable retroviral transfection of ovine endothelial cells with green fluorescent protein for cardiovascular tissue engineering,�?? Tissue Eng. 9, 137-41 (2003).
[CrossRef] [PubMed]

Other

Identification of a commercial product is made only to facilitate experimental reproducibility and to adequately describe experimental procedure. In no case does it imply endorsement by NIST or imply that it is necessarily the best product for the experimental procedure.

<a href= "http://www.olympusmicro.com/primer/techniques/fluorescence/fluorotable2.html">http://www.olympusmicro.com/primer/techniques/fluorescence/fluorotable2.html</a>

Supplementary Material (1)

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Figures (4)

Fig. 1.
Fig. 1.

Experimental apparatus for collinear OCM/ CFM. BP: band pass filter; CL: collimating lens; CM: cold mirror; Demod: Demodulator; SOAS: semiconductor optical amplifier source; DBS: dichroic beam splitter; LP: Long pass filter; NDF: Neutral density filter; NF: Notch filter; PM: Polarization maintaining 50/50 coupler; Pol: In-line polarizer; PS: Polarization splitter; PZTFM: Piezoelectic fiber

Fig. 2.
Fig. 2.

OCM (A.) and CFM (B.) images 145 µm from surface of PCL cultured for 10 weeks with fetal chick osteoblasts.

Fig. 3.
Fig. 3.

Scanning electron micrograph of PCL scaffold. PCL was co-extruded with poly-ethyleneoxide (PEO) at 0.50 mass fraction. The scaffold was immersed in solvent to dissolve the PEO, and the remaining PCL was subject to a 30 min anneal at 75 °C.

Fig. 4.
Fig. 4.

(0.72 MB) Movie of merged and registered OCM and CFM images of the cultured PCL scaffold.

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