Advertisement

Transmission Electron Microscopy of Tissue-Polymer Constructs

  • Paul V. Hatton
Protocol
  • 958 Downloads
Part of the Methods in Molecular Biology™ book series (MIMB, volume 238)

Abstract

Numerous publications have described the histology of tissue-engineered constructs and tissue-biomaterial interfaces observed with the light microscope (1, 2, 3, 4, 5). Indeed, this approach has become a routine method for the evaluation of the biological quality of engineered tissues. Additional microscopical techniques, including immunohistochemistry and confocal microscopy, have also been described. These methods have enabled the localization of important biological molecules in the tissue-engineered construct. However, all of these techniques are limited to describing relatively gross structural features, and they give little insight into the ultrastructural relationships between cells and biomaterials within a construct. Scanning electron microscopy (SEM) has been applied more recently to the study of constructs, but it too provides only limited information and not the “fine detail” of cell-biomaterial interaction (6,7). Despite the limitations noted here, relatively little research has been reported in which the transmission electron microscope (TEM) has been used to study the ultrastructure of cell-polymer constructs. This may be partly because of the difficulties associated with specimen preparation when living tissues and materials are both present. However, these difficulties are not insurmountable. Studies of the cell-biomaterial interface using electron microscopy have been reported in the general field of biomaterials and medical devices (8, 9, 10, 11). Here, they have enabled researchers to gain a greater understanding of the response of cells and tissues to biomaterial surfaces.

Keywords

Transmission Electron Microscope Study Propylene Oxide Sodium Cacodylate Buffer Diamond Knife Biomaterial Surface 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Puelacher, W. C., Kim, S. W., Vacanti, J. P., Schloo, B., Mooney, D., and Vacanti, C. A. (1994) Tissue-engineered growth of cartilage: the effect of varying the concentration of chondrocytes seeded onto synthetic polymer matrices. Int. J. Oral Maxillofac. Surg. 23, 49–53.CrossRefGoogle Scholar
  2. 2.
    Rotter, N., Aigner, J., Naumann, A., Planck, H., Hammer, C., Burmester, G., et al. (1998) Cartilage reconstruction in head and neck surgery: comparison of resorbable polymer scaffolds for tissue engineering of human septal cartilage. J. Biomed. Mater. Res. 42, 47–56.CrossRefGoogle Scholar
  3. 3.
    Martin, I., Vunjak-Novakovic, G., Yang, J., Langer, R., and Freed, L. E. (1999) Mammalian chondrocytes expanded in the presence of fibroblast growth factor 2 maintain the ability to differentiate and regenerate three-dimensional cartilaginous tissue. Exp. Cell Res. 253, 681–688.CrossRefGoogle Scholar
  4. 4.
    Holy, C. E., Shoichet, M. S., and Davies, J. E. (2000) Engineering three-dimensional bone tissue in vitro using biodegradable scaffolds: investigating initial cell-seeding density and culture period. J. Biomed. Mater. Res. 51, 376–382.CrossRefGoogle Scholar
  5. 5.
    Salata, L. A., Hatton, P. V., Devlin, A. J., Craig, G. T., and Brook, I. M. (2001) In vitro and in vivo evaluation of e-PTFE and alkali-cellulose membranes for guided bone regeneration. Clin. Oral Implants Res. 12, 62–68.CrossRefGoogle Scholar
  6. 6.
    He, Q., Li, Q., Chen, B., and Wang, Z. (2002) Repair of flexor tendon defects of rabbit with tissue engineering method. Chin. J. Traumatol. 5, 200–208Google Scholar
  7. 7.
    Caterson, E. J., Li, W. J., Nesti, L. J., Albert, T., Danielson, K., and Tuan, R. S. (2002) Polymer/alginate amalgam for cartilage-tissue engineering. Ann. NY Acad. Sci. 961, 134–138.CrossRefGoogle Scholar
  8. 8.
    Bakker, D., van Blitterswijk, C.A., Hesseling, S. C., Daems, W. T., and Grote, J. J. (1990) Tissue/biomaterial interface characteristics of four elastomers. A transmission electron microscopical study. J. Biomed. Mater. Res. 24, 277–293.CrossRefGoogle Scholar
  9. 9.
    Hatton, P. V., Craig, G. T., and Brook, I. M. (1992) Characterization of the interface between bone and glass-ionomer (polyalkenoate) cement using transmission electron microscopy and X-ray microanalysis. Advances in Biomaterials 10, 331–336. (Doherty, P. J., ed.), Elsevier Science Publishers B.V.Google Scholar
  10. 10.
    Hatton, P. V., Walsh, J. and Brook, I. M. (1995) The response of cultured bone cells to resorbable polyglycolic acid and reinforced membranes for use in orbital floor repair. Clinical Materials 17, 71–80.CrossRefGoogle Scholar
  11. 11.
    Da Rocha Barros, V. M., Salata, L.A., Van Noort, R., and Hatton, P. V. (2002) In vivo bone tissue response to a canasite glass-ceramic. Biomaterials, 23, 2895–2900.Google Scholar
  12. 12.
    Glauert, A. M. (1975) Practical Methods in Electron Microscopy (Glauert, A. M., ed), Elsevier, Amsterdam, The Neterlands.Google Scholar
  13. 13.
    Hayat, M. A. (1989) Principles and Techniques of Electron Microscopy: Biological Applications (Hayat, M. A., ed.), MacMillan Press, London.Google Scholar
  14. 14.
    van Dorp, A. G., Verhoeven, M. C., Koerten, H. K., van Blitterswijk, C. A., and Ponec, M. (1999) Bilayered biodegradable poly(ethylene glycol)/poly(butylene terephthalate) copolymer (Polyactive) as substrate for human fibroblasts and keratinocytes. J. Biomed. Mater. Res. 47, 292–300.CrossRefGoogle Scholar
  15. 15.
    Marra, K. G., Szem, J. W., Kumta, P. N., DiMilla, P.A., and Weiss, L.E. (1999) In vitro analysis of biodegradable polymer blend/hydroxyapatite composites for bone tissue engineering. J. Biomed. Mater. Res. 47, 324–335.CrossRefGoogle Scholar
  16. 16.
    Springer, I. N., Fleiner, B., Jepsen, S., and Acil, Y. (2001) Culture of cells gained from temporomandibular joint cartilage on non-absorbable scaffolds. Biomaterials 22, 2569–2577.CrossRefGoogle Scholar
  17. 17.
    X Muller, L. L. and Jacks, T. J. (1975) Rapid chemical dehydration of samples for electron microscopic examinations. J. Histochem. Cytochem. 23, 107–112.Google Scholar
  18. 18.
    Y Conway, K. and Kiernan, J. A. (1999) Chemical dehydration of specimens with 2,2-dimethoxypropane (DMP) for paraffin processing of animal tissues: practical and economic advantages over dehydration in ethanol. Biotech. Histochem. 74, 20–26.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Paul V. Hatton
    • 1
  1. 1.Centre for Biomaterials and Tissue Engineering, School of Clinical DentistryUniversity of SheffieldSheffieldUK

Personalised recommendations