Hostname: page-component-78c5997874-8bhkd Total loading time: 0 Render date: 2024-11-15T11:16:50.375Z Has data issue: false hasContentIssue false
  • English
  • Français

A Collagen/DBP Sponge System Designed for in Vitro Analysis of Chondroinduction

Published online by Cambridge University Press:  15 February 2011

Shuichi Mizuno ,
Chris Lycette ,
Charlene Quinto  and
Julie Glowacki
Shuichi Mizuno
Affiliation:
Orthopedic Research, Brigham and Women's Hospital, Harvard Medical School, 75 Francis St., Boston, MA 02115
Chris Lycette
Affiliation:
Orthopedic Research, Brigham and Women's Hospital, Harvard Medical School, 75 Francis St., Boston, MA 02115
Charlene Quinto
Affiliation:
Orthopedic Research, Brigham and Women's Hospital, Harvard Medical School, 75 Francis St., Boston, MA 02115
Julie Glowacki
Affiliation:
Orthopedic Research, Brigham and Women's Hospital, Harvard Medical School, 75 Francis St., Boston, MA 02115
Get access

Abstract

In response to subcutaneous implants of demineralized bone powder (DBP), cells are attracted to the DBP, are converted to chondroblasts, and produce a cartilage matrix that is resorbed and replaced by bone. In order to define the cellular mechanisms of this induction, we developed a collagen sponge model for simulating the in vivo environment and for promoting the ingrowth and viability of cells cultured in them in vitro. Reconstituted pepsin–digested type I collagen from bovine hide was neutralized. Rat DBP (75–250 εm) was added into the collagen mixture (20 mg/ml). In order to simulate the connective tissue environment, modified chondroitin sulfate, heparan sulfate, or hyaluronic acid was added into the mixture. Aliquots (0.2 ml) were placed in 3/8 inch diameter molds and freeze-dried. Human dermal fibroblasts were cultured from minced fresh tissue and inoculated at 1.5 × 105 cells/sponge. Fifteen hours later, some sponges were transferred to medium which contained growth factors (PDGF or TGF-β). At intervals, samples were examined histologically. The inoculated cells attached to the collagen fibers and migrated into the sponge. Eventually the sponges contracted and acquired an oval shape. Cells on or near DBP were ovoid or stellate in shape. Cell morphology was modulated by glycosaminoglycan composition of the sponge. Increasing doses of PDGF or TGF-β promoted cellularity within the sponges. In conclusion, this system simulates the in vivo environment but allows accessibility for analysis. This three-dimensional matrix culture system will enable investigation of mechanisms of chondroinduction by morphogenic material.

Type
Research Article
Information
MRS Online Proceedings Library (OPL) , Volume 252: Symposium T – Tissue-Inducing Biomaterials , 1991 , 133
Copyright
Copyright © Materials Research Society 1992

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

1. Urist, M.R., Science 150, 893 (1965)Google Scholar
2. Reddi, A.H. and Huggins, C.B., Proc Natl Acad Sci, USA 69, 1601 (1972)Google Scholar
3. Glowacki, J. and Mulliken, J.B., Clin Plast Surg 12, 233 (1985)Google Scholar
4. Glowacki, J., J Cell Biol 107, 1072a (1988)Google Scholar
5. Urist, M.R. et al., Proc Natb Acad Sci, USA 81, 371 (1984)Google Scholar
6. Sampath, T.K. et al., Proc Natl Acad Sci, USA 84, 7109 (1987)CrossRefGoogle Scholar
7. Seyedin, S.M. et al., J Biol Chem 261, 5693 (1986)Google Scholar
8. Wozney, J.M. et al., Science 242, 1528, (1988)CrossRefGoogle Scholar
9. Glowacki, J. and Mulliken, J., Clin Plast Surg 12, 233 (1985)CrossRefGoogle Scholar
10. Strivastava, S. and Gorham, S. D., Biomaterials 11, 162 (1990)Google Scholar
11. Strivastava, S. et al., Biomaterials 11, 155 (1990)Google Scholar
12. Doillon, C.J. et al., Biomaterials B 8, 195 (1990)Google Scholar
13. Yamagata, M. et al., J Biol Chem 261, 13526 (1986)Google Scholar
14. Yamagata, M. et al., J Biol Chem 264, 8012 (1989)Google Scholar
15. Turley, E.A. et al., J Cell Biol 112, 1041 (1991)Google Scholar