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Physiological Cultured Skin Substitutes for Wound Healing

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Abstract

Physiological dermal and dermal-epidermal skin analogs have been developed in our laboratory using a novel technology for three-dimensional tissue culture. Human neonatal dermal fibroblasts are seeded on a biodegradable mesh made of polyglycolic or polyglactic acid (PGA/PGL). As the fibroblasts proliferate, they stretch across the mesh openings and secrete growth factors and human dermal matrix proteins, including collagen types I & III and elastin. This process forms a metabolically active, three-dimensional dermal tissue around the mesh scaffolding. The mesh fibers are hydrolyzed over time and is completely resorbed in vivo within four to eight weeks. Multiple sheets of the PGA/PGL-dermal analog are grown simultaneously in a closed, continuous media-flow system, also developed in our laboratory. After attaining confluence, the dermal sheets may be seeded with keratinocytes, to create a living dermal-epidermal composite tissue.

Pre-clinical studies in mini-pigs and athymic mice have shown that the dermal analog “takes” and vascularizes rapidly in full-thickness wounds, and provides a viable dermal base for both meshed split-thickness skin grafts and cultured keratinocyte sheets. There has been no evidence of rejection. The dermal analog is now being studied clinically beneath thin meshed skin autografts in patients with severe bums. In vivo pre-clinical studies of the dermal-epidermal composite, as a complete skin replacement, are also in progress. This technology has potential for applications in severe burns and other full-thickness skin injuries.

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References

  1. Butler, M. Serum-free Media. p. 91–108. In: W.G. Thilly (ed.), Mammalian Cell Technology, Butterworths, Boston, MA, 1986.

    Google Scholar 

  2. Hayashi, I. and G.H. Sato. Replacement of Serum by Hormones Permits Growth of Cells in a Defined Medium, Nature, 259: 132–134, 1976.

    Article  CAS  Google Scholar 

  3. Griffiths, J.B. Serum and Growth Factors in Cell Culture Media-An Introductory Review, Develop. biol. Standard., 66: 155–160, 1985.

    Google Scholar 

  4. Maurer, H.R. Towards Chemically-defined, Serum-free Media for Mammalian Cell Culture. p. 13–31. In: R.I. Freshney (ed.), Animal Cell Culture: A Practical Approach, IRL Press, Washington, D.C., 1986

    Google Scholar 

  5. Chandler, J.P. Cultivation of Mammalian Cells in Serum-free Medium, ABL, 18–28, January, 1990.

    Google Scholar 

  6. Griffiths, B. Perfusion Systems for Cell Cultivation. p. 217–250. In: A.S. Lubiniecki (ed.), Large-Scale Mammalian Cell Culture Technology, Marcel Dekker, Inc., New York, N.Y., 1990.

    Google Scholar 

  7. Feder, J. Development of Large-Scale Mammalian Cell Perfusion Culture Systems. p. 454–483. In: R.E. Spier and J.B. Griffiths (ed.), Modern Approaches to Animal Cell Technology, Butterworth, London, 1987.

    Chapter  Google Scholar 

  8. Feder, J. The Development of Large-Scale Animal Cell Perfusion Culture Systems. p. 125–152. In: A. Mizrahi (ed.), Upstream Processes: Equipment and Techniques, Alan R. Liss, Inc., New York, N.Y., 1988.

    Google Scholar 

  9. Oka, M.S. and R.G. Rupp. Large-Scale Animal Cell Culture: A Biological Perspective. p. 71–92. In: A.S. Lubiniecki (ed.), Large-Scale Mammalian Cell Culture Technology, Marcel Dekker, Inc., New York, N.Y., 1990.

    Google Scholar 

  10. Glacken, M.W., Fleischaker, R.J., and A.J. Sinskey. Large-scale Production of Mammalian Cells and Their Products: Engineering Principles and Barriers to Scale-up. Annals of the New York Academy of Sciences, 413: 355–372, 1983.

    Article  CAS  Google Scholar 

  11. Van Brunt, I. Artificial Organs From Culture, Bio/Technology, 9: 136–137, 1991.

    Google Scholar 

  12. Naughton, G.K., Jacob, L., and Naughton, B.A., A Physiological Skin Model for in vitro Toxicity Studies. In: Alternative Methods in Toxicology, Vol. 7. In Vitro Toxicology: New Directions, A.M. Goldberg, Editor, Mary Ann Liebert Publishers, New York, 1989, pp. 183–189.

    Google Scholar 

  13. Naughton, B.A., Sibanda, B., and Naughton, G.K. Long-Term Liver Cell Cultures as Potential Substrates for Toxicity Assessment. In: In Vitro Toxicology: Mechanisms and New Technology, A.M. Goldberg, Editor, Mary Ann Liebert Publishers, New York, 1991, pp. 193–202.

    Google Scholar 

  14. Naughton, B.A., Preti, R.A., and Naughton, G.K., Hematopoiesis on Nylon Mesh Templates. I. Long-term Culture of Rat Bone Marrow Cells, J. Med. 18:219–250, 1987.

    CAS  Google Scholar 

  15. Naughton, B.A., and Naughton G.K., Modulation of Long-term Bone Marrow Culture by Stromal Support Cells, In: Cellular and Molecular Controls of Hematopoiesis, D. Orlic, ed., Ann. N.Y. Acad. Sci. 554:125–140, 1989.

    CAS  Google Scholar 

  16. Naughton, B.A., Jacob, L., and Naughton G.K., A Three-dimensional Culture System for the Growth of Hematopoietic Cells, Progs. Clin., Biol. Res., Vol. 333: Bone marrow processing and purging, S. Gross, A.P. Gee, and D.A. Worthington-White, eds, 435–445, 1990.

    Google Scholar 

  17. Horvath, B.E. Mammalian Cell Culture Scale-Up: Is Bigger Better?, Bio/Technology, 7: 468–469, 1989.

    Google Scholar 

  18. Tharakan, J.P., Galllagher, S.L., and P.C. Chau. Hollow-Fiber Bioreactors for Mammalian Cell Culture. p. 153–184. In: A. Mizrahi (ed.), Upstream Processes: Equipment and Techniques, Alan R. Liss, Inc., New York, N.Y., 1988.

    Google Scholar 

  19. Knazek, R.A. Solid Tissue Masses Formed In Vitro from Cells Cultured on Artificial Capillaries, Federation Proceedings, 33(8): 1978–1981, 1974.

    CAS  Google Scholar 

  20. Knight, P. Hollow Fiber Bioreactors for Mammalian Cell Culture, Bio/Technology, 7: 459–461, 1989.

    CAS  Google Scholar 

  21. Gallagher, S.L., Tharakan, J.T., and P.C. Chau. An Intercalated-Spiral Wound Hollow Fiber Bioreactor for the Culture of Mammalian Cells, Biotechnology Techniques, 1(2): 91–96, 1987.

    Article  CAS  Google Scholar 

  22. Tharakan, J.P. and P.C. Chau. A Radial Flow Hollow Fiber Bioreactor for the Large-Scale Culture of Mammalian Cells, Biotechnology and Bioengineering, 18: 329–342, 1986.

    Article  Google Scholar 

  23. Lim, F. and R.D. Moss. Microencapsulation of Living Cells and Tissues, Journal of Pharmaceutical Sciences, 70(4): 351–354, 1981.

    Article  CAS  Google Scholar 

  24. Nilsson, K., Scheirer, W., Merten, O.W. Ostberg, L., Liehl, E., Katinger, H.W.D. and K. Mosbach. Entrapment of Animal Cells for Production of Monoclonal Antibodies and Other Biomolecules, Nature, 302: 629–630, 1983.

    Article  CAS  Google Scholar 

  25. Lim, F. Microencapsulation of Living Mammalian Cells, Advances in Biotechnological Processes, 7:185–197, 1988.

    CAS  Google Scholar 

  26. Tyler, J.E. Microencapsulation of Mammalian Cells. p. 343–361. In: A.S. Lubiniecki (ed.), Large-Scale Mammalian Cell Culture Technology, Marcel Dekker, Inc., New York, N.Y., 1990.

    Google Scholar 

  27. Runstadler Jr., P.W., Tung, A.S., Hayman, E.G., Ray, N.G., Sample, J.V.G., and D.E. DeLucia. Continuous Culture with Macroporous Matrix, Fluidized Bed Systems. p. 363–391. In: A.S. Lubiniecki (ed.), Large-Scale Mammalian Cell Culture Technology, Marcel Dekker, Inc., New York, N.Y., 1990.

  28. Seaver, S.S. Culture Method Affects Antibody Secretion of Hybridoma Cells. p. 49–71. In S.S. Seaver (ed.), Commercial Production of Monoclonal Antibodies: A Guide for Scale-up, Marcel Dekker, Inc., New York, N.Y., 1987.

    Google Scholar 

  29. Fleischmajer, R., P. Cantard; Collagen and Microfibril Formation in Three-Dimensional Mesh Dermal Fibroblast Cultures, J. Invest. Derm., 96(4):540, 1991.

    Google Scholar 

  30. Audus, K.L., Bartel, R.L., Hidalgo, I.J., Borchardt, R.T.; The Use of Cultured Epithelial and Endothelial Cells for Drum Transport and Metabolism Studies, Pharm Res, 1990, 7:435–451

    Article  CAS  Google Scholar 

  31. Slivka, S.R., Landeen, L.L., Donelly, T., Zimber, M., Naughton, G.K., Bartel, R.L.; Characterization of a Three-Dimensional Co-culture of Neonatal Human Fibroblasts and Keratinocytes. J. Invest. Derm., 96(4):554, 1991.

    Google Scholar 

  32. Borenfreund, D., Puerner, J.; A Simple Quantitative Procedure Using Monolayer Cultures for Cytotoxicity Assays (HTD/NR-90). T. Tissl. Cult. Meth. 9(1):7–9, 1984.

    Article  Google Scholar 

  33. Hansbrough, J.F., Cooper, M.L., Cohen, R. Spielvogel, R., Greenleaf, G., Bartel, R., and Naughton, G. Evaluation of a Biodegradable Matrix Containing Cultured Human Fibroblasts as a Dermal Replacement Beneath Meshed Skin Grafts on Athymic Mice, Surgery 110: 1991.

  34. Cooper, M.L., Hansbrough, J.F., Spielvogel, R.L., Cohen, R., Bartel, R.L., Naughton, G. In Vivo Optimization of a Living Dermal Substitute Employing Cultured Human Fibroblasts on a Biodegradable Polyglycolic Acid or Polyglactin Mesh, Biomaterials, 12: 243–248, 1991.

    Article  CAS  Google Scholar 

  35. Cohen, R., Zimber, M., Hansbrough, J.F., Fung, Y.C., Debes, J., Skalak, R. Tear Strength Properties of a Novel Cultured Dermal Tissue Model, Presented at Biomedical Engineering Conference, October 11-14, 1991, Charlottesville, VA.

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Authors and Affiliations

  1. Advanced Tissue Sciences, La Jolla, CA, USA

    Craig Halberstadt, Phil Anderson, Ronnda Bartel, Ron Cohen & Gail Naughton

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  1. Craig Halberstadt
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  2. Phil Anderson
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  3. Ronnda Bartel
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  4. Ron Cohen
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  5. Gail Naughton
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Halberstadt, C., Anderson, P., Bartel, R. et al. Physiological Cultured Skin Substitutes for Wound Healing. MRS Online Proceedings Library 252, 323–330 (1991). https://doi.org/10.1557/PROC-252-323

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