Fibroblastic Subpopulations in Uninjured and Wounded Rabbit Oral Mucosa

Abstract

Fibroblast cultures derived from uninjured and reparative rabbit buccal mucosa were compared in terms of extracellular glycosaminoglycan (GAG) content and cellular response to interleukin-1 (IL-1). Under identical growth conditions, proliferation of both cell lines was the same. Both lines incorporated [³H]-glucosamine into GAG in cellular, pericellular, and medium fractions, with the majority of incorporated label residing in the medium. Dermatan sulfate (DS) was the predominant GAG in the medium fraction of both normal and wound fibroblast cultures; however, the two cell lines differed in the identity of the medium fraction's secondary GAG: chondroitin sulfate (CS) for normal fibroblasts and hyaluronic acid (HA) for wound-derived cells. The GAG content of the pericellular matrix for all cultures was the same regardless of the tissue of origin: heparan sulfate (HS) accompanied by a very small amount of CS. Exposure to IL-1 produced limited but highly specific effects: It was not mitogenic for either cell line but did cause a quantitative change (increase) in overall incorporation into GAG for medium and pericellular fractions for both cell lines. Further, IL-1 induced a qualitative change in GAG composition for normal mucosal fibroblastic medium fractions by causing the synthesis/release of heparan sulfate (HS) and a variant form of DS. These data support the hypothesis that different fibroblastic substrains can populate a given oral site as a function of variables such as injury andlor healing status.

Get full access to this article

View all access options for this article.

References

Bertolami, C.N. (1982): Oral Wound Repair. In: Clinical Dentistry, Vol. 3, J.W. Clark , Ed., Philadelphia: Harper & Row, pp. 1-13.

Bordin, S. ; Page, R.C. ; and Narayanan, A.S. (1984): Heterogeneity of Normal Diploid Fibroblasts: Isolation and Characterization of One Phenotype, Science 223:171-173.

Botstein, G.R. ; Sherer, G.K. ; and Leroy, E.C. (1982): Fibroblast Selection in Scleroderma, Arthritis Rheum 25:189-195.

Brinckerhoff, C.E. ; McMillan, R.M. ; Fahey, J.V. ; and Harris, E.D., Jr. (1979): Collagenase Production by Synovial Fibroblasts Treated with Phorbol Myristate Acetate, Arthritis Rheum 22:1109-1116.

Brinckerhoff, C.E. and Nagel, J.E. (1981): Collagenase Production by Cloned Populations of Rabbit Synovial Fibroblasts, Coll Rel Res 1:433-444.

Bronson, R.E. ; Argenta, J.G. ; and Bertolami, C.N. (1988): Interleukin-1-induced Changes in Extracellular Glycosaminoglycan Composition of Cutaneous Scar-derived Fibroblasts in Culture , Coll Rel Res 8:199-208.

Bronson, R.E. ; Bertolami, C.N. ; and Siebert, E.P. (1987): Modulation of Fibroblast Growth and Glycosaminoglycan Synthesis by Interleukin-1, Coll Rel Res 7:323-332.

Burton, K. (1956): A Study of the Conditions and Mechanism of the Diphenylamine Reaction for the Colorimetric Estimation of DNA, Biochem J 62:315-322.

Cappelletti, R. ; Del Rosso, M. ; and Chiarugi, V.P. (1979a): Rapid Multisample Separation of the Five Most Widespread Animal Glycosaminoglycans, Anal Biochem 93:37-40.

10.

Cappelletti, R. ; DEL Rosso, M. ; and Chiarugi, V.P. (1979b): A New Electrophoretic Method for the Complete Separation of All Known Glycosaminoglycans in a Monodimensional Run, Anal Biochem 99:311-315.

11.

Diegelmann, R.F. ; Cohen, I. ; and McCoy, B. (1979): Growth Kinetics and Collagen Synthesis of Normal Skin, Normal Scar and Keloid Fibroblasts in vitro, Cell Physiol 98:341-346.

12.

Dinarello, C.A. (1984): Interleukin-1 and the Pathogenesis of the Acute-Phase Response, New Eng J Med 311:1413-1418.

13.

Goldstein, S. ; Niewiarowski, S. ; and Singal, D.P. (1975): Pathological Implications of Cell Aging in vitro , Fed Proc 34 :56-63.

14.

Grotendorst, G.R. ; Pencev, D. ; Martin, G.R. ; and Sodek, J. (1984): Molecular Mediators of Tissue Repair. In: Soft and Hard Tissue Repair; Biological and Clinical Aspects, T.K. Hunt , R.B. Heppenstall , E. Pines , and D. Rovee , Eds., New York: Praeger Publishers, pp. 20-40.

15.

Hamerman, D. ; Smith, C. ; Keiser, H. ; and Craig, R. (1982): Glycosaminoglycans Produced by Human Synovial Cell Cultures, Coll Rel Res 2:313-329.

16.

Hassell, T.M. and Gilbert, G.H. (1983): Phenytoin Sensitivity of Fibroblasts as the Basis for Susceptibility to Gingival Enlargement, Am J Pathol 112:218-223.

17.

Hassell, T.M. ; Page, R.C. ; Narayanan, A.S. ; and Cooper, C.G. (1976): Diphenyl-hydantoin (Dilantin) Gingival Hyperplasia: Drug-induced Abnormality of Connective Tissue, Proc Natl Acad Sci USA 73:2909-2912.

18.

Hassell, T.M. and Stanek, E.J., III (1983): Evidence that Healthy Human Gingiva Contains Functionally Heterogeneous Fibroblast Subpopulations, Arch Oral Biol 28:617-625.

19.

Laato, M. and Heino, J. (1988): Interleukin-1 Modulates Collagen Accumulation by Rat Granulation Tissue Cells both in vivo and in vitro, Experientia 44:32-34.

20.

Libby, P. ; Wyler, D.J. ; Janicka, M.W. ; and Dinarello, C.A. (1985): Differential Effects of Human Interleukin-1 on Growth of Human Fibroblasts and Vascular Smooth Muscle Cells, Arteriosclerosis 5:186-191.

21.

Mizel, S.B. ; Oppenheim, J.J. ; and Rosenstreich, D.L. (1978): Characterization of Lymphocyte-activating Factor (LAF) Produced by the Macrophage Cell Line, P388D1. Enhancement of LAF Production by Activated T-Lymphocytes, J Immunol 120 :1497-1503.

22.

Ohya, T. and Kaneko, Y. (1970): A Novel Hyaluronidase from Streptomyces, Biochim Biophys Acta 198:607-609.

23.

Roden, L. ; Baker, J.R. ; Cifonelli, J.A. ; and Mathews, M. (1972): Isolation and Characterization of Connective Tissue Polysaccharidcs, Meth Enzymol 78:73-140.

24.

Ross, R. (1968): The Fibroblast and Wound Repair, Biol Rev 43 :51-94.

25.

Savage, K.E. and Swann, D.A. (1985): A Comparison of Glycosaminoglycan Synthesis by Human Fibroblasts from Normal Skin, Normal Scar and Hypertrophic Scar, J Invest Dermatol 84 :521-526.

26.

Schmidt, J.A. ; Mizel, S.B. ; Cohen, D. ; and Green, I. (1982): Interleukin 1, A Potential Regulator of Fibroblast Proliferation, J Immunol 128:2177-2182.

27.

Shoshan, S. (1981): Wound Healing , Int Rev Connect Tissue Res 9:1-26.

28.

Stiles, C.D. ; Capone, G.T. ; Scher, C.D. ; Antoniades, H.N. ; Van Wyk, J.J. ; and Pledger, W.J. (1979): Dual Control of Cell Growth by Somatomedins and Platelet-derived Growth Factor, Proc Natl Acad Sci USA 76:1279-1283.

29.

Toole, B.P. (1976): Morphogenetic Role of Glycosaminoglycans (Acid Mucopolysaccharides) in Brain and Other Tissues. In: Neuronal Recognition , S.H. Barondes , Ed., New York: Plenum Press, pp. 275-329.

30.

Toole, B.P. (1982): Glycosaminoglycans in Morphogcnesis. In: Cell Biology of the Extracellular Matrix, E.D. Hay , Ed., New York: Plenum Press , pp. 259-294.

31.

Whiteside, T.L. ; Worrall, J.G. ; Prince, R.K. ; Buck-Ingham, R.B. ; and Rodnan, G. (1985): Soluble Mediators From Mononuclear Cells Increase the Synthesis of Glycosaminoglycan by Dermal Fibroblast Cultures Derived From Normal Subjects and Progressive Systemic Sclerosis Patients, Arthritis Rheum 28:188-197.

32.

Yamagata, T. ; Saito, H. ; Habuchi, O. ; and Suzuki, S. (1968): Purification and Properties of Bacterial Chondroitinases and Chondrosulfatases, J Biol Chem 243:1523-1535.