Production of Matrix-degrading Enzymes and Inhibition of Osteoclast-like Cell Differentiation by Fibroblast-like Cells from the Periodontal Ligament of Human Primary Teeth

Abstract

Clinically, the most apparent difference between the primary and permanent dentitions is the physiologic loss of the primary tooth by root resorption. Root resorption is associated with loss of integrity of the periodontal ligament (PDL), followed by recruitment of resorptive cells that remove root structure. We therefore cultured primary dentition PDL fibroblasts (PPDL cells) to investigate in vitro their production of matrix metalloproteinases (MMPs) and tissue inhibitors of MMP (TIMPs), and the effects of soluble factors produced by these cells on osteoclast-like cell differentiation. These studies demonstrate that PPDL cells in vitro have a heterogeneous morphology, and they constitutively synthesize 92-kDa gelatinase, 72-kDa gelatinase, and 53/57-kDa procollagenase as well as TIMP-1, -2, and a third inhibitor of matrix metalloproteinase, as determined by substrate gel zymography and immunoblot analysis. Compared with PDL cells from the permanent dentition, PPDL cells generally produced a greater amount of collagenase but similar amounts of the gelatinases and inhibitors. PPDL cells were treated with pro-inflammatory cytokines to determine their effect on the expression of matrix-degrading enzymes and inhibitors. Interleukin-la and tumor necrosis factor-a enhanced the constitutive expression of proteinases but not that of inhibitors in PPDL cells. Conditioned media from PPDL cell lines inhibited the differentiation of osteoclastlike cells in mouse bone marrow cultures. These findings indicate that PPDL cells may modulate the cascade of root resorption both by their regulated production of proteinases and inhibitors and by synthesis of unknown soluble factor(s) that may regulate osteoclast development.

Keywords

root resorption periodontal ligament fibroblast matrix metalloproteinase tissue inhibitor of metalloproteinase osteoclast.

References

Adams AM , Soames JV , Searle RF (1993). Cultural and morphological characteristics of human periodontal ligament cells in vitro. Arch Oral Biol 38:657-662.

Andreasen JO (1988). Review of root resorption systems and models. Etiology of root resorption and the homeostatic mechanisms of the periodontal ligament. In: Biological mechanisms of tooth eruption and root resorption. Davidovitch Z , editor. Birmingham: EBSCO Media, pp. 9-22.

Aukhil I. , Simpson DM , Schaberg TV (1983). An experimental study of new attachment procedure in beagle dogs. J Periodont Res 18:643-654.

Banda MJ , Howard EW , Herron GS , Apodaca G. (1992). Secreted inhibitors of metalloproteinases (IMPs) that are distinct from TIMP. Matrix (Suppl 1):294-298.

Birkedal-Hansen H. , Lin HY , Birkedal-Hansen B. , Windsor LJ , Pierson MC (1992). Degradation of collagen fibrils by live cells: role of expression and activation of procollagenase. Matrix (Suppl 1):368-374.

Brunette DM , Kanoza RJ , Marmary T. , Chan J. , Melcher AH (1977). Interactions between epithelial and fibroblast-like cells in cultures derived from monkey periodontal ligament. J Cell Sci 27:127-140.

Davidovitch Z. , Lynch P. , Shanfeld JL (1988). Immunohistochemical localization of interleukins in dental and paradental cells during tooth eruption and root resorption in kittens. In: Biological mechanisms of tooth eruption and root resorption. Davidovitch Z , editor. Birmingham: EBSCO Media, pp. 355-364.

Dayer JM , Beutler B. , Cerami A. (1985). Cachectin/tumor necrosis factor stimulates collagenase and prostaglandin E2 production by human synovial cells and dermal fibroblasts . J Exp Med 162:2163-2168.

Delaisse JM , Eeckhout Y. , Vaes G. ( 1988). Bone-resorbing agents affect the production and distribution of procollagenase as well as the activity of collagenase in bone tissue. Endocrinology 123:264-276.

10.

Fisher S. , Werb Z. ( 1991). Techniques for studying the catabolism of extracellular matrix components. In: Extracellular matrix molecules. A practical approach. Haralson MA , Hassell JR , editors. Oxford: IPL Press, pp. 261-286.

11.

Giniger MS , Norton L. , Sousa S. , Lorenzo JA , Bronner F. (1991). A human periodontal ligament fibroblast clone releases a bone resorption inhibition factor in vitro. J Dent Res 70:99-101.

12.

Gorski JP , Marks SC Jr (1992). Current concepts of the biology of tooth eruption . Crit Rev Oral Biol Med 3:185-206.

13.

Herron GS , Banda MJ , Clark EJ , Gavrilovic J. , Werb Z. ( 1986). Secretion of metalloproteinases by stimulated capillary endothelial cells. II. Expression of collagenase and stromelysin activities is regulated by endogenous inhibitors. J Biol Chem 261:2814-2818.

14.

Kapila S. , Lee C. , Richards D. (1995). Characterization and identification of proteinases and proteinase inhibitors synthesized by temporomandibular joint disc cells . J Dent Res 74:1228-1236.

15.

Kapila YL , Kapila S. , Johnson PW (1996). Fibronectin and fibronectin fragments modulate the expression of proteinases and proteinase inhibitors in human periodontal ligament cells. Matrix Biol 15:251-261.

16.

Karring T. , Nyman S. , Lindhe J. , Sirirat M. (1984). Potentials for root resorption during periodontal wound healing. J Clin Periodontol 11:41-52.

17.

Meikle MC , Heath JK , Atkinson SJ , Hembry RM , Reynolds JJ (1989). Molecular biology of stressed connective tissues at sutures and hard tissues, in vitro. In: The biology of tooth movement. Norton LA , Burstone CJ , editors. Boca Raton, FL: CRC Press, Inc., pp. 71-85.

18.

Minkin C. (1982). Bone acid phosphatase: tartrate-resistant acid phosphatase as a marker of osteoclast function. Calcif Tissue Int 34:285-290.

19.

Morita H. , Yamashiya H. , Shimizu M. , Sasaki S. (1970). The collagenolytic activity during root resorption of bovine deciduous tooth. Arch Oral Biol 15:503-508.

20.

Nicholson GC , Moseley JM , Sexton PM , Mendelsohn FA , Martin TJ (1986). Abundant calcitonin receptors in isolated rat osteoclasts. J Clin Invest 78:355-360.

21.

Nijweide PJ , van Iperen-van Gent AS , Kawilarang-de Haas EW , van der Plas A. , Wassenaar AM (1982). Bone formation and calcification by isolated osteoblast-like cells. J Cell Biol 93:318-323.

22.

Okamura T. , Shimokawa H. , Takagi Y. , Ono H. , Sasaki S. (1993). Detection of collagenase mRNA in odontoclasts of bovine root-resorbing tissue by in situ hybridization. Calcif Tissue Int 52:325-330.

23.

Otsuka K. , Pitaru S. , Overall CM , Aubin JE , Sodek J. (1988). Biochemical comparison of fibroblast populations from different periodontal tissues: characterization of matrix protein and collagenolytic enzyme synthesis. Biochem Cell Biol 66:167-176.

24.

Pfeilschifter J. , Chenu C. , Bird A. , Mundy GR , Roodman GD (1989). Interleukin-1 and tumor necrosis factor stimulate the formation of human osteoclastlike cells in vitro. J Bone Miner Res 4:113-118.

25.

Postlethwaite AE , Lachman LB , Mainardi CL , Kang AH ( 1983). Interleukin 1 stimulation of collagenase production by cultured fibroblasts. J Exp Med 157:801-806.

26.

Richards D. , Rutherford RB (1990). Interleukin-1 regulation of procollagenase mRNA and protein in periodontal fibroblasts in vitro. J Periodont Res 25:222-229.

27.

Rifas L. , Halstead LR , Peck WA , Avioli LV , Welgus HG (1989). Human osteoblasts in vitro secrete tissue inhibitor of metalloproteinases and gelatinase but not interstitial collagenase as major cellular products. J Clin Invest 84:686-694.

28.

Rowe DJ , Leung WW , Del Carlo DL (1996). Osteoclast inhibition by factors from cells associated with regenerative tissue. J Periodont 67:414-421.

29.

Rygh P. ( 1977). Orthodontic root resorption studies by electron microscopy . Angle Orthod 47:1-16.

30.

Sakamoto S. , Sakamoto M. (1984). Osteoblast collagenase: collagenase synthesis by clonally derived mouse osteogenic (MC3T3-E1) cells. Biochem Int 9:51-58.

31.

Sasaki T. , Takahashi N. , Watanabe C. , Suzuki H. , Higashi S. , Suda T. ( 1988). Cytodifferentiation of odontoclasts in physiologic root resorption of human deciduous teeth. In: Biological mechanisms of tooth eruption and root resorption. Davidovitch, Z , editor. Birmingham: EBSCO Media, pp. 321-328.

32.

Takahashi N. , Yamana H. , Yoshiki S. , Roodman GD , Mundy GR , Jones SJ , et al. (1988). Osteoclast-like cell formation and its regulation by osteotropic hormones in mouse bone marrow cultures. Endocrinology 122:1373-1382.

33.

Tronstad L. (1988). Root resorption-a multidisciplinary problem in dentistry. In: Biological mechanisms of tooth eruption and root resorption. Davidovitch, Z , editor. Birmingham: EBSCO Media, pp. 293-301.

34.

Werb Z. ( 1989). Proteinase and matrix degradation. In: Textbook of rheumatology. 3rd ed. Kelly WN , Harris ED Jr , Ruddy S , Sledge CB , editors. Philadelphia: Saunders, pp. 300-321.