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The Expression of the Fibrillar Collagen Genes During Fracture Healing: Heterogeneity of the Matrices and Differentiation of the Osteoprogenitor Cells

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Abstract

The cells that express the genes for the fibrillar collagens, types I, II, III and V, during callus development in rabbit tibial fractures healing under stable and unstable mechanical conditions were localized. The fibroblast-like cells in the initial fibrous matrix express types I, III and V collagen mRNAs. Osteoblasts, and osteocytes in the newly formed membranous bone under the periosteum, express the mRNAs for types I, III and V collagens, but osteocytes in the mature trabeculae express none of these mRNAs. Cartilage formation starts at 7 days in calluses forming under unstable mechanical conditions. The differentiating chondrocytes express both types I and II collagen mRNAs, but later they cease expression of type I collagen mRNA. Both types I and II collagens were located in the cartilaginous areas. The hypertrophic chondrocytes express neither type I, nor type II, collagen mRNA. Osteocalcin protein was located in the bone and in some cartilaginous regions. At 21 days, irrespective of the mechanical conditions, the callus consists of a layer of bone; only a few osteoblasts lining the cavities now express type I collagen mRNA.

We suggest that osteoprogenitor cells in the periosteal tissue can differentiate into either osteoblasts or chondrocytes and that some cells may exhibit an intermediate phenotype between osteoblasts and chondrocytes for a short period. The finding that hypertrophic chondrocytes do not express type I collagen mRNA suggests that they do not transdifferentiate into osteoblasts during endochondral ossification in fracture callus.

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References cited

  • Andrikopoulos K, Suzuki HR, Solursh M, Ramirez F (1992) Localization of pro-α2(V) collagen transcripts in the tissues of the developing mouse embryo. Develop Dyn 195: 113-120.

    Google Scholar 

  • Ashhurst DE (1986) The influence of mechanical conditions on the healing of experimental fractures in the rabbit: a microscopical study. Philos Trans R Soc Lond 313: 271-302.

    Google Scholar 

  • Ashhurst DE (1990) Collagens synthesized by healing fractures. Clin Orthop Rel Res 255: 273-283.

    Google Scholar 

  • Ashhurst DE, Hogg J, Perren SM (1982) A method for making reproducible fractures of the rabbit tibia. Injury 14: 236-242.

    Google Scholar 

  • Benya PD, Brown PD (1986) Modulation of the chondrocyte phenotype in vitro. In Kuettner K, ed. Articular Cartilage Biochemistry. New York: Raven Press, pp. 219-230.

    Google Scholar 

  • Beresford WA (1981) Chondroid Bone, Secondary Cartilage and Metaplasia. Baltimore, Munich: Urban & Schwarzenberg, 454 pp.

    Google Scholar 

  • Bland YS, Ashhurst DE (1996) Changes in the content of the fibrillar collagens and the expression of their mRNAs in the menisci of the rabbit knee joint during development and ageing. Histochem J 28: 265-274.

    Google Scholar 

  • Bland YS, Critchlow MA, Ashhurst DE (1991) Digoxigenin as a probe label for in situ hybridization on skeletal tissues. Histochem J 23: 415-418.

    Google Scholar 

  • Cancedda FD, Gentili C, Manduca P, Cancedda R (1992) Hypertrophic chondrocytes undergo further differentiation in culture. J Cell Biol 117: 427-435.

    Google Scholar 

  • Carter DR, Blenman PR, Beaupre GS (1988) Correlations between mechanical stress history and tissue differentiation in initial fracture healing. J Orthop Res 6: 736-748.

    Google Scholar 

  • Critchlow MA, Bland YS, Ashhurst DE (1994) The effects of age on the response of rabbit periosteal osteoprogenitor cells to exogenous transforming growth factor-β2. J Cell Sci 107: 499-516.

    Google Scholar 

  • Critchlow MA, Bland YS, Ashhurst DE (1995) The expression of collagen mRNAs in normally developing neonatal rabbit long bones and after treatment of neonatal and adult rabbit tibia with transforming growth factor-α2. Histochem J 27: 505-515.

    Google Scholar 

  • Hughes SS, Hicks DG, O'Keefe RJ, Hurwitz SR, Crabb ID, Krasinskas AM, Loveys L, Puzas JE, Rosier RN (1995) Shared phenotypic expression of osteoblasts and chondrocytes in fracture callus. J Bone Min Res 10: 533-544.

    Google Scholar 

  • Hulth A, Johnell O, Lindberg L, Paulsson M, Heinegard D (1990) Demonstration of blood-vessel like structures in cartilaginous callus by anti-laminin and anti-heparan sulfate proteoglycan antibodies. Clin Orthop Rel Res 254: 289-293.

    Google Scholar 

  • Hunziker EB, Schenk RK (1984) Cartilage ultrastructure after high pressure freezing, freeze substitution, and low temperature embedding. II. Intracellular matrix ultrastructure — preservation of proteoglycans in their native state. J Cell Biol 98: 277-282.

    Google Scholar 

  • Hunziker E, Schenk RK (1989) Physiological mechanisms adopted by chondrocytes in regulating longitudinal bone growth in rats. J Physiol 414: 55-71.

    Google Scholar 

  • Kosher RA, Kulyk WM, Gay SW (1986) Collagen gene expression during limb cartilage differentiation. J Cell Biol 102: 1151-1156.

    Google Scholar 

  • Lane JM, Suda M, von der Mark K, Timpl R (1986) Immunofluorescent localization of structural collagen types in endochondral fracture repair. J Orthop Res 4: 381-389.

    Google Scholar 

  • Lee FY-I, Choi YW, Behrens FF, Defouw DO, Einhorn TA (1998) Programmed removal of chondrocytes during endochondral fracture healing. J Orthop Res 16: 144-150.

    Google Scholar 

  • Lian JB, Mckee MD, Todd AM, Gerstenfeld LC (1993) Induction of bone-related proteins, osteocalcin and osteopontin, and their matrix ultrastructural localization with development of chondrocyte hypertrophy in vitro. J Cell Biochem 52: 206-219.

    Google Scholar 

  • Mäkelä JR, Raassina M, Virta A, Vuorio E (1988) Human proα1(I) collagen: cDNA sequence for the C-propeptide domain. Nucleic Acids Res 16: 349.

    Google Scholar 

  • Metsäranta M, Kujala UM, Pellimemi L, Sterman H, Aho H, Vuorio E (1996) Evidence for insufficient chondrocytic differentiation during repair of full-thickness defects of articular cartilage. Matrix Biol 15: 39-47.

    Google Scholar 

  • Mizoguchi I, Nakamura M, Takahashi I, Sasano Y, Kagayama M, Mitani H (1993) Presence of chondroid bone on rat mandibular condylar cartilage. An immunohistochemical study. Anat Embryol 187: 9-15.

    Google Scholar 

  • Page M, Hogg J, Ashhurst DE (1986) The effects of mechanical stability on the macromolecules of the connective tissue matrices produced during fracture healing. I. The collagens. Histochem J 18: 251-265.

    Google Scholar 

  • Roach HI (1997) New aspects of endochondral ossification in the chick: chondrocyte apoptosis, bone formation by former chondrocytes, and acid phosphatase activity in endochondral bone matrix. J Bone Min Res 12: 795-805.

    Google Scholar 

  • Sandberg M, Vuorio E (1987) Localization of types I, II and III collagen mRNAs in developing human tissues by in situ hybridization. J Cell Biol 104: 1077-1084.

    Google Scholar 

  • SandbergM, Aro H, Multimäki P, Aho H, Vuorio E (1989) In situ localization of collagen production by chondrocytes and osteoblasts in fracture callus. J Bone Joint Surg 71A: 69-77.

    Google Scholar 

  • Sandell LJ, Sugai JV, Trippel SB (1994) Expression of collagens I, II, X amd XI and aggrecan mRNAs by bovine growth plate chondrocytes in situ. J Orthop Res 12: 1-14.

    Google Scholar 

  • Scammell BE & Roach HI (1996) A new role for the chondrocyte in fracture repair: endochondral ossification includes direct bone formation by former chondrocytes. J Bone Min Res 11: 737-745.

    Google Scholar 

  • Schenk R, Willenegger H (1967) Morphological findings in primary fracture healing. Symp Biol Hung 7: 75-86.

    Google Scholar 

  • Silver MH, Foidart J-M, Pratt RM (1981) Distribution of fibronectin and collagen during mouse limb and palate development. Differentiation 18: 141-149.

    Google Scholar 

  • Simmons DJ, Kahn AJ (1979) Cell lineage in fracture healing in chineric bone grafts. Calcif Tissue Int 27: 247-253.

    Google Scholar 

  • Singley CT, Solursh M (1981) The spatial distribution of hyaluronic acid and mesenchymal condensation in the embryonic chick wing. Develop Biol 84: 102-120.

    Google Scholar 

  • Stafford HJ, Roberts MT, Oni OOA, Hay J, Gregg P (1994) Localization of bone-forming cells during fracture healing by osteocalcin immunocytochemistry: an experimental study of the rabbit tibia. J Orthop Res 12: 29-39.

    Google Scholar 

  • Strauss PG, Closs EI, Schmidt J, Erfle V (1990) Gene expression during osteogenic differentiation in mandibular condyles in vitro. J Cell Biol 110 1369-1378.

    Google Scholar 

  • Thesingh CW, Groot CG, Wassenaar AM (1991) Transdifferentiation of hypertrophic chondrocytes into osteoblasts in murine fetal metatarsal bones induced by co-cultured cerebrum. Bone Mineral 12: 25-40.

    Google Scholar 

  • Yamazaki M, Majeska RJ, Yoshioka H, Moriya H, Einhorn TA (1997) Spatial and temporal expression of fibril-forming minor collagen genes (types V and XI) during fracture healing. J Orthop Res 15: 757-764.

    Google Scholar 

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Authors and Affiliations

  1. Department of Anatomy, St. George's Hospital Medical School, ooting, London, SW17 0RE, UK

    Yvette S. Bland, Matthew A. Critchlow & Doreen E. Ashhurst

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  1. Yvette S. Bland
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  2. Matthew A. Critchlow
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  3. Doreen E. Ashhurst
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Bland, Y.S., Critchlow, M.A. & Ashhurst, D.E. The Expression of the Fibrillar Collagen Genes During Fracture Healing: Heterogeneity of the Matrices and Differentiation of the Osteoprogenitor Cells. Histochem J 31, 797–809 (1999). https://doi.org/10.1023/A:1003954104290

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