Expression of β1 integrins during periosteal chondrogenesis

Abstract Objective The interactions between integrins and extracellular matrix proteins are known to modulate cell behavior, and may be involved in regulating cartilage formation and repair. The purpose of this study was to determine the patterns and localization of expression of the β1 integrins during cartilage formation by periosteum, which is used to repair articular cartilage. Design Periosteal explants from 2-month-old rabbit medial proximal tibiae were cultured in agarose suspension for 0 to 6 weeks, with 10ng/ml transforming growth factor-β1 added for the first 2 days of culture. Integrin expressions were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and localized by immunohistochemistry. Results Normal periosteum expressed the α1, α3, α5, β1 subunits at low levels, and the proteins for all but the α3 subunits were identified by immunohistochemistry in the periosteum. Significant two- to five-fold up-regulation of the mRNA expression of the α1, α3, α5 and β1 integrin subunits during the early proliferative stage of chondrogenesis was observed. The initial change was a five-fold increase in α5 expression on day 2 and a two-fold increase in α3 expression. On day 5, α1 expression was up-regulated (four-fold). β1 expression was broadly up-regulated (three to four-fold) from day 5 to 14. In the early stage of chondrocyte differentiation, after day 14, α1 expression was down-regulated, while there was upregulation of α3 (three-fold), α5 (three-fold) and β1 (four-fold) expressions. Thereafter, α1 expression was down-regulated, while α3, α5 and β1 expressions were up-regulated again during matrix synthesis. Immunohistochemistry confirmed this late decrease in α1 levels and increase in α3, α5 and β1 levels in chondrocytes. Conclusions These observations indicate that the β1 integrins play an important role in the process of chondrogenesis in periosteum.

[1]  J. W. Tawil,et al.  Molecular cloning of the rat integrin alpha 1-subunit: a receptor for laminin and collagen , 1990, The Journal of cell biology.

[2]  R. Loeser Growth factor regulation of chondrocyte integrins. Differential effects of insulin-like growth factor 1 and transforming growth factor beta on alpha 1 beta 1 integrin expression and chondrocyte adhesion to type VI collagen. , 1997, Arthritis and rheumatism.

[3]  E. Ruoslahti,et al.  Amino acid sequence of the human fibronectin receptor , 1987, The Journal of cell biology.

[4]  W. Kulyk,et al.  Promotion of embryonic chick limb cartilage differentiation by transforming growth factor-beta. , 1989, Developmental biology.

[5]  S A Newman,et al.  The mechanism of precartilage mesenchymal condensation: a major role for interaction of the cell surface with the amino-terminal heparin-binding domain of fibronectin. , 1989, Developmental biology.

[6]  D. Rosen,et al.  Purification and characterization of two cartilage-inducing factors from bovine demineralized bone. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[7]  R. Loeser Growth factor regulation of chondrocyte integrins: Differential effects of insulin-like growth factor 1 and transforming growth factor β on α1β1 integrin expression and chondrocyte adhesion to type VI collagen , 1997 .

[8]  C. J. Campbell The healing of cartilage defects. , 1969, Clinical orthopaedics and related research.

[9]  R Cancedda,et al.  Changes in the expression of collagen genes show two stages in chondrocyte differentiation in vitro , 1988, The Journal of cell biology.

[10]  O. Korkala Periosteal primary resurfacing of joint surface defects of the patella due to injury. , 1988, Injury.

[11]  J. Lauritzen,et al.  Glued periosteal grafts in the knee. , 1985, Acta orthopaedica Scandinavica.

[12]  J. Lupton,et al.  Rapid competitive PCR determination of relative gene expression in limiting tissue samples. , 1996, Clinical chemistry.

[13]  S. O’Driscoll,et al.  Relationship of donor site to chondrogenic potential of periosteum in vitro , 1994, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[14]  D. Salter,et al.  Integrin expression by human articular chondrocytes. , 1992, British journal of rheumatology.

[15]  A. Caplan,et al.  Morphology of bone development and bone remodeling in embryonic chick limbs. , 1986, Bone.

[16]  B. Powers,et al.  Neochondrogenesis in free intra-articular, periosteal, and perichondrial autografts in horses. , 1989, American journal of veterinary research.

[17]  V. Hoikka,et al.  Reconstruction of the patellar articulation with periosteal grafts. 4-year follow-up of 13 cases. , 1990, Acta orthopaedica Scandinavica.

[18]  S W O'Driscoll,et al.  Periosteum responds to dynamic fluid pressure by proliferating in vitro , 1999, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[19]  S A Newman,et al.  Role of transforming growth factor-beta in chondrogenic pattern formation in the embryonic limb: stimulation of mesenchymal condensation and fibronectin gene expression by exogenenous TGF-beta and evidence for endogenous TGF-beta-like activity. , 1991, Developmental biology.

[20]  M. Bolander,et al.  Gene digging , 1998, Molecular biotechnology.

[21]  M. Shakibaei,et al.  Changes in integrin expression during chondrogenesis in vitro: an immunomorphological study. , 1995, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[22]  A. Ham A HISTOLOGICAL STUDY OF THE EARLY PHASES OF BONE REPAIR , 1930 .

[23]  F M Watt,et al.  Regulation of development and differentiation by the extracellular matrix. , 1993, Development.

[24]  R. Cancedda,et al.  Regulated expression of fibronectin, laminin and related integrin receptors during the early chondrocyte differentiation. , 1997, Journal of cell science.

[25]  K. Sekiguchi,et al.  Up-Regulation of Integrin α5β1 Expression by Interleukin-6 in Rabbit Corneal Epithelial Cells , 1995 .

[26]  Y. Takada,et al.  Molecular cloning and expression of the cDNA for alpha 3 subunit of human alpha 3 beta 1 (VLA-3), an integrin receptor for fibronectin, laminin, and collagen , 1991, The Journal of cell biology.

[27]  R. Salter,et al.  The induction of neochondrogenesis in free intra-articular periosteal autografts under the influence of continuous passive motion. An experimental investigation in the rabbit. , 1984, The Journal of bone and joint surgery. American volume.

[28]  V. Ritsilä,et al.  Chondrogenesis in repair of articular cartilage defects by free periosteal grafts in rabbits. , 1982, Acta orthopaedica Scandinavica.

[29]  Stuart A. Newman,et al.  Role of transforming growth factor-β in chondrogenic pattern formation in the embryonic limb: Stimulation of mesenchymal condensation and fibronectin gene expression by exogenenous TGF-β and evidence for endogenous TGF-β-like activity , 1991 .

[30]  D. Boettiger,et al.  Beta 1 integrins mediate chondrocyte interaction with type I collagen, type II collagen, and fibronectin. , 1993, Experimental cell research.

[31]  S W O'Driscoll,et al.  Durability of regenerated articular cartilage produced by free autogenous periosteal grafts in major full-thickness defects in joint surfaces under the influence of continuous passive motion. A follow-up report at one year. , 1988, The Journal of bone and joint surgery. American volume.

[32]  K. von der Mark,et al.  Changes in the patterns of collagens and fibronectin during limb-bud chondrogenesis. , 1980, Journal of embryology and experimental morphology.

[33]  D. Salter,et al.  Altered electrophysiological responses to mechanical stimulation and abnormal signalling through alpha5beta1 integrin in chondrocytes from osteoarthritic cartilage. , 2000, Osteoarthritis and cartilage.

[34]  L. Rome,et al.  Cloning and sequence of the cDNA encoding the rat oligodendrocyte integrin β1 subunit , 1995 .

[35]  Virgil L. Woods,et al.  Integrin expression by human articular chondrocytes. , 1994, Arthritis and rheumatism.

[36]  G. Stein,et al.  Concepts of osteoblast growth and differentiation: basis for modulation of bone cell development and tissue formation. , 1992, Critical reviews in oral biology and medicine : an official publication of the American Association of Oral Biologists.

[37]  Y. Miura Enhancement of periosteal chondrogenesis in vitro , 1994 .

[38]  R. Loeser,et al.  Expression of beta 1 integrins by cultured articular chondrocytes and in osteoarthritic cartilage. , 1995, Experimental cell research.

[39]  S. Goodman,et al.  Localization of beta 1-integrins in human cartilage and their role in chondrocyte adhesion to collagen and fibronectin. , 1993, Experimental cell research.

[40]  Richard O. Hynes,et al.  Integrins: Versatility, modulation, and signaling in cell adhesion , 1992, Cell.

[41]  M. Bolander,et al.  An effective method of completely removing contaminating genomic DNA from an RNA sample to be used for PCR , 1997, Molecular biotechnology.

[42]  W. Horton,et al.  Transforming growth factor‐beta and fibroblast growth factor act synergistically to inhibit collagen II synthesis through a mechanism involving regulatory DNA sequences , 1989, Journal of cellular physiology.

[43]  三浦 恭志 Enhancement of periosteal chondrogenesis in vitro dose-response for transforming growth factor-beta 1 (TGF-β1) , 1996 .

[44]  R. Salter,et al.  The chondrogenic potential of free autogenous periosteal grafts for biological resurfacing of major full-thickness defects in joint surfaces under the influence of continuous passive motion. An experimental investigation in the rabbit. , 1986, The Journal of bone and joint surgery. American volume.

[45]  S. O’Driscoll,et al.  Articular cartilage regeneration using periosteum. , 1999, Clinical orthopaedics and related research.

[46]  B. Toole,et al.  Temporal and spatial transitions in collagen types during embryonic chick limb development. , 1973, Developmental biology.

[47]  R. Briesewitz,et al.  Expression of native and truncated forms of the human integrin alpha 1 subunit. , 1993, The Journal of biological chemistry.

[48]  K. Ostergaard,et al.  Expression of alpha and beta subunits of the integrin superfamily in articular cartilage from macroscopically normal and osteoarthritic human femoral heads. , 1998, Annals of the rheumatic diseases.

[49]  A. Caplan,et al.  Morphological and histochemical events during first bone formation in embryonic chick limbs. , 1986, Bone.

[50]  A. Poole,et al.  Chondrogenesis in periosteal explants. An organ culture model for in vitro study. , 1994, The Journal of bone and joint surgery. American volume.

[51]  F. M. Schultz,et al.  The enhancement of periosteal chondrogenesis in organ culture by dynamic fluid pressure , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[52]  S. Albelda,et al.  Integrins and other cell adhesion molecules , 1990, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[53]  L. Rome,et al.  Cloning and sequence of the cDNA encoding the rat oligodendrocyte integrin beta 1 subunit. , 1995, Gene.

[54]  S. O’Driscoll,et al.  Enhancement of periosteal chondrogenesis in vitro. Dose-response for transforming growth factor-beta 1 (TGF-beta 1). , 1994, Clinical orthopaedics and related research.

[55]  S. O’Driscoll,et al.  Localization of chondrocyte precursors in periosteum. , 2001, Osteoarthritis and cartilage.

[56]  Campbell Cj The healing of cartilage defects. , 1969 .