Pulsed Electromagnetic Fields Increase Growth Factor Release by Nonunion Cells

The mechanisms involved in pulsed electromagnetic field stimulation of nonunions are not known. Animal and cell culture models suggest endochondral ossification is stimulated by increasing cartilage mass and production of transforming growth factor-beta 1. For the current study, the effect of pulsed electromagnetic field stimulation on cells from human hypertrophic (n = 3) and atrophic (n = 4) nonunion tissues was examined. Cultures were placed between Helmholtz coils, and an electromagnetic field (4.5-ms bursts of 20 pulses repeating at 15 Hz) was applied to ½ of them 8 hours per day for 1, 2, or 4 days. There was a time-dependent increase in transforming growth factor-beta 1 in the conditioned media of treated hypertrophic nonunion cells by Day 2 and of atrophic nonunion cells by Day 4. There was no effect on cell number, [3H]-thymidine incorporation, alkaline phosphatase activity, collagen synthesis, or prostaglandin E2 and osteocalcin production. This indicates that human nonunion cells respond to pulsed electromagnetic fields in culture and that transforming growth factor-beta 1 production is an early event. The delayed response of hypertrophic and atrophic nonunion cells (> 24 hours) suggests that a cascade of regulatory events is stimulated, culminating in growth factor synthesis and release.

[1]  B. Boyan,et al.  Treatment of resting zone chondrocytes with transforming growth factor-beta 1 induces differentiation into a phenotype characteristic of growth zone chondrocytes by downregulating responsiveness to 24,25-(OH)2D3 and upregulating responsiveness to 1,25-(OH)2D3. , 1998, Bone.

[2]  R. Diegelmann,et al.  Use of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins. , 1971, Biochemistry.

[3]  B D Boyan,et al.  Surface roughness modulates the local production of growth factors and cytokines by osteoblast-like MG-63 cells. , 1996, Journal of biomedical materials research.

[4]  J. Heckman,et al.  Initial effects of partially purified bone morphogenetic protein on the expression of glycosaminoglycan, collagen, and alkaline phosphatase in nonunion cell cultures. , 1992, Clinical orthopaedics and related research.

[5]  B. Boyan,et al.  Effects of vitamin D metabolites on collagen production and cell proliferation of growth zone and resting zone cartilage cells in vitro , 1989, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[6]  L. Bonewald,et al.  Direct effects of transforming growth factor-beta on chondrocytes are modulated by vitamin D metabolites in a cell maturation-specific manner. , 1993, Endocrinology.

[7]  D. Baksi.  Fracture healing. , 1980, Journal of the Indian Medical Association.

[8]  L. Bonewald,et al.  Vitamin D3 metabolites regulate LTBP1 and latent TGF‐β1 expression and latent TGF‐β1 incorporation in the extracellular matrix of chondrocytes , 1999, Journal of cellular biochemistry.

[9]  A. Economides,et al.  Morphogenesis of digits in the avian limb is controlled by FGFs, TGFbetas, and noggin through BMP signaling. , 1998, Developmental biology.

[10]  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.

[11]  M Misakian,et al.  Biological, physical, and electrical parameters for in vitro studies with ELF magnetic and electric fields: a primer. , 1993, Bioelectromagnetics.

[12]  C. Rubin,et al.  Electric fields modulate bone cell function in a density‐dependent manner , 1993, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[13]  L. Bonewald,et al.  Characterization and autoregulation of latent transforming growth factor beta (TGF beta) complexes in osteoblast-like cell lines. Production of a latent complex lacking the latent TGF beta-binding protein. , 1994, The Journal of biological chemistry.

[14]  L. Raisz,et al.  Biphasic effects of prostaglandin E2 on bone formation in cultured fetal rat calvariae: interaction with cortisol. , 1990, Endocrinology.

[15]  L. Bonewald,et al.  Growth plate chondrocytes store latent transforming growth factor (TGF)‐β1 in their matrix through latent TGF‐β1 binding protein‐1 , 1998 .

[16]  B. Boyan,et al.  Characterization of rat calvarial nonunion defects. , 1990, Acta anatomica.

[17]  L. Bonewald,et al.  Stimulation of matrix vesicle enzyme activity in osteoblast-like cells by 1,25(OH)2D3 and transforming growth factor beta (TGF beta). , 1992, Bone and mineral.

[18]  F. P. Magee,et al.  Combined magnetic fields increase insulin-like growth factor-II in TE-85 human osteosarcoma bone cell cultures. , 1995, Endocrinology.

[19]  L. Bonewald,et al.  Latent transforming growth factor-beta is produced by chondrocytes and activated by extracellular matrix vesicles upon exposure to 1,25-(OH)2D3. , 1994, The Journal of biological chemistry.

[20]  Z. Schwartz,et al.  Osteochondral progenitor cells in acute and chronic canine nonunions , 1999, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[21]  R. Aaron,et al.  Acceleration of experimental endochondral ossification by biophysical stimulation of the progenitor cell pool , 1996, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[22]  B D Boyan,et al.  Effect of titanium surface roughness on proliferation, differentiation, and protein synthesis of human osteoblast-like cells (MG63). , 1995, Journal of biomedical materials research.

[23]  C. Brighton,et al.  Electrical stimulation induces the level of TGF-beta1 mRNA in osteoblastic cells by a mechanism involving calcium/calmodulin pathway. , 1997, Biochemical and biophysical research communications.

[24]  J. Trowsdale,et al.  Alkaline phosphatases. , 1990, Biochemical Society transactions.

[25]  J. Kanczler,et al.  Pulsed electromagnetic fields simultaneously induce osteogenesis and upregulate transcription of bone morphogenetic proteins 2 and 4 in rat osteoblasts in vitro. , 1998, Biochemical and biophysical research communications.

[26]  S. Pollack,et al.  Treatment of recalcitrant non-union with a capacitively coupled electrical field. A preliminary report. , 1985, The Journal of bone and joint surgery. American volume.

[27]  J. Kirschvink,et al.  Uniform magnetic fields and double-wrapped coil systems: improved techniques for the design of bioelectromagnetic experiments. , 1992, Bioelectromagnetics.

[28]  M A Stuchly,et al.  Design and fabrication of well confined uniform magnetic field exposure systems. , 1994, Bioelectromagnetics.

[29]  L. Bonewald,et al.  Stimulation of matrix vesicle enzyme activity in osteoblast-like cells by 1,25(OH)2D3 and transforming growth factor β (TGFβ) , 1992 .

[30]  P. Shaman,et al.  Tibial Nonunion Treated With Direct Current, Capacitive Coupling, or Bone Graft , 1995, Clinical orthopaedics and related research.

[31]  M E Bolander,et al.  Transforming growth factor-beta and the initiation of chondrogenesis and osteogenesis in the rat femur , 1990, The Journal of cell biology.

[32]  K. Ono,et al.  Transforming growth factor-beta 1 stimulates chondrogenesis and inhibits osteogenesis in high density culture of periosteum-derived cells. , 1993, Endocrinology.

[33]  H. Cameron,et al.  The Canadian Experience with Pulsed Magnetic Fields in the Treatment of Ununited Tibial Fractures , 1986, Clinical orthopaedics and related research.

[34]  F. P. Magee,et al.  IGF‐II receptor number is increased in TE‐85 osteosarcoma cells by combined magnetic fields , 1995, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

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

[36]  L. Bonewald,et al.  Localization of 1,25-(OH)2D3-responsive alkaline phosphatase in osteoblast-like cells (ROS 17/2.8, MG 63, and MC 3T3) and growth cartilage cells in culture. , 1989, The Journal of biological chemistry.

[37]  D. Carnes,et al.  Epithelial cell lines that induce bone formation in vivo produce alkaline phosphatase-enriched matrix vesicles in culture. , 1992, Clinical orthopaedics and related research.

[38]  C. Bassett,et al.  Modification of fracture repair with selected pulsing electromagnetic fields. , 1982, The Journal of bone and joint surgery. American volume.

[39]  B. Boyan,et al.  Localization of vitamin D3-responsive alkaline phosphatase in cultured chondrocytes. , 1988, The Journal of biological chemistry.

[40]  J. McPherson,et al.  Cartilage-inducing factor-A. Apparent identity to transforming growth factor-beta. , 1986, The Journal of biological chemistry.

[41]  G. Mundy,et al.  Effects of Transforming Growth Factor-β on Osteoblastic Osteosarcoma Cells* , 1987 .

[42]  L. Bonewald,et al.  Effects of combining transforming growth factor beta and 1,25-dihydroxyvitamin D3 on differentiation of a human osteosarcoma (MG-63). , 1992, The Journal of biological chemistry.

[43]  B. Boyan,et al.  The effect of prostaglandin E2 on costochondral chondrocyte differentiation is mediated by cyclic adenosine 3',5'-monophosphate and protein kinase C. , 1998, Endocrinology.

[44]  B. Boyan,et al.  Titanium surface roughness alters responsiveness of MG63 osteoblast‐like cells to 1α,25‐(OH)2D3 , 1998 .

[45]  A. Caplan,et al.  The in vitro chondrogenic response of limb-bud mesenchyme to a water-soluble fraction prepared from demineralized bone matrix. , 1985, Differentiation; research in biological diversity.

[46]  B. Boyan,et al.  Pulsed electromagnetic field stimulation of MG63 osteoblast‐like cells affects differentiation and local factor production , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[47]  M. Nagai,et al.  Pulsating Electromagnetic Field Stimulates mRNA Expression of Bone Morphogenetic Protein-2 and -4 , 1994, Journal of dental research.

[48]  L. Bonewald,et al.  Dual role for the latent transforming growth factor-beta binding protein in storage of latent TGF-beta in the extracellular matrix and as a structural matrix protein , 1995, The Journal of cell biology.

[49]  J. Stuyck,et al.  Treatment of nonunion using pulsed electromagnetic fields: a retrospective follow-up study. , 1990, Acta orthopaedica Belgica.

[50]  G. Mundy,et al.  Effects of transforming growth factor-beta on osteoblastic osteosarcoma cells. , 1987, Endocrinology.

[51]  L. Bonewald,et al.  Effects of transforming growth factor β on bone nodule formation and expression of bone morphogenetic protein 2, osteocalcin, osteopontin, alkaline phosphatase, and type I collagen mRNA in long‐term cultures of fetal rat calvarial osteoblasts , 1994 .

[52]  C A Bassett,et al.  Fundamental and practical aspects of therapeutic uses of pulsed electromagnetic fields (PEMFs). , 1989, Critical reviews in biomedical engineering.

[53]  B. Boyan,et al.  Treatment of resting zone chondrocytes with 24,25-dihydroxyvitamin D3 [24,25-(OH)2D3] induces differentiation into a 1,25-(OH)2D3-responsive phenotype characteristic of growth zone chondrocytes. , 1995, Endocrinology.