Related Bone Morphogenetic Proteins Research Articles

Bone modeling: biomechanics, molecular mechanisms, and clinical perspectives

Abstract Bone modeling is a mechanically mediated adaptive process for changing a bone's size, shape, or position. Site specific, anabolic and catabolic modeling events are manifestations of overload hypertrophy and disuse atrophy, respectively. Catabolic bone modeling at the periodontal ligament (PDL) surface is the rate-limiting step in tooth movement. In initiating tooth movement, connective tissue growth factor (CTGF) is expressed in osteoblasts and osteocytes, and osteopontin (Opn) is expressed by osteoclasts and osteocytes. Undermining resorption occurs by recruiting osteoclasts to the site of maximal PDL compression and a paravascular osteogenic response is initiated in widened areas of the PDL: (1) preosteoblast formation at ∼10 hours, (2) peak DNA synthesis at ∼20 hours, (3) maximum rate of mitosis at ∼30 hours, and (4) initiation of bone formation at ∼48 hours. At other skeletal sites, additional genes have been linked to mechanical activation of bone: glutamate/aspartate transporter (GLAST), nitric oxide synthetase (NOS), prostaglandin G/H synthetase (PGHS-2), Msx1 , and c-fos . An intricate series of endocrine, paracrine, and autocrine factors has been described. Parathyroid hormone (PTH) enhances expression of insulin-like growth factor I (IGF-I) and the estrogen receptor beta (ER-β), which are both modulators of mechanical loading. Transforming growth factor beta (TGF-β) and the related bone morphogenetic proteins (BMPs) help control anabolic modeling. Catabolic modeling is mediated by PGE2, IL-1β, and other inflammatory cytokines. Osteoclast differentiation and activation is controlled by genes related to tumor necrosis factor (TNF) and its receptor (TNFR): colony stimulating factor 1 (CSF-1), osteoprotegerin (OPG), receptor activator of nuclear factor (RANK), and RANK ligand (RANKL). Orthodontic aspects of bone modeling are reviewed relative to morphology, biomechanics, neurology, molecular control mechanisms, and the periodontal adaptation to loading. Clinical correlations discussed are orthodontic analgesics, inflammatory cytokines, surgical enhancement of tooth movement, distraction osteogenesis, closure of atrophic defects, periodontal compromise, traumatized periodontium, and systemic disease.

Cartilage-derived morphogenetic proteins and cartilage morphogenesis.

Cartilage morphogenesis is a prerequisite for skeletal development and maintenance. The morphogenesis of cartilage determines the shape of bones, and joints including articular cartilage, ligaments, and tendon. This article reviews the recent advances in cartilage-derived morphogenetic proteins (CDMPs) and related bone morphogenetic proteins (BMPs). Cartilage-derived morphogenetic proteins (CDMPs) are related to BMPs and are critical for cartilage and joint morphogenesis. Cartilage morphogenesis is a multistep cascade that includes factors for initiation, promotion, and maintenance of cartilage phenotype. The extracellular matrix of cartilage consists of a constellation of macromolecules such as collagens, proteoglycans, and glycoproteins. Morphogens bind to extracellular matrix components and assemble a morphogenetic scaffold. Recent advances in CDMPs may aid in articular cartilage repair and regeneration.

Osteogenic protein-1 and related bone morphogenetic proteins regulate dendritic growth and the expression of microtubule-associated protein-2 in rat sympathetic neurons

Osteogenic protein-1 (OP-1) is expressed in the developing nervous system and it has been found to induce dendritic growth in sympathetic neurons. To further characterize this phenomenon, the effects of OP-1 were compared to those of other members of the bone morphogenetic protein (BMP) family of growth factors. Recombinant human OP-1, BMP-6, BMP-2 and the Drosophila 60A protein induced dendritic growth in rat sympathetic neurons in a concentration-dependent manner with EC 50-values of 1.8, 1.0, 1.7 and 2.7 ng/ml, respectively. In contrast, BMP-3 and cartilage-derived morphogenetic protein-2 (CDMP-2) as well as other classes of growth factors were inactive at concentrations up to 50 ng/ml. The dendritic growth induced by OP-1, BMP-6, BMP-2 and 60A was accompanied by increased expression of microtubule-associated protein-2 (MAP2) without changes in the expression of the phosphorylated forms of the M and H neurofilament subunits. These results suggest that several members of the BMP family have the capacity to regulate the morphological development of sympathetic neurons and that they may act by induction of specific cytoskeletal proteins.

Bone Morphogenetic Proteins Induce Differentiation in Astrocyte Lineage Cells

Serum-free mouse embryo (SFME) cells express Glial Fibrillary Acidic Protein (GFAP), a specific marker of the astrocyte lineage, when treated with either Transforming Growth Factor Beta (TGF-beta) or calf serum. We examined the effects of the related Bone Morphogenetic Proteins (BMPs) which are expressed in the developing murine nervous system. Treatment with the heterodimers BMP-2/6 and 2/7 followed by the homodimers BMP-2, 4, 5, 6, and 7 induced higher levels of GFAP in these cells than either TGF-beta 1 or activin when tested at the same concentration. The BMP-induced cells resembled classically described astrocytes and were characterized by antibody markers as type 1 and type 2. In addition, these astrocytes also showed increased levels of the cell adhesion molecules CD44 and neural cell adhesion molecule (N-CAM), both known to be expressed by this cell type. These data clearly demonstrate that the BMPs function as differentiation factors as well as regulators of adhesion molecule expression for cells of the astrocyte lineage and suggest a key role in glial development in the nervous system.

Osteogenin and recombinant bone morphogenetic protein 2B are chemotactic for human monocytes and stimulate transforming growth factor beta 1 mRNA expression.

Subcutaneous implantation of demineralized bone matrix initiates a sequence of developmental events, which culminate in endochondral bone formation. During early stages of development of matrix-induced implants, ED1, Ia-positive monocytes-macrophages were observed, suggesting that in the initial phases of the endochondral bone formation cascade, the bone-inductive protein osteogenin and related bone morphogenetic proteins (BMPs) might serve as potent chemoattractants to recruit circulating monocytes. In this investigation, we demonstrate that at concentrations of 10-100 fg/ml (0.3-3 fM), native bovine osteogenin and recombinant human BMP-2B (rhBMP-2B) induce the directed migration of human blood monocytes in vitro. This chemotactic response was associated with expression of BMP binding sites (receptors) on monocytes. About 750 receptors per cell were detected with an apparent dissociation constant of 200 pM. Both osteogenin and rhBMP-2B at higher concentrations (0.1-30 ng/ml) stimulated mRNA expression for an additional regulatory molecule, type beta 1 transforming growth factor (TGF-beta 1) in human monocytes. TGF-beta 1, in turn, is known to induce a cascade of events leading to matrix generation. Monocytes stimulated by TGF-beta are known to secrete a number of chemotactic and mitogenic cytokines that recruit endothelial and mesenchymal cells and promote their synthesis of collagen and associated matrix constituents. TGF-beta 1 in concert with these other cytokines and matrix components regulates chemotaxis, mesenchymal proliferation, differentiation, angiogenesis, and controlled synthesis of extracellular matrix. Our results demonstrate that osteogenin and related BMPs through their profound effects on monocyte recruitment and cytokine synthesis may promote additional successive steps in the endochondral bone formation cascade.

Initiation of Bone Regeneration in Adult Baboons by Osteogenin, a Bone Morphogenetic Protein

Osteogenin, and related bone morphogenetic proteins, induce endochondral bone differentiation through a cascade of events which include formation of cartilage, hypertrophy and calcification of the cartilage, vascular invasion, differentiation of osteoblasts, and formation of bone. These events have been studied in a postnatal model of bone development in rodents. Information concerning the morphogenetic potential of osteogenin in primates is a prerequisite for potential clinical application in man. The efficacy of allogeneic osteogenin in primates was investigated in both extraskeletal and skeletal sites in 19-Chacma baboons (Papio ursinus). Osteogenin was isolated from demineralized baboon bone matrix and purified by chromatography on heparin-Sepharose, hydroxyapatite, and Sephacryl S-200. Protein fractions with a molecular mass range of 26-42 kDa induced cartilage and bone differentiation in the subcutaneous space of rats. Final purification to homogeneity was obtained by electroendosmotic elution from a preparative sodium dodecyl sulphate (SDS) polyacrylamide gel, resulting in a single band on a SDS-polyacrylamide gel with an apparent molecular mass of 30-34 kDa, with biological activity in rats. The osteoinductive potential of osteogenin in primates was tested first in intramuscular sites in baboons and found to be active. The bone regeneration potential was investigated in nonhealing calvarial defects surgically prepared in adult male baboons. Baboon osteogenin induced complete regeneration of the cranial wound. These findings in adult primates establish a primary role for osteogenin in initiation and promotion of osteogenesis, and imply a potential therapeutic application based on cell biology of extracellular matrix-cell interactions.

Natural bovine osteogenin and recombinant human bone morphogenetic protein-2B are equipotent in the maintenance of proteoglycans in bovine articular cartilage explant cultures.

Osteogenin and related bone morphogenetic proteins are members of the transforming growth factor-beta superfamily, and were isolated by their ability to induce cartilage and bone formation in vivo. The influence of osteogenin, purified from bovine bone, and of recombinant human bone morphogenetic protein-2B (BMP-2B) has been examined in bovine articular cartilage explants. Both differentiation factors stimulated in a dose-dependent manner the synthesis of proteoglycans and decreased their rate of degradation. At a dose of 30 ng/ml, proteoglycan synthesis was increased to levels observed with either 20 ng/ml insulin-like growth factor I, 10 ng/ml transforming growth factor-beta, or 20% fetal bovine serum. This increase of biosynthetic rates above basal medium levels was observed in young, adolescent, and adult tissues. Analysis of the size of the newly synthesized proteoglycans, the glycosaminoglycan chain size, and the glycosaminoglycan type of explants treated with osteogenin or BMP-2B were very comparable to each other, and to proteoglycans isolated from cartilage treated with either insulin-like growth factor I or fetal bovine serum. These results demonstrate that osteogenin and BMP-2B alone are capable of stimulating and maintaining the chondrocyte phenotype in vitro.

Growth and Morphogenetic Factors in Bone Induction: Role of Osteogenin and Related Bone Morphogenetic Proteins in Craniofacial and Periodontal Bone Repair

Bone has considerable potential for repair as illustrated by the phenomenon of fracture healing. Repair and regeneration of bone recapitulate the sequential stages of development. It is well known that demineralized bone matrix has the potential to induce new bone formation locally at a heterotopic site of implantation. The sequential development of bone is reminiscent of endochondral bone differentiation during bone development. The collagenous matrix-induced bone formation is a prototype model for matrix-cell interactions in vivo. The developmental cascade includes migration of progenitor cells by chemotaxis, attachment of cells through fibronectin, proliferation of mesenchymal cells, and differentiation of bone. The bone inductive protein, osteogenin, was isolated by heparin affinity chromatography. Osteogenin initiates new bone formation and is promoted by other growth factors. Recently, the genes for osteogenin and related bone morphogenetic proteins were cloned and expressed. Recombinant osteogenin is osteogenic in vivo. The future prospects for bone induction are bright, and this is an exciting frontier with applications in oral and orthopaedic surgery.

Spontaneous multidrug transport in human glioma cells is regulated by transforming growth factors type ?

The multidrug transporting cell membrane molecule P-glycoprotein can be spontaneously expressed in human glioma cells. Transcripts of mdr genes were detected in glial tumor cells by polymerase chain reaction and Northern blotting, expression of P-glycoprotein was analyzed by immunocytochemistry and functional activity by cytofluorometry of fluorescent probe transport. In vitro treatment of glioma cells with vincristine induced coordinate over-expression of both mdr1 and mdr3 genes associated with very high P-glycoprotein-mediated multidrug transport, resistant to the inhibitory activity of chemosensitizers like verapamil. The physiological modulators of multidrug transport are as yet unknown. We therefore initiated a screening program to analyze the effects of cytokines on multidrug transport. We observed, that transforming growth factors (TGF)-beta 1, -beta 2, and -beta 1.2-but not the related bone morphogenetic protein (BMP) 2--inhibited multidrug transport. Interestingly, BMP 2 antagonized the TGF-beta induced inhibition of multidrug transport.