https://www.researchgate.net/file.PostFileLoader.html?id=526892a1d4c118a61d4360bd&assetKey=AS%3A272155479085058%401441898330516

Porosity of 3D biomaterial scaffolds and osteogenesis.

Since their isolation in the early 1990s, members of the Hedgehog family of intercellular signaling proteins have come to be recognized as key mediators of many fundamental processes in embryonic development. Their activities are central to the growth, patterning, and morphogenesis of many different regions within the body plans of vertebrates and insects, and most likely other invertebrates. In some contexts, Hedgehog signals act as morphogens in the dose-dependent induction of distinct cell fates within a target field, in others as mitogens regulating cell proliferation or as inducing factors controlling the form of a developing organ. These diverse functions of Hedgehog proteins raise many intriguing questions about their mode of operation. How do these proteins move between or across fields of cells? How are their activities modulated and transduced? What are their intracellular targets? In this article we review some well-established paradigms of Hedgehog function inDrosophila and vertebrate development and survey the current understanding of the synthesis, modification, and transduction of Hedgehog proteins. Embryological studies over much of the last century that relied primarily on the physical manipulation of cells within the developing embryo or fragments of the embryo in culture, provided many compelling examples for the primacy of cell–cell interactions in regulating invertebrate and vertebrate development. The subsequent identification of many of the signaling factors that mediate cellular communication has led to two general conclusions. First, although there are many important signals, most of these fall into a few large families of secreted peptide factors: theWnt (Wodarz and Nusse 1998), fibroblast growth factor (Szebenyi and Fallon 1999), TGFsuperfamily (Massague and Chen 2000), plateletderived growth factor (Betsholtz et al. 2001), ephrin (Bruckner and Klein 1998), and Hedgehog families. Second, parallel studies in invertebrate and vertebrate systems have shown that although the final outcome might look quite different (e.g., a fly vs. a mouse), there is a striking conservation in the deployment of members of the same signaling families to regulate development of these seemingly quite different organisms. This review focuses on one of the most intriguing examples of this phenomenon, that of the Hedgehog family. As with many of the advances in our understanding of the genetic regulation of animal development, hedgehog (hh) genes owe their discovery to the pioneering work of Nusslein-Volhard and Wieschaus (1980). In their screen for mutations that disrupt the Drosophila larval body plan, these authors identified several that cause the duplication of denticles (spiky cuticular processes that decorate the anterior half of each body segment) and an accompanying loss of naked cuticle, characteristic of the posterior half of each segment (see Fig. 1). The ensuing appearance of a continuous lawn of denticles projecting from the larval cuticle evidently suggested the spines of a hedgehog to the discoverers, hence the origin of the name of one of these genes. Other loci identified by mutants with this phenotype included armadillo, gooseberry, and wingless (wg). In contrast, animals mutant for the aptly named naked gene showed the converse phenotype, with denticle belts replaced by naked cuticle in every segment. On the basis of these mutant phenotypes, Nusslein-Volhard and Wieschaus (1980) proposed that these so-called segment-polarity genes regulate pattern within each of the segments of the larval body, individual genes acting within distinct subregions of the emerging segmental pattern. The first important breakthrough in unraveling how segment-polarity genes act came in the mid-1980s with the cloning of two members of the class, wingless and engrailed (en). Wg was shown to be the ortholog of the vertebrate proto-oncogene int1 (subsequently renamed Wnt1 and the founder member of the Wnt family of secreted peptide factors; Rijsewijk et al. 1987), whereas the sequence of en revealed that it encodes a homeodomaincontaining transcription factor (Fjose et al. 1985; Poole et al. 1985). Intriguingly, the two genes were found to be expressed in adjacent narrow stripes of cells in each segment (Martinez Arias et al. 1988). A close spatial relationship between Wnt1 and En expression domains was also reported in the primordial midbrain and hindbrain of the vertebrate embryo (McMahon et al. 1992). AnalyWe dedicate this review to the memory of our dear friend and colleague Rosa Beddington, whose encouragement led to our initial collaboration. 3Corresponding authors. E-MAIL p.w.ingham@sheffield.ac.uk; FAX 0114-222-288. E-MAIL amcmahon@biosun.harvard.edu; FAX (617) 496-3763. Article and publication are at http://www.genesdev.org/cgi/doi/10.1101/ gad.938601.

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Hedgehog signaling in animal development: paradigms and principles.

Biomedical applications of collagen

Biomaterial developments for bone tissue engineering

National High Blood Pressure Education Program.

In this study we describe a principle for the accomplishment of bone regeneration based on the hypothesis that different cellular components in the tissue have varying rates of migration into a wound area during healing By a mechanical hindrance, using a membrane technique, fibroblasts and other soft connective-tissue cells are prevented from entering the bone defect so that the presumably slower-migrating cells with osteogenic potential are allowed to repopulate the defect Defects of standard size were created bilaterally through the mandibular angles of rats On one side of the jaw the defect was covered with Teflon membranes, whereas the defect on the other side served as control Histologic analysis after healing demonstrated that on the test (membrane) side, half the number of animals showed complete bone healing after 3 weeks and all animals showed complete healing after 6 weeks Little or no sign of healing was evident on the control side even after an observation period of 22 weeks

Healing of bone defects by guided tissue regeneration.

The whereabouts of a morphogen: direct evidence for short- and graded long-range activity of hedgehog signaling peptides.

Sonic hedgehog ( Shh ), a member of the mammalian Hedgehog

(Hh) family, plays a key role during embryogenesis and organogenesis. Tooth

development, odontogenesis, is governed by sequential and reciprocal

epithelial-mesenchymal interactions. Genetic removal of Shh activity from the

dental epithelium, the sole source of Shh during tooth development, alters

tooth growth and cytological organization within both the dental epithelium

and mesenchyme of the tooth. In this model it is not clear which aspects of

the phenotype are the result of the direct action of Shh on a target tissue

and which are indirect effects due to deficiencies in reciprocal signalings

between the epithelial and mesenchymal components. To distinguish between

these two alternatives and extend our understanding of Shh9s actions in

odontogenesis, we have used the Cre-loxP system to remove Smoothened

(Smo) activity in the dental epithelium. Smo, a seven-pass membrane protein is

essential for the transduction of all Hh signals. Hence, removal of Smo

activity from the dental epithelium should block Shh signaling within dental

epithelial derivatives while preserving normal mesenchymal signaling. Here we

show that Shh-dependent interactions occur within the dental epithelium

itself. The dental mesenchyme develops normally up until birth. In contrast,

dental epithelial derivatives show altered proliferation, growth,

differentiation and polarization. Our approach uncovers roles for Shh in

controlling epithelial cell size, organelle development and polarization.

Furthermore, we provide evidence that Shh signaling between ameloblasts and

the overlying stratum intermedium may involve subcellular localization of

 Patched 2 and Gli1 mRNAs, both of which are targets of Shh

signaling in these cells.

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Shh signaling within the dental epithelium is necessary for cell proliferation, growth and polarization

Cyst-like cavities in the jaw bone often heal incompletely owing to ingrowth of connective tissue, thus preventing osteogenesis from occurring. In the present study, a new membrane technique has been utilized in an attempt to improve bone healing. By means of an inert, porous membrane, placed in close contact with the bone surface, a secluded space is created which can only be repopulated by cells from the adjacent bone. Thus, osteogenesis is able to occur without interference from other tissue types. Through-and-through bone defects were produced bilaterally (1) in edentulous areas of monkey (n = 5) mandibles, and (2) in conjunction with apicectomy of the lateral maxillary incisors, also in monkeys (n = 7). On one side, the defects were covered buccally as well as lingually/palatally with expanded PTFE membranes, whereas the defects on the other side served as controls (no membrane). In the mandible, complete bone healing was seen at all test sites after a healing period of 3 months. On the control side, 3 experimental sites showed bone discontinuity with a transosseous core of connective tissue, whereas some bone healing had occurred lingually at 2 sites, but with massive soft tissue ingrowth from the buccal side. In the maxillary periapical defects, all the membrane-covered defects had healed with bone closure after 3 months but with a minute portion of connective tissue, probably derived from the periodontal ligament, around the tooth apices. None of the control defects (no membrane) healed spontaneously, but all were filled with connective tissue to varying degrees.(ABSTRACT TRUNCATED AT 250 WORDS)

Healing of maxillary and mandibular bone defects using a membrane technique. An experimental study in monkeys.

Dentin may be regarded as a mineralized connective tissue. In its composition as well as its mode of formation, dentin exhibits several similarities with bone, but also definite differences. The dentin organic phase, the matrix, determines its morphology and is believed to be instrumental in the formation of the mineral phase. A fibrous web of collagen type I dominates the organic matrix. Also, minor amounts of other collagen types may be present. The noncollagenous proteins (NCPs), which constitute about 10% of the matrix, fall into several categories: phosphoproteins, Gla-proteins of the osteocalcin type as well as matrix Gla-protein, proteoglycans, different acidic glycoproteins, and serum proteins. Some of these NCPs have unique chemical compositions that give them specific properties. Dentinogenesis occurs by two simultaneous processes: the formation of a collagenous web in predentin, which is followed by the formation of the inorganic phase at the mineralization front. The composition of the predentin organic matrix differs from that of dentin, as some NCP components are secreted extracellularly just in advance of the mineralization front. In addition, some constituents of predentin seem to be metabolized. The NCPs may be important to several processes during dentinogenesis. Much evidence indicates that noncollagenous components in the matrix are instrumental in mineral formation. New data show that polyanionic NCPs, such as phosphoprotein and proteoglycans, when immobilized on a solid support, induce apatite formation under physiological conditions. These data indicate that polyanionic NCPs may function as mineral nucleators in vivo. They may also act as size and rate regulators for crystallization and promote calcium ion diffusion in the tissue. In addition, NCPs may regulate collagen fibrillogenesis.

Anders Linde

Papers

Healing of bone defects by guided tissue regeneration.

The whereabouts of a morphogen: direct evidence for short- and graded long-range activity of hedgehog signaling peptides.

Shh signaling within the dental epithelium is necessary for cell proliferation, growth and polarization

Healing of maxillary and mandibular bone defects using a membrane technique. An experimental study in monkeys.

Dentin matrix proteins: Composition and possible functions in calcification