Critical Reviews in Oral Biology & Medicine

The Official Publication of the American Association of Oral Biologists

A Publication of the International/American Associations for Dental Research
Table of Contents for Volume 10, 1999

Volume 10, Issue 1
Volume 10, Issue 2
Volume 10, Issue 3
Volume 10, Issue 4

Volume 10, Issue 1

• Byers MR, Narhi MV.
• Department of Anesthesiology, University of Washington, Seattle 98195-6540, USA.
• Pages 4-39

Recent research has shown that peripheral mechanisms of pain are much more complex than previously thought, and they differ for acutely injured normal tissues compared with chronic inflammation or neuropathic (nerve injury) pain. The purpose of the present review is to describe uses of dental injury models as experimental tools for understanding the normal functions of polymodal nociceptive nerves in healthy tissues, their neuroinflammatory interactions, and their roles in healing. A brief review of normal dental innervation and its interactions with healthy pulp tissue will be presented first, as a framework for understanding the changes that occur after injury. Then, the different types of dental injury that allow gradation of the extent of tissue damage will be described, along with the degree and duration of inflammation, the types of reactions in the trigeminal ganglion and brainstem, and the type of healing. The dental injury models have some unique features compared with neuroinflammation paradigms that affect other peripheral tissues such as skin, viscera, and joints. Peripheral inflammation models can all be contrasted to nerve injury studies that produce a different kind of neuroplasticity and neuropathic pain. Each of these models provides different insights about the normal and pathologic functions of peripheral nerve fibers and their effects on tissue homeostasis, inflammation, and wound healing. The physical confinement of dental pulp and its innervation within the tooth, the high incidence of polymodal A-delta and C-fibers in pulp and dentin, and the somatotopic organization of the trigeminal ganglion provide some special advantages for experimental design when dental injury models are used for the study of neuroinflammatory interactions.

• Franceschi RT.
• Department of Periodontics/Prevention/Geriatrics, School of Dentistry, University of Michigan, Ann Arbor 48109-1078, USA
• Pages 40-57

Bone formation is a carefully controlled developmental process involving morphogen-mediated patterning signals that define areas of initial mesenchyme condensation followed by induction of cell-specific differentiation programs to produce chondrocytes and osteoblasts. Positional information is conveyed via gradients of molecules, such as Sonic Hedgehog that are released from cells within a particular morphogenic field together with region-specific patterns of hox gene expression. These, in turn, regulate the localized production of bone morphogenetic proteins and related molecules which initiate chondrocyte- and osteoblast-specific differentiation programs. Differentiation requires the initial commitment of mesenchymal stem cells to a given lineage, followed by induction of tissue-specific patterns of gene expression. Considerable information about the control of osteoblast-specific gene expression has come from analysis of the promoter regions of genes encoding proteins like osteocalcin that are selectively expressed in bone. Both general and tissue-specific transcription factors control this promoter. Osf2/Cbfa1, the first osteoblast-specific transcription factor to be identified, is expressed early in the osteoblast lineage and interacts with specific DNA sequences in the osteocalcin promoter essential for its selective expression in osteoblasts. The OSF2/CBFA1 gene is necessary for the development of mineralized tissues, and its mutation causes the human disease, cleidocranial dysplasia. Committed osteoprogenitor cells already expressing Osf2/Cbfa1 must synthesize a collagenous ECM before they will differentiate. A cell:ECM interaction mediated by integrin-type cell-surface receptors is essential for the induction of osteocalcin and other osteoblast-related proteins. This interaction stimulates the binding of Osf2/Cbfa1 to the osteocalcin promoter through an as-yet-undefined mechanism.

• Kamer AR, Krebs L, Hoghooghi SA, Liebow C.
• Department of Oral and Maxillofacial Surgery, School of Dental Medicine, State University of New York at Buffalo, 14214, USA.
• Pages 58-78

The study of signal transduction pathways for mechanisms of apoptosis and proliferation has significantly advanced our understanding of human cancer, subsequently leading to more effective treatments. Discoveries of growth factors and oncogenes, especially those that function through phosphorylation on tyrosine residues, have greatly benefited our appreciation of the biology of cancer. The regulation of proliferation and apoptosis through phosphorylation via tyrosine kinases and phosphatases is discussed, as well as the contributions of other systems, such as serine and threonine kinases and phosphatases. Receptors with seven-transmembrane domains, steroid hormones, genes, and "death domains" will also be discussed. This review attempts to compare the regulation of the growth of normal tissues and cancers with an effort to highlight the current knowledge of these factors in the growth regulation of oral/oropharyngeal cancers. Despite the strides made in our understanding of growth regulation in human cancers, the study of oral/oropharyngeal cancer specifically lags behind. More research must be done to further our understanding of oral cancer biology, if we are to develop better, more effective treatment protocols.

• Ganss B, Kim RH, Sodek J.
• Medical Research Council Group in Periodontal Physiology, Faculty of Dentistry, University of Toronto, Ontario, Canada.
• Pages 79-98

The search for a protein nucleator of hydroxyapatite crystal formation has been a focus for the isolation and characterization of the major non-collagenous proteins in bone. Of the proteins characterized to date, bone sialoprotein (BSP) has emerged as the only bona fide candidate for nucleation. BSP is a highly glycosylated and sulphated phosphoprotein that is found almost exclusively in mineralized connective tissues. Characteristically, polyglutamic acid and arginine-glycine-aspartate (RGD) motifs with the ability to bind hydroxyapatite and cell-surface integrins, respectively, have been conserved in the protein sequence. Expression of the BSP gene, which is induced in newly formed osteoblasts, is up-regulated by hormones and cytokines that promote bone formation and down-regulated by factors that suppress bone formation. Thus, BSP has the biophysical and chemical properties of a nucleator, and its temporo-spatial expression coincides with de novo mineralization in bone and cementum. Moreover, BSP has been associated with mineral crystal formation in several pathologies, including breast carcinomas. However, the ability of BSP to mediate cell attachment and to signal through the RGD motif points to alternate functions for BSP which need further investigation. In combination, the hydroxyapatite-binding polyglutamic acid sequences and the RGD provide bi-functional entities through which BSP may mediate the targeting and attachment of normal and metastasizing cells to the bone surface.

• Radford DR, Challacombe SJ, Walter JD.
• Department of Prosthetic Dentistry, Guy's, King's and St. Thomas' Dental Institute, Guy's Dental Hospital, University of London, United Kingdom.
• Pages 99-116

The aim of this paper is to review our understanding of the mechanisms and clinical significance of adhesion of C. albicans to denture-base materials in relation to denture plaque and denture-related stomatitis. Earlier reports in the literature of a 65% prevalence level of denture-related stomatitis seem to be exaggerated. More recent studies indicate that denture-related stomatitis is considerably less common, particularly in normal healthy subjects. The etiology of the condition is discussed in this review, and although much of the literature supports the view that the condition is strongly associated with C. albicans, this is not always so. In some subjects, the cause appears to be related to a non-specific plaque. This review also considers the role of denture plaque in the pathogenesis of denture-related stomatitis, the sequential development of denture plaque, and its colonization by Candida organisms. Designing controlled in vivo studies is difficult, and as a consequence, many investigators have had to resort to in vitro studies. The majority of these studies have attempted to investigate the hydrophobicity of C. albicans, relating the surface free-energy of denture-base materials, particularly acrylic resin, to that of the organism. Surprisingly little work has been directed at surface roughness and how it affects retention of organisms. Further, no attention has been paid to the properties and character of the surface, other than average surface roughness, as it affects adhesion. A comparison of results from in vitro studies on the effect on adhesion of pre-coating the surfaces of denture-base materials with saliva has produced equivocal conclusions. This is largely due to little standardization of experimental protocols between studies, particularly in the collection and handling of the saliva used. In conclusion, the review strongly supports the suggestion that adherence of C. albicans to denture-base materials in vitro is related to the hydrophobicity of the organism. The clinical significance of the observation and the mechanisms for the development and maturation of denture plaque are yet to be understood. There is a clear need for further investigation of other factors that may moderate the adhesion of organisms and subsequent colonization of denture-base materials.

Volume 10, Issue 2

• Yeung MK.
• Department of Pediatric Dentistry, University of Texas Health Science Center at San Antonio, 78284, USA.
• Pages 120-38

Members of the genus Actinomyces are predominant primary colonizers of the oral cavity and play an important role in initiating plaque development. These bacteria have evolved unique mechanisms that favor colonization and persistence in this micro-environment. The expression of cell-surface fimbriae is correlated with the ability of these bacteria to adhere to specific receptors on the tooth and mucosal surfaces, and to interact with other plaque bacteria. The elaboration of sialidase is thought to enhance fimbriae-mediated adherence by unmasking the fimbrial receptors on mammalian cells. The presence of certain cell-associated or extracellular enzymes, including those involved in sucrose or urea metabolism, may provide the means for these bacteria to thrive under conditions when other growth nutrients are not available. Moreover, these enzyme activities may influence the distribution of other plaque bacteria and promote selection for Actinomyces spp. in certain ecological niches. The recent development of a genetic transfer system for Actinomyces spp. has allowed for studies the results of which demonstrate the existence of multiple genes involved in fimbriae synthesis and function, and facilitated the construction of allelic replacement mutants at each gene locus. Analyses of these mutants have revealed a direct correlation between the synthesis of assembled fimbriae and the observed adherence properties. Further genetic analysis of the various enzyme activities detected from strains of Actinomyces should allow for an assessment of the role of these components in microbial ecology, and their contribution to the overall success of Actinomyces spp. as a primary colonizer and a key player in oral health and disease.

• Polverini PJ, Nor JE.
• Department of Oral Medicine/Pathology/Surgery, University of Michigan School of Dentistry, Ann Arbor 48109-1078, USA.
• Pages 139-52

The term apoptosis, also known as programmed cell death (PCD), was coined by developmental biologists a number of years ago to describe a form of cell death characterized by several unique morphological and biochemical features. Genetic studies of the round worm Caeneorhabditis elegans, a simple multicellular organism, first revealed apoptosis to be an integral part of the developmental program. Subsequently, the importance of apoptosis in higher organisms was demonstrated in several eukaryotic systems. [n mammals, apoptosis is widespread during embryogenesis and in adult tissues. It is required for normal tissue homeostasis and for clonal selection in the immune system. In both developing and adult organisms, apoptosis plays a central role in reinforcing appropriate cellular patterns and in regulating cell number by eliminating cells that are harmful or no longer needed. It is becoming increasingly clear that disruption in the apoptosis pathway can contribute to the development of a number of developmental, inflammatory, degenerative, and neoplastic diseases. The effector arm of the apoptotic program includes members of the Bcl-2 gene family that function as either death agonists or death antagonists. These proteins participate in an elaborate genetically controlled biochemical pathway that functions to maintain tissue and organ homeostasis and serve as a critical defense mechanism to guard against malignant transformation. Cancer is the result of a series of genetic lesions that include activation of oncogenes and inactivation or loss of tumor suppressor genes. Several groups of investigators have observed that deregulated expression of oncogenes can subvert apoptotic pathways, resulting in prolonged cell survival. In pathological settings such as cancer, members of the Bcl-2 gene family are able to synergize with oncogenes and tumor suppressor genes to transform cells. In this review, we describe the process of apoptosis in mammalian cells and define the role and biochemical pathways through which the Bcl-2 gene family induce and/or protect cells from apoptosis. Last, we will discuss the evidence which suggests that alterations in this pathway may play a central role in tumorigenesis by allowing genetically damaged cells normally destined for elimination to persist, predisposing them to additional mutations and driving them to malignancy.

• Lingen MW.
• Department of Pathology and the Cardinal Bernardin Cancer Center, Loyola University Medical Center, Maywood, Illinois 60153, USA.
• Pages 153-64

Squamous cell carcinoma is an aggressive malignancy that often develops as multiple independent lesions throughout the mucosa of the upper aerodigestive tract. Therefore, the comprehensive treatment of this disease must not only address the initial primary neoplasm, but also prevent the progression of the premalignant lesions lurking throughout the rest of the mucosal surfaces. The need to treat these lesions has resulted in a search for chemopreventive agents that can halt or even reverse their malignant progression. The biologic and molecular mechanisms by which most chemopreventive agents act have remained unclear and controversial. Recent work from several laboratories has demonstrated that some drugs may act in part by inhibiting the ability of tumors to induce blood vessel growth. Angiogenesis, the growth of new blood vessels from pre-existing ones, is absolutely required for solid neoplasms to grow beyond 2-3 mm in diameter. Therefore, chemopreventive agents that act to inhibit angiogenesis may provide a very powerful modality by which one may limit the growth of both pre-malignant lesions and small nests of tumor cells. This review will outline the basic changes that occur in tumor cells that result in the switch from an anti-angiogenic to an angiogenic phenotype. In addition, it will discuss the mechanisms by which some chemopreventive agents, presently under clinical investigation, inhibit tumor angiogenesis. Finally, this paper will present a rationale for the use of multiple anti-angiogenic agents as a means of developing new chemopreventive protocols that result in reduced patient toxicity while maintaining similar clinical efficacies.

• Krebsbach PH, Kuznetsov SA, Bianco P, Robey PG.
• Department of Oral Medicine, Pathology, and Oncology, University of Michigan School of Dentistry, Ann Arbor 48109-1078, USA
• Pages 165-81

The bone marrow stroma consists of a heterogeneous population of cells that provide the structural and physiological support for hematopoietic cells. Additionally, the bone marrow stroma contains cells with a stem-cell-like character that allows them to differentiate into bone, cartilage, adipocytes, and hematopoietic supporting tissues. Several experimental approaches have been used to characterize the development and functional nature of these cells in vivo and their differentiating potential in vitro. In vivo, presumptive osteogenic precursors have been identified by morphologic and immunohistochemical methods. In culture, the stromal cells can be separated from hematopoietic cells by their differential adhesion to tissue culture plastic and their prolonged proliferative potential. In cultures generated from single-cell suspensions of marrow, bone marrow stromal cells grow in colonies, each derived from a single precursor cell termed the colony-forming unit-fibroblast. Culture methods have been developed to expand marrow stromal cells derived from human, mouse, and other species. Under appropriate conditions, these cells are capable of forming new bone after in vivo transplantation. Various methods of cultivation and transplantation conditions have been studied and found to have substantial influence on the transplantation outcome. The finding that bone marrow stromal cells can be manipulated in vitro and subsequently form bone in vivo provides a powerful new model system for studying the basic biology of bone and for generating models for therapeutic strategies aimed at regenerating skeletal elements.

• Cheifetz S.
• MRC Group in Periodontal Physiology, University of Toronto, Ontario, Canada.
• Pages 182-98

Members of the TGF-beta superfamily signal through receptor complexes comprised of type I and type II receptors. These receptors, which are serine/threonine kinases, form two new classes of transmembrane receptor kinases. The activity of both of the kinases is necessary for signal transduction in response to ligand binding. Bone morphogenetic proteins (BMPs), which are members of the TGF-beta superfamily, bind to multiple type I and type II receptors. There is growing evidence to support the hypothesis that the BMP receptors are differentially regulated during development and that they have both unique and overlapping functions. Thus, the nature and distribution of the BMP receptors, which are reviewed here in the context of the development of limbs and teeth, appear to be critical in the control of the diverse activities of BMPs.

• Melvin JE.
• Center for Oral Biology, Rochester Institute for Biomedical Sciences, University of Rochester Medical Center, New York 14642, USA.
• Pages 199-209

Fluid and electrolyte transport is driven by transepithelial Cl- movement. The opening of Cl- channels in the apical membrane of salivary gland acinar cells initiates the fluid secretion process, whereas the activation of Cl- channels in both the apical and the basolateral membranes of ductal cells is thought to be necessary for NaCl re-absorption. Saliva formation can be evoked by sympathetic and parasympathetic stimulation. The composition and flow rate vary greatly, depending on the type of stimulation. As many as five classes of Cl- channels with distinct gating mechanisms have been identified in salivary cells. One of these Cl- channels is activated by intracellular Ca2+, while another is gated by cAMP. An increase in the intracellular free Ca2+ concentration is the dominant mechanism triggering fluid secretion from acinar cells, while cAMP may be required for efficient NaCl re-absorption in many ductal cells. In addition to cAMP- and Ca(2+)-gated Cl- channels, agonist-induced changes in membrane potential and cell volume activate different Cl- channels that likely play a role in modulating fluid and electrolyte movement. In this review, the properties of the different types of Cl- currents expressed in salivary gland cells are described, and functions are proposed based on the unique properties of these channels.

• Turner JT, Landon LA, Gibbons SJ, Talamo BR.
• Department of Pharmacology, University of Missouri-Columbia School of Medicine, 65212, USA.
• Pages 210-24

The effects of ATP on salivary glands have been recognized since 1982. Functional and pharmacological studies of the P2 nucleotide receptors that mediate the effects of ATP and other extracellular nucleotides have been supported by the cloning of receptor cDNAs, by the expression of the receptor proteins, and by the identification in salivary gland cells of multiple P2 receptor subtypes. Currently, there is evidence obtained from pharmacological and molecular biology approaches for the expression in salivary gland of two P2X ligand-gated ion channels, P2Z/P2X7 and P2X4, and two P2Y G protein-coupled receptors, P2Y1 and P2Y2. Activation of each of these receptor subtypes increases intracellular Ca2+, a second messenger with a key role in the regulation of salivary gland secretion. Through Ca2+ regulation and other mechanisms, P2 receptors appear to regulate salivary cell volume, ion and protein secretion, and increased permeability to small molecules that may be involved in cytotoxicity. Some localization of the various salivary P2 receptor subtypes to specific cells and membrane subdomains has been reported, along with evidence for the co-expression of multiple P2 receptor subtypes within specific salivary acinar or duct cells. However, additional studies in vivo and with intact organ preparations are required to define clearly the roles the various P2 receptor subtypes play in salivary gland physiology and pathology. Opportunities for eventual utilization of these receptors as pharmacotherapeutic targets in diseases involving salivary gland dysfunction appear promising.

• Molsted K.
• Copenhagen Cleft Palate Center, Speech and Hearing Institute, Hellerup, Denmark.
• Pages 225-39

In the last 40 years, great progress has been made toward a better understanding of many aspects of the cleft lip and palate defect, but there is still a long way to go before there is agreement on the optimal treatment procedures. With regard to the primary operations, it can be stated, in a somewhat simplified form, that there are two main schools of thought in cleft treatment. One advocates early closure of the lip and palate, a procedure which imparts a high priority to early speech function. The other recommends delayed closure of the hard palate, thereby according a high priority to the growth of the maxilla. A number of intercenter and multicenter studies have been carried out recently in an effort to elucidate which procedures give the best result, both esthetically and functionally. The results are ambiguous, and this has led a number of researchers to suggest that the randomized clinical trial is the only way to resolve the ambiguity. The fact that it has proved difficult to identify the optimal procedures in the field of cleft lip and palate treatment need not only be due to a less than optimal research design; a contributory factor might also be the great variability in craniofacial morphology and in the response to treatment in patients who have exactly the same cleft lip and palate diagnosis. Intensive research has made it possible to state categorically that clefts occur due to many different factors in an interplay between genetics and environment. Therefore, it is not likely that a single gene can be responsible for clefting. Since scar tissue presents many problems-for instance, impairment of growth-the reduction or prevention of scar formation has long been a desirable goal. The discovery that a fetus can heal without scar formation has led to many animal experiments. The timing of the surgical intervention on fetuses is critical, since late-gestation fetuses heal with adult-like scarring. There are still many unsolved problems connected with fetal surgery, and at present prenatal surgery for repair of cleft lip and palate is not ethically defensible in humans. On the other hand, it appears that there are considerable possibilities for the reduction of human scarring after surgery with the introduction of various wound-healing medications.

Volume 10, Issue 3

• Loesche WJ.
• University of Michigan School of Dentistry, Ann Arbor 48109, USA.
• Pages 245-75

Over the last 100 years, methods of surgical periodontal treatment have enjoyed a history of success in improving oral health. The paradigm of care is based on the "non-specific plaque hypothesis"--that is, the overgrowth of bacterial plaques cause periodontal disease, and the suppression of this overgrowth reduces disease risk. The central feature of this approach to care is the removal of inflamed gingival tissue around the teeth to reduce periodontal pocket depth, thereby facilitating plaque removal by the dentist and by the patient at home. Over the last 30 years, with the recognition that periodontal disease(s) is caused by specific bacteria and that specific antimicrobial agents can reduce or eliminate the infection, a second paradigm has developed. This new paradigm, the "specific plaque hypothesis", focuses on reducing the specific bacteria that cause periodontal attachment loss. The contrast between the two paradigms can be succinctly stated as follows: The antimicrobial therapy reduces the cause, while the surgical therapy reduces the result of the periodontal infection. The specific plaque hypothesis has two important implications. First, with the increasing attention to evidence-based models for prevention, treatment, outcome assessment, and reimbursement of care, increasing attention and financial effort will be channeled into effective preventive and treatment methods. Second, the recent observations that periodontal infections increase the risk of specific systemic health problems, such as cardiovascular disease, argue for the prevention and elimination of these periodontal infections. This review highlights some of the evidence for the specific plaque hypothesis, and the questions that should be addressed if antimicrobial agents are to be used responsively and effectively.

• Baum BJ, O'Connell BC.
• Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 20892, USA.
• Pages 276-83

Considerable progress has occurred recently in transferring foreign genes to different tissues in vivo. Gene transfer to salivary glands has mirrored progress in the general field. Most salivary studies have utilized replication-deficient, recombinant adenoviruses as gene transfer vectors in rat models. These vectors are able to transduce almost all rat salivary epithelial cell types and lead to relatively high levels of transgene expression. Additionally, successful, though quite modest, gene transfer to salivary glands has been achieved with retroviral vectors and with different plasmid conjugates (liposomes; nonrecombinant adenoviruses). Salivary gland gene transfer has been used for two potential clinical goals: (i) the repair of hypofunctional gland parenchyma, and (ii) the production of secretory transgene products for either systemic or upper gastrointestinal tract pharmaceutical use. Gene transfer can also be used as a powerful tool to alter cellular phenotype in vivo and probe cell biological questions. The current spectrum of studies demonstrates the potential broad and profound influence gene transfer can make on salivary physiology and pathophysiology.

• Watson EL.
• Department of Oral Biology, University of Washington, Health Sciences Center, Seattle 98195-7132, USA.
• Pages 284-306

Regulated exocytosis, which occurs in response to stimuli, is a two-step process involving the docking of secretory granules (SGs) at specific sites on the plasma membrane (PM), with subsequent fusion and release of granule contents. This process plays a crucial role in a number of tissues, including exocrine glands, chromaffin cells, platelets, and mast cells. Over the years, our understanding of the proteins involved in vesicular trafficking has increased dramatically. Evidence from genetic, biochemical, immunological, and functional assays supports a role for ras-like monomeric GTP-binding proteins (smgs) as well as heterotrimeric GTP-binding protein (G-protein) subunits in various steps of the vesicular trafficking pathway, including the transport of secretory vesicles to the PM. Data suggest that the function of GTP-binding proteins is likely related to their localization to specific cellular compartments. The presence of both G-proteins and smgs on secretory vesicles/granules implicates a role for these proteins in the final stages of exocytosis. Molecular mechanisms of exocytosis have been postulated, with the identification of a number of proteins that modify, regulate, and interact with GTP-binding proteins, and with the advent of approaches that assess the functional importance of GTP-binding proteins in downstream, exocytotic events. Further, insight into vesicle targeting and fusion has come from the characterization of a SNAP receptor (SNARE) complex composed of vesicle, PM, and soluble membrane trafficking components, and identification of a functional linkage between GTP-binding and SNARES.

• Maeda T, Ochi K, Nakakura-Ohshima K, Youn SH, Wakisaka S.
• Department of Oral Anatomy, Niigata University School of Dentistry, Japan.
• Pages 307-27

The periodontal ligament receives a rich sensory nerve supply and contains many nociceptors and mechanoreceptors. Although its various kinds of mechanoreceptors have been reported in the past, only recently have studies revealed that the Ruffini endings--categorized as low-threshold, slowly adapting, type II mechanoreceptors--are the primary mechanoreceptors in the periodontal ligament. The periodontal Ruffini endings display dendritic ramifications with expanded terminal buttons and, furthermore, are ultrastructurally characterized by expanded axon terminals filled with many mitochondria and by an association with terminal or lamellar Schwann cells. The axon terminals of the periodontal Ruffini endings have finger-like projections called axonal spines or microspikes, which extend into the surrounding tissue to detect the deformation of collagen fibers. The functional basis of the periodontal Ruffini endings has been analyzed by histochemical techniques. Histochemically, the axon terminals are reactive for cytochrome oxidase activity, and the terminal Schwann cells have both non-specific cholinesterase and acid phosphatase activity. On the other hand, many investigations have suggested that the Ruffini endings have a high potential for neuroplasticity. For example, immunoreactivity for p75-NGFR (low-affinity nerve growth factor receptor) and GAP-43 (growth-associated protein-43), both of which play important roles in nerve regeneration/development processes, have been reported in the periodontal Ruffini endings, even in adult animals (though these proteins are usually repressed or down-regulated in mature neurons). Furthermore, in experimental studies on nerve injury to the inferior alveolar nerve, the degeneration of Ruffini endings takes place immediately after nerve injury, with regeneration beginning from 3 to 5 days later, and the distribution and terminal morphology returning to almost normal at around 14 days. During regeneration, some regenerating Ruffini endings expressed neuropeptide Y, which is rarely observed in normal animals. On the other hand, the periodontal Ruffini endings show stage-specific configurations which are closely related to tooth eruption and the addition of occlusal forces to the tooth during postnatal development, suggesting that mechanical stimuli due to tooth eruption and occlusion are a prerequisite for the differentiation and maturation of the periodontal Ruffini endings. Further investigations are needed to clarify the involvement of growth factors in the molecular mechanisms of the development and regeneration processes of the Ruffini endings.

• Heyeraas KJ, Berggreen E.
• Department of Physiology, University of Bergen, Norway.
• Pages 328-36

Tissue pressure is the hydrostatic pressure in the interstitial fluid which surrounds the pulpal cells. This pressure outside the vessels is normally considerably lower than the blood pressure inside the vessels. The dental pulp has a relatively low interstitial compliance due to its enclosure between rigid dentin walls. Accordingly, even a modest increase in pulpal fluid volume will raise the tissue pressure, which may compress blood vessels, leading to ischemia and necrosis. Inflammation may lead to an increase in both interstitial fluid volume and blood volume in the low-compliant pulp and thereby increase the tissue pressure. However, the increased tissue pressure may, in turn, initiate increased lymph flow and absorption of fluid into capillaries in nearby non-inflamed tissue. Both of these latter factors will transport fluid out of the affected area and subsequently out of the tooth and consequently lower the tissue pressure. Increased tissue pressure, whether caused by increased blood volume or increased capillary filtration, will promote outward flow of fluid through exposed dentin tubules and thereby help to protect the pulp against entry of harmful substances. It seems physiologically beneficial, therefore, for the pulp to have a high tissue pressure, which promptly increases when blood flow increases due to its low compliance.

• Rojas AI, Ahmed AR.
• Department of Oral Medicine and Diagnostic Sciences, Harvard School of Dental Medicine, Boston, Massachusettes 02115, USA
• Pages 337-58

Cell adhesion molecules have been recognized to play a major role in a variety of physiological and pathological phenomena. They determine the specificity of cell-cell binding and the interactions between cells and extracellular matrix proteins. Some of them may also function as receptors that trigger intracellular pathways and participate in cellular processes like migration, proliferation, differentiation, and cell death. The receptors that mediate adhesion between epithelial cells that are discussed in this review include integrins, selectins, the immunoglobulin superfamily members, and cadherins. The intent of this review is to inform the reader about recent advances in cellular and molecular functions of certain receptors, specifically those that are considered important in cell adhesion. We have deliberately not provided all-inclusive detailed information on every molecule, but instead, have presented a generalized overview in order to give the reader a global perspective. This information will be useful in enhancing the reader's understanding of the molecular pathology of diseases and recognizing the potential role of these receptors and ligands as therapeutic agents.

• Cannon RD, Chaffin WL.
• Department of Oral Sciences and Orthodontics, Faculty of Dentistry, University of Otago, Dunedin, New Zealand.
• Pages 359-83

Candida albicans is a commensal yeast normally present in small numbers in the oral flora of a large proportion of humans. Colonization of the oral cavity by C. albicans involves the acquisition and maintenance of a stable yeast population. Micro-organisms are continually being removed from the oral cavity by host clearance mechanisms, and so, in order to survive and inhabit this eco-system, C. albicans cells have to adhere and replicate. The oral cavity presents many niches for C. albicans colonization, and the yeast is able to adhere to a plethora of ligands. These include epithelial and bacterial cell-surface molecules, extracellular matrix proteins, and dental acrylic. In addition, saliva molecules, including basic proline-rich proteins, adsorbed to many oral surfaces promote C. albicans adherence. Several adhesins present in the C. albicans cell wall have now been partially characterized. Adherence involves lectin, protein-protein, and hydrophobic interactions. As C. albicans cells evade host defenses and colonize new environments by penetrating tissues, they are exposed to new adherence receptors and respond by expressing alternative adhesins. The relatively small number of commensal Candida cells in the oral flora raises the possibility that strategies can be devised to prevent oral colonization and infection. However, the variety of oral niches and the complex adherence mechanisms of the yeast mean that such a goal will remain elusive until more is known about the contribution of each mechanism to colonization.

• Kingsmill VJ.
• Department of Conservative Dentistry, St Bartholomew's and the Royal London School of Medicine and Dentistry, UK.
• Pages 384-404

Following tooth loss, the mandible shows an extensive loss of bone in some individuals. This may pose a significant problem in the prosthodontic restoration of function and esthetics. The many factors which have been proposed as being responsible for the inter-individual variation in post-extraction remodeling mean that a perfunctory analysis of the literature, in which well-controlled, relevant studies are scarce, may not provide the whole story. This article reviews the local and systemic factors which may play a role in the post-extraction remodeling of the mandible. Since severe residual ridge resorption may occur even when the bone status in the rest of the skeleton is good and vice versa, it is concluded that local functional factors are of paramount significance. It is now essential to determine how they can be modified and applied to help maintain ridge height and quality in our aging, edentulous population.

• Imfeld T.
• Department of Preventive Dentistry, Periodontology and Cariology, School of Dentistry, University of Zurich, Switzerland.
• Pages 405-19

The world market for chewing gum is estimated to be 560,000 tons per year, representing approximately US $5 billion. Some 374 billion pieces of chewing gum are sold worldwide every year, representing 187 billion hours of gum-chewing if each piece of gum is chewed for 30 minutes. Chewing gum can thus be expected to have an influence on oral health. The labeling of sugar-substituted chewing gum as "safe for teeth" or "tooth-friendly" has been proven beneficial to the informed consumer. Such claims are allowed for products having been shown in vivo not to depress plaque pH below 5.7, neither during nor for 30 minutes after the consumption. However, various chewing gum manufacturers have recently begun to make distinct health promotion claims, suggesting, e.g., reparative action or substitution for mechanical hygiene. The aim of this critical review--covering the effects of the physical properties of chewing gum and those of different ingredients both of conventional and of functional chewing gum--is to provide a set of guidelines for the interpretation of such claims and to assist oral health care professionals in counseling patients.

Volume 10, Issue 4

• Bartlett JD, Simmer JP.
• Department of Biomineralization, Forsyth Institute, Boston, Massachusetts 02115, USA
• Pages 425-41

For almost three decades, proteinases have been known to reside within developing dental enamel. However, identification and characterization of these proteinases have been slow and difficult, because they are present in very small quantities and they are difficult to purify directly from the mineralizing enamel. Enamel matrix proteins such as amelogenin, ameloblastin, and enamelin are cleaved by proteinases soon after they are secreted, and their cleavage products accumulate in the deeper, more mature enamel layers, while the full-length proteins are observed only at the surface. These results suggest that proteinases are necessary for "activating" enamel proteins so the parent proteins and their cleavage products may perform different functions. A novel matrix metalloproteinase named enamelysin (MMP-20) was recently cloned from tooth tissues and was later shown to localize primarily within the most recently formed enamel. Furthermore, recombinant porcine enamelysin was demonstrated to cleave recombinant porcine amelogenin at virtually all of the sites that have previously been described in vivo. Therefore, enamelysin is at least one enzyme that may be important during early enamel development. As enamel development progresses to the later stages, a profound decrease in the enamel protein content is observed. Proteinases have traditionally been assumed to degrade the organic matrix prior to its removal from the enamel. Recently, a novel serine proteinase named enamel matrix serine proteinase-1 (EMSP1) was cloned from enamel organ epithelia. EMSP1 localizes primarily to the early maturation stage enamel and may, therefore, be involved in the degradation of proteins prior to their removal from the maturing enamel. Other, as yet unidentified, proteinases and proteinase inhibitors are almost certainly present within the forming enamel and await discovery.

• Dotto GP.
• Cutaneous Biology Research Center, Massachusetts General Hospital and Harvard Medical School, Charlestown 02129, USA.
• Pages 442-57

Self-renewing epithelia are characterized by a high turnover rate and a fine balance between growth and differentiation. Such a balance is influenced by many exogenous factors, including gradients of diffusible molecules, cell/substrate adhesion contacts, and direct cell-cell communication. The inter-connection between these various extracellular signals and underlying intracellular pathways is clearly of great interest. Primary keratinocytes of either human or murine origin provide an ideal experimental system to elucidate early signaling events involved in the control of epithelial differentiation. Relative to established cell lines, use of a primary system eliminates the possibility of alterations in critical regulatory events which may occur during prolonged propagation in culture. Primary keratinocytes are easily grown in large numbers, and their differentiation can be induced under well-defined culture conditions. The ensuing rapid and homogeneous response is amenable to careful biochemical analysis. Gene transfer technology (transient transfections, adenoviral and retroviral vectors), together with the use of keratinocytes derived from gene knockout and transgenic mice, makes it possible to assess the specific contribution of individual genes to the control of the differentiation process. This review focuses on the significant progress that has been made over the last few years in our understanding of the specific signals that trigger keratinocyte differentiation, the underlying signaling pathways, and how they impinge on specific transcription and cell-cycle control mechanisms associated with the onset of keratinocyte differentiation. Recent developments and future directions in this important area of research will be highlighted.

• Battino M, Bullon P, Wilson M, Newman H.
• Institute of Biochemistry, Faculty of Medicine, University of Ancona, Italy.
• Pages 458-76

In recent years, there has been a tremendous expansion in medical and dental research concerned with free radicals, reactive oxygen species, and anti-oxidant defense mechanisms. This review is intended to provide a critical, up-to-date summary of the field, with particular emphasis on its implications for the application of "anti-oxidant therapy" in periodontal disease. We have reviewed the nomenclature, mechanisms of actions, features, and sources of most common free radicals and reactive oxygen species, as well as analyzed the typical biological targets for oxidative damage. Based on a review of direct and indirect anti-oxidant host defenses, particularly in relation to the key role of polymorphonuclear neutrophils in periodontitis, we review current evidence for oxidative damage in chronic inflammatory periodontal disease, and the possible therapeutic effects of anti-oxidants in treating and/or preventing such pathology, with special attention to vitamin E and Co-enzyme Q.

• Iwamoto M, Enomoto-Iwamoto M, Kurisu K.
• Department of Oral Anatomy & Developmental Biology, Osaka University Faculty of Dentistry, Suita, Japan.
• Pages 477-86

Recent advances in developmental and molecular biology during embryogenesis and organogenesis have provided new insights into the mechanism of bone formation. Members of the hedgehog gene family were initially characterized as patterning factors in embryonic development, but recently they have been shown to regulate skeletal formation in vertebrates. The amino terminal fragment of Sonic hedgehog (Shh-N), which is an active domain of Shh, has the ability to induce ectopic cartilage and bone formation in vivo. Shh-N stimulates chondrogenic differentiation in cultures of chondrogenic cell line cells in vitro and inhibits chondrogenesis in primary limb bud cells. These findings suggest that the regulation of chondrogenesis by hedgehog proteins depends on the cell populations being studied. Indian hedgehog (Ihh) is prominently expressed in developing cartilage. Ectopic expression of Ihh decreases type X collagen expression and induces the up-regulation of parathyroid hormone-related peptide (PTHrp) gene expression in perichondrium cells. A negative feedback loop consisting of Ihh and PTHrp, induced by Ihh, appears to regulate the rate of chondrocyte maturation. The direct actions of Shh and Ihh on stimulation of osteoblast differentiation are evidenced by the findings that these factors stimulate alkaline phosphatase activity in cultures of pluripotent mesenchymal cell line cells and osteoblastic cells and that these cells express putative receptors of hedgehog proteins. In conclusion, hedgehog proteins seem to be significantly involved in skeletal formation through multiple actions on chondrogenic mesenchymal cells, chondrocytes, and osteogenic cells.

• Takahashi N, Yamada T.
• Department of Oral Biochemistry, Tohoku University School of Dentistry, Sendai, Japan
• Pages 487-503

Actinomyces are among the predominant bacteria in the oral microflora. This review discusses the glucose and lactate metabolism of Actinomyces naeslundii and its ecological significance in dental plaque. This bacterium has the Embden-Meyerhof-Parnas (EMP) pathway as the main route to degrade glucose. The EMP pathway-derived metabolic intermediates, phosphoenolpyruvate (PEP) and pyruvate, are further converted into different end-products, depending on the environment. Under anaerobic conditions in the absence of bicarbonate, the pyruvate is converted into lactate by a lactate dehydrogenase. In the presence of bicarbonate, the PEP is combined with bicarbonate and then converted into succinate through the succinate pathway, while the pyruvate is converted into formate and acetate through the pyruvate formate-lyase pathway. Under aerobic conditions, the pyruvate liberates acetate and CO2 through a pathway initiated by a pyruvate dehydrogenase. A. naeslundii strains also degrade lactate, aerobically, to acetate and CO2 through the conversion of lactate into pyruvate by a NAD-independent lactate dehydrogenase. These strains also synthesize glycogen from a glycolytic intermediate, glucose 6-phosphate. Besides atmospheric conditions and bicarbonate, the intracellular reduction-oxidation potential, carbohydrate concentration, and environmental pH also modulate the metabolism of A. naeslundii. Some of the phosphorylating enzymes involved in A. naeslundii metabolism--e.g., GTP/polyphosphate (PPn)-dependent glucokinase, pyrophosphate (PPi)-dependent phosphofructokinase, UDP-glucose pyrophosphorylase, and GDP/IDP-dependent PEP carboxykinase--are unique to A. naeslundii and have not been found in other oral bacteria. The utilization of PPn and PPi as phosphoryl donors, together with glycogen synthesis and lactate utilization, could contribute to the efficient energy metabolism found in A. naeslundii. Through this flexible and efficient metabolic capacity, A. naeslundii can adapt to fluctuating environments and compete with other bacteria in dental plaque. Further, this bacterium may modify the dental plaque environment and promote the microbial population shifts in dental plaque.

• Stohler CS.
• Department of Biologic and Materials Sciences, and Center for Human Growth and Development, The University of Michigan, Ann Arbor 48109-1078, USA.
• Pages 504-18

Many structural, behavioral, and pharmacological interventions imply that favorable treatment effects in musculoskeletal pain states are mediated through the correction of muscle function. The common theme of these interventions is captured in the popular idea that structural or psychological factors cause muscle hyperactivity, muscle overwork, muscle fatigue, and ultimately pain. Although symptoms and signs of motor dysfunction can sometimes be explained by changes in structure, there is strong evidence that they can also be caused by pain. This new understanding has resulted in a better appreciation of the pathogenesis of symptoms and signs of the musculoskeletal pain conditions, including the sequence of events that leads to the development of motor dysfunction. With the improved understanding of the relationship between pain and motor function, including the inappropriateness of many clinical assumptions, a new literature emerges that opens the door to exciting therapeutic opportunities. Novel treatments are expected to have a profound impact on the care of musculoskeletal pain and its effect on motor function in the not-too-distant future.