Nucleation of apatite crystals in vitro by self-assembled dentin matrix protein 1

He, Gen; Dahl, Tom; Veis, Arthur; George, Anne

doi:10.1038/nmat945

Cited by 496 publications

(513 citation statements)

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“…Also, spherical particles containing substructures have been observed for the formation of calcium phosphate in the presence of dentin [42]. In their study, He et al found that the amorphous calcium phosphate particles were initially formed and then at later times HAP needles formed within the amorphous matrix by solid-state nucleation.…”

Section: Calcium Phosphate Characterizationmentioning

confidence: 99%

The nucleation and growth of calcium phosphate by amelogenin

Tarasevich

Howard

Larson

et al. 2007

Journal of Crystal Growth

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The nucleation processes involved in calcium phosphate formation in tooth enamel are not well understood but are believed to involve proteins in the extracellular matrix. The ability of one enamel protein, amelogenin, to promote the nucleation and growth of calcium phosphate was studied in an in vitro system involving metastable supersaturated solutions. It was found that recombinant amelogenin (rM179 and rp(H)M180) promoted the nucleation of calcium phosphate compared to solutions without protein. The amount of calcium phosphate increased with increasing supersaturation of the solutions and increasing protein concentrations up to 6.5 μg/mL. At higher protein concentrations, the amount of calcium phosphate decreased. The kinetics of nucleation was studied in situ and in real time using a quartz crystal microbalance (QCM) and showed that the protein reduced the induction time for nucleation compared to solutions without protein. This work shows a nucleation role for amelogenin in vitro which may be promoted by the association of amelogenin into nanosphere templates, exposing charged functionality at the surface.

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Section: Calcium Phosphate Characterizationmentioning

confidence: 99%

The nucleation and growth of calcium phosphate by amelogenin

Tarasevich

Howard

Larson

et al. 2007

Journal of Crystal Growth

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“…DMP1 was detected at high levels by Northern analysis in fetal bovine brain and cultured long bone as well as in odontoblasts (6). Most recently, DMP1 was localized by immunohistochemistry in human lung cancer tissue (7).DMP1 is a small protein that has been postulated to have a high affinity for hydroxylapatite (HA) (8,9). This coupled with its localization in mineralized tissues suggested that DMP1 could have an important role in mineralization.…”

mentioning

confidence: 99%

“…DMP1 is a small protein that has been postulated to have a high affinity for hydroxylapatite (HA) (8,9). This coupled with its localization in mineralized tissues suggested that DMP1 could have an important role in mineralization.…”

mentioning

confidence: 99%

In Vitro Effects of Dentin Matrix Protein-1 on Hydroxyapatite Formation Provide Insights into in Vivo Functions

Tartaix

Doulaverakis

George

et al. 2004

Journal of Biological Chemistry

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Dentin matrix protein-1 (DMP1) is a mineralized tissue matrix protein synthesized by osteoblasts, hypertrophic chondrocytes, and ameloblasts as well as odontoblasts. DMP1 is believed to have multiple in vivo functions, acting both as a signaling molecule and a regulator of biomineralization. Using a cell-free system in vitro, we evaluated the action of DMP1 in the regulation of hydroxylapatite (HA) formation and crystal growth. The non-phosphorylated recombinant protein acted as an HA nucleator, increasing the amount of mineral formed in a gelatin gel HA growth system relative to protein-free controls. The recombinant protein phosphorylated in vitro had no detectable effect on HA formation and growth. In contrast, phosphorylated bovine DMP1 expressed in marrow stromal cells with an adenovirus vector containing 29.7 phosphates/mol was an effective inhibitor of HA formation and growth. The native full-length protein appeared to be absent or present in only small amounts in the extracellular matrix of bones and teeth. However, two highly phosphorylated fragments representing the N-and C-terminal portions of DMP1 have been identified, apparently arising from proteolytic cleavage of four X-Asp bonds. The highly phosphorylated C-terminal 57-kDa fragment (containing 42 phosphates/mol), like the non-phosphorylated DMP1, was an HA nucleator. These data suggest that, in its native form, DMP1 inhibits mineralization, but when cleaved or dephosphorylated, it initiates mineralization. These in vitro data are consistent with the findings in the DMP1 knockout mouse.Dentin matrix protein-1 (DMP1) 1 is an acidic, phosphorylated, integrin-binding extracellular matrix protein first identified by screening a rat cDNA library (1). Northern blot analysis in the rat originally suggested that the DMP1 message was odontoblast-specific, with in situ hybridization showing DMP1 mRNA expression to be restricted to those fully differentiated odontoblasts engaged in active dentin matrix formation, with transient expression by ameloblasts (1, 2). More recent in situ hybridization studies show a much broader pattern of expression of DMP1, with expression associated with a number of mineralizing tissues, including bone and cementum (3). In rat and chicken bone, immunohistochemical detection showed an association of DMP1 with osteocytes, but not with osteoblasts (4). In the mouse, hypertrophic chondrocytes express more DMP1 than other cell types (5). DMP1 was detected at high levels by Northern analysis in fetal bovine brain and cultured long bone as well as in odontoblasts (6). Most recently, DMP1 was localized by immunohistochemistry in human lung cancer tissue (7).DMP1 is a small protein that has been postulated to have a high affinity for hydroxylapatite (HA) (8,9). This coupled with its localization in mineralized tissues suggested that DMP1 could have an important role in mineralization. DMP1 is a member of the SIBLING (small integrin-binding ligand, Nlinked glycoprotein) family of proteins (10, 11). Like other members of the SIBLING family...

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“…He et al [41] used a double-diffusion method to study the growth of HA on self-assembled recombinant DMP-1 that was adsorbed on glass slides. This study did not employ a gel, but calcium and phosphate were electro-diffused to the adsorbed protein site from opposite directions using an electrophoretic device.…”

Section: Gel Precipitation Studiesmentioning

confidence: 99%

Diffusion Systems for Evaluation of Biomineralization

Silverman

Boskey

2004

Calcif Tissue Int

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A variety of in vitro study methods have been used to elucidate the roles of matrix molecules in biomineralization processes. Among these, gel diffusion-precipitation studies have proved to be an effective tool. This methodology is uniquely capable of characterizing the effects of matrix molecules on mineralization while only using very small quantities of material. Furthermore, gel methods have been extended for use as a mineralization assay system to characterize modified matrix molecules and synthetic analogues. Here we discuss the advantages and limitations of gelatin, agar, agarose, and other systems for studying the mechanisms of biomineralization. KeywordsHydroxyapatite; Biomineralization; Gel diffusion; Gelatin; Agarose; Agar In Vitro StudiesThe functions of vertebrate and non-vertebrate mineralized tissue matrix biomolecules, such as osteopontin [1][2][3][4], osteonectin [5,6], amelogenin [7,8], bone sialoprotein [2,3,6,9], and a variety of shell proteins [10,11] generally have been demonstrated in a three-step process. First, molecules that may influence the biomineralization process are either isolated from mineralized tissue, purified, and characterized, or cloned and expressed in recombinant forms. Their spatial and temporal distribution in mineralizing tissue, and their presence or absence from diseased or non-mineralizing tissue, as assessed by immunolocalization or in situ hybridization, suggests their role in mineral formation. Second, the molecule being studied can be associated with the biomineralization process in vivo via tissue studies on genetically modified or diseased organisms where expression of the substance of interest is altered, prevented, or enhanced. Third, in vitro mineralization studies may be used to provide insight into the mechanism of action of matrix proteins and to elucidate a molecule's function in vivo [12]. While many different techniques have been used in the in vitro studies, gel precipitation methods, used for over a century for the growth of large crystals [13], have recently been introduced to help elucidate how non-collagenous proteins and other biomolecules act to control biomineralization processes.The interplay of in vivo and in vitro studies allows characterization of a molecule's role in complex living tissue, where we have limited experimental control, against a backdrop of information from well-controlled, simpler precipitation studies. The cell-free in vitro studies allow characterization of an individual compound or mixture's effects on nucleation, growth kinetics, inhibition, and crystal composition and habit, as well as quantitative comparisons We review here the recent application of crystal formation in gels to probe mineralization mechanisms with emphasis placed on hydroxyapatite formation and growth, although the techniques described are applicable to other types of biomineralization.Most in vitro studies of matrix proteins involved in hydroxyapatite (HA) formation have been done in solution, as opposed to gels. The solution studies charact...

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Nucleation of apatite crystals in vitro by self-assembled dentin matrix protein 1

Cited by 496 publications

References 28 publications

The nucleation and growth of calcium phosphate by amelogenin

The nucleation and growth of calcium phosphate by amelogenin

In Vitro Effects of Dentin Matrix Protein-1 on Hydroxyapatite Formation Provide Insights into in Vivo Functions

Diffusion Systems for Evaluation of Biomineralization

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