RESUMO
Without evidence for an organic framework, biological and biochemical processes observed during amelogenesis provided limited information on how extracellular matrix proteins control the development of the complex fibrous architecture of human enamel. Over a decade ago, amelogenin nanoribbons were first observed from recombinant proteins during in vitro mineralization experiments in our laboratory. In enamel from mice lacking the enzyme kallikrein 4 (KLK4), we later uncovered ribbon-like protein structures that matched the morphology, width, and thickness of the nanoribbons assembled by recombinant proteins. Interestingly, similar structures had already been described since the 1960s, when enamel sections from various mammals were demineralized and stained for transmission electron microscopy analysis. However, at that time, researchers were not aware of the ability of amelogenin to form nanoribbons and instead associated the filamentous nanostructures with possible imprints of mineral ribbons in the gel-like matrix of developing enamel. Further evidence for the significance of amelogenin nanoribbons for enamel development was stipulated when recent mineralization experiments succeeded in templating and orienting the growth of apatite ribbons along the protein nanoribbon framework. This article provides a brief historical review of the discovery of amelogenin nanoribbons in our laboratory in the context of reports by others on similar structures in the developing enamel matrix.
Assuntos
Proteínas do Esmalte Dentário , Nanotubos de Carbono , Amelogênese , Amelogenina , Animais , Esmalte Dentário , Proteínas do Esmalte Dentário/genética , CamundongosRESUMO
The nanofibrous nature and its intricate structural organization are the basis for the extraordinary ability of sound enamel to outlive masticatory forces at minimal failure rates. Apatite nanofibers of several hundreds of micrometers to possibly millimeters in length originate during the secretory stage of amelogenesis as 2-nm-thin and 15-nm-wide ribbons that develop and grow in length under the guidance of a dynamic mixture of specialized proteins, the developing enamel matrix (DEM). A critical role in the unidirectional and oriented growth of enamel mineral ribbons has been attributed to amelogenin, the major constituent of the DEM. This review elaborates on recent studies on the ability of ribbon-like assemblies of amelogenin to template the formation of an amorphous calcium phosphate precursor that transforms into apatite mineral ribbons similar to the ones observed in developing enamel. A mechanistic model of the biological processes that drive biomineralization in enamel is presented in the context of a comparative analysis of enamel mouse models and earlier structural data of the DEM emphasizing a regulatory role of the matrix metalloproteinase 20 in mineral deposition and the involvement of a process-directing agent for the templated mineral growth directed by amelogenin nanoribbons.
Assuntos
Proteínas do Esmalte Dentário , Nanotubos de Carbono , Amelogênese , Amelogenina , Animais , Esmalte Dentário , Metaloproteinase 20 da Matriz , CamundongosRESUMO
OBJECTIVES: Acetate and lactate are important cariogenic acids produced by oral bacteria. They produced different residual dentin structures in artificial lesions of similar depth. We evaluated if such lesions responded in the same way to a polymer-induced-liquid-precursor (PILP) remineralization. DESIGN: Dentin blocks obtained from human third molars, divided into 6 groups (n=3). Blocks were demineralized with acetate (66h) or lactate (168h) buffer at pH 5.0 to create 140µm target lesion depths. A-DEM and L-DEM groups received no remineralization. Other groups were remineralized for 14days. 100µg/mL polyaspartate was added into the remineralizing buffer for A-PIL and L-PIL, whereas A-CAP and L-CAP were treated with the same solution but without polyaspartate. Cross-sectioned blocks were examined for shrinkage and AFM-topography. Line profiles of reduced elastic modulus (Er) were obtained by AFM-based nanoindentation across the lesion. Ultrastructures were examined with TEM. RESULTS: A-PIL and L-PIL recovered in shrinkage to the original height of the dentin and it appeared normal with tubules, with increases in Er at both outer flat and inner sloped zones. At the sloped zone, acetate lesions lost more Er but recovery rate after PILP was not statistically different from lactate lesions. A-CAP and L-CAP showed surface precipitates, significantly less recovery in shrinkage or Er as compared to PILP groups. TEM-ultrastructure of PILP groups showed similar structural and mineral components in the sloped zone for lesions produced by either acid. CONCLUSIONS: The PILP process provided significant recovery of both structure and mechanical properties for artificial lesions produced with acetate or lactate.
Assuntos
Dentina/química , Polímeros/química , Desmineralização do Dente/induzido quimicamente , Remineralização Dentária/métodos , Acetatos , Módulo de Elasticidade , Humanos , Técnicas In Vitro , Ácido Láctico , Microscopia de Força Atômica , Microscopia Eletrônica de Transmissão , Dente Serotino , Peptídeos/farmacologia , Propriedades de SuperfícieRESUMO
Dentinogenesis imperfecta type II (DGI-II) lacks intrafibrillar mineral with severe compromise of dentin mechanical properties. A Dspp knockout (Dspp-/-) mouse, with a phenotype similar to that of human DGI-II, was used to determine if poly-L-aspartic acid [poly(ASP)] in the "polymer-induced liquid-precursor" (PILP) system can restore its mechanical properties. Dentin from six-week old Dspp-/- and wild-type mice was treated with CaP solution containing poly(ASP) for up to 14 days. Elastic modulus and hardness before and after treatment were correlated with mineralization from Micro x-ray computed tomography (Micro-XCT). Transmission electron microscopy (TEM)/Selected area electron diffraction (SAED) were used to compare matrix mineralization and crystallography. Mechanical properties of the Dspp-/- dentin were significantly less than wild-type dentin and recovered significantly (P < 0.05) after PILP-treatment, reaching values comparable to wild-type dentin. Micro-XCT showed mineral recovery similar to wild-type dentin after PILP-treatment. TEM/SAED showed repair of patchy mineralization and complete mineralization of defective dentin. This approach may lead to new strategies for hard tissue repair.
RESUMO
OBJECTIVES: We studied artificial dentin lesions in human teeth generated by lactate and acetate buffers (pH 5.0), the two most abundant acids in caries. The objective of this study was to determine differences in mechanical properties, mineral density profiles and ultrastructural variations of two different artificial lesions with the same approximate depth. METHODS: 0.05M (pH 5.0) acetate or lactate buffer was used to create 1) 180µm-deep lesions in non-carious human dentin blocks (acetate 130h; lactate 14days); (2) demineralized, â¼180µm-thick non-carious dentin discs (3 weeks). We performed nanoindentation to determine mechanical properties across the hydrated lesions, and micro X-ray computed tomography (MicroXCT) to determine mineral profiles. Ultrastructure in lesions was analyzed by TEM/selected area electron diffraction (SAED). Demineralized dentin discs were analyzed by small angle X-ray scattering (SAXS). RESULTS: Diffusion-dominated demineralization was shown based on the linearity between lesion depths versus the square root of exposure time in either solution, with faster kinetics in acetate buffer. Nanoindentation revealed lactate induced a significantly sharper transition in reduced elastic modulus across the lesions. MicroXCT showed lactate demineralized lesions had swelling and more disorganized matrix structure, whereas acetate lesions had abrupt X-ray absorption near the margin. At the ultrastructural level, TEM showed lactate was more effective in removing minerals from the collagenous matrix, which was confirmed by SAXS analysis. CONCLUSIONS: These findings indicated the different acids yielded lesions with different characteristics that could influence lesion formation resulting in their distinct predominance in different caries activities, and these differences may impact strategies for dentin caries remineralization.
Assuntos
Acetatos/farmacocinética , Dentina/ultraestrutura , Ácido Láctico/farmacocinética , Desmineralização do Dente , Acetatos/química , Fenômenos Biomecânicos , Módulo de Elasticidade , Dureza , Humanos , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Ácido Láctico/química , Microscopia Eletrônica de Transmissão , Dente Serotino , Espalhamento a Baixo Ângulo , Microtomografia por Raio-XRESUMO
Enamel is unique. It is the only epithelial-derived mineralized tissue in mammals and has a distinct micro- and nanostructure with nanofibrous apatite crystals as building blocks. It is synthesized by a highly specialized cell, the ameloblast, which secretes matrix proteins with little homology to any other known amino acid sequence, but which is composed of a primary structure that makes it competent to self-assemble and control apatite crystal growth at the nanometer scale. The end-product of ameloblast activity is a marvel of structural engineering: a material optimized to provide the tooth with maximum biting force, withstanding millions of cycles of loads without catastrophic failure, while also protecting the dental pulp from bacterial attack. This review attempts to bring into context the mechanical behavior of enamel with the developmental process of amelogenesis and structural development, since they are linked to tissue function, and the importance of controlling calcium phosphate mineralization at the nanometer scale. The origins of apatite nanofibers, the development of a stiffness gradient, and the biological processes responsible for the synthesis of a hard and fracture-resistant dental tissue are discussed with reference to the evolution of enamel from a fibrous composite to a complex, tough, and damage-tolerant coating on dentin.
Assuntos
Ameloblastos/fisiologia , Esmalte Dentário/fisiologia , Ameloblastos/metabolismo , Amelogênese/fisiologia , Apatitas/farmacologia , Fenômenos Biomecânicos , Fosfatos de Cálcio/farmacologia , Cristalização , Esmalte Dentário/ultraestrutura , Proteínas do Esmalte Dentário/farmacologia , Humanos , Nanofibras/ultraestrutura , Calcificação de Dente/fisiologiaRESUMO
The recent discovery of conditions that induce nanoribbon structures of amelogenin protein in vitro raises questions about their role in enamel formation. Nanoribbons of recombinant human full-length amelogenin (rH174) are about 17 nm wide and self-align into parallel bundles; thus, they could act as templates for crystallization of nanofibrous apatite comprising dental enamel. Here we analyzed the secondary structures of nanoribbon amelogenin by x-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) and tested if the structural motif matches previous data on the organic matrix of enamel. XRD analysis showed that a peak corresponding to 4.7 Å is present in nanoribbons of amelogenin. In addition, FTIR analysis showed that amelogenin in the form of nanoribbons was comprised of ß-sheets by up to 75%, while amelogenin nanospheres had predominantly random-coil structure. The observation of a 4.7-Å XRD spacing confirms the presence of ß-sheets and illustrates structural parallels between the in vitro assemblies and structural motifs in developing enamel.
Assuntos
Amelogenina/química , Esmalte Dentário/química , Nanopartículas/química , Motivos de Aminoácidos , Cloreto de Cálcio/química , Humanos , Concentração de Íons de Hidrogênio , Microscopia de Força Atômica , Nanosferas/química , Fosfatos/química , Cloreto de Potássio/química , Compostos de Potássio/química , Proteínas Recombinantes , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Difração de Raios X/métodosRESUMO
Electromechanical coupling, a phenomenon present in collagenous materials, may influence cell-extracellular matrix interactions. Here, electromechanical coupling has been investigated via piezoresponse force microscopy in transparent and opaque membranes consisting of helical-like arrays of aligned type I collagen fibrils self-assembled from acidic solution. Using atomic force microscopy, the transparent membrane was determined to contain fibrils having an average diameter of 76 ± 14 nm, whereas the opaque membrane comprised fibrils with an average diameter of 391 ± 99 nm. As the acidity of the membranes must be neutralized before they can serve as cell culture substrates, the structure and piezoelectric properties of the membranes were measured under ambient conditions before and after the neutralization process. A crimp structure (1.59 ± 0.37 µm in width) perpendicular to the fibril alignment became apparent in the transparent membrane when the pH was adjusted from acidic (pH = 2.5) to neutral (pH = 7) conditions. In addition, a 1.35-fold increase was observed in the amplitude of the shear piezoelectricity of the transparent membrane. The structure and piezoelectric properties of the opaque membrane were not significantly affected by the neutralization process. The results highlight the presence of an additional translational order in the transparent membrane in the direction perpendicular to the fibril alignment. The piezoelectric response of both membrane types was found to be an order of magnitude lower than that of collagen fibrils in rat tail tendon. This reduced response is attributed to less-ordered molecular assembly than is present in D-periodic collagen fibrils, as evidenced by the absence of D-periodicity in the membranes.
Assuntos
Colágeno Tipo II/química , Colágeno Tipo I/química , Técnicas Eletroquímicas , Membranas Artificiais , Animais , Concentração de Íons de Hidrogênio , Microscopia de Força Atômica , RatosRESUMO
Assembling artificial collagenous tissues with structural, functional, and mechanical properties which mimic natural tissues is of vital importance for many tissue engineering applications. While the electro-mechanical properties of collagen are thought to play a role in, for example, bone formation and remodeling, this functional property has not been adequately addressed in engineered tissues. Here the electro-mechanical properties of rat tail tendon are compared with those of dried isoelectrically focused collagen hydrogels using piezoresponse force microscopy under ambient conditions. In both the natural tissue and the engineered hydrogel D-periodic type I collagen fibrils are observed, which exhibit shear piezoelectricity. While both tissues also exhibit fibrils with parallel orientations, Fourier transform analysis has revealed that the degree of parallel alignment of the fibrils in the tendon is three times that of the dried hydrogel. The results obtained demonstrate that isoelectrically focused collagen has similar structural and electro-mechanical properties to that of tendon, which is relevant for tissue engineering applications.
Assuntos
Colágeno/farmacologia , Dessecação , Eletroquímica/métodos , Hidrogéis/farmacologia , Tendões/efeitos dos fármacos , Tendões/fisiologia , Animais , Fenômenos Biomecânicos , Análise de Fourier , Focalização Isoelétrica , Microscopia de Força Atômica , Ratos , Cauda , Tendões/ultraestruturaRESUMO
The developing enamel matrix is a highly dynamic system mainly composed of the full-length amelogenin and its proteolytic cleavage products. In this study, size, zeta-potential, and the isoelectric points of nanoparticles of the recombinant full-length human amelogenin (rH174) and two proteolytic products (rH163 and rH146) were analyzed by dynamic light-scattering and electrokinetic measurements. We tested the hypothesis that zeta-potential may be used as a control parameter in directing the self-assembly of amelogenins. Extensive aggregation of amelogenin molecules with the particle size reaching about one micron occurred at a mildly acidic to neutral pH, and coincided with the red shift of the internal fluorescence. Zeta-potential was between +/- 15 mV in the same pH range, indicating that amelogenin aggregation occurred when surface potentials were minimal. This suggests that electrostatic interactions may be another crucial factor, aside from hydrophobic interaction, in the aggregation and hierarchical assembly of spherical particles of amelogenins into supramolecular structures of a higher order.
Assuntos
Amelogenina/química , Humanos , Concentração de Íons de Hidrogênio , Hidrólise , Focalização Isoelétrica , Cinética , Potenciais da Membrana , Nanosferas , Tamanho da Partícula , Proteínas Recombinantes , Espalhamento de Radiação , Propriedades de SuperfícieRESUMO
The objective of this article is to critically evaluate the methods that are used to assess outcomes of remineralization of dentin. Currently, the most used assessment methods fall either into quantitative analysis of the mineral content of the remineralized structures or dry measurements of their mechanical properties. Properties obtained from the dehydrated organic dentin matrix may not reflect the true mechanical behavior of the remineralized tissue under physiological and hydrated conditions. Here we seek to clarify the biomechanical aspects of remineralization of dentin, pointing out the effects of hydration and dehydration on the mechanical properties of treated tissues. We also emphasize that a more appropriate endpoint to evaluate the effectiveness of remineralization in dentin should be associated with the recovery of the mechanical properties of the hydrated tissue, which is presumed to correlate well with its overall functionality.
Assuntos
Análise do Estresse Dentário/métodos , Dentina/química , Dentina/fisiologia , Remineralização Dentária , Fenômenos Biomecânicos , Dessecação , Elasticidade , Colágenos Fibrilares/química , Testes de Dureza , Humanos , Microrradiografia , Microscopia Eletrônica de Transmissão , Modelos Químicos , Avaliação de Resultados em Cuidados de Saúde/métodos , Análise Espectral , Termogravimetria , ÁguaRESUMO
Dentin and bone derive their mechanical properties from a complex arrangement of collagen type-I fibrils reinforced with nanocrystalline apatite mineral in extra- and intrafibrillar compartments. While mechanical properties have been determined for the bulk of the mineralized tissue, information on the mechanics of the individual fibril is limited. Here, atomic force microscopy was used on individual collagen fibrils to study structural and mechanical changes during acid etching. The characteristic 67 nm periodicity of gap zones was not observed on the mineralized fibril, but became apparent and increasingly pronounced with continuous demineralization. AFM-nanoindentation showed a decrease in modulus from 1.5 GPa to 50 MPa during acid etching of individual collagen fibrils and revealed that the modulus profile followed the axial periodicity. The nanomechanical data, Raman spectroscopy and SAXS support the hypothesis that intrafibrillar mineral etches at a substantially slower rate than the extrafibrillar mineral. These findings are relevant for understanding the biomechanics and design principles of calcified tissues derived from collagen matrices.
Assuntos
Colágeno/química , Dente/química , Apatitas/química , Fenômenos Biomecânicos , Dentina/química , Dureza , Humanos , Microscopia de Força Atômica/métodos , Nanopartículas/química , Espalhamento de Radiação , Análise Espectral Raman , Estresse Mecânico , Propriedades de Superfície , Dente/patologia , Raios XRESUMO
Dental enamel forms through a protein-controlled mineralization and enzymatic degradation with a nanoscale precision that new engineering technologies may be able to mimic. Recombinant full-length human amelogenin (rH174) and a matrix-metalloprotease (MMP-20) were employed in a pH-stat titration system that enabled a continuous supply of calcium and phosphate ions over several days, mimicking the initial stages of matrix processing and crystallization in enamel in-vitro. Effects on the self-assembly and crystal growth from a saturated aqueous solution containing 0.4 mg/ml rH174 and MMP-20 with the weight ratio of 1:1000 with respect to rH174 were investigated. A transition from nanospheres to fibrous amelogenin assemblies was facilitated under conditions that involved an interaction between rH174 and the proteolytic cleavage products. Despite continuous titration, the levels of calcium exhibited a consistent trend of decreasing, thereby indicating its possible role in the protein self-assembly. This study suggests that mimicking enamel formation in-vitro requires the synergy between the aspects of matrix self-assembly, proteolysis and crystallization.
RESUMO
Dentin is a mesenchymal tissue, and, as such, is based on a collagenous matrix that is reinforced by apatite mineral. Collagen fibrils show piezoelectricity, a phenomenon that is used by piezoresponse force microscopy (PFM) to obtain high-resolution images. We applied PFM to image human dentin with 10-nm resolution, and to test the hypothesis that zones of piezoactivity, indicating the presence of collagen fibrils, can be distinguished in dentin. Piezoelectricity was observed by PFM in the dentin intertubular matrix, while the peritubular dentin remained without response. High-resolution imaging of chemically treated intertubular dentin attributed the piezoelectric effect to individual collagen fibrils that differed in the signal strength, depending on the fibril orientation. This study supports the hypothesis that peritubular dentin is a non-collagenous tissue and is thus an exception among mineralized tissues that derive from the mesenchyme.
Assuntos
Dentina/fisiologia , Dentina/ultraestrutura , Eletrofisiologia , Colágenos Fibrilares/análise , Colágenos Fibrilares/ultraestrutura , Humanos , Microscopia de Força Atômica , Dente SerotinoRESUMO
High-resolution studies of dental tissues are of considerable interest for biomedical engineering and clinical applications. In this paper, we demonstrate the application of piezoresponse force microscopy (PFM) to nanoscale imaging of internal structure of human teeth by monitoring the local mechanical response to an electrical bias applied via a conductive tip. It is shown that PFM is capable of detecting dissimilar components of dental tissues, namely, proteins and calcified matrix, which have resembling morphology but different piezoelectric properties. It is demonstrated that collagen fibrils revealed in chemically treated intertubular dentin exhibit high piezoelectric activity and can be visualized in PFM with spatial resolution of 10 nm. Evidence of the presence of protein inclusions of 100-200 nm wide and several micrometers long in tooth enamel has been obtained. Furthermore, it is found that the peritubular dentin and intertubular dentin exhibit different piezoelectric behavior suggesting different concentration of collagen fibrils. The obtained results demonstrate a high potential of PFM in providing an additional insight into the structure of dental tissues. It is suggested that the PFM approach can be used to study the structure of a wide range of biological materials by monitoring their electromechanical behavior at the nanoscale.
Assuntos
Microscopia de Varredura por Sonda/instrumentação , Microscopia de Varredura por Sonda/métodos , Proteínas/ultraestrutura , Dente/ultraestrutura , Adulto , Humanos , Proteínas/análise , Dente/químicaRESUMO
OBJECTIVE: To study the mechanisms which promote the interactions of amelogenin proteins with the forming mineral to establish suitable conditions for the biomimetic synthesis of enamel in vitro. DESIGN: Saturated calcium phosphate solutions were used in conjunction with recombinant amelogenin proteins to induce mineral formation on glass-ceramics substrates containing oriented fluoroapatite crystals (FAP). The height of mineral layers formed on these substrates within 24 h was measured by atomic force microscopy (AFM). EXPERIMENTAL VARIABLES: The effect of protein concentration, pH and degree of saturation (DS) on the growth of apatite mineral was evaluated. Mineralization experiments were performed at 0, 0.4 and 1.6 mg/ml amelogenin concentrations. Mineralization solutions were used at pH values of 6.5, 7.4, 8.0 and 8.8 and DS of calcium and phosphate between 9 and 13. OUTCOME MEASURE: Height and morphology of mineralized layer formed on glass-ceramic substrates as determined from AFM measurements. RESULTS: Homogeneous nucleation and crystal growth of thin layers on the FAP were observed, when calcium and phosphate ions were added. The height of these layers grown on (001) planes of FAP was strongly dependent on the protein concentration and pH. At concentrations of 0 and 0.4 mg/ml crystal grew 5-15 nm on the FAP, while they grew approximately to 200 nm at 1.6 mg/ml. The enhanced crystal growth was observed only at pH 6.5, 7.4 and 8.0, while layers only 20 nm thick were obtained at pH 8.8. An increase in DS resulted in uncontrolled growth of calcium phosphate mineral covering large areas of the substrate. CONCLUSIONS: Protein concentration, pH and the saturation of the mineralizing solution need to be considered carefully to provide suitable conditions for amelogenin-guided growth of apatite crystals.
Assuntos
Amelogênese , Apatitas/química , Cristalização , Proteínas do Esmalte Dentário/farmacologia , Proteínas do Esmalte Dentário/fisiologia , Amelogenina , Materiais Biomiméticos/química , Fosfatos de Cálcio/química , Humanos , Concentração de Íons de Hidrogênio , Microscopia de Força Atômica , Concentração Osmolar , Proteínas Recombinantes/farmacologiaRESUMO
The formation of aligned fibrous apatite crystals in enamel is predominantly attributed to the involvement of amelogenin proteins. We developed a model to study interactions of matrix proteins with apatite mineral in vitro and tested the hypothesis that amelogenin solubility affects the ability to induce protein-guided mineralization. Crystal growth experiments were performed on fluoroapatite (FAP) glass-ceramics in mineralizing solutions containing recombinant full-length amelogenin (rH174) at different concentrations. Using atomic force microscopy, we observed that mineral precipitated randomly on the substrate, but also formed thin layers (height, 10 nm) on FAP within 24 hrs. This growth pattern was unaffected when 0.4 mg/mL of rH174 was added. In contrast, crystals grew on FAP at a rate up to 20 times higher, at 1.6 mg/mL protein. Furthermore, nanospheres and mineral bound specifically to FAP and aligned in strings approximately parallel to the c-axis of FAP, leading us to the conclusion that amelogenin proteins indeed control direction and rate of growth of apatite in enamel.
Assuntos
Apatitas/química , Cerâmica/química , Proteínas do Esmalte Dentário/química , Vidro/química , Amelogenina , Materiais Biomiméticos/química , Precipitação Química , Cristalização , Humanos , Microscopia de Força Atômica , Proteínas Recombinantes , Solubilidade , Análise Espectral RamanRESUMO
It is widely held that the hardness and modulus of dentin increase in proportion to the mineral concentration. To test this belief, we measured hardness and modulus of normal dentin and an altered form of dentin without gap-zone mineralization in wet and dry conditions by AFM nanoindentation to determine if the modulus and hardness scale linearly with mineral concentration. Mineral concentrations in the mid-coronal location of the normal and altered dentins were 44.4 vol% and 30.9 vol%, respectively. Surrounding the pulp of the altered dentin was a region of higher mineralization, 40.5 vol%. The indentation modulus of normal dentin was 23.9 (SD = 1.1) GPa dry and 20.0 (SD = 1.0) GPa wet. In mid-coronal regions of the altered dentin, the indentation modulus was 13.8 (SD = 2.0) GPa dry and 5.7 (SD = 1.4) GPa wet. In the more mineralized regions of the altered dentin, the modulus was 20.4 (SD = 1.8) GPa dry and 5.3 (SD = 0.8) GPa wet; the properties of the altered wet dentin did not correlate with mineral concentration. The results of this study raise doubt as to whether mineral concentration alone is a sufficient endpoint for assessing the success or failure of remineralization approaches in restorative dentistry.
Assuntos
Colágeno/química , Dentina/anatomia & histologia , Adolescente , Adulto , Fenômenos Biomecânicos , Polpa Dentária/ultraestrutura , Dentina/química , Dentinogênese Imperfeita/metabolismo , Dentinogênese Imperfeita/patologia , Dessecação , Elasticidade , Feminino , Dureza , Humanos , Processamento de Imagem Assistida por Computador , Modelos Lineares , Masculino , Microscopia de Força Atômica , Minerais/química , Nanotecnologia , Tomografia Computadorizada por Raios X , Água/químicaRESUMO
The dentino-enamel junction (DEJ) constitutes a structurally unique interphase uniting two mineralized tissues with very different matrix composition and physical properties. Its excellent biomechanical properties have drawn interest as a biomimetic model for joining dissimilar materials. In order to characterize the functional width of the DEJ, nanoscratching experiments were performed on human third molars. Friction coefficients of enamel, of dentin, and at the DEJ were obtained with a nanoscratch tester attached to an atomic force microscope (AFM). Normal loads in the range of 50 to 600 microN were applied to a spherical diamond indenter (r = 10 microm), which was driven 10 microm across the sample surface, recording the lateral force. Imaging with an AFM facilitated exact positioning of the scratches. The friction coefficient of intertubular dentin was 0.31 +/- 0.05, significantly above the coefficient of enamel of 0.14 +/- 0.02. The increased friction of dentin is attributed to the higher content of organic phases. Scratches performed across the interphase between enamel and dentin showed a sharp monotonic change in the friction coefficient. The average width of the slope between the friction coefficients of dentin and enamel was 2.0 +/- 1.1 microm and is assumed to represent the functional width of the dentino-enamel junction. The effect of the scalloped structure of the DEJ on its functional width as determined by mechanical testing is discussed.
Assuntos
Esmalte Dentário/química , Esmalte Dentário/ultraestrutura , Dentina/química , Dentina/ultraestrutura , Microscopia de Força Atômica/métodos , Fenômenos Químicos , Físico-Química , Humanos , Estresse MecânicoRESUMO
Most restorative materials are bonded to caries-affected dentin that has altered structure. We tested the hypothesis that hydrated dentin of the transparent zone did not have increased hardness or elastic modulus. Nanoindentation by modified AFM was used to determine site-specific elastic modulus and hardness for components of hydrated dentin from 8 carious and non-carious human teeth. Indentations in intertubular dentin were made at intervals from pulp through the affected layers (subtransparent, transparent, and discolored zones). The values of intertubular dentin increased slightly from near the pulp into the transparent zone, then remained constant or decreased slightly through transparent dentin (E, 18.3 GPa; H, 0.8 GPa; confirming the hypothesis), and decreased markedly through the discolored region. Peritubular dentin values were unaltered in transparent dentin, and intratubular mineral had values between those of normal peritubular and intertubular dentin. Superficial areas contained distorted tubules without peritubular dentin or intratubular mineral.