RESUMO
CeO2-x nanorods are functional mimics of natural haloperoxidases. They catalyze the oxidative bromination of phenol red to bromophenol blue and of natural signaling molecules involved in bacterial quorum sensing. Laboratory and field tests with paint formulations containing 2 wt% of CeO2-x nanorods show a reduction in biofouling comparable to Cu2 O, the most typical biocidal pigment.
RESUMO
The fundamental mechanisms of biomineralization and their translation into innovative synthetic approaches have yielded promising perspectives for the fabrication of biomimetic and bioinspired organic-inorganic hybrid materials. In siliceous sponges, the enzyme silicatein catalyzes the polycondensation of molecular precursors to nano-structured SiO2 that is deposited on self-assembled filaments consisting of the two silicatein isoforms (silicatein-α and -ß) and the scaffold protein silintaphin-1. Due to its broad substrate specificity silicatein is also able to convert in vitro various other precursors to non-biogenic materials (e.g., hydrolysis of titanium bis(ammonium lactato)-dihydroxide [TiBALDH] and subsequent polycondensation to titania [TiO2]). In the present approach, silicatein was bioengineered to carry a protein tag (Arg-tag) that confers binding affinity to TiO2. Then, by combining Arg-tagged silicatein-α with silicatein-ß and silintaphin-1, self-assembled branched hybrid protein microfilaments were fabricated. Upon subsequent incubation with TiBALDH the filaments were decorated with TiO2 and assayed for photocatalytic activity through photodegradation of the dye methylene blue. This is the first approach that considers concomitant application of two silicatein isoforms for the synthesis of bioinspired organic-inorganic hybrid materials. It is also the first time that the biocatalytic activity of the enzymes has been combined with both the structure-providing properties of silintaphin-1 and a TiO2 affinity protein tag to fabricate self-assembled branched protein filaments as template for a silicatein-synthesized TiO2 photocatalyst. The TiO2-decorated filaments might be explored as a practical alternative to approaches where biotemplates have to be laboriously isolated from their original biological source prior to TiO2 immobilization.
Assuntos
Catepsinas/química , Nanopartículas Metálicas/química , Titânio/química , Marcadores de Afinidade , Catálise , Eletroforese em Gel de Poliacrilamida , Processos FotoquímicosRESUMO
We report the fabrication of a novel type of artificial small diameter blood vessels, termed biomimetic tissue-engineered blood vessels (bTEBV), with a modular composition. They are composed of a hydrogel scaffold consisting of two negatively charged natural polymers, alginate and a modified chitosan, N,O-carboxymethyl chitosan (N,O-CMC). Into this biologically inert scaffold two biofunctionally active biopolymers are embedded, inorganic polyphosphate (polyP) and silica, as well as gelatin which exposes the cell recognition signal, Arg-Gly-Asp (RGD). These materials can be hardened by exposure to Ca(2+) through formation of Ca(2+) bridges between the polyanions, alginate, N,O-CMC, and polyP (alginate-Ca(2+)-N,O-CMC-polyP). The bTEBV are formed by pressing the hydrogel through an extruder into a hardening solution, containing Ca(2+). In this universal scaffold of the bTEBV biomaterial, polycations such as poly(L-Lys), poly(D-Lys) or a His/Gly-tagged RGD peptide (three RGD units) were incorporated, which promote the adhesion of endothelial cells to the vessel surface. The mechanical properties of the biopolymer material (alginate-Ca(2+)-N,O-CMC-polyP-silica) revealed a hardness (elastic modulus) of 475 kPa even after a short incubation period in CaCl2 solution. The material of the artificial vascular grafts (bTEBVs with an outer size 6 mm and 1.8 mm, and an inner diameter 4 mm and 0.8 mm, respectively) turned out to be durable in 4-week pulsatile flow experiments at an alternating pressure between 25 and 100 mbar (18.7 and 75.0 mm Hg). The burst pressure of the larger (smaller) vessels was 850 mbar (145 mbar). Incorporation of polycationic poly(L-Lys), poly(D-Lys), and especially the His/Gly-tagged RGD peptide, markedly increased the adhesion of human, umbilical vein/vascular endothelial cells, EA.HY926 cells, to the surface of the hydrogel. No significant effect of the polyP samples on the clotting of human plasma is measured. We propose that the metabolically degradable polymeric scaffold bTEBV is a promising biomaterial for future prosthetic vascular grafts.
Assuntos
Implantes Absorvíveis , Materiais Biocompatíveis/química , Prótese Vascular , Células Endoteliais/citologia , Oligopeptídeos/farmacologia , Alginatos/química , Coagulação Sanguínea/efeitos dos fármacos , Cloreto de Cálcio/farmacologia , Linhagem Celular Transformada , Quitosana/química , Módulo de Elasticidade , Ácido Glucurônico/química , Ácidos Hexurônicos/química , Células Endoteliais da Veia Umbilical Humana/citologia , Humanos , Hidrogéis/química , Teste de Materiais , Polifosfatos/química , Dióxido de Silício , Resistência à Tração , Engenharia Tecidual , Alicerces Teciduais , Enxerto VascularRESUMO
BACKGROUND: Laccases are copper-containing enzymes that catalyze the oxidation of a wide variety of phenolic substrates. METHODS: We describe the first poriferan laccase from the marine demosponge Suberites domuncula. RESULTS: This enzyme comprises three characteristic multicopper oxidase homologous domains. Immunohistological studies revealed that the highest expression of the laccase is in the surface zone of the animals. The expression level of the laccase gene is strongly upregulated after exposure of the animals to the bacterial endotoxin lipopolysaccharide. To allow the binding of the recombinant enzyme to ferromagnetic nanoparticles, a recombinant laccase was prepared which contained in addition to the His-tag, a Glu-tag at the N-terminus of the enzyme. The recombinant laccase was enzymatically active. The apparent Michaelis constant of the enzyme is 114 µM, using syringaldazine as substrate. Exposure of E. coli to the nanoparticles, coated with Glu-tagged laccase, and to the mediator 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) in the presence of lignin, as the oxidizable substrate, resulted in an almost complete inhibition of colony formation. Quantitative studies of the effect of the laccase-coated iron oxide nanoparticles were performed using E. coli grown in suspension in reaction tubes within a magnetic nanoparticle separator. CONCLUSIONS: This newly designed magnetic nanoparticle separator allowed a removal of the nanoparticles after terminating the reaction. Using this system, a strong dose-dependent inhibition of the growth of E. coli by the laccase iron oxide nanoparticles was determined. GENERAL SIGNIFICANCE: From our data we conclude that the sponge laccase is involved in the anti-bacterial defense of the sponge organism.
Assuntos
Antibacterianos/metabolismo , Lacase/metabolismo , Proteínas Recombinantes/metabolismo , Suberites/enzimologia , Sequência de Aminoácidos , Animais , Biocatálise , Relação Dose-Resposta a Droga , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Compostos Férricos/química , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Hidrazonas/metabolismo , Cinética , Lacase/classificação , Lacase/genética , Lignina/metabolismo , Lipopolissacarídeos/farmacologia , Dados de Sequência Molecular , Nanopartículas/química , Nanopartículas/toxicidade , Oxirredução , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de Aminoácidos , Suberites/genética , Especificidade por Substrato , Regulação para Cima/efeitos dos fármacosRESUMO
Polymeric silica is formed from ortho-silicate during a sol-gel formation process, while biosilica is the product of an enzymatically driven bio-polycondensation reaction. Both polymers have recently been described as a template that induces an increased expression of the genes encoding bone morphogenetic protein 2 (BMP-2) and osteoprotegerin in osteoblast-related SaOS-2 cells; simultaneously or subsequently the cells respond with enhanced hydroxyapatite formation. In order to assess whether the biocompatible polymeric silica/biosilica can serve as a morphogenetically active matrix suitable for three-dimensional (3D) cell growth, or even for 3D cell bioprinting, SaOS-2 cells were embedded into a Na-alginate-based hydrogel. Four different gelatinous hydrogel matrices were used for suspending SaOS-2 cells: (a) the hydrogel alone; (b) the hydrogel with 400 µM ortho-silicate; (c) the hydrogel supplemented with 400 µM ortho-silicate and recombinant silicatein to allow biosilica synthesis to occur; and (d) the hydrogel with ortho-silicate and BSA. The SaOS-2 cells showed an increased growth if silica/biosilica components were present in the hydrogel. Likewise intensified was the formation of hydroxyapatite nodules in the silica-containing hydrogels. After an incubation period of 2 weeks, cells present in silica-containing hydrogels showed a significantly higher expression of the genes encoding the cytokine BMP-2, the major fibrillar structural protein collagen 1 and likewise of carbonic anhydrase. It is concluded that silica, and to a larger extent biosilica, retains its morphogenetic/osteogenic potential after addition to Na-alginate-based hydrogels. This property might qualify silica hydrogels to be also used as a matrix for 3D cell printing.
Assuntos
Alginatos/química , Materiais Biocompatíveis/química , Osso e Ossos/fisiologia , Hidrogéis/química , Dióxido de Silício/química , Alicerces Teciduais/química , Bioimpressão , Proteína Morfogenética Óssea 2/química , Anidrases Carbônicas/química , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular , Colágeno/química , Citocinas/metabolismo , Durapatita/química , Humanos , Osteoblastos/metabolismo , Osteogênese , Polímeros/química , Proteínas Recombinantes/química , Silicatos/químicaRESUMO
Multicomponent particles have emerged in recent years as new compartmentalized colloids with two sides of different chemistry or polarity that have opened up a wide field of unique applications in medicine, biochemistry, optics, physics and chemistry. A drawback of particles containing a ZnO hemisphere is their low stability in biological environment due to the amphoteric properties of Zn2+. Therefore we have synthesized monodisperse Au@ZnO Janus particles by seed-mediated nucleation and growth whose ZnO domain was coated selectively with a thin SiO2 layer as a protection from the surrounding environment that imparts stability in aqueous media while the Au domain remained untouched. The thickness of the SiO2 layer could be precisely controlled. The SiO2 coating of the oxide domain allows biomolecule conjugation (e.g. antibodies, proteins) in a single step for converting the photoluminescent and photocatalytic active Janus nanoparticles into multifunctional efficient vehicles for cell targeting. The SiO2-coated functionalized nanoparticles were stable in buffer solutions and other aqueous systems. Biocompatibility and potential biomedical applications of the Au@ZnO@SiO2 Janus particles were assayed by a cell viability analysis by co-incubating the Au@ZnO@SiO2 Janus particles with epithelia cells and compared to those of uncoated ZnO.
RESUMO
Biomimetic materials have been gaining increasing importance in tissue engineering since they may provide regenerative alternatives to the use of autologous tissues for transplantation. In the present study, we applied for bioprinting of a functionalized three-dimensional template, N,O-carboxymethyl chitosan (N,O-CMC), mimicking the physiological extracellular matrix. This polymer, widely used in tissue engineering, has been provided with functional activity by integration of polyphosphate (polyP), an osteogenically acting natural polymer. The two polymers, N,O-CMC and polyP, are linked together via Ca2+ bridges. This N,O-CMC + polyP material was proven to be printable and durable. The N,O-CMC + polyP printed layers and tissue units retain their properties to induce SaOS-2 bone-like cells to biomineralization. Subsequent in vivo experiments revealed a strong regeneration-inducing activity of the material in the rat calvarial defect model. In turn, N,O-CMC + polyP represents a promising hybrid material useful as a potential custom-designed scaffold for alternative tissue-engineering solutions.
RESUMO
Bioprinting/3D cell printing procedures for the preparation of scaffolds/implants have the potential to revolutionize regenerative medicine. Besides biocompatibility and biodegradability, the hardness of the scaffold material is of critical importance to allow sufficient mechanical protection and, to the same extent, allow migration, cell-cell, and cell-substrate contact formation of the matrix-embedded cells. In the present study, we present a strategy to encase a bioprinted, cell-containing, and soft scaffold with an electrospun mat. The electrospun poly(ϵ-caprolactone) (PCL) nanofibers mats, containing tetraethyl orthosilicate (TEOS), were subsequently incubated with silicatein. Silicatein synthesizes polymeric biosilica by polycondensation of ortho-silicate that is formed from prehydrolyzed TEOS. Biosilica provides a morphogenetically active matrix for the growth and mineralization of osteoblast-related SaOS-2 cells in vitro. Analysis of the microstructure of the 300-700 nm thick PCL/TEOS nanofibers, incubated with silicatein and prehydrolyzed TEOS, displayed biosilica deposits on the mats formed by the nanofibers. We conclude and propose that electrospun PCL nanofibers mats, coated with biosilica, may represent a morphogenetically active and protective cover for bioprinted cell/tissue-like units with a suitable mechanical stability, even if the cells are embedded in a softer matrix.
Assuntos
Materiais Biocompatíveis , Calcificação Fisiológica/efeitos dos fármacos , Nanofibras/química , Poliésteres/química , Dióxido de Silício/química , Alicerces Teciduais/química , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Biotecnologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Humanos , Nanotecnologia , Osteoclastos/citologia , Osteoclastos/metabolismoRESUMO
Silicateins, a group of proteins forming the proteinaceous axial filaments of the inorganic biosilica spicules of the siliceous sponges, are unique in their dual function to exhibit both structure-guiding (providing the structural platform for the biosilica product) and structure-forming activities (enzymatic function: biosilica synthesis from ortho-silicate). The primary translation product of the silicatein gene comprises a signal peptide, a pro-peptide and, separated by an autocatalytic cleavage site glutamine/aspartic acid [Q/D], the sequence of the mature silicatein protein. In order to dissect the biocatalytic, structure-forming activity of silicatein from its structure-guiding function, two mutated genes were constructed based on the silicatein-α gene of the demosponge Suberites domuncula. (i) A gene encoding for a non-processed silicatein that was mutated, by replacing Q/D [Gln (Q)/Asp (D)] by Q/Q, at the cleavage site within the primary translation product between the pro-peptide and the mature enzyme of wild type silicatein. (ii) A gene encoding for a mature enzymatically-active silicatein in which the S-stretch was replaced by a Q-stretch. The enzymatic activity of the mutated protein was significantly enhanced in the presence of the sponge-specific silicatein-interacting protein, silintaphin-1. The two recombinant proteins were applied for micro-contact printing. Using this technique, parallel layers (diameter 10 µm) of the enzymatically inactive, non-processed silicatein were printed onto a gold surface and used as a structure-guiding template for coating with the soluble enzymatically active silicatein. The experiments revealed that after enzymatic reaction with an ortho-silicate substrate a biosilica mantle is formed that can act as a light waveguide.
RESUMO
The enzyme silicatein has been bioengineered to carry a thiol-bearing Au-affinity tag (Cys-tag) for direct immobilization on gold carriers in shortest time without the need for prior surface functionalization. Through microcontact printing, defined silicatein micropatterns were created on gold surfaces, facilitating the subsequent enzymatically controlled synthesis of photocatalytically active TiO(2).
Assuntos
Catepsinas/química , Ouro/química , Titânio/química , Raios Ultravioleta , Catálise , Catepsinas/genética , Catepsinas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Propriedades de SuperfícieRESUMO
In nature, mineralization of hard tissues occurs due to the synergistic effect of components present in the organic matrix of these tissues, with templating and catalytic effects. In Suberites domuncula, a well-studied example of the class of demosponges, silica formation is mediated and templated by an axial proteinaceous filament with silicatein-α, one of the main components. But so far, the effect of other organic constituents from the proteinaceous filament on the catalytic effect of silicatein-α has not been studied in detail. Here we describe the synthesis of core-shell TiO(2)@SiO(2) and TiO(2)@ZrO(2) nanofibers via grafting of silicatein-α onto a TiO(2) nanowire backbone followed by a coassembly of silintaphin-1 through its specifically interacting domains. We show for the first time a linker-free, one-step funtionalization of metal oxides with silicatein-α using glutamate tag. In the presence of silintaphin-1 silicatein-α facilitates the formation of a dense layer of SiO(2) or ZrO(2) on the TiO(2)@protein backbone template. The immobilization of silicatein-α onto TiO(2) probes was characterized by atomic force microscopy (AFM), optical light microscopy, and high-resolution transmission electron microscopy (HRTEM). The coassembly of silicatein-α and silintaphin-1 may contribute to biomimetic approaches that pursue a controlled formation of patterned biosilica-based biomaterials.
Assuntos
Materiais Biomiméticos/química , Catepsinas/química , Suberites/química , Titânio/química , Zircônio/química , Animais , Enzimas Imobilizadas/química , Ácido Glutâmico/química , Nanofios/químicaRESUMO
The metabolic energy state of sponge tissue in vivo is largely unknown. Quantitative bioluminescence-based imaging was used to analyze the ATP distribution of Suberites domuncula (Olivi 1792) tissue, in relation to differences between the cortex and the medulla. This method provides a quantitative picture of the ATP distribution closely reflecting the in vivo situation. The obtained data suggest that the highest ATP content occurs around channels in the sponge medulla. HPLC reverse-phase C-18, used for measurement of ATP content, established a value of 1.62 µmol ATP g⻹ dry mass in sponge medulla, as opposed to 0.04 µmol ATP g⻹ dry mass in the cortex, thus indicating a specific and defined energy distribution. These results correlate with the mitochondria localization, determined using primary antibodies against cytochrome oxidase c subunit 1 (COX1) (immunostaining), as well as with the distribution of arginine kinase (AK), essential for cellular energy metabolism (in situ hybridization with AK from S. domuncula; SDAK), in sponge sections. The highest energy consumption seemed to occur in choanocytes, the cells that drive the water through the channel system of the sponge body. Taken together, these results showed that the majority of energetic metabolism in S. domuncula occurs in the medulla, in the proximity of aqueous channels.
Assuntos
Trifosfato de Adenosina/metabolismo , Metabolismo Energético/fisiologia , Mitocôndrias/fisiologia , Especificidade de Órgãos/fisiologia , Suberites/citologia , Animais , Arginina Quinase/metabolismo , Cromatografia Líquida de Alta Pressão , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Hibridização In Situ , Suberites/metabolismoRESUMO
The skeleton of siliceous sponges consists of amorphous biogenous silica (biosilica). Biosilica formation is driven enzymatically by means of silicatein(s). During this unique process of enzymatic polycondensation, skeletal elements (spicules) that enfold a central proteinaceous structure (axial filament), mainly comprising silicatein, are formed. However, only the concerted action of silicatein and other proteins can explain the genetically controlled diversity of spicular morphotypes, from simple rods with pointed ends to intricate structures with up to six rays. With the scaffold protein silintaphin-1, a first silicatein interactor that facilitates the formation of the axial filament and, consequently, of the growing spicule was discovered. In this study, a new interactor has been identified by both a conventional yeast two-hybrid library screening and a newly established pull-down assay. For the latter approach, silicatein-α has been bioengineered to carry a Glu tag, which confers binding affinity to hydroxyapatite. After immobilization on a solid-phase matrix (hydroxyapatite), the Glu-tagged silicatein was used as bait for the identification of interactors. Both approaches revealed a 15 kDa polypeptide, and its identity was confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Colocalization of silintaphin-2 and silicatein-α within the axial filament and on the spicule surface was shown by immunohistological analyses. Subsequent autoradiography demonstrated the Ca(2+) binding affinity of this silicatein interactor. These findings indicate that both proteins operate in concert during spiculogenesis. Besides binding of calcium, silintaphin-2 shares several structural features with certain acidic, secreted extracellular matrix proteins that facilitate tissue mineralization in Metazoa. Hence, silintaphin-2 might mediate signal transduction during spiculogenesis or may play a more direct role during biosilica formation, in concert with silicatein.
Assuntos
Catepsinas/metabolismo , Mapeamento de Interação de Proteínas/métodos , Suberites/metabolismo , Sequência de Aminoácidos , Animais , Autorradiografia , Cálcio/metabolismo , Dados de Sequência Molecular , Ligação Proteica , Transporte Proteico , Dióxido de Silício/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Técnicas do Sistema de Duplo-HíbridoRESUMO
Since its discovery, numerous biotechnological approaches have aimed to explore the silica-polymerizing catalytic activity of the enzyme silicatein. In vivo, silicatein catalyzes polymerization of amorphous silica nanospheres from soluble precursors. In vitro, it directs the formation of nanostructured biosilica. This is of interest for various applications that strive to benefit from both the advantages of the biological system (i.e., silica synthesis under physiological conditions) and the cell mineralization-stimulating effect of biosilica. However, so far immobilization of silicatein has been hampered by the complex multistep procedure required. In addition, the chemical surface modifications involved not only restrict the choice of carrier materials but also render application of silicatein to hydroxyapatite (HA) of mineralized tissue impossible. Here we describe the bioengineering of silicatein, adapted for application in the fields of bone regeneration, tissue engineering, and dental care. Inspired by Glu-rich sequences of mammalian proteins that confer binding affinity to HA, a novel protein-tag was developed, the Glu-tag. Following expression of Glu-tagged silicatein, the HA-binding capacity of the enzyme is demonstrated in combination with synthetic and dental HA. Furthermore, immobilized Glu-tagged silicatein catalyzes synthesis of biosilica coatings on both synthetic HA nanofibrils and dental HA. Hence, Glu-tagged silicatein reveals a considerable biomedical potential with regenerative and prophylactic implementations.
Assuntos
Bioengenharia/métodos , Catepsinas/metabolismo , Durapatita/síntese química , Polímeros/síntese química , Dióxido de Silício/síntese química , Animais , Catepsinas/isolamento & purificação , Cristalização , Enzimas Imobilizadas/metabolismo , Ácido Glutâmico/metabolismo , Nanofibras/ultraestrutura , Ligação Proteica , Suberites/enzimologia , Sus scrofaRESUMO
Biosilicification in sponges is initiated by formation of proteinaceous filaments, predominantly consisting of silicateins. Silicateins enzymatically catalyze condensation of silica nanospheres, resulting in symmetric skeletal elements (spicules). In order to create tailored biosilica structures in biomimetic approaches it is mandatory to elucidate proteins that are fundamental for the assembly of filaments. Silintaphin-1 is a core component of modularized filaments and also part of a spicule-enfolding layer. It bears no resemblance to other proteins, except for the presence of an interaction domain that is fundamental for its function as scaffold/template. In the presence of silicatein silintaphin-1 facilitates the formation of in vitro filaments. Also, it directs the assembly of gamma-Fe(2)O(3) nanoparticles and surface-immobilized silicatein to rod-like biocomposites, synthetic spicules. Thus, silintaphin-1 will contribute to biomimetic approaches that pursue a controlled formation of patterned biosilica-based materials. Its combination with gamma-Fe(2)O(3) nanoparticles and immobilized silicatein will furthermore inspire future biomedical applications and clinical diagnostics.
Assuntos
Materiais Biomiméticos/metabolismo , Calcificação Fisiológica , Catepsinas/metabolismo , Proteínas/metabolismo , Suberites/enzimologia , Sequência de Aminoácidos , Animais , Anticorpos/farmacologia , Calcificação Fisiológica/efeitos dos fármacos , Catepsinas/ultraestrutura , DNA Complementar/genética , Enzimas Imobilizadas/metabolismo , Compostos Férricos/metabolismo , Dados de Sequência Molecular , Ligação Proteica/efeitos dos fármacos , Transporte Proteico/efeitos dos fármacos , Proteínas/química , Proteínas Recombinantes/metabolismo , Regeneração/efeitos dos fármacos , Ácido Silícico/farmacologia , Suberites/anatomia & histologia , Suberites/citologia , Suberites/efeitos dos fármacos , Técnicas do Sistema de Duplo-HíbridoRESUMO
In a previous study (Schröder et al., J Biomed Mater Res B Appl Biomater 75:387-392, 2005) we demonstrated that human SaOS-2 cells, when cultivated on bio-silica matrices, respond with an increased hydroxyapatite deposition. In the present contribution we investigate if silica-based components (Na-silicate, tetraethyl orthosilicate [TEOS], silica-nanoparticles) (1) change the extent of biomineralization in vitro (SaOS-2 cells) and (2) cause an alteration of the expression of the genes amelogenin, ameloblastin, and enamelin, which are characteristic for an early stage of osteogenesis. We demonstrate that the viability of SaOS-2 cells was not affected by the silica-based components. If Na-silicate or TEOS was added together with ss-glycerophosphate, an organic phosphate donor, a significant increase in biomineralization was measured. Finally, expression levels of the amelogenin, ameloblastin, and enamelin genes were determined in SaOS-2 cells during exposure to the silica-based components. After exposure for 2 days, expression levels of amelogenin and enamelin strongly increased in response to the silica-based components, while no significant change was seen for ameloblastin. In contrast, exposure of SaOS-2 cells to ss-glycerophosphate resulted in increased expression of all three genes. We conclude that the levels of the structural molecules of the enamel matrix, amelogenin and enamelin, increase in the presence of silica-based components and substantially contribute to the extent of hydroxyapatite crystallite formation. These results demonstrate that silica-based components augment hydroxyapatite deposition in vitro and suggest that enzymatically synthesized bio-silica (via silicatein) might be a promising route for tooth reconstruction in vivo.