Biomimetic Remineralization Research Articles

Self-Etch Adhesive as a Carrier for ACP Nanoprecursors to Deliver Biomimetic Remineralization.

Lab biomineralization should be carried out in an actual clinical practice. This study evaluated self-etch adhesive as a carrier for amorphous calcium phosphate (ACP) nanoprecursors to continuously deliver biomimetic remineralization of self-assembly type I collagen and demineralized dentin. Si-containing ACP particles (Si-ACP) stabilized with polyaspartic acid (PAsp) were synthesized and characterized by transmission electron microscopy (TEM), scanning electron microscopy-energy-dispersive X-ray spectroscopy, Fourier transform infrared analysis, X-ray powder diffractometry, and X-ray phototelectron spectroscopy. The biomimetic remineralization of single-layer reconstituted type I collagen fibrils and demineralized dentin was analyzed by using two one-bottle self-etch dentin adhesives (Clearfil S3 Bond (S3), Kurraray-Noritake; Adper Easy One (AEO), 3 M ESPE) as a carrier loaded (or not, in the case of the control) with 25 wt % of Si-ACP particles. In vitro cytotoxicity assessed by the Cell Counting Kit-8 indicated that the Si-ACP particles had no adverse effect on cell viability. The capacity for Ca and P ions release from cured Si-ACP-containing adhesives (S3, AEO) was evaluated by inductively coupled plasma-atomic emission spectrometry, revealing the successively increasing release of Ca and P ions for 28 days. The intra- and extrafibrillar remineralization of type I collagen and demineralized dentin was confirmed by TEM and selected-area electron diffraction when the adhesives were used as a carrier loaded with Si-ACP particles. Therefore, we propose self-etch adhesive as a novel carrier for ACP nanoprecursors to continuously deliver biomimetic remineralization.

Biomimetic Remineralization of Carious Lesions by Self-Assembling Peptide

Caries is the most common disease in the world. Great efforts have been undertaken for prevention and to identify a regenerative treatment solution for dental caries. Self-assembling β-sheet forming peptides have previously shown to form 3-dimensional fiber networks supporting tissue regeneration. In particular, the self-assembling peptide P11-4 has shown potential in the treatment and prevention of dental caries. It has previously been shown that application of monomeric P11-4 solution to early carious lesions can increase net mineral gain by forming de novo hydroxyapatite crystals. The hypothesis for the mode of action was that monomeric self-assembling peptide P11-4 diffuses into the subsurface lesion body and assembles therein into higher order fibrils, facilitating mineralization of the subsurface volume by mimicking the natural biomineralization of the tooth enamel, and it remains within the lesion body as a scaffold built-in by the newly formed hydroxyapatite. The aim of the present study was to investigate the mechanism of action of the self-assembling peptide P11-4 supporting mineralization of carious enamel. By various analytical methods, it could be shown that the self-assembling peptide P11-4 diffuses into the subsurface lesion, assembles into higher formed aggregates throughout the whole volume of the lesion, and supports nucleation of de novo hydroxyapatite nanocrystals and consequently results in increased mineral density within the subsurface carious lesion. The results showed that the application of self-assembling peptide P11-4 can facilitate the subsurface regeneration of the enamel lesion by supporting de novo mineralization in a similar mode of action as has been shown for the natural formation of dental enamel.

Oriented and Ordered Biomimetic Remineralization of the Surface of Demineralized Dental Enamel Using HAP@ACP Nanoparticles Guided by Glycine

Achieving oriented and ordered remineralization on the surface of demineralized dental enamel, thereby restoring the satisfactory mechanical properties approaching those of sound enamel, is still a challenge for dentists. To mimic the natural biomineralization approach for enamel remineralization, the biological process of enamel development proteins, such as amelogenin, was simulated in this study. In this work, carboxymethyl chitosan (CMC) conjugated with alendronate (ALN) was applied to stabilize amorphous calcium phosphate (ACP) to form CMC/ACP nanoparticles. Sodium hypochlorite (NaClO) functioned as the protease which decompose amelogenin in vivo to degrade the CMC-ALN matrix and generate HAP@ACP core-shell nanoparticles. Finally, when guided by 10 mM glycine (Gly), HAP@ACP nanoparticles can arrange orderly and subsequently transform from an amorphous phase to well-ordered rod-like apatite crystals to achieve oriented and ordered biomimetic remineralization on acid-etched enamel surfaces. This biomimetic remineralization process is achieved through the oriented attachment (OA) of nanoparticles based on non-classical crystallization theory. These results indicate that finding and developing analogues of natural proteins such as amelogenin involved in the biomineralization by natural macromolecular polymers and imitating the process of biomineralization would be an effective strategy for enamel remineralization. Furthermore, this method represents a promising method for the management of early caries in minimal invasive dentistry (MID).

Intrafibrillar mineralization of polyacrylic acid-bound collagen fibrils using a two-dimensional collagen model and Portland cement-based resins.

The biomimetic remineralization of apatite-depleted dentin is a potential method for enhancing the durability of resin-dentin bonding. To advance this strategy from its initial proof-of-concept design, we sought to investigate the characteristics of polyacrylic acid (PAA) adsorption to desorption from type I collagen and to test the mineralization ability of PAA-bound collagen. Portland cement and β-tricalcium phosphate (β-TCP) were homogenized with a hydrophilic resin blend to produce experimental resins. The collagen fibrils reconstituted on nickel (Ni) grids were mineralized using different methods: (i) group I consisted of collagen treated with Portland cement-based resin in simulated body fluid (SBF); (ii) group II consisted of PAA-bound collagen treated with Portland cement-based resin in SBF; and (iii) group III consisted of PAA-bound collagen treated with β-TCP-doped Portland cement-based resin in deionized water. Intrafibrillar mineralization was evaluated using transmission electron microscopy. We found that a carbonyl-associated peak at pH 3.0 increased as adsorption time increased, whereas a hydrogen bond-associated peak increased as desorption time increased. The experimental resins maintained an alkaline pH and the continuous release of calcium ions. Apatite was detected within PAA-bound collagen in groups II and III. Our results suggest that PAA-bound type I collagen fibrils can be mineralized using Portland cement-based resins.

Research advances in dentin biomimetic remineralization

With similar characteristic as nature dentin in structure and function, regenerated dentin is considered to be the ideal material for detected dentin restoration. Accordingly, dentin regeneration is a key issue in oral medicine and biomaterial science. Biomimetic mineralizatioin and remineralization are thus considered as strategies in dentin regeneration. This review is about the research advances of dentin biomimetic remineralization.

In vitro remineralization of hybrid layers using biomimetic analogs.

Resin-dentin bond degradation is a major cause of restoration failures. The major aim of the current study was to evaluate the impact of a remineralization medium on collagen matrices of hybrid layers of three different adhesive resins using nanotechnology methods. Coronal dentin surfaces were prepared from freshly extracted premolars and bonded to composite resin using three adhesive resins (FluoroBond II, Xeno-III-Bond, and iBond). From each tooth, two central slabs were selected for the study. The slabs used as controls were immersed in a simulated body fluid (SBF). The experimental slabs were immersed in a Portland cement-based remineralization medium that contained two biomimetic analogs (biomineralization medium (BRM)). Eight slabs per group were retrieved after 1, 2, 3, and 4 months, respectively and immersed in Rhodamine B for 24 h. Confocal laser scanning microscopy was used to evaluate the permeability of hybrid layers to Rhodamine B. Data were analyzed by analysis of variance (ANOVA) and Tukey's honest significant difference (HSD) tests. After four months, all BRM specimens exhibited a significantly smaller fluorescent area than SBF specimens, indicating a remineralization of the hybrid layer (P≤0.05). A clinically applicable biomimetic remineralization delivery system could potentially slow down bond degradation.

Influence of niobium pentoxide addition on the properties of glass ionomer cements

Objective: To determine the influence of niobium pentoxide (Nb2O5) addition on the physical and chemical properties of glass ionomer cements (GICs).Materials and methods: Five, 10 or 20 wt.% of Nb2O5 were incorporated into commercial GICs (Maxxion R, Vitro Molar, Vitro Fil R) and one group of each GIC remained without Nb2O5 (control groups). The GICs were evaluated by Knoop hardness, compressive strength, acid erosion, particle size and radiopacity. Data were analyzed by two-way ANOVA followed by Tukey's test.Results: The addition of 10% and 20% reduced the microhardness of two GICs (p < .05). Compressive strength showed no difference among groups (p > .05). Nb2O5 did not influence Maxxion R and Vitro Fil R regarding the acid erosion test (p > .05). Vitro Molar increased its acid erosion with 10% of Nb2O5 (p < .05). Maxxion R presented 15.78 μm, while Vitro Molar and Vitro Fil R showed 5.14 μm and 6.18 μm, respectively. As the Nb2O5 concentration increased, the radiopacity increased for all groups. Vitro Molar and Vitro Fil R did not present significant difference to at least 1 mm aluminum (p > .05).Conclusion: The addition of 5 wt.% Nb2O5 did not affect the tested physical and chemical properties of the GICs and improved the radiopacity of one of the cements. These materials are therefore suitable for further testing of biomimetic remineralization properties.

Glutaraldehyde-induced remineralization improves the mechanical properties and biostability of dentin collagen

The purpose of this study was to induce a biomimetic remineralization process by using glutaraldehyde (GA) to reconstruct the mechanical properties and biostability of demineralized collagen. Demineralized dentin disks (35% phosphoric acid, 10s) were pretreated with a 5% GA solution for 3min and then cultivated in a calcium phosphate remineralization solution. The remineralization kinetics and superstructure of the remineralization layer were evaluated by Raman spectroscopy, transmission electron microscopy, scanning electron microscopy and nanoindentation tests. The biostability was examined by enzymatic degradation experiments. A significant difference was found in dentin remineralization process between dentin with and without GA pretreating. GA showed a specific affinity to dentin collagen resulting in the formation of a cross-linking superstructure. GA pretreating could remarkably shorten remineralization time from 7days to 2days. The GA-induced remineralized collagen fibrils were well encapsulated by newly formed hydroxyapatite mineral nanocrystals. With the nano-hydroxyapatite coating, both the mechanical properties (elastic modulus and hardness) and the biostability against enzymatic degradation of the collagen were significantly enhanced, matching those of natural dentin. The results indicated that GA cross-linking of dentin collagen could promote dentin biomimetic remineralization, resulting in an improved mechanical properties and biostability. It may provide a promising tissue-engineering technology for dentin repair.

Biomineralization and Biomaterial Considerations in Dentin Remineralization

ABSTRACTDentin is a composite hard tissue, comprising of inorganic and organic matrices, and regulated by many proteins during development. The demineralization of dentin results from the loss of inorganic matrix [mainly hydroxyapatite (HAP)], but the organic matrix (mainly type I collagen) will sustain for a period of time after demineralization. Over the past decade, there has been a growing interest on the remineralization of demineralized dentin, primarily in connection with minimally invasive caries management. More and more biomaterials and methods are currently being evaluated to achieve newer approaches for the remineralization of demineralized dentin. These strategies are mostly based on biomimetic approaches and aim to achieve the characteristics of natural hard tissue. This article will present a complete review on the basic compositions and properties of dentin, which formed the basis for the biomimetic remineralization of demineralized dentin.How to cite this articleZhang X, Xiao Z, Wang H, Kishen A. Biomineralization and Biomaterial Considerations in Dentin Remineralization. J Oper Dent Endod 2016;1(1):7-12.

In vitro biomimetic remineralization of dentin collagen by phosphorylated chitosan/amorphous calcium phosphate compound and tripolyphosphate

In vitro biomimetic remineralization of dentin collagen by phosphorylated chitosan/amorphous calcium phosphate compound and tripolyphosphate

Biomimetic Remineralization of Human Enamel in the Presence of Polyamidoamine Dendrimers in vitro

Poly(amidoamine) (PAMAM) dendrimers, known as artificial proteins, have unique and well-defined molecular size and structure. It has previously been used to mimic protein-crystal interaction during biomineralization. In this study, generation 4.5 (4.5G) PAMAM with carboxylic acid (PAMAM-COOH) was synthesized and utilized to remineralize the surface of etched enamel in vitro. Using confocal laser scanning microscopy, Fourier transform infrared spectroscopy, X-ray diffraction analysis and scanning electron microscopy we observed that 4.5G PAMAM-COOH can be absorbed on the etched enamel surface and that it can induce the formation of hydroxyapatite crystals with the same orientation as that of the enamel prisms on longitudinal and transversal enamel surfaces. The self-assembly behavior of PAMAM in the mineralization solution was also investigated and the result showed that 4.5G PAMAM can assemble to microribbon structure similar to the behavior of amelogenins. Therefore, we concluded that 4.5G PAMAM-COOH assemblies can act as the organic template on enamel surface and in mineralization solution to control the nucleation site and morphology of new-grown crystals to form the biomimetic structure of human enamel, which may open a new way for repairing damaged enamel.

Influence of phosphoproteins’ biomimetic analogs on remineralization of mineral-depleted resin–dentin interfaces created with ion-releasing resin-based systems

ObjectiveThe study aimed at evaluating the remineralization of acid-etched dentin pre-treated with primers containing biomimetic analogs and bonded using an ion-releasing light-curable resin-based material. MethodsAn experimental etch-and-rinse adhesive system filled with Ca2+, PO43−-releasing Ca-Silicate micro-fillers was created along with two experimental primers containing biomimetic analogs such as sodium trimetaphosphate (TMP) and/or polyaspartic acid (PLA). Dentin specimens etched with 37% H3PO4 were pre-treated with two different aqueous primers containing the polyanionic biomimetic analogs or deionized water and subsequently bonded using the experimental resin-based materials. The specimens were sectioned and analyzed by AFM/nanoindentation to evaluate changes in the modulus of elasticity (Ei) across the resin–dentin interface at different AS storage periods (up to 90 days). Raman cluster analysis was also performed to evaluate the chemical changes along the interface. The phosphate uptake by the acid-etched dentin was evaluated using the ATR-FTIR. Additional resin–dentin specimens were tested for microtensile bond strength. SEM examination was performed after de-bonding, while confocal laser microscopy was used to evaluate the interfaces ultramorphology and micropermeability. ResultsBoth biomimetic primers induced phosphate uptake by acid-etched dentin. Specimens created with the ion-releasing resin in combination with the pre-treatment primers containing either PLA and TMA showed the greatest recovery of the Ei of the hybrid layer, with no decrease in μTBS (p>0.05) after 3-month AS storage. The ion-releasing resin applied after use of the biomimetic primers showed the greatest reduction in micropermeability due to mineral precipitation; these results were confirmed using SEM. SignificanceThe use of the ion-releasing resin-based system applied to acid-etched dentin pre-treated with biomimetic primers containing analogs of phosphoproteins such as poly-l-aspartic acid and/or sodium trimetaphosphate provides a suitable bonding approach for biomimetic remineralization of resin–dentin interfaces.

Methods for biomimetic remineralization of human dentine: a systematic review.

This study aimed to review the laboratory methods on biomimetic remineralization of demineralized human dentine. A systematic search of the publications in the PubMed, TRIP, and Web of Science databases was performed. Titles and abstracts of initially identified publications were screened. Clinical trials, reviews, non-English articles, resin-dentine interface studies, hybrid layer studies, hybrid scaffolds studies, and irrelevant studies were excluded. The remaining papers were retrieved with full texts. Manual screening was conducted on the bibliographies of remaining papers to identify relevant articles. A total of 716 studies were found, and 690 were excluded after initial screening. Two articles were identified from the bibliographies of the remaining papers. After retrieving the full text, 23 were included in this systematic review. Sixteen studies used analogues to mimic the functions of non-collagenous proteins in biomineralization of dentine, and four studies used bioactive materials to induce apatite formation on demineralized dentine surface. One study used zinc as a bioactive element, one study used polydopamine, and another study constructed an agarose hydrogel system for biomimetic mineralization of dentine. Many studies reported success in biomimetic mineralization of dentine, including the use of non-collagenous protein analogues, bioactive materials, or elements and agarose hydrogel system.

Leucine‐rich amelogenin peptide (LRAP) as a surface primer for biomimetic remineralization of superficial enamel defects: An in vitro study

This study was carried out to obtain more information about the assembly of hydroxyapatite bundles formed in the presence of Leucine-Rich Amelogenin Peptide (LRAP) and to evaluate its effect on the remineralization of enamel defects through a biomimetic approach. One or 2 mg/mL LRAP solutions containing 2.5 mM of Ca(+2) and 1.5 mM phosphate were prepared (pH = 7.2) and stored at 37 °C for 24 h. The products of the reaction were studied using atomic force microscopy (AFM), transmission electron microscopy (TEM), and selected area electron diffraction (SAED). Vickers surface microhardness recovery (SMR%) of acid-etched bovine enamel, with or without LRAP surface treatment, were calculated to evaluate the influence of peptide on the lesion remineralization. Distilled water and 1 or 2 mg/mL LRAP solution (pH = 7.2) were applied on the lesions and the specimens were incubated in mineralization solution (2.5mM Ca(+2) , 1.5mM PO4 (-3) , pH = 7.2) for 24 h. One-way ANOVA and Tukey's multi-comparison tests were used for statistical analysis. The pattern of enamel surface repair was studied using FE-SEM. AFM showed the formation of highly organized hierarchical structures, composed of hydroxyapatite (HA) crystals, similar to the dental enamel microstructure. ANOVA procedure showed significant effect of peptide treatment on the calculated SMR% (p < 0.001). Tukey's test revealed that peptide treated groups had significantly higher values of SMR%. In conclusion, LRAP is able to regulate the formation of HA and enhances the remineralization of acid-etched enamel as a surface treatment agent.

Biomimetic remineralization of demineralized dentine using scaffold of CMC/ACP nanocomplexes in an in vitro tooth model of deep caries.

Currently, it is still a tough task for dentists to remineralize dentine in deep caries. The aim of this study was to remineralize demineralized dentine in a tooth model of deep caries using nanocomplexes of carboxymethyl chitosan/amorphous calcium phosphate (CMC/ACP) based on mimicking the stabilizing effect of dentine matrix protein 1 (DMP1) on ACP in the biomineralization of dentine. The experimental results indicate that CMC can stabilize ACP to form nanocomplexes of CMC/ACP, which is able to be processed into scaffolds by lyophilization. In the single-layer collagen model, ACP nanoparticles are released from scaffolds of CMC/ACP nanocomplexes dissolved and then infiltrate into collagen fibrils via the gap zones (40 nm) to accomplish intrafibrillar mineralization of collagen. With this method, the completely demineralized dentine was partially remineralized in the tooth mode. This is a bottom-up remineralizing strategy based on non-classical crystallization theory. Since nanocomplexes of CMC/ACP show a promising effect of remineralization on demineralized dentine via biomimetic strategy, thereby preserving dentinal tissue to the maximum extent possible, it would be a potential indirect pulp capping (IPC) material for the management of deep caries during vital pulp therapy based on the concept of minimally invasive dentistry (MID).

Use of multifunctional phosphorylated PAMAM dendrimers for dentin biomimetic remineralization and dentinal tubule occlusion

Phosphorylated poly(amidoamine) dendrimers can induce biomimetic remineralization of demineralized dentin as analogs of non-collagenous proteins in the presence of polyacrylic acid, an amorphous calcium phosphate stabilizing agent.

An Electrophoresis-Aided Biomineralization System for Regenerating Dentin- and Enamel-Like Microstructures for the Self-Healing of Tooth Defects

It is the challenging and desirable aim of biomineralization studies to regenerate tooth tissue microstructures and to accelerate the speed of available biomimetic remineralization protocols for potential use in clinical dentistry. In this study, we developed a novel electrophoresis-aided calcium and phosphate agarose hydrogel system that can be used to quickly regenerate tooth tissue microstructure. After remineralization for 12 h, a demineralized dentin collagen matrix was remineralized with intrafibrillar and interfibrillar hydroxyapatites that mimicked the structure of the original calcified dentin collagen matrix. The precipitated hydroxyapatites were densely packed, occluded the exposed dentinal tubules, and regularly and homogeneously distributed along the collagen fibrils in a “string-of-beads” structure within the dentin collagen matrix. Needle-like hydroxyapatite crystals were densely packed, with their c-axis parallel to one another, to form the enamel-like tissue that precipitated onto the rem...

Biomimetic remineralization of demineralized enamel with nano-complexes of phosphorylated chitosan and amorphous calcium phosphate.

Remineralization of enamel plays a crucial role in the progression of carious process and the management of early caries lesion. Based on the influence of phosphorylated proteins in biomineralization, the objective of this study was to synthesize nano-complexes of phosphorylated chitosan and amorphous calcium phosphate (Pchi-ACP), and evaluate their ability to remineralize enamel subsurface lesions invitro. Pchi was synthesized using a previously established chemical method. The biomimetic remineralizing solution containing nano-complexes of Pchi-ACP was prepared by adding CaCl2 and K2HPO4 into Pchi-ACP solution (0.5% w/v) in sequence. The final concentrations of calcium and phosphate ions were 10 and 6mM, respectively. The nano-complexes of Pchi-ACP were characterized by Fourier-transform infrared (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and selected area electron diffraction (SAED). During testing the enamel lesions were treated with Pchi-ACP and fluoridated remineralizing solutions, respectively. The remineralizing of enamel lesions was examined with field emission electron microscope (FE-SEM) and Micro-CT. ACP was stabilized by Pchi to form nano-complexes that were soluble in water. The size of Pchi-ACP nano-complexes particles was determined to be less than 50nm. XRD and SAED results confirmed their amorphous phases. FE-SEM and Micro-CT results showed that the remineralizing effect of Pchi-ACP on enamel lesions was similar to that of fluoride. However, the remineralizing rate of Pchi-ACP treatment was significantly higher than that of fluoride treatment (P<0.05). This study highlighted the potential of nanoparticles functionalized with a natural analogue involved in biomineralization, to remineralize early enamel caries.

Biomimetic mineralization of a single-layer reconstituted type I collagen model induced by sodium tripolyphosphate and polyacrylic acid

To investigate the functions of sodium tripolyphosphate (STTP) and polyacrylic acid (PAA) in the process of collagen biomimetic mineralization. This would allow future applications to other collagen matrices such as bone collagen or 3-D collagen scaffolds. Glass cover slips and transmission electron microscopy (TEM) grids were coated with reconstituted typeIcollagen fibrils. Mineralization of the reconstituted collagens was demonstrated with scanning electron microscopy (SEM) and TEM using a Portland cement-containing resin composite and a phosphate-containing fluid in the presence of PAA and STTP. The rest were immersed in a biomimetic remineralization medium without PAA and/or STTP (control). In the presence of PAA and STTP in the mineralization medium, intrafibrillar mineralization based on the non-classical crystallisation pathway could be identified. Mineral phases were evident within the collagen fibrils as early as 12 h after the initially-formed amorphous calcium phosphate nanoprecursors were transformed into apatite nanocrystals. Collagens at 72 h were heavily mineralized with periodically arranged intrafibrillar apatite platelets. Conversely, only large mineral spheres with no preferred association with collagen fibrils were observed in the absence of biomimetic analogues in the medium (control). Intrafibrillar apatite deposition can be achieved via biomimetic mineralization system containing PAA and STTP when amorphous calcium phosphate precursor is stabilized.

Current Strategies in Dentin Remineralization

Partial caries removal procedures are used clinically in an attempt to conserve tooth structure and prevent pulp damage. Within this approach, the caries-infected dentin is removed, and the partially demineralized caries-affected dentin is preserved and sealed with materials that enhance remineralization. Fluoride-releasing glass ionomers have been commonly used for this purpose. Recent studies have shown potential for other cements and bioactive adhesive materials to promote dentin repair through various strategies. These strategies include ion releasing of Ca-P and the guided tissue remineralization or biomimetic remineralization of dentin. The latter is potentially useful in the remineralization of the demineralized acid-etched dentin that is incompletely infiltrated by adhesives in dentin bonding. The purpose of this Critical Appraisal is to provide the clinician with a summary of current literature that clarifies understanding of the process of dentin remineralization and to describe current strategies in this area.