Salivary opiorphin levels in denture-related traumatic ulcers.

OBJECTIVE: Opiorphin is a pentapeptide secreted in saliva and has a strong analgesic effect. Salivary opiorphin has been shown to increase in orofacial pain and may act as a pain reliever in pain caused by denture-related ulcers. The current study aimed to evaluate the salivary opiorphin levels in traumatic ulcers caused by ill-fitting dentures and demonstrate whether there is any correlation between trauma-related pain levels and salivary opiorphin levels. MATERIALS AND METHODS: Twenty-two individuals with new full dentures and a complaint of pain due to ill-fitting were included in this study. Patients were asked to rate their level of pain on a visual analog scale (VAS). Then, saliva specimens were collected at the first visit with the complaint of pain and 7 days following the denture adjustment. RESULTS: The average saliva opiorphin level before and after denture adjustment were 19.29 ± 5.44 and 15.78 ± 3.95 ng/mL, respectively. A dependent (paired) t-test determined that the mean salivary opiorphin level differed statistically significantly before and after the adjustment of the dentures. CONCLUSIONS: The findings show that salivary opiorphin levels increase in pain associated with denture-related traumatic ulcers. Adjusting the dentures resulted in pain relief and a statistically significant reduction in opiorphin levels.

In vitro caries-preventive effect of a mineralization-promoting peptide combined with fluoride gel on sound primary teeth.

BACKGROUND: Mineralization-promoting peptide-3 (MPP3) is a new biomimetic remineralization agent. AIM: To assess the remineralization efficiency of MPP3, either alone or in combination with fluoride gel. DESIGN: The samples were divided into four groups: control, 1.23% fluoride gel, 10% MPP3 gel, and 1.23% fluoride gel + 10% MPP3. Following the application of remineralization agents (4 min), the samples remained in a pH-cycling model (37°C, 4 weeks). Microhardness, microcomputed tomography (micro-CT), polarized light microscopy (PLM), and field emission scanning electron microscopy (FE-SEM) analysis were conducted. RM-ANOVA, one-way ANOVA, and intraclass correlation coefficient (ICC) were used for statistical analysis, and a significance level of p < .05 was employed. RESULTS: Mineralization-promoting peptide 3 and fluoride gel + MPP3 increased the microhardness of the enamel compared with initial values in each group (p < .05). Mineralization-promoting peptide 3 successfully maintained the mineral density of enamel, although the cariogenic pH-cycling and PLM results indicated that the lesion depth (µm) was significantly lower in the fluoride gel + MPP3 group (27.0336 ± 12.53650) than in the control group (37.3907 ± 12.76002, p < .05). CONCLUSION: The combined use of MPP3 with fluoride gel enhanced the caries-protective and mineralization-promoting effects of fluoride. Mineralization-promoting peptide 3 may be a potential agent that can be employed to improve the physical properties of enamel.

Biocatalytic synthesis and ordered self-assembly of silica nanoparticles via a silica-binding peptide.

Achieving scalable and economic methods for manufacturing ordered structures of nanoparticles is an ongoing challenge. Ordered structures of SiO2 nanoparticles have gained increased attention due to the great potential they offer in filtering, separation, drug delivery, optics, electronics, and catalysis. Biomolecules, such as peptides and proteins, have been demonstrated to be useful in the synthesis and self-assembly of inorganic nanostructures. Herein, we describe a simple Stöber-based method wherein both the synthesis and the self-assembly of SiO2 nanoparticles can be facilitated by a silica-binding peptide (SiBP). We demonstrate that the SiBP acts as a multirole agent when used alone or in combination with a strong base catalyst (NH3). When used alone, SiBP catalyzes the hydrolysis of precursor molecules in a dose-dependent manner and produces 17-20 nm SiO2 particles organized in colloidal gels. When used in combination with NH3, the SiBP produces smaller and more uniformly distributed submicrometer particles. The SiBP also improves the long-range self-assembly of the as-grown particles into an opal-like structure by changing the surface charge, without any need for further modification or processing of the particles. The results presented here provide a biomimetic route to the single-step synthesis and assembly of SiO2 nanoparticles into colloidal gels or opal-like structures.

Effects of a mineralization-promoting peptide on the physical and chemical properties of mineral trioxide aggregate.

Mineral trioxide aggregate (MTA) has been used widely in dentistry due to its sealing ability and biocompatibility. Delayed setting time is one of the major limitations of MTA. Various additives have been studied to further improve the properties of MTA with varied degrees of success. In this study, we have investigated the effect of a calcium phosphate mineralization promoting-peptide (MPP3) on the physical and chemical properties of MTA in comparison with Na2HPO4. Based on the reported effects of MPP3 on calcium-phosphate mineralization reaction, our hypothesis was that MPP3 may also show beneficial effects on the calcium-silicate mineralization system of MTA. Na2HPO4 was used for comparison since its setting accelerant effect on MTA has been well documented. The cements were prepared by mixing with distilled water, 0.40 mM MPP3 solution, 15% Na2HPO4 solution, and a combination of MPP3 and Na2HPO4 solution. Initial and final setting times were measured via Vicat needle. Microhardness values were measured via Vickers indenter at 1,3,7, and 28 days after hydration. Compressive strength after setting was measured via universal testing machine. Morphological and compositional analyses were performed via FESEM imaging, XRD and Raman spectroscopy. The microhardness data was evaluated via repeated-measures ANOVA. Setting time and compressive strength data were evasluated via one-way ANOVA. Initial setting time was reduced to ∼3 min in the Na2HPO4 containing groups but remained at ∼5 min in the control and MPP3 groups. Final setting times were significantly reduced in all groups compared to the control group. The reduction in the final setting times in the Na2HPO4 containing groups were significantly higher compared to the MPP3 group. Microhardness was significantly higher in the MPP3 group at all time points. No statistically significant difference in compressive strength was observed among the groups. FESEM analysis showed presence of ettringite crystals in the MPP3 group, and NaBiO3 crystals in the Na2HPO4 containing groups. XRD analysis showed a broadening of peaks at 2θ = 32° in the Na2HPO4 containing groups, possibly due to presence of NaBiO3. Raman spectroscopy showed statistically higher ettringite content in the MPP3 containing groups. Our findings indicate that MPP3 is a beneficial additive to eliminate some of the drawbacks associated with MTA with no detrimental effects on mechanical properties and without resulting in phases that potentially cause discoloration, such as NaBiO3. We propose that the reduced final setting time and increased microhardness by MPP3 may be associated with the increased ettringite content. Future studies, where wider range of MPP3 concentrations are studied may help elucidate and optimize the beneficial effects of MPP3 observed in this study.

Peptide-assisted pre-bonding remineralization of dentin to improve bonding.

Bonding with dentin is a complex process involving physical and chemical adhesion where the adhesive must be able to penetrate and envelop collagen fibers. Acid etching clears the dentin of debris, which prevents adhesives to interact with dentin. However, it also demineralizes the outermost surface of dentin and exposes collagen fibers. The mineral-free collagen is susceptible to collapse after drying and to proteolytic or microbial attack, ultimately impairing the bonding with dentin. To address this, we have attempted a pre-bonding rapid remineralization approach to recover the mineral content of etched dentin. We have used a mineralization-promoting peptide and high calcium/phosphate concentration to achieve this in a clinically applicable timeframe. Partial remineralization was confirmed via SEM and XRD analyses. The mechanical properties and the stability of the partially remineralized dentin were investigated via microhardness, collagen hydrolysis and shrinkage tests. The bonding properties were investigated via shear bond strength (SBS) and microleakage tests. Pre-bonding remineralization of dentin with peptide for 10 min significantly increased the stiffness, resistance to hydrolysis and reduced shrinkage due to drying. SBS was increased with both an etch&rinse and a self-etch adhesive. However, pre-bonding remineralization resulted in reduced microleakage only with the etch&rinse adhesive. The described method is readily applicable to clinic since it is expected to add only 10 min to the procedure. Future in situ and/or in vivo studies will help to confirm the benefits observed in this in vitro study and allow optimize the parameters of the method.

Silicon nitride ceramic for all-ceramic dental restorations.

Silicon nitride (Si3N4) is one of the promising ceramics for dental restoration due to providing significant benefits during the application. This study aimed to investigate the potential use of Si3N4 for all-ceramic dental restorations by characterizing some critical properties as color shade, mechanical resistance, shear-bond strength and radiolucency. For our study, porous Si3N4 ceramic was produced by partial sintering process with limited amounts of sintering additives and low temperature. A commercial ZrO2 ceramic was prepared according to manufacturer's instructions and results were compared with Si3N4. Si3N4 is an attractive ceramic for dental applications with good mechanical properties even in porous form, it has additional advantages over the conventional ceramics used as restorative material, such as, inherent antibacterial/anti-infective activity, radiolucency, and lower hardness. It is expected that Si3N4 will become popular in dental applications as well.

In vitro remineralization of primary teeth with a mineralization-promoting peptide containing dental varnish.

Mineralization-promoting peptides are attractive candidates for new remineralization systems. In previous studies, peptides have been applied as aqueous solutions, which is not a clinically relevant form. OBJECTIVE: This study aims to investigate the efficiency of a mineralization-promoting peptide, applied in varnish, on remineralizing artificial caries on primary teeth. METHODOLOGY: 55 primary molars were collected. Specimens were immersed in a demineralizing solution for 7 days and then, divided into 7 groups: Baseline: No-remineralization, Placebo: Blank colophony, F: Colophony 5% fluoride, P: Colophony 10% peptide, P+F: Colophony 5% fluoride and 10% peptide, Embrace: Embrace™ varnish, Durashield: Durashield™ varnish. A mixture of 35% w/v colophony varnishes were prepared in ethanol and applied accordingly. Specimens were immersed in a remineralization solution for 4 weeks and it was evaluated using PLM and SEM. Lesion depth reduction was examined by one-way ANOVA. RESULTS: There was no significant difference in mean lesion depths between baseline (147.04 ± 10.18 µm) and placebo groups (139.73 ± 14.92 µm), between F (120.95 ± 12.23 µm) and Durashield (113.47 ± 14.36 µm) groups and between P (81.79 ± 23.15 µm) and Embrace (90.26 ± 17.72 µm) groups. Lesion depth for the P+F group (66.95±10.59 µm) was significantly higher compared to all other groups. All groups contained samples with subsurface demineralized regions. Number of subsurface demineralized regions were higher in fluoride-containing groups. CONCLUSIONS: We conclude that the mineralization-promoting peptide (MPP3) is effective in this in vitro study and the peptide shows benefits over fluoride as it yields less subsurface demineralized regions.

Tear Opiorphin Levels in Ocular Pain Caused by Corneal Foreign Body.

PURPOSE: Opiorphin is an endogenous inhibitor of enkephalin-degrading enzymes. It has a strong analgesic effect in chemical and mechanical pain models. We aimed to evaluate the tear opiorphin levels in ocular pain caused by corneal foreign bodies and demonstrate whether there is any correlation with pain levels obtained from the Visual Analog Scale (VAS) score and tear opiorphin level. METHODS: Thirty-two healthy individuals and 34 individuals diagnosed with corneal foreign bodies were included in this study. Tear opiorphin levels were measured by the ELISA method using a commercially available kit. The difference in tear opiorphin levels between the patient and control groups were evaluated using the Mann-Whitney U test. The correlation between VAS scores and tear opiorphin levels was evaluated using the Spearman rank correlation coefficient. RESULTS: The median values of tear opiorphin levels of the patient and control groups were 134 pg/mL (86.86-296.25) and 109.80 pg/mL (66.15-191.49), respectively. The Mann-Whitney U test showed a statistically significant difference in tear opiorphin levels between patient and control groups (P < 0.05). No ocular pain was reported in the control group. The median VAS score of the patient group was 6 points (1-9). No correlation was found between VAS scores and tear opiorphin levels in the patient group. CONCLUSIONS: The cornea is the most densely innervated tissue, and the highest opiorphin concentrations have been observed in tear. It is, therefore, expected that the stimulation or damage to the nerve endings in cornea would cause an increase in opiorphin secretion as a pain relief mechanism.

Accelerated Calcium Phosphate Mineralization by Peptides with Adjacent Oppositely Charged Residues.

Calcium phosphate mineralizing peptides are of special importance for dental and orthopedic applications, such as caries remineralization and improved osteointegration. Uncovering the mechanism of action for such peptides is an ongoing challenge with the aim of a better fundamental understanding of biomineralization processes and developing optimized peptides for clinical use. It has recently been reported that "adjacent oppositely charged residue" motifs are found abundantly in cation binding, inorganic surface binding, or biomineralization-related proteins and may play a key role in the biomineralization events. Despite their medical importance, the role of these motifs has not yet been investigated on calcium phosphate mineral systems. To investigate this, we have designed peptides with different structural properties and different numbers of adjacent oppositely charged residues. We have evaluated their effects on in vitro calcium phosphate mineralization kinetics and mineral properties. The kinetics of the mineralization increased proportionally with an increasing number of adjacent oppositely charged residues. Two peptides with relatively high structural stability and two adjacent oppositely charged residues resulted in faster mineralization and more crystalline mineral compared to a peptide with a higher structural degree of freedom that contained only acidic residues. The fastest mineralization and the highest mineral crystallinity were obtained with a peptide containing the highest number of adjacent oppositely charged residues and highest structural degree of freedom. Our findings and observations from previously identified natural or designed peptides indicate that, in addition to structural instability, adjacent oppositely charged residues play a role in the cation binding, inorganic surface binding, and biomineralization of peptides and require further investigation. Lastly, the peptide identified in this study is an agent with potential medical applications involving the treatment of mineralized tissues.

In vitro remineralization of primary teeth with a mineralization-promoting peptide containing dental varnish

Abstract Mineralization-promoting peptides are attractive candidates for new remineralization systems. In previous studies, peptides have been applied as aqueous solutions, which is not a clinically relevant form. Objective This study aims to investigate the efficiency of a mineralization-promoting peptide, applied in varnish, on remineralizing artificial caries on primary teeth. Methodology 55 primary molars were collected. Specimens were immersed in a demineralizing solution for 7 days and then, divided into 7 groups: Baseline: No-remineralization, Placebo: Blank colophony, F: Colophony 5% fluoride, P: Colophony 10% peptide, P+F: Colophony 5% fluoride and 10% peptide, Embrace: Embrace™ varnish, Durashield: Durashield™ varnish. A mixture of 35% w/v colophony varnishes were prepared in ethanol and applied accordingly. Specimens were immersed in a remineralization solution for 4 weeks and it was evaluated using PLM and SEM. Lesion depth reduction was examined by one-way ANOVA. Results There was no significant difference in mean lesion depths between baseline (147.04 ± 10.18 µm) and placebo groups (139.73 ± 14.92 µm), between F (120.95 ± 12.23 µm) and Durashield (113.47 ± 14.36 µm) groups and between P (81.79 ± 23.15 µm) and Embrace (90.26 ± 17.72 µm) groups. Lesion depth for the P+F group (66.95±10.59 µm) was significantly higher compared to all other groups. All groups contained samples with subsurface demineralized regions. Number of subsurface demineralized regions were higher in fluoride-containing groups. Conclusions We conclude that the mineralization-promoting peptide (MPP3) is effective in this in vitro study and the peptide shows benefits over fluoride as it yields less subsurface demineralized regions.

Transient Heat Transfer in Dental Implants for Thermal Necrosis-Aided Implant Removal: A 3D Finite Element Analysis.

Removal of osseointegrated but otherwise failed (mechanical failure, mispositioning, esthetics, etc) dental implants is a traumatic process resulting in loss of healthy bone and complicating the treatment process. The traumatic effects of implant removal can be reduced by weakening the implant-bone attachment. Thermal necrosis-aided implant removal has been proposed as a minimally invasive method toward this end. In this method, an electrocautery tip is contacted to the implant to increase the temperature to 47°C and generate a limited and controlled thermal necrosis at the bone-implant interface. So far, no controlled studies have been performed to investigate the optimal clinical parameters for this method. In this study, we aimed to investigate, using finite element analysis method, the optimal settings to achieve intentional thermal necrosis on 3 implant systems, at 5 W and 40 W device power and with different size tips. The temperature increase of the implants at 40 W power was very sudden (< 0.5 seconds) and as the bone reached 47°C, the implants were at unacceptable temperatures. At 5 W power, temperature increase of the implants happened at manageable durations (< 1 second). Moreover, the temperature increase was even slower with larger implants and larger tip sizes. Therefore, low power settings must be used for thermal necrosis-aided implant removal. Also, the size of the implant and the tip must be taken into consideration in deciding the duration of contact with the electrocautery tip and the implant.

Salivary opiorphin in dental pain: A potential biomarker for dental disease.

OBJECTIVES: Opiorphin is a recently discovered peptide shown to inhibit the enkephalin-degrading enzymes and prolong the effects of enkephalins. Although opiorphin is found in high concentrations in saliva, the relationship between salivary opiorphin and orofacial pains is not yet fully understood. We aimed to determine salivary opiorphin concentrations in dental pain related to symptomatic irreversible pulpitis (SIP), and symptomatic apical periodontitis (SAP). DESIGN: 39 patients participated in this study. The participants were categorized into SIP and SAP based on their diagnosis. All the patients were treated with root canal treatment. Saliva specimens were collected, and pain levels were recorded at pre-treatment, 7 days post-treatment and 30 days post-treatment. Saliva opiorphin levels were measured using a commercially available ELISA kit. Pre-treatment and post-treatment opiorphin levels were evaluated using repeated measures ANOVA. Correlations between VAS scores, opiorphin levels and age were evaluated using Spearman's Rank Correlation. RESULTS: The average saliva opiorphin level pre-treatment, 7 days post-treatment and 30 days post-treatment were 31.28 ± 7.10 ng/ml, 20.41 ± 2.67 ng/ml and 18.61 ± 2.05 ng/ml respectively. Significantly higher pre-treatment opiorphin levels were observed in the SIP group compared to the SAP group. A strong correlation was observed between the pre-treatment pain levels and the saliva opiorphin concentrations. CONCLUSIONS: Our findings indicate that saliva opiorphin levels increase in inflammation related dental pain. The level of salivary opiorphin is strongly correlated with the reported level of pain. The extent of the inflammation (pulpal vs. periodontal) also affects the opiorphin level.

Cementomimetics-constructing a cementum-like biomineralized microlayer via amelogenin-derived peptides.

Cementum is the outer-, mineralized-tissue covering the tooth root and an essential part of the system of periodontal tissue that anchors the tooth to the bone. Periodontal disease results from the destructive behavior of the host elicited by an infectious biofilm adhering to the tooth root and left untreated, may lead to tooth loss. We describe a novel protocol for identifying peptide sequences from native proteins with the potential to repair damaged dental tissues by controlling hydroxyapatite biomineralization. Using amelogenin as a case study and a bioinformatics scoring matrix, we identified regions within amelogenin that are shared with a set of hydroxyapatite-binding peptides (HABPs) previously selected by phage display. One 22-amino acid long peptide regions referred to as amelogenin-derived peptide 5 (ADP5) was shown to facilitate cell-free formation of a cementum-like hydroxyapatite mineral layer on demineralized human root dentin that, in turn, supported attachment of periodontal ligament cells in vitro. Our findings have several implications in peptide-assisted mineral formation that mimic biomineralization. By further elaborating the mechanism for protein control over the biomineral formed, we afford new insights into the evolution of protein-mineral interactions. By exploiting small peptide domains of native proteins, our understanding of structure-function relationships of biomineralizing proteins can be extended and these peptides can be utilized to engineer mineral formation. Finally, the cementomimetic layer formed by ADP5 has the potential clinical application to repair diseased root surfaces so as to promote the regeneration of periodontal tissues and thereby reduce the morbidity associated with tooth loss.

In vitro labeling of hydroxyapatite minerals by an engineered protein.

Biological and biomimetic synthesis of inorganics have been a major focus in hard tissue engineering as well as in green processing of advanced materials. Among the minerals formed by organisms, calcium phosphate mineralization is studied extensively to understand the formation of mineral-rich tissues. Herein, we report an engineered fusion protein that not only targets calcium phosphate minerals but also allows monitoring of biomineralization. To produce the bi-functional fusion protein, nucleotide sequence encoding combinatorially selected hydroxyapatite-binding peptides (HABP) was genetically linked to the 3' end of the open reading frame of green fluorescence protein (GFPuv) and successfully expressed in Escherichia coli. The fluorescence and binding activities of the bi-functional proteins were characterized by, respectively, using fluorescence microscopy and quartz crystal microbalance spectroscopy. The utility of GFPuv-HABP fusion protein was assessed for both time-wise monitoring of mineralization and the visualization of the mineralized tissues. We used an alkaline phosphatase-based reaction to control phosphate release, thereby mimicking biological processes, to monitor calcium phosphate mineralization. The increase in mineral amount was observed using the fusion protein at different time points. GFPuv-HABP1 was also used for efficient fluorescence labeling of mineralized regions on the extracted human incisors. Our results demonstrate a simple and versatile application of inorganic-binding peptides conjugated with bioluminescence proteins as bi-functional bioimaging molecular probes that target mineralization, and which can be employed to a wide range of biomimetic processing and cell-free tissue engineering.

Biofunctionalization of materials for implants using engineered peptides.

Uncontrolled interactions between synthetic materials and human tissues are a major concern for implants and tissue engineering. The most successful approaches to circumvent this issue involve the modification of the implant or scaffold surfaces with various functional molecules, such as anti-fouling polymers or cell growth factors. To date, such techniques have relied on surface immobilization methods that are often applicable only to a limited range of materials and require the presence of specific functional groups, synthetic pathways or biologically hostile environments. In this study we have used peptide motifs that have been selected to bind to gold, platinum, glass and titanium to modify surfaces with poly(ethylene glycol) anti-fouling polymer and the integrin-binding RGD sequence. The peptides have several advantages over conventional molecular immobilization techniques; they require no biologically hostile environments to bind, are specific to their substrates and could be adapted to carry various active entities. We successfully imparted cell-resistant properties to gold and platinum surfaces using gold- and platinum-binding peptides, respectively, in conjunction with PEG. We also induced a several-fold increase in the number and spreading of fibroblast cells on glass and titanium surfaces using quartz and titanium-binding peptides in conjunction with the integrin ligand RGD. The results presented here indicate that control over the extent of cell-material interactions can be achieved by relatively simple and biocompatible surface modification procedures using inorganic binding peptides as linker molecules.

Self assembled bi-functional peptide hydrogels with biomineralization-directing peptides.

A peptide-based hydrogel has been designed that directs the formation of hydroxyapatite. MDG1, a twenty-seven residue peptide, undergoes triggered folding to form an unsymmetrical beta-hairpin that self-assembles in response to an increase in solution ionic strength to yield a mechanically rigid, self supporting hydrogel. The C-terminal portion of MDG1 contains a heptapeptide (MLPHHGA) capable of directing the mineralization process. Circular dichroism spectroscopy indicates that the peptide folds and assembles to form a hydrogel network rich in beta-sheet secondary structure. Oscillatory rheology indicates that the hydrogel is mechanically rigid (G' 2500Pa) before mineralization. In separate experiments, mineralization was induced both biochemically and with cementoblast cells. Mineralization-domain had little effect on the mechanical rigidity of the gel. SEM and EDXS show that MDG1 gels are capable of directing the formation of hydroxapatite. Control hydrogels, prepared by peptides either lacking the mineral-directing portion or reversing its sequence, indicated that the heptapeptide is necessary and its actions are sequence specific.

Molecular biomimetics: GEPI-based biological routes to technology.

In nature, the viability of biological systems is sustained via specific interactions among the tens of thousands of proteins, the major building blocks of organisms from the simplest single-celled to the most complex multicellular species. Biomolecule-material interaction is accomplished with molecular specificity and efficiency leading to the formation of controlled structures and functions at all scales of dimensional hierarchy. Through evolution, Mother Nature developed molecular recognition by successive cycles of mutation and selection. Molecular specificity of probe-target interactions, e.g., ligand-receptor, antigen-antibody, is always based on specific peptide molecular recognition. Using biology as a guide, we can now understand, engineer, and control peptide-material interactions and exploit them as a new design tool for novel materials and systems. We adapted the protocols of combinatorially designed peptide libraries, via both cell surface or phage display methods; using these we select short peptides with specificity to a variety of practical materials. These genetically engineered peptides for inorganics (GEPI) are then studied experimentally to establish their binding kinetics and surface stability. The bound peptide structure and conformations are interrogated both experimentally and via modeling, and self-assembly characteristics are tested via atomic force microscopy. We further engineer the peptide binding and assembly characteristics using a computational biomimetics approach where bioinformatics based peptide-sequence similarity analysis is developed to design higher generation function-specific peptides. The molecular biomimetic approach opens up new avenues for the design and utilization of multifunctional molecular systems in a wide-range of applications from tissue engineering, disease diagnostics, and therapeutics to various areas of nanotechnology where integration is required among inorganic, organic and biological materials. Here, we describe lessons from biology with examples of protein-mediated functional biological materials, explain how novel peptides can be designed with specific affinity to inorganic solids using evolutionary engineering approaches, give examples of their potential utilizations in technology and medicine, and, finally, provide a summary of challenges and future prospects.

Regulation of in vitro calcium phosphate mineralization by combinatorially selected hydroxyapatite-binding peptides.

We report selection and characterization of hydroxyapatite-binding heptapeptides from a peptide-phage library and demonstrate the effects of two peptides, with different binding affinities and structural properties, on the mineralization of calcium phosphate mineral. In vitro mineralization studies carried out using one strong- and one weak-binding peptide, HABP1 and HABP2, respectively, revealed that the former exhibited a drastic outcome on mineralization kinetics and particle morphology. Strong-binding peptide yielded significantly larger crystals, as observed by electron microscopy, in comparison to those formed in the presence of a weak-binding peptide or in the negative control. Molecular structural studies carried out by circular dichroism revealed that HABP1 and HABP2 differed in their secondary structure and conformational stability. The results indicate that sequence, structure, and molecular stability strongly influence the mineralization activity of these peptides. The implication of the research is that the combinatorially selected short-sequence peptides may be used in the restoration or regeneration of hard tissues through their control over of the formation of calcium phosphate biominerals.

Materials specificity and directed assembly of a gold-binding peptide.

Adsorption studies of a genetically engineered gold-binding peptide, GBP1, were carried out using a quartz-crystal microbalance (QCM) to quantify its molecular affinity to noble metals. The peptide showed higher adsorption onto and lower desorption from a gold surface compared to a platinum substrate. The material specificity, that is, the preferential adsorption, of GBP1 was also demonstrated using gold and platinum micropatterned on a silicon wafer containing native oxide. The biotinylated three-repeat units of GBP1 were preferentially adsorbed onto gold regions delineated using streptavidin-conjugated quantum dots (SAQDs). These experiments not only demonstrate that an inorganic-binding peptide could preferentially adsorb onto a metal (Au) rather than an oxide (SiO2) but also onto one noble metal (Au) over another (Pt). This result shows the utility of an engineered peptide as a molecular erector in the directed immobilization of a nanoscale hybrid entity (SAQDs) over selected regions (Au) on a fairly complex substrate (Au and Pt micropatterned regions on silica). The selective and controlled adsorption of inorganic-binding peptides may have significant implications in nano- and nanobiotechnology, where they could be genetically tailored for specific use in the development of self-assembled molecular systems.