Performance Assessment of Three Similar Dental Restorative Composite Materials via Raman Spectroscopy Supported by Complementary Methods Such as Hardness and Density Measurements.

(1) Background: A widespread problem in oral health is cavities produced by cariogenic bacteria that consume fermentable carbohydrates and lower pH to 5.5-6.5, thus extracting Ca2+ and phosphate ions (PO43-) from teeth. Dental restorative materials based on polymers are used to fill the gaps in damaged teeth, but their properties are different from those of dental enamel. Therefore, a question is raised about the similarity between dental composites and natural teeth in terms of density and hardness. (2) Methods: We have used Raman spectroscopy and density and microhardness measurements to compare physical characteristics of several restorative dental composites at different polymerization intervals. (3) Results: XRVHerculite®, Optishade®, and VertiseFlow® showed the very different characteristics of the physical properties following four polymerization intervals. Of the three composites, OptiShade showed the highest polymerization rate. (4) Conclusions: Only fully polymerized composites can be used in teeth restoring, because incomplete polymerization would result in cracks, pitting, and lead finally to failure.

Preliminary Assessment of In Vivo Raman Spectroscopy Technique for Bone Quality Evaluation of Augmented Maxillary Sinus Floor.

(1) Background: In oral surgery, bone regeneration is achieved through various types of bone grafts or bone substitutes and its success is usually analyzed by micro-computed tomography and histomorphometry. The aim of this study was to evaluate the usefulness of Raman spectroscopy as an alternative to other techniques for bone quality evaluation during a standard oral surgery procedure. (2) Methods: The preliminary evaluation of bone augmentation during maxillary sinus floor elevation oral surgery was performed by Raman spectroscopy for several (five) patients during and after the surgery and the results were compared with postoperative data from histomorphometry, EDX and SEM analysis. (3) Results: After analyzing all the results for the bone samples according to the four methods (Raman, EDX, SEM and Histology) that were used in our study, the obtained result of the investigation provided a good augmentation process for three of the patients and partly successful augmentation process for two of the patients. The primary evaluation using Raman spectroscopy (in vivo and ex vivo investigation) was confirmed by histological results, thus having a first step for validation of Raman as a new method of imaging for dentistry. (4) Conclusions: Our results show that Raman spectroscopy could provide fast and reliable insight on bone condition during augmentation of the maxillary sinus floor. We emphasize the advantages and drawbacks of the proposed techniques as its accuracy could increase by performing larger size clinical trials. Using the Raman mapping, the method can serve as an alternative to histology.

Raman Spectroscopy as Spectral Tool for Assessing the Degree of Conversion after Curing of Two Resin-Based Materials Used in Restorative Dentistry.

(1) Background: The treatment of dental cavities and restoration of tooth shape requires specialized materials with specific clinical properties, including being easy to model, light-cured, having a natural color, reduced shrinkage, a hardness similar to hydroxyapatite, and no leakage. The dimensional stability of resin composite materials is affected by polymerization shrinkage, degree of conversion (number of π carbon bonds converted into σ ones), thermal contraction and expansion, and interactions with an aqueous environment. (2) Methods: The materials used in our investigation were two composite resins with similar polymer matrices, but different filler (micro/nano filler). To evaluate the properties of samples, we employed the pycnometer technique (pycnometer from Paul Marienfeld Gmbh, Lauda-Königshofen, Germany), RAMAN spectroscopy technique (MiniRam Equipment from B&W Tek Inc., Plainsboro Township, NJ, USA; 785 nm laser source), SEM and EDX (FEI Inspect S.). (3) Results: The size of the filler plays an important role in the polymerization: for the pycnometric results, the larger particle filler (Sample 1) seems to undergo a rapid polymerization during the 45 s curing, while the nanoparticle filer (Sample 2) needs additional curing time to fully polymerize. This is related to a much larger porosity, as proved by SEM images. The lower degree of conversion, as obtained by Raman spectroscopy, in the same geometry means that the same volume is probed for both samples, but Sample 1 is more porous, which means less amount of polymer is probed for Sample 1. (4) Conclusions: For the two composites, we obtained a degree of conversion of 59% for Sample 1 and 93% for Sample 2, after 45 s of curing.

Raman Spectroscopy: In Vivo Application for Bone Evaluation in Oral Reconstructive (Regenerative) Surgery.

The aim of this study was to evaluate the quality of the bone, revealing the different phases for calcified tissues independent of the medical history of the patient in relation to periodontitis by means of in vivo Raman spectroscopy. Raman spectroscopy measurements were performed in vivo during surgery and then ex vivo for the harvested bone samples for the whole group of patients (ten patients). The specific peaks for the Raman spectrum were traced for reference compounds (e.g., calcium phosphates) and bone samples. The variation in the intensity of the spectrum in relation to the specific bone constituents' concentrations reflects the bone quality and can be strongly related with patient medical status (before dental surgery and after a healing period). Moreover, bone sample fluorescence is related to collagen content, enabling a complete evaluation of bone quality including a "quasi-quantification" of the healing process similar to the bone augmentation procedure. A complete evaluation of the processed spectra offers quantitative/qualitative information on the condition of the bone tissue. We conclude that Raman spectroscopy can be considered a viable investigation method for an in vivo and quick bone quality assessment during oral and periodontal surgery.

3D Superparamagnetic Scaffolds for Bone Mineralization under Static Magnetic Field Stimulation.

We reported on three-dimensional (3D) superparamagnetic scaffolds that enhanced the mineralization of magnetic nanoparticle-free osteoblast cells. The scaffolds were fabricated with submicronic resolution by laser direct writing via two photons polymerization of Ormocore/magnetic nanoparticles (MNPs) composites and possessed complex and reproducible architectures. MNPs with a diameter of 4.9 ± 1.5 nm and saturation magnetization of 30 emu/g were added to Ormocore, in concentrations of 0, 2 and 4 mg/mL. The homogenous distribution and the concentration of the MNPs from the unpolymerized Ormocore/MNPs composite were preserved after the photopolymerization process. The MNPs in the scaffolds retained their superparamagnetic behavior. The specific magnetizations of the scaffolds with 2 and 4 mg/mL MNPs concentrations were of 14 emu/g and 17 emu/g, respectively. The MNPs reduced the shrinkage of the structures from 80.2 ± 5.3% for scaffolds without MNPs to 20.7 ± 4.7% for scaffolds with 4 mg/mL MNPs. Osteoblast cells seeded on scaffolds exposed to static magnetic field of 1.3 T deformed the regular architecture of the scaffolds and evoked faster mineralization in comparison to unstimulated samples. Scaffolds deformation and extracellular matrix mineralization under static magnetic field (SMF) exposure increased with increasing MNPs concentration. The results are discussed in the frame of gradient magnetic fields of ~3 × 10-4 T/m generated by MNPs over the cells bodies.

3D Biomimetic Magnetic Structures for Static Magnetic Field Stimulation of Osteogenesis.

We designed, fabricated and optimized 3D biomimetic magnetic structures that stimulate the osteogenesis in static magnetic fields. The structures were fabricated by direct laser writing via two-photon polymerization of IP-L780 photopolymer and were based on ellipsoidal, hexagonal units organized in a multilayered architecture. The magnetic activity of the structures was assured by coating with a thin layer of collagen-chitosan-hydroxyapatite-magnetic nanoparticles composite. In vitro experiments using MG-63 osteoblast-like cells for 3D structures with gradients of pore size helped us to find an optimum pore size between 20-40 µm. Starting from optimized 3D structures, we evaluated both qualitatively and quantitatively the effects of static magnetic fields of up to 250 mT on cell proliferation and differentiation, by ALP (alkaline phosphatase) production, Alizarin Red and osteocalcin secretion measurements. We demonstrated that the synergic effect of 3D structure optimization and static magnetic stimulation enhances the bone regeneration by a factor greater than 2 as compared with the same structure in the absence of a magnetic field.

Laser-direct writing by two-photon polymerization of 3D honeycomb-like structures for bone regeneration.

A major limitation of existing 3D implantable structures for bone tissue engineering is that most of the cells rapidly attach on the outer edges of the structure, restricting the cells penetration into the inner parts and causing the formation of a necrotic core. Furthermore, these structures generally possess a random spatial arrangement and do not preserve the isotropy on the whole volume. Here, we report on the fabrication and testing of an innovative 3D hierarchical, honeycomb-like structure (HS), with reproducible and isotropic arhitecture, that allows in 'volume' migration of osteoblasts. In particular, we demonstrate the possibility to control the 3D spatial cells growth inside these complex architectures by adjusting the free spaces inside the structures. The structures were made of vertical microtubes arranged in a mulitlayered configuration, fabricated via laser direct writing by two photons polymerization of the IP-L780 photopolymer. In vitro tests performed in MG-63 osteoblast-like cells demonstrated that the cells migration inside the 3D structures is conducted by the separation space between the microtubes layers. Specifically, for layers separation between 2 and 10 µm, the cells gradually penetrated between the microtubes. Furthermore, these structures induced the strongest cells osteogenic differentiation and mineralization, with ALP activity 1.5 times stronger, amount of calcified minerals 1.3 times higher and osteocalcin secretion increased by 2.3 times compared to the other structures. On the opposite, for layers separation less than 2 µm and above 10 µm, the cells were not able to make interconnections and exhibited poor mineralization ability.

Mechanical, Corrosion and Biological Properties of Room-Temperature Sputtered Aluminum Nitride Films with Dissimilar Nanostructure.

Aluminum Nitride (AlN) has been long time being regarded as highly interesting material for developing sensing applications (including biosensors and implantable sensors). AlN, due to its appealing electronic properties, is envisaged lately to serve as a multi-functional biosensing platform. Although generally exploited for its intrinsic piezoelectricity, its surface morphology and mechanical performance (elastic modulus, hardness, wear, scratch and tensile resistance to delamination, adherence to the substrate), corrosion resistance and cytocompatibility are also essential features for high performance sustainable biosensor devices. However, information about AlN suitability for such applications is rather scarce or at best scattered and incomplete. Here, we aim to deliver a comprehensive evaluation of the morpho-structural, compositional, mechanical, electrochemical and biological properties of reactive radio-frequency magnetron sputtered AlN nanostructured thin films with various degrees of c-axis texturing, deposited at a low temperature (~50 °C) on Si (100) substrates. The inter-conditionality elicited between the base pressure level attained in the reactor chamber and crystalline quality of AlN films is highlighted. The potential suitability of nanostructured AlN (in form of thin films) for the realization of various type of sensors (with emphasis on bio-sensors) is thoroughly probed, thus unveiling its advantages and limitations, as well as suggesting paths to safely exploit the remarkable prospects of this type of materials.

Electrically stimulated osteogenesis on Ti-PPy/PLGA constructs prepared by laser-assisted processes.

This work describes a versatile laser-based protocol for fabricating micro-patterned, electrically conductive titanium-polypyrrole/poly(lactic-co-glycolic)acid (Ti-PPy/PLGA) constructs for electrically stimulated (ES) osteogenesis. Ti supports were patterned using fs laser ablation in order to create high spatial resolution microstructures meant to provide mechanical resistance and physical cues for cell growth. Matrix Assisted Pulsed Laser Evaporation (MAPLE) was used to coat the patterned Ti supports with PPy/PLGA layers acting as biocompatible surfaces having chemical and electrical properties suitable for cell differentiation and mineralization. In vitro biological assays on osteoblast-like MG63 cells showed that the constructs maintained cell viability without cytotoxicity. At 24 h after cell seeding, electrical stimulation with currents of 200 µA was applied for 4 h. This treatment was shown to promote earlier onset of osteogenesis. More specifically, the alkaline phosphatase activity of the stimulated cultures reached the maximum before that of the non-stimulated ones, i.e. controls, indicating faster cell differentiation. Moreover, mineralization was found to occur at an earlier stage in the stimulated cultures, as compared to the controls, starting with Day 6 of cell culture. At later stages, calcium levels in the stimulated cultures were higher than those in control samples by about 70%, with Ca/P ratios similar to those of natural bone. In all, the laser-based protocol emerges as an efficient alternative to existing fabrication technologies.