Photochemical Ligation to Ultrasensitive DNA Detection with Upconverting Nanoparticles.

In this work, we explore a photochemical ligation reaction to covalently modify oligonucleotide-conjugated upconverting nanoparticles (UCNPs) in the presence of a specific target DNA sequence. The target sequence acts as a hybridization template, bringing together a biotinylated photoactivatable oligonucleotide probe and the oligonucleotide probe that is attached to UCNPs. The illumination of the UCNPs by NIR light to generate UV emission internally or illuminating the photoactivatable probe directly by an external UV light promotes the photochemical ligation reaction, yielding covalently biotin functionalized UCNPs that can be selectively captured in streptavidin-coated microwells. Following this strategy, we developed a DNA sensor with a limit of detection of 1 × 10-18 mol per well (20 fM). In addition, we demonstrate the possibility to create UCNP patterns on the surface of solid supports upon NIR illumination that are selectively formed under the presence of the target oligonucleotide.

Temperature controlled fluorescence on Au@Ag@PNIPAM-PTEBS microgels: effect of the metal core size on the MEF extension.

In this work, we present a novel method to produce thermoresponsive, monodisperse microgels which display temperature-dependent photoluminescence. The system is based on bimetallic cores of Au@Ag encapsulated within thermoresponsive poly(N-isopropylacrylamide) microgels and coated with a photoluminescent polymer (poly[2-(3-thienyl)ethoxy-4-butylsulfonate] (PTEBS) using the Layer-by-Layer technique. The electromagnetic radiation used to excite the PTEBS induces a local electromagnetic field on the surface of the bimetallic cores that enhances the excitation and emission rates of the PTEBS, yielding a metal enhanced fluorescence (MEF). This effect was studied as a function of the bimetallic core size and the separation distance between the PTEBS and the bimetallic cores. Our results permit evaluation of the effect that the metallic core size of colloidal particles exerts on the MEF for the first time, and prove the relevance of the metallic cores to extend the effect far away from the metallic surface.

Membranes over the lateral window in sinus augmentation procedures: a two-arm and split-mouth randomized clinical trials.

OBJECTIVE: This study evaluates whether or not, among other factors, membrane-coverage of antrostomy defects improves implant survival in sinus augmentation procedures. MATERIALS AND METHODS: We performed a two-arm and split-mouth randomized controlled clinical trial on 104 and 5 patients respectively. In the two-arm study, antrostomy defects were membrane-covered in 66 procedures and uncovered in 69, before placing a total of 265 implants that were followed up for 1 year. In the split-mouth study, following bilateral sinus augmentation, antrostomy defects were membrane-covered on one side and left uncovered on the contra-lateral. Bone biopsies from each sinus were histologically analysed 6 months later. RESULTS: In the two-arm study, implant survival rates were similar (p = 0.08) in the membrane-covered (96.1%) and uncovered (94.2%) groups. In the split-mouth study, bone augmentation was similar in both groups (p = 0.52). Delayed implant placement (p = 0.04), thick Schneider's membrane (≥2 mm) (p < 0.01), treatment for hypertension (p = 0.04) and non-smoking (p = 0.01) seemed to be associated with lower risk of implant failure. CONCLUSIONS: Implant survival in sinus lifting procedures could be influenced significantly by timing of implant placement, Schneider's membrane thickness, antihypertensive treatment and smoking habits, but not by antrostomy membrane coverage.

Osteoinduction in the palatal submucosa by injecting BMP-2 on 2 different carriers.

In this work, we investigated the ability of injected recombinant human bone morphogenetic protein 2 (rhBMP-2) on brushite cement (a ß-tricalcium phosphate-based biomaterial) and collagen gel as carriers to induce osteogenic differentiation in the palatal submucosa of 10-day-old rats. This was part of a broader study aiming to create bone in the palatal submucosa at cleft palate edges in the search for a minimally invasive treatment. Thirteen treated animals, 7 with rhBMP-2/brushite cement and 6 with rhBMP-2/collagen gel, were injected with 5 to 10 µL of each biomaterial in the right palatal submucosa at the level between the second and third rugae. The contralateral site was uninjected and served as the control. Six weeks after injection, both brushite cement and collagen gel were histologically unrecognizable in all treated animals. New bone structures such as ossicles of woven bone were not detected. However, an augmentation in the thickness of the palatal fibromucosa was observed at the injection site of all palates. In addition, immunolabeling for osteopontin, proliferating cell nuclear antigen, and TUNEL revealed intense osteogenic induction at the injection site with both constructs, which was negative in the control site from the same specimens; no differences regarding cell proliferation and death were observed. The present study confirms the feasibility of generating osteogenic cells in the palatal submucosa by injecting low doses of rhBMP-2 in these 2 biomaterials, together with their inability to form bone.

Synthesis of thermosensitive microgels with a tunable magnetic core.

In this work, we describe a new methodology for the preparation of monodisperse and thermosensitive microgels with magnetic core. In order to produce such a material, hydrophobic magnetic Fe(3)O(4) nanoparticles were prepared by two methods: thermal decomposition and coprecipitation. The surface of these nanoparticles was modified by addition of 3-butenoic acid, and after that these nanoparticles were dispersed in water and submitted to free radical polymerization at 70 °C in the presence of N-isopropylacrylamide (NIPAM) and bisacrylamide. The result of this reaction was monodisperse microgels with a magnetic core. By varying the amount of 3-butenoic acid, it was possible to obtain hybrid microgels with different magnetic core sizes and different architectures.

Vertical bone augmentation with 3D-synthetic monetite blocks in the rabbit calvaria.

INTRODUCTION: Long-term success of osteointegrated dental implants requires sufficient volume of healthy bone at the recipient sites. However, this is frequently lacking as a result of trauma, tooth loss, or infection. Onlay autografting is amongst the most predictable techniques for craniofacial vertical bone augmentation, however, complications related to donor site morbidity are common and alternatives to onlay autografts are desirable. AIM: To develop and evaluate a new synthetic onlay block for vertical bone augmentation. MATERIAL AND METHODS: Sixteen synthetic monetite monolithic discs-shaped blocks were prepared using a 3D-printing technique. The blocks were computer-designed, and had a diameter of 9.0 mm, a thickness of either 4.0 mm (n = 8) or 3.0 mm (n = 8) and one 0.5-mm wide central hole that enabled their surgical fixation with osteosynthesis screws. The blocks were randomly allocated to each side of the calvaria (right or left) of eight New Zealand rabbits and fixed with screws to achieve vertical bone augmentation. Eight weeks after the surgical intervention, the animals were sacrificed and the calvaria were retrieved for histological analysis. The following parameters were analysed: the interaction between the graft and the original bone surface, the amount of bone ingrowth within the graft and the gain in bone height achieved by the procedure. Wilcoxon t-test was used to evaluate significant differences between the two types of monetite bone block grafts. RESULTS: The blocks were easy to handle and no damage or fracture was registered while being screw-fixated to the calvarial bone. As a result, the surgical procedure was easy and quick. After a healing of 8 weeks, the synthetic blocks were strongly fused to the calvarial bone surface. Upon histological observation, the monetite blocks appeared to be infiltrated by newly formed bone, without histological signs of necrosis, osteolysis or foreign body reaction. Histomorphometry revealed that bone augmentation occurred within and over the monetite block. The 4.0- and 3.0-mm high blocks were filled with newly formed bone with 35% and 41% of their respective volumes. These observations indicated that craniofacial bone augmentations of at least 4 mm could be achieved with synthetic monetite blocks. CONCLUSION: Within the limits of our study, this novel material may be able to eliminate the need for autologous bone transplantation for the augmentation of large vertical bone defects.

Matrix tablets based on a novel poly (magnesium acrylate) hydrogel for the treatment of inflammatory bowel diseases.

The objective of this work was to evaluate the potential use of a new polymer (PAMgA) in the development sustained release matrix tablets for the treatment of bowel inflammatory diseases. For this purpose, budesonide, a highly lipophilic compound, was used as model drug. Tablets with two reticulation grades of PAMgA (PAMgA 5 and 40) and with 9 mg of budesonide were developed and characterized. All the studies were carried out using biorelevant media (FaSSGF and FaSSIF). Swelling and erosion of PAMgA tablets was influenced by the reticulation grade of the polymer and the biorelevant media assayed, being water uptake higher for PAMgA 40 tablets in intestinal fluid, whereas PAMgA 5 showed more intense erosion in this biorelevant medium. Budesonide was released slowly from PAMgA tablets, both in gastric and intestinal environment, following Super case II transport kinetics (relaxation-controlled delivery), with a lag time of around 1-2 h. When the dissolution medium was changed sequentially throughout the trial, 75% of the budesonide dose was released in a sustained manner between 4 and 20 h of testing from PAMgA tablets, showing a more controlled budesonide release than Entocort® and Budenofalk® (commercially available sustained release formulations of budesonide). In conclusion, PAMgA polymer allows controlling the release of highly lipophilic drugs as budesonide, being an useful excipient for the development of sustained release matrix tablets.

Platelet-rich plasma may prevent titanium-mesh exposure in alveolar ridge augmentation with anorganic bovine bone.

OBJECTIVE: Bone augmentation with the titanium-mesh (Ti-mesh) technique is susceptible to a large rate of complications such as morbidity of bone graft donor site, and mesh exposure to the oral cavity. The purpose of this study was to evaluate the effectiveness of anorganic bovine bone (ABB) in alveolar bone augmentation with the Ti-mesh technique. In addition, we investigated the effect of platelet-rich plasma (PRP) in preventing mesh exposure by using it to cover the Ti-mesh. PATIENTS AND METHODS: Patients included in the clinical trial were randomly allocated by a blinded assistant into two groups. The 30 patients recruited for this study underwent 43 alveolar bone augmentation with the Ti-mesh technique using ABB as graft material in all of them. In 15 patients, the Ti-meshes were covered with PRP (PRP group) whereas in the other 15 the Ti-meshes were not (control group). After 6 months, patients were called for clinical, radiographic, and histological evaluation, and implant placement surgery. A total of 97 implants were placed in the augmented bone and their evolution was followed up for a period of 24 months. RESULTS: Significant differences were found between the two study groups in terms of complications and bone formation. In the control group, 28.5% of the cases suffered from mesh exposure, while in the PRP group, no exposures were registered. Radiographic analysis revealed that bone augmentation was higher in the PRP group than in the control group. Overall, 97.3% of implants placed in the control group and 100% of those placed in the PRP group were successful during the monitoring period. We suggest that the positive effect of PRP on the Ti-mesh technique is due to its capacity to improve soft tissue healing, thereby protecting the mesh and graft material secured beneath the gingival tissues. CONCLUSIONS: Alveolar bone augmentation using ABB alone in the Ti-mesh technique is sufficient for implant rehabilitation. Besides, covering the Ti-meshes with PRP was a determining factor in avoiding mesh exposure. Ti-mesh exposure provoked significant bone loss, but in most cases it did not affect the subsequent placement of implants.

Effect of combining platelet-rich plasma with anorganic bovine bone on vertical bone regeneration: early healing assessment in rabbit calvariae.

PURPOSE: The purpose of this study was to evaluate the combination of anorganic bovine bone allograft (ABB) with platelet-rich plasma (PRP) when used in vertical bone augmentation. MATERIALS AND METHODS: Sixteen healthy 6-month-old female New Zealand rabbits were used in this study. Rabbits were randomly divided into two groups of eight animals each. Created calvarial defects were grafted with ABB or PRP in the first group whereas, in the second group, one cylinder was filled with PRP+ABB and the contralateral cylinder was left ungrafted to be filled by autologous blood (control cylinder). Six weeks after intervention, animals were sacrificed and biopsies were taken. Densitometric, histologic, and histomorphometric analyses were performed to evaluate bone mineral density, vertical bone augmentation, and remaining graft volume, respectively. Statistical analyses were performed with the Mann-Whitney test, using a significance level of P < .05. RESULTS: Six weeks after rabbit calvariae were augmented, data analysis revealed that the mixture of PRP and ABB produced twice the vertical bone volume of ABB alone (P < .0001). CONCLUSION: Based upon this study of 16 rabbits, it appears that the combination of ABB with PRP resulted in increased vertical bone augmentation when compared with autologous blood in similarly sized created defects.

Controlled Release of Highly Hydrophilic Drugs from Novel Poly(Magnesium Acrylate) Matrix Tablets.

The potential of a new poly(magnesium acrylate) hydrogel (PAMgA) as a pharmaceutical excipient for the elaboration of matrix tablets for the extended release of highly hydrophilic drugs was evaluated. The polymer was synthetized with two different crosslinking degrees that were characterized by FTIR and DSC. Their acute oral toxicity was determined in a mouse model, showing no toxicity at doses up to 10 g/kg. Matrix tablets were prepared using metformin hydrochloride as a model drug and the mechanisms involved in drug release (swelling and/or erosion) were investigated using biorrelevant media. This new hydrogel effectively controlled the release of small and highly hydrophilic molecules as metformin, when formulated in matrix tablets for oral administration. The rate of metformin release from PAMgA matrices was mainly controlled by its diffusion through the gel layer (Fickian diffusion). The swelling capacity and the erosion of the matrix tablets influenced the metformin release rate, that was slower at pH 6.8, where polymer swelling is more intensive, than in gastric medium, where matrix erosion is slightly more rapid. The crosslinking degree of the polymer significantly influenced its swelling capacity in acid pH, where swelling is moderate, but not in intestinal fluid, where swelling is more intense.

Silicon-calcium phosphate ceramics and silicon-calcium phosphate cements: Substrates to customize the release of antibiotics according to the idiosyncrasies of the patient.

Bone substitutes based on calcium phosphates can be classified in two major groups: ceramics and cements. Both are biomaterials with excellent biocompatibility that have been studied as local delivery systems for drugs. This study aims to evaluate drug-release kinetics in silicon beta-tricalcium phosphate ceramics (Si-ß-TCP) and in silicon calcium phosphate cements (Si-CPCs). We want to investigate if the differences in composition and in structure of the Si-ß-TCP and the Si-CPC may influence for drug loading and in its release kinetics from the biomaterial. The results obtained indicate that all drug-loaded materials were efficient to tailor drug release kinetics and inhibited the growth of Staphylococcus aureus. The cements prepared with high concentrations of silicon (80% Si-CPC) present zero-order release kinetics, independent of the drug concentration loaded. Si-ß-TCP and Si-CPC offer a simple technology that could serve to personalize the delivery of bioactive molecules according to each patient's needs in the treatment of bone conditions, not only limited to prophylaxis, but also for the treatment of bone infection.

Contribution of resonance energy transfer to the luminescence quenching of upconversion nanoparticles with graphene oxide.

Upconversion nanoparticles (UCNP) are increasingly used due to their advantages over conventional fluorophores, and their use as resonance energy transfer (RET) donors has permitted their application as biosensors when they are combined with appropriate RET acceptors such as graphene oxide (GO). However, there is a lack of knowledge about the design and influence that GO composition produces over the quenching of these nanoparticles that in turn will define their performance as sensors. In this work, we have analysed the total quenching efficiency, as well as the actual values corresponding to the RET process between UCNPs and GO sheets with three different chemical compositions. Our findings indicate that excitation and emission absorption by GO sheets are the major contributor to the observed luminescence quenching in these systems. This challenges the general assumption that UCNPs luminescence deactivation by GO is caused by RET. Furthermore, RET efficiency has been theoretically calculated by means of a semiclassical model considering the different nonradiative energy transfer rates from each Er3+ ion to the GO thin film. These theoretical results highlight the relevance of the relative positions of the Er3+ ions inside the UCNP with respect to the GO sheet in order to explain the RET-induced efficiency measurements.

Ultrafast photochemistry produces superbright short-wave infrared dots for low-dose in vivo imaging.

Optical probes operating in the second near-infrared window (NIR-II, 1,000-1,700 nm), where tissues are highly transparent, have expanded the applicability of fluorescence in the biomedical field. NIR-II fluorescence enables deep-tissue imaging with micrometric resolution in animal models, but is limited by the low brightness of NIR-II probes, which prevents imaging at low excitation intensities and fluorophore concentrations. Here, we present a new generation of probes (Ag2S superdots) derived from chemically synthesized Ag2S dots, on which a protective shell is grown by femtosecond laser irradiation. This shell reduces the structural defects, causing an 80-fold enhancement of the quantum yield. PEGylated Ag2S superdots enable deep-tissue in vivo imaging at low excitation intensities (<10 mW cm-2) and doses (<0.5 mg kg-1), emerging as unrivaled contrast agents for NIR-II preclinical bioimaging. These results establish an approach for developing superbright NIR-II contrast agents based on the synergy between chemical synthesis and ultrafast laser processing.

Effect of platelet-rich plasma on sinus lifting: a randomized-controlled clinical trial.

OBJECTIVE: The combination of anorganic bovine bone (ABB) with platelet-rich plasma (PRP) has been widely used in bone regeneration procedures although its benefits are still unclear. The purpose of this study was to evaluate whether or not PRP improves the efficacy of ABB in sinus floor augmentation. In addition, we have investigated the effect of residual bone height and tobacco on implant survival in sinus augmentation procedures. PATIENT AND METHODS: Eighty-seven patients recruited for this study underwent 144 sinus floor augmentation procedures using ABB alone or ABB plus PRP (ABB+PRP) in a randomized clinical trial. A total of 286 implants were placed in the augmented bone, and their evolution was followed up for a period of 24 months. In order to investigate on a histological level and any adjunctive effects, we performed an ancillary study in five edentulous patients with a symmetrical severely resorbed maxilla. In these patients, a bilateral sinus augmentation was randomly performed using ABB or ABB+PRP in a split-mouth design, and after 6 months, bone biopsies were taken from the implant sites for histological and histomorphometric analysis. RESULTS: Overall, 96.2% of ABB and 98.6% of ABB+PRP implant success were obtained during the monitoring period and differences were not found between sites grafted with and without PRP in the 87 patients studied. Densitometry assessments and graft resorption were similar in both experimental groups. However, the histological and histomorphometrical analysis in the five edentulous patients revealed that bone augmentation was significantly higher in sites treated with ABB+PRP (p

Control of upconversion luminescence by gold nanoparticle size: from quenching to enhancement.

Metallic nanostructures have the potential to modify the anti-Stokes emission of upconverting nanoparticles (UCNPs) by coupling their plasmon resonance with either the excitation or the emission wavelength of the UCNPs. In this regard gold nanoparticles (AuNPs) have often been used in sensors for UCNP luminescence quenching or enhancement, although systematic studies are still needed in order to design optimal UCNP-AuNP based biosensors. Amidst mixed experimental evidence of quenching or enhancement, two key factors arise: the nanoparticle distance and nanoparticle size. In this work, we synthesize AuNPs of different sizes to assess their influence on the luminescence of UCNPs. We find that strong luminescence quenching due to resonance energy transfer is preferentially achieved for small AuNPs, peaking at an optimal size. A further increase in the AuNP size is accompanied by a reduction of luminescence quenching due to an incipient plasmonic enhancement effect. This enhancement counterbalances the luminescence quenching effect at the biggest tested AuNP size. The experimental findings are theoretically validated by studying the decay rate of the UCNP emitters near a gold nanoparticle using both a classical phenomenological model and the finite-difference time-domain method. Results from this study establish general guidelines to consider when designing sensors based on UCNPs-AuNPs as donor-quencher pairs, and suggest the potential of plasmon-induced luminescence enhancement as a sensing strategy.

Ion-specific and reversible wetting of imidazolium-based minigels.

Cross-linked imidazolium-based [poly(ViEtIm +Br -)] microparticles were synthesized, and their wetting properties were studied by optical microscopy, after addition of aqueous solutions of sodium halides. Particle wetting showed ion specificity due to counterion binding, described by Desnoyer's model. The interaction between anions and the microparticles allowed exchanging halogenides between them in a reversible way. A salt-independent characteristic wetting time was found as well as a decreasing power law with salt concentration, for the network diffusion coefficient. It modified the polymer network elasticity as ion concentration increased, making the network softer.

Silicon bioceramic loaded with vancomycin stimulates bone tissue regeneration.

Porous ceramics doped with silicon and pure ß-TCP were analyzed in terms of internal microstructure, cell behavior, and the percentage of newly formed bone. Additionally the materials were tested to determine which of the two had better properties to load and release vancomycin hydrochloride. Internal pore distribution and porosity were determined through high pressure mercury porosimetry and the specific surface area was measured by the Brunauer Emmet-Teller method. The proliferation and viability of the human osteoblast-like cell line MG-63 was studied to validate both materials. The materials were tested on eight New Zealand rabbits which created defects, 10 mm in diameter, in the calvaria bone. After 8 and 12 weeks a histological and histomorphometric analysis was performed. Si-ß-TCP showed a higher porosity and specific surface area. The cytocompatibility test revealed acceptable results in terms of proliferation and viability whereas the percentage of new bone was higher in Si-ß-TCP with a two-time study being statistically significant with 12 weeks of healing (p < 0.05).The vancomycin loaded within the ceramic scaffolds were burst released and the material had the ability to inhibit bacterial growth. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2307-2315, 2018.

Sensors and bioassays powered by upconverting materials.

In recent years, considerable efforts have been done to better understand the peculiar emission properties of upconverting materials due to their widespread applications in different and important technological fields such as upconversion-based photoactivated cancer therapies, photoactivated drug-delivery, magnetic resonance imaging contrast agents, bioimaging. However, one of the most promising applications of upconverting materials concerns the field of sensing, due to their unique emission properties. In fact, the minimal autofluorescence, blinking, photo-bleaching, and high photostability makes them an excellent alternative to organic dyes or quantum dots. This article reviews the state-of-the-art, design, and sensing strategies of upconversion-based sensing platforms, with special attention to upconverting nanoparticles, as well as how the incorporation of these materials into pre-existing diagnostic tests and bioassays have improved their capabilities for the detection of different kinds of analytes.

Antibiotic Release from Calcium Phosphate Materials in Oral and Maxillofacial Surgery. Molecular, Cellular and Pharmaceutical Aspects.

Calcium phosphate materials (CPM) are widely used in dentistry and maxillofacial surgery. The presence of microbial biofilms and external infections is responsible for the failure of many procedures of dental implants and bone grafts. In an attempt to reduce the percentage of these infectious processes antibiotics have been associated with CPM improving certain conditions. For instance, antibiotics administered orally or intravenously have less effect and the blood flow in relation to this is poor near implants and grafts. Tissue engineering (TE) has employed CPM as a local drug delivery vehicle to be more effective and efficient in bone infections. This review is presented to describe current antibiotics used and the physical and chemical properties of scaffolds.

Oligonucleotide Sensor Based on Selective Capture of Upconversion Nanoparticles Triggered by Target-Induced DNA Interstrand Ligand Reaction.

We present a sensor that exploits the phenomenon of upconversion luminescence to detect the presence of specific sequences of small oligonucleotides such as miRNAs among others. The sensor is based on NaYF4:Yb,Er@SiO2 nanoparticles functionalized with ssDNA that contain azide groups on the 3' ends. In the presence of a target sequence, interstrand ligation is possible via the click-reaction between one azide of the upconversion probe and a DBCO-ssDNA-biotin probe present in the solution. As a result of this specific and selective process, biotin is covalently attached to the surface of the upconversion nanoparticles. The presence of biotin on the surface of the nanoparticles allows their selective capture on a streptavidin-coated support, giving a luminescent signal proportional to the amount of target strands present in the test samples. With the aim of studying the analytical properties of the sensor, total RNA samples were extracted from healthy mosquitoes and were spiked-in with a specific target sequence at different concentrations. The result of these experiments revealed that the sensor was able to detect 10-17 moles per well (100 fM) of the target sequence in mixtures containing 100 ng of total RNA per well. A similar limit of detection was found for spiked human serum samples, demonstrating the suitability of the sensor for detecting specific sequences of small oligonucleotides under real conditions. In contrast, in the presence of noncomplementary sequences or sequences having mismatches, the luminescent signal was negligible or conspicuously reduced.