Latest Trends in Surface Modification for Dental Implantology: Innovative Developments and Analytical Applications.

An increase in the world population and its life expectancy, as well as the ongoing concern about our physical appearance, have elevated the relevance of dental implantology in recent decades. Engineering strategies to improve the survival rate of dental implants have been widely investigated, focusing on implant material composition, geometry (usually guided to reduce stiffness), and interface surrounding tissues. Although efforts to develop different implant surface modifications are being applied in commercial dental prostheses today, the inclusion of surface coatings has gained special interest, as they can be tailored to efficiently enhance osseointegration, as well as to reduce bacterial-related infection, minimizing peri-implantitis appearance and its associated risks. The use of biomaterials to replace teeth has highlighted the need for the development of reliable analytical methods to assess the therapeutic benefits of implants. This literature review considers the state-of-the-art strategies for surface modification or coating and analytical methodologies for increasing the survival rate for teeth restoration.

Novel Utilization of Therapeutic Coatings Based on Infiltrated Encapsulated Rose Bengal Microspheres in Porous Titanium for Implant Applications.

Despite the increasing progress achieved in the last 20 years in both the fabrication of porous dental implants and the development of new biopolymers for targeting drug therapy, there are important issues such as bone resorption, poor osseointegration, and bacterial infections that remain as critical challenges to avoid clinical failure problems. In this work, we present a novel microtechnology based on polycaprolactone microspheres that can adhere to porous titanium implant models obtained by the spacer holder technique to allow a custom biomechanical and biofunctional balance. For this purpose, a double emulsion solvent evaporation technique was successfully employed for the fabrication of the microparticles properly loaded with the antibacterial therapeutic agent, rose bengal. The resulting microspheres were infiltrated into porous titanium substrate and sintered at 60 °C for 1 h, obtaining a convenient prophylactic network. In fact, the sintered polymeric microparticles were demonstrated to be key to controlling the drug dissolution rate and favoring the early healing process as consequence of a better wettability of the porous titanium substrate to promote calcium phosphate nucleation. Thus, this joint technology proposes a suitable prophylactic tool to prevent both early-stage infection and late-stage osseointegration problems.

Exploring polar hydrophobicity in organized media for extracting oligopeptides: application to the extraction of opiorphin in human saliva.

Supramolecular solvents (dubbed SUPRAS) are gaining momentum as extractants of compounds of interest from complex matrixes such as foodstuff and biological and environmental samples. However, their powerful extraction mechanism, based on multiligand ability for solute binding, fails when applied to very polar compounds, hindering their applicability to the extraction of highly polar metabolites. In this work, we introduce the synthesis, characterization, and application of a new kind of SUPRAS formed by heptafluorobutyric acid (HFBA). The polar hydrophobicity of this perfluorinated acid results in a SUPRAS, which coacervates at acidic pHs, that shows a great capability to extract amino acids and oligopeptides (recoveries in the range 81-105%) with nonpolar alkyl, cyclic or aromatic side chain substituents (with log D > -3.62). To further demonstrate the potential of this novel SUPRAS, an analytical methodology for the determination of opiorphin in real saliva samples was developed and fully validated. The HFBA-based SUPRAS was synthetized in situ from 950 µL of stabilized saliva, by the addition of 150 µL of HFBA and 400 µL of HCl 37% (v/v). The resulting SUPRAS was directly injected into a LC-MS/MS system for further quantification. Quantitative recoveries in the range of 87-110% were obtained with relative standard deviations below 20%. The HFBA-based SUPRAS is, therefore, capable of efficiently extracting opiorphin from saliva samples and shows a high potential for the determination of several amino acids and oligopeptides from biological samples.

Effects of Dietary Zn/Se and α-Tocopherol Supplementation on Metabolic Milieu, Haemogram and Semen Traits of Breeding Stallions.

Trace element status and metabolic milieu are sometimes overlooked in common veterinary clinical practice across animal species. The evaluation of requirements of trace elements, in fact, may be useful to prevent the perturbation of tissue-specific metabolic impair. In particular, essential trace elements in the diet play key roles within sub-cellular metabolic patterns with macro effects at the systemic level, like blood cell stability and semen quality. This effect was studied in breeding stallions, in which semen quality and haemogram are important for reproduction. A case-control feeding trial involved 40 stallions (age: 8-21 years; body weight, BW: 510-531 kg) of one stud centre, allotted to two experimental groups (n = 20 control, CON vs. n = 20 supplemented, SUPPL100), following a matched-pairs approach based on age. Supplemented stallions (SUPPL100) received a mixed mineral and vitamin supplement of Zn/Se and α-tocopherol (α-TOH) (100 g/day stallion) to compound feed, fed as control diet to horses of the control group (CON). Horses resulted deficient in circulating α-TOH and Zn at the start, though clinically healthy. After supplementation, different plasmatic levels of α-TOH, Zn and Se were found between groups. Circulating basophils (BASO) and mean cell haemoglobin concentration (MCHC) were affected by the dietary treatment (p < 0.05). Plasmatic Se affected monocyte count, haematocrit, mean cell volume and mean cell haemoglobin concentration. Semen traits were not affected by the dietary treatment per se, except for mobile/progressive sperm cells (%) of stallions aged > 13 years marginal circulating levels of α-TOH (p = 0.04). Ameliorating the micromineral status showed to improve the haemogram of stallions in view of circulating levels of Cu. Semen quality appeared to be strongly dependent on animal effects.

Hexanol-Based Supramolecular Solvents Tool for the Determination of 11 Illicit Phenethylamines in Oral Fluid by LC-MS/MS.

Monitoring of new phenethylamine designer drugs in oral fluid (OF) is a crucial aim in workplace testing and driving under the influence of drug programs. In this study a simple and very quick method for the quantification of 11 illicit drugs in OF, which gave negative results to immunoassay tests, is proposed. Sample treatment and extraction of analytes were simultaneously achieved by applying supramolecular solvents (SUPRAS) tool. Chromatographic separation and compounds quantification were carried out by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Efficacy of cleaning-up/extraction of this SUPRAS approach was fully confirmed by recovery and matrix effect results. The entire analytical procedure was validated following the international guidelines. The SUPRAS extraction coupled with LC-MS/MS resulted in powerful tool for the control of phenethylamines abuse, with rapid run time and minimal sample preparation. The use of this methodology could be easily extended to monitoring of other drugs of abuse.

Variation of Hematochemical Profile and Vitamin E Status in Feral Giara Horses From Free Grazing in the Wild to Hay Feeding During Captivity.

Wildlife protection and management are important priorities for landscape identity and biodiversity preservation. Feeding practices of fauna confined in facilities during temporary captivity are fundamental to support animal health and natural behavior. Appropriate provision of feedstuffs appears to be necessary to support the best practices in respect of animal species-specific natural diet. This investigation explored the variation of the metabolic profile by means of selected metabolite and respective circulating levels in a group feral Giara horses undergoing the change of the diet, moving from natural free grazing in the wild to temporary captivity. Six Giara horses (4 mares and 2 stallions; estimated age: 2.5-3 years; body weight: 163-170 kg) were captured to monitor the serological reaction to equine infectious anemia (EIA; screening at Coggins test). Animals were sheltered in a wildlife rescue center for a duration of 4 weeks, and all received the same hay-based diet (ad libitum). On 0 and 28 days of captivity, blood serum alpha-tocopherol (α-TOH) concentration was determined alongside selected metabolites (liver enzymes, total protein and fractions, cholesterol, triglycerides, and macrominerals and trace elements). Comparative feces quality and composition were also assessed. Both serum samples (0 vs. 28 days) displayed α-TOH levels below (<2 µg/mL) adequacy established for the domestic horse. Initial levels markedly (P = .020) decreased after the 4 weeks of captivity (Δ = -32.5%). Vitamin E status and ALT levels varied significantly, but serum protein fractions did not point to significant variations before and after captivity. All horses tested negative to EIA. Monitoring of vitamin E status of wild and feral herbivores may be recommendable in the context of adequate feeding practices during captivity to prevent potential deficiency or excessive depletion.

The metabolic profile of Asinara (albino) and Sardo donkeys (pigmented) (Equus asinus L., 1758) points to unequivocal breed assignment of individuals.

This study pointed to explore if variations in circulating levels of metabolites in the blood stream of no. 25 feral donkeys occur in view of the different coat color between specimens of Asinara (albino, no. 8) vs. Sardo (dun-grey, no. 17) breed. All individuals involved in this investigation are living in the nature, at Mediterranean latitudes and roam in the same areas all over the National Park of Capo Caccia, where they feed on spontaneous vegetation sources. The study was conducted during the positive photoperiod of the boreal hemisphere (peak in the month of June, 2019) to maximize the effect of exposure to the natural sun radiation and thus elicit the coping ability of albino (Asinara) in comparison with pigmented donkeys (Sardo). The biochemical profile of all donkeys was used in a Discriminant Analysis (DA) to explore if circulating levels of metabolites could point to metabolic markers for breed assignment of individuals following a canonical discriminant analysis (CANDISC). The biochemical investigation included also the determination of the circulating Vitamin E (alpha tocopherol, α-TOH), as an essential biologically active compound involved in antioxidant mechanisms, and its respective status (circulating α-TOH to total triglycerides and total cholesterol ratio). In the CANDISC, the distance between the two breeds was not significant. However, it pointed to different metabolites (UREA, total protein, total triglycerides, Zn) capable of describing biochemical patterns on each respective breed (Asinara vs. Sardo). The multivariate analysis DA carried out using 22 metabolites correctly assigned individuals to the two breeds in the 100% of cases. In view of such metabolic background, circulating α-TOH found in the bloodstream of Asinara vs. Sardo donkeys under free grazing conditions turned out to reach similar values (2.114 vs. 1.872 µg/ml, respectively, p = 0.676). It is worth noting that significant differences were observed as to circulating lactate dehydrogenase (LDH, p = 0.022) levels, in association with increased creatine phosphokinase (CPK, p = 0.076), both above the upper limit of the physiological range reported in other donkey breeds, and found in the totality of Asinara (albino) donkeys solely, still apparently clinically healthy.

Restricted Access Volatile Supramolecular Solvents for Single-Step Extraction/Cleanup of Benzimidazole Anthelmintic Drugs in Milk Prior to LC-MS/MS.

In this work, a restricted access volatile supramolecular solvent (RAM-VOL-SUPRAS) directly synthesized in milk is proposed for the first time for the simultaneous extraction and cleanup of benzimidazole anthelmintic drugs in milk meant for human consumption. The RAM-VOL-SUPRAS was formed by the self-assembly and coacervation of hexanol in tetrahydrofuran induced by the water content in milk. Benzimidazoles legislated by the European Union were quantitatively extracted (80-110%), and proteins were precipitated by the action of THF and the amphiphile; extraction of carbohydrates was avoided by a size exclusion mechanism, and lipids were removed during hexanol evaporation. The analytical methodology was fully validated according to Commission Decision 2002/657/EC. Method detection limits from 0.03 to 0.14 µg L-1 were well below the maximum residue limits legislated in milk for these drugs, with interday precisions at maximum residue levels below 13%. This novel methodology guarantees a rapid and reliable tool for daily and routine laboratory analyses in the field of food quality control.

SUPRAS extraction approach for matrix-independent determination of amphetamine-type stimulants by LC-MS/MS.

Monitoring of amphetamine-type stimulant (ATS) confronts clinical labs with a high number of samples involving a variety of biological matrices. Liquid chromatography-tandem mass spectrometry (LC-MS/MS), routinely used for confirmation of ATS abuse, requires of laborious and matrix-dependent sample treatment methods, this increasing analysis time and cost. In this work, a universal and single-step sample treatment, based on supramolecular solvents (SUPRAS), was proposed for simplifying ATS confirmation in seven biological matrices. The SUPRAS was synthesized in situ in the sample (900 µL of basified oral fluid, urine, serum, sweat or breast milk or 50 mg of digested hair or fingernails) by the addition of hexanol (200 µL) and tetrahydrofuran (900 µL). The mixture was vortex-shaken and centrifuged and the SUPRAS extract was subsequently analyzed by positive ion mode electrospray LC-MS/MS. The method was fully validated for amphetamine (AMP), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), N-ethyl-3,4-methylenedioxyamphetamine (MDEA) and N-methyl-3,4-methylenedioxyamphetamine (MDMA). Maximum ion suppression or enhancement was 9% and 7%, respectively, and extraction recoveries (87-111%) and within- (0.1-6.7%) and between-day (0.3-9.7%) CVs were all within required values. The lower limits of quantification (LLOQ) for biological fluids (5 ng/mL), and hair and fingernails (100 ng/g) were all well below the cut-offs established by worldwide organizations. Confirmation of MDA was carried out in five urine samples that tested positive for ATS by immunoassay. The SUPRAS-LC-MS/MS methodology succeeded in developing a hitherto unexplored and universal tool for quantifying ATS in a comprehensive pool of biological matrices of interest in forensic and clinical samples.

Blood serum retinol levels in Asinara white donkeys reflect albinism-induced metabolic adaptation to photoperiod at Mediterranean latitudes.

Previous works on albinism form of Asinara white donkeys (Equus asinus) identified the mutation leading to the peculiar phenotype spread to all specimens of the breed. Inbreeding naturally occurred under geographic isolation, on Asinara Island, in the Mediterranean Sea. Albino individuals can be more susceptible to develop health problems when exposed to natural sun radiation. Alternative metabolic pathways involved in photoprotection were explored in this trial. Nutrition-related metabolites are believed to contribute to the conservation of Asinara donkeys, in which melanin, guaranteeing photoprotection, is lacking. Biochemical profiles with particular focus on blood serum ß-carotene and retinol levels were monitored. Identical natural grazing conditions for both Asinara (albino) and Sardo (pigmented) donkey breeds were assured on same natural pastures throughout the experimental period. A comparative metabolic screening, with emphasis on circulating retinol and nutrient-related metabolites between the two breeds, was carried out over one year. Potential intra- and interspecimen fluctuations of metabolites involved in photoprotection were monitored, both during negative and positive photoperiods. Differences (p = .064) between blood serum concentrations of retinol from Asinara versus Sardo breed donkeys (0.630 vs. 0.490 µg/ml, respectively) were found. Retinol levels of blood serum turned out to be similar in the two groups (0.523 vs. 0.493 µg/ml, respectively, p = .051) during the negative photoperiod, but markedly differed during the positive one (0.738 vs. 0.486, respectively, p = .016). Blood serum ß-carotene levels displayed to be constantly around the limit of sensitivity in all animals of both breeds. Variations in blood serum concentrations of retinol in Asinara white donkeys can reflect the need to cope with seasonal exposure to daylight at Mediterranean latitudes, as an alternative to the lack of melanin. These results may suggest that a pulsed mobilization of retinol from body stores occurs to increase circulating levels during positive photoperiod.