Experimental scatter of the fatigue response of additively manufactured components: a statistical method based on the Profile Likelihood.

The fatigue response of additively manufactured (AM) specimens is mainly driven by manufacturing defects, like pores and lack of fusion defects, which are mainly responsible for the large variability of fatigue data in the S-N plot. The analysis of the results of AM tests can be therefore complex: for example, the influence of a specific factor, e.g. the building direction, can be concealed by the experimental variability. Accordingly, appropriate statistical methodologies should be employed to safely and properly analyze the results of fatigue tests on AM specimens. In the present paper, a statistical methodology for the analysis of the AM fatigue test results is proposed. The approach is based on shifting the experimental failures to a reference number of cycles starting from the estimated P-S-N curves. The experimental variability of the fatigue strength at the reference number of cycles is also considered by estimating the profile likelihood function. This methodology has been validated with literature datasets and has proven its effectiveness in dealing with the experimental scatter typical of AM fatigue test results.

Simultaneous quantification of Gemcitabine and Irinotecan hydrochloride in rat plasma by using high performance liquid chromatography-diode array detector.

In this manuscript we aimed at the simultaneous separation and quantification of Gemcitabine and Irinotecan hydrochloride (injected both as single components and in combination) from Sprague Dawley rat plasma by using a validated method obtained through the use of a High Performance Liquid Chromatography (HPLC)-diode array detector (DAD). Gemcitabine and Irinotecan hydrochloride were detected and quantified using a Zorbax Extend C-18 column (250 mm × 4.6 mm; 5 µm particle size) in gradient elution mode. The chromatographic analyses were carried out in 15 min. The analytical mode was calibrated and validated in the concentration range from 0.1 to 18 µg/mL both for Gemcitabine and Irinotecan hydrochloride. Sprague Dawley rat plasma was used to perform the analysis. 3-methylxanthine was the internal standard. The weighted-matrix matched standard curves of Gemcitabine and Irinotecan hydrochloride showed a good linearity up to 18 µg/mL. Parallelism tests were also performed to evaluate whether the over-range samples could be analyzed after dilution without affecting the analytical performance. The intra- and inter-day precision (RSD%) values of Gemcitabine and Irinotecan hydrochloride were ≤7.14% and ≤11.5%, respectively. The intra- and inter-day trueness (Bias%) values were in the range from -11.5% to 1.70% for both drugs. The analytical mode performance was further tested after collecting Sprague Dawley rat plasma following a single-dose administration of chemotherapeutics or their association. The validated HPLC-DAD method allowed the simultaneous quantification of Gemcitabine and Irinotecan hydrochloride in the rat plasma, besides the evaluation of the pharmacokinetic parameters and drug delivery.

Micro-CT evaluation of several glide path techniques and ProTaper Next shaping outcomes in maxillary first molar curved canals.

AIM: To evaluate the ability of ProGlider instruments, PathFiles and K-files to maintain canal anatomy during glide path preparation using X-ray computed micro-tomography (micro-CT). METHODOLOGY: Forty-five extracted maxillary first permanent molars were selected. Mesio-buccal canals were randomly assigned (n = 15) to manual K-file, PathFile or ProGlider groups for glide path preparation. Irrigation was achieved with 5% NaOCl and 10% EDTA. After glide path preparation, each canal was shaped with ProTaper Next X1 and X2 to working length. Specimens were scanned (isotropic voxel size 9.1 µm) for matching volumes and surface areas and post-treatment analyses. Canal volume, surface area, centroid shift, canal geometry variation through ratio of diameter ratios and ratio of cross-sectional areas were assessed in the apical and coronal levels and at the point of maximum canal curvature. One-way factorial anovas were used to evaluate the significance of instrument in the various canal regions. RESULTS: Post-glide path analysis revealed that instrument factor was significant at the apical level for both the ratio of diameter ratios and the ratio of cross-sectional areas (P < 0.001), with an improved maintenance of root canal geometry by ProGlider and PathFile. At the coronal level and point of maximum canal curvature, ProGlider demonstrated a tendency to pre-flare the root canal compared with K-file and PathFile. PathFile and ProGlider demonstrated a significantly lower centroid shift compared with K-file at the apical level (P = 0.023). Post-shaping analysis demonstrated a more centred preparation of ProGlider, compared with PathFile and K-files, with no significant differences for other parameters. CONCLUSIONS: Use of ProGlider instruments led to less canal transportation than PathFiles and K-files.

Longevity of class 2 direct restorations in root-filled teeth: A retrospective clinical study.

OBJECTIVES: To evaluate retrospectively the longevity of endodontically treated teeth restored with direct resin composite without cusp coverage, with or without the insertion of fibre posts. The null hypothesis was that direct restorations with fibre posts perform better than those without fibre posts. METHODS: Patients recruited for this study were treated in the Department of Cariology and Operative Dentistry, University of Turin, between 2008 and 2011. In total, 247 patients with 376 root treated posterior teeth, restored with direct resin composite, were recalled for a control visit. Only second-class cavities were considered. Two groups were defined based on the absence (Group A) or presence (Group B) of fibre post. Failures and complications, such as periodontal failure, endodontic failure, tooth extraction, root fracture, post fracture, post debonding, replacement of restoration, crown displacement, and coronal-tooth fracture, were noted. Functional restoration quality was evaluated following the modified USPHS criteria. Data were evaluated statistically with ANOVA. RESULTS: Group A consisted of 128 patients with 178 restorations (88 premolars, 90 molars) with a median follow-up of 34.44 months. Group B consisted of 119 patients with 198 teeth (92 premolars, 106 molars) with a median follow-up of 35.37 months. Direct restorations with fibre posts were statistically significantly more functional (95.12% success) than those without fibre posts (80% success) because of less marginal discolouration, better marginal integrity, and higher restoration integrity. CONCLUSIONS: The null hypothesis was accepted because direct post-endodontic restorations with fibre posts performed better than restorations without posts after 3 years of masticatory function. CLINICAL SIGNIFICANCE: An evaluation of the longevity of post endodontic direct restoration would seem to enhance the fiber post insertion within a composite restoration to reduce clinical failures.

Bisphosphonate-polyaspartamide conjugates as bone targeted drug delivery systems.

Poly-hydroxy-aspartamide was used as a backbone to synthesize bisphosphonate derivatives thus achieving macromolecular carriers to be potentially used as targeting agents for bone drug delivery. Molecules bearing bisphosphonate groups, such as aminobisphosphonate (ABP) and neridronate (NRD), have been conjugated to polyaspartamide (α,ß-poly(N-2-hydroxyethyl)-dl-aspartamide, PHEA), with or without a spacer (succinic acid or 6-aminocaproic acid) thus obtaining PHEA-succinate-ABP and PHEA-caproylcarbamate-ABP and PHEA-ABP and PHEA-NRD, respectively. Bisphosphonate-polymer conjugates were physico-chemically characterized using size exclusion chromatography and 1H-NMR; and their in vitro and ex vivo affinity for bone tissue has been further tested using the hydroxylapatite and rabbit bone binding assays, respectively. In vivo studies were carried out using rats to evaluate the biodistribution features of bisphosphonate-polymer conjugates in comparison with the starting PHEA. In vivo findings evidenced a suitable selectivity of bisphosphonate-polymer conjugates toward the bone tissues also as a function of time.

Fibre post adaptation and bond strength in oval canals.

AIM: To evaluate ex vivo the bond strength and adaptation of fibre posts with oval and circular cross sections luted in oval canals with post spaces prepared using dedicated drills or ultrasonic tips. METHODOLOGY: Forty extracted premolars with oval canals were root filled, then randomly divided into four groups according to the post space preparation device and the shape of the luted fibre post: dedicated drill + round post, dedicated drill + oval post, ultrasonic tip + round post and ultrasonic tip + oval post. Posts were cemented with a self-adhesive cement (RelyX Unicem 2; 3M ESPE). Samples were sectioned in 1-mm-thick slices and observed under a microscope, and the area occupied by the post within the post space area was calculated. Bond strength was then measured using a push-out test, and the failure modes were evaluated with a stereomicroscope at 40× magnification. Fibre post adaptation and push-out test results were evaluated by analysis of variance (P < 0.05). RESULTS: Fibre posts, both round and oval, were better adapted to the apical region of the post space (P = 0.001). In oval canals, the bond strength was significantly higher in coronal regions, when the post space was prepared with a dedicated drill and an oval post was luted (P < 0.0001). Adhesive failures between cement and post were the most frequent type of failure in all groups. CONCLUSIONS: Circular and oval posts achieved similar adaptation to oval canals, but the use of ultrasonic tips and round posts resulted in reduced bond strength values.

Comparative evaluation of the enamel margins roughness obtained with different finishing devices.

AIM: The aim of this in vitro study was to compare the surface roughness of enamel margins resulting by the use of rotating, sonic and ultrasonic devices for cavity margins finishing. METHODS: Forty-eight anterior intact teeth were selected for this study. Each item was sectioned 1 mm below the CEJ, perpendicular to the long axis of the tooth, with the carborundum separating disk mounted on the high-speed handpiece. With the same bur the crown was separated into two parts along the midline vertically. At the end 96 "half-crowns" were obtained. The samples were divided into 6 groups of 8 samples each, according to enamel margin's finishing technique (A and B: diamond ultrasonic tip; C: multisteel ultrasonic tip; D: fine diamond rotating bur; E: ultra-fine diamond rotating bur; F: sonicflex prep). The surface roughness evaluation of the enamel of each sample has been carried out by using a profilometer. The statistical analysis was performed with a balanced hierarchical ANOVA. RESULTS AND CONCLUSION: The results of this in vitro study showed that the enamel roughness obtained with sonic and ultrasonic devices was significantly higher than roughness obtained with rotating burs. Within the sonic and ultrasonic tips, the multisteel ones gave better results, that were comparable to diamond sonic device.

Gemcitabine and tamoxifen-loaded liposomes as multidrug carriers for the treatment of breast cancer diseases.

The effects of a lipid composition on the physico-chemical and technological properties of a multidrug carrier (MDC) containing both gemcitabine (GEM) and tamoxifen (TMX), as well as its in vitro antitumoral activity on different breast cancer cell lines, were investigated. In particular, the following three different liposomal formulations were prepared: DPPC/Chol/DSPE-mPEG2000 (6:3:1 molar ratio, formulation A), DPPC/Chol/DOTAP (6:3:1 molar ratio, formulation B) and DPPC/Chol/DPPG (6:3:1 molar ratio, formulation C). The colloidal systems were obtained by the TLE technique and the extrusion process allowed us to obtain vesicles having mean sizes of 150-200 nm, while the surface charges varied between 50 mV and -30 mV. Formulation A showed the best encapsulation efficiency between the two compounds and the presence of TMX influenced the release profile of GEM (hydrophilic compound) as a consequence of its effect on the fluidity of the bilayer. An MDC of formulation A was used to effectuate the in vitro cytotoxicity experiments (MTT-test) on MCF-7 and T47D cells. The liposomal MDC provided the best results with respect to the single drug tested in the free form or entrapped in the same liposomal formulation. The CLSM experiments showed a great degree of cell interaction of liposomal MDC after just 6h.

Drug delivery applications with ethosomes.

Ethosomes are specially tailored vesicular carriers able to efficiently deliver various molecules with different physicochemical properties into deep skin layers and across the skin. This paper reviews the unique characteristics of the ethosomal carriers, focusing on work carried out with drug containing ethosomal systems in animal models and in clinical studies. The paper concludes with a discussion on the safety of the ethosomal system applications.

Novel PEG-coated niosomes based on bola-surfactant as drug carriers for 5-fluorouracil.

Innovative niosomes made up of α,ω-hexadecyl-bis-(1-aza-18-crown-6) (bola), Span 80® and cholesterol (2:5:2 molar ratio) are proposed as suitable delivery systems for the administration of 5-fluorouracil (5-FU), an antitumoral compound largely used in the treatment of breast cancer. The bola-niosomes, after sonication procedure, showed mean sizes of ~200 nm and a loading capacity of ~40% with respect to the amount of 5-FU added during the preparation. Similar findings were achieved with PEG-coated bola-niosomes (bola, Span 80(R), cholesterol, DSPE-mPEG2000, 2:5:2:0.1 molar ratio respectively). 5-FU-loaded PEG-coated and uncoated bola-niosomes were tested on MCF-7 and T47D cells. Both bola-niosome formulations provided an increase in the cytotoxic effect with respect to the free drug. Confocal laser scanning microscopy studies were carried out to evaluate both the extent and the time-dependent bola-niosome-cell interaction. In vivo experiments on MCF-7 xenograft tumor SCID mice models showed a more effective antitumoral activity of the PEGylated niosomal 5-FU at a concentration ten times lower (8 mg/kg) than that of the free solution of the drug (80 mg/kg) after a treatment of 30 days.

Retinoids: new use by innovative drug-delivery systems.

BACKGROUND: Retinoids represent an old class of bioactives used in the treatment of different skin pathologies (such as acne and psoriasis) and in the treatment of many tumors. Unfortunately, they present several side effects, i.e., burning of skin and general malaise after systemic administration and they are very unstable after exposition to light. METHODS: One of the most promising new approaches for reducing the side effects of retinoids while improving their pharmacological effect is the use of drug-delivery devices. This review explains the current status of retinoid drug transport, which has been developing over the last few years, explaining the modification of their biopharmaceutical properties in detail after encapsulation/inclusion in vesicular and polymeric systems. RESULTS/CONCLUSION: Different colloidal and micellar systems containing retinoid drugs have been realized furnishing important potential advancements in traditional therapy.

Lipoamino acid prodrugs of paclitaxel: synthesis and cytotoxicity evaluation on human anaplastic thyroid carcinoma cells.

Lipophilic derivatives of the anticancer drug paclitaxel (PTX) were prepared by means of its conjugation to lipoamino acid (LAA) residues, with the aim of increasing drug accumulation in tumor cells. PTX was linked to the methyl esters of norleucine (C6) or 2-aminodecanoic acid (C10). A succinic acid group was used as a spacer to link the 2'-hydroxyl group of PTX and the LAA residue, respectively by means of an ester and an amide bond. The in vitro anticancer activity of the prodrugs was tested on a human thyroid anaplastic cancer cell line (ARO). The intracellular uptake kinetics of free PTX and its prodrugs was assessed by HPLC. PTX-LAA prodrugs showed a noticeable cytotoxic activity against ARO cells at shorter incubation time (12 h) and lower doses (0.01-0.1 microM) than PTX. Intracellular accumulation experiments indicated an improvement of drug concentration inside these cells, related to the block of the cellular expulsion by means of multi drug resistance efflux complex and improved physicochemical features that allowed the greater passive cellular membrane permeation. The enhanced activity of PTX-LAA prodrugs, in terms of potency and onset of the effect, as well as the interesting intracellular accumulation data suggest that these compounds can be further tested as possible alternatives to PTX for the treatment of resistant cancer cells.

Colloidal carriers for the enhanced delivery through the skin.

BACKGROUND: The skin is the largest organ of our body and acts as a protective barrier with sensory and immunological functions. Its peculiar structure influences the passage of bioactives and only its modulation can facilitate the drug dermal/transdermal diffusion. In the past few years research in this field has assured better use of this application area. METHODS: One of the most promising approaches is the use of drug delivery devices; this review explains the state of the art of drug transport through the skin by means of vesicular (classic liposomes, Transfersomes, niosomes and ethosomes) and particulate systems. RESULTS/CONCLUSION: Colloidal drug delivery systems are important in the field of drug delivery systems as their different characteristics make them suitable for various purposes.

In vitro skin permeation of sunscreen agents from O/W emulsions.

The effects of different emulsifiers on the in vitro permeation through human skin of two sunscreen agents [octylmethoxycinnamate (OMC) and butylmethoxydibenzoylmethane (BMBM)] were investigated from O/W emulsions. The test formulations were prepared using the same oil and aqueous phase ingredients and the following emulsifier and coemulsifier systems: Emulgade SE((R)) (ceteareth-12 and ceteareth-20 and cetearyl alcohol and cetyl palmitate) and glycerylmonostearate (emulsion 1); Brij 72((R)) (steareth-2), Brij 721((R)) (steareth-21) and cetearyl alcohol (emulsion 2); Phytocream((R)) (potassium palmitoyl-hydrolysed wheat protein and glyceryl stearate and cetearyl alcohol) and glycerylmonostearate (emulsion 3); Montanov 68((R)) (cetearyl glucoside and cetearyl alcohol) (emulsion 4); Xalifin-15((R)) (C(15-20) acid PEG-8 ester) and cetearyl alcohol (emulsion 5). The cumulative amount of OMC that permeated in vitro through human skin after 22 h from the formulations being tested decreased in the order 3 > 1 congruent with 4 > 5 > 2 and was about nine-fold higher from emulsion 3 compared with that from emulsion 2. As regards BMBM, no significant difference was observed as regards its skin permeation from emulsions 1, 3, 4 and 5, whereas formulation 2 allowed significantly lower amounts of BMBM to permeate the skin. In vitro release experiments of OMC and BMBM from emulsions 1-6 through cellulose acetate membranes showed that only emulsions 4 and 5 provided pseudo-first-order release rates only for OMC. The results of this study suggest that the type of emulsifying systems used to prepare an O/W emulsion may strongly affect sunscreen skin permeation from these formulations. Therefore, the vehicle effects should be carefully considered in the formulation of sunscreen products.

Polyaspartylhydrazide copolymer-based supramolecular vesicular aggregates as delivery devices for anticancer drugs.

In this paper we report on three different hydrophilic copolymers based on alpha,beta-polyaspartylhydrazide (PAHy) bearing butyric groups in the side chain (C 4) (PAHy-C 4) or a combination of butyric groups and positive charged residues ((carboxypropyl)trimethylammonium chloride, CPTACl) (PAHy-C 4-CPTA) that were synthesized and used for the preparation of new supramolecular vesicular aggregates (SVAs) containing gemcitabine as an antitumor drug. Gemcitabine-loaded SVAs containing synthesized PAHy derivatives were characterized from the physicochemical and technological point of view and the in vitro toxicity and anticancer activity on two different human cancer cell lines, i.e., CaCo-2 (human colon carcinoma) and ARO (human anaplastic thyroid carcinoma) cells, were also evaluated. Moreover, considering that carrier-cell interaction is an important factor to achieve an improvement of anticancer drug activity, confocal laser scanning microscopy and flow cytometric experiments were carried out on the two different cancer cell lines.

Influence of modified cyclodextrins on solubility and percutaneous absorption of celecoxib through human skin.

We evaluated the ability of two modified cyclodextrins, hydroxypropyl-beta-cyclodextrin (HP-beta-Cyd) and 2,6-di-O-methyl-beta-cyclodextrin (DM-beta-Cyd), to influence the percutaneous absorption through isolated human stratum corneum and epidermis (SCE) of celecoxib (CCB). Previous studies demonstrated that DM-beta-Cyd includes the drug, producing a significant increase of water solubility (0.5 mg/ml at 25 degrees C) and dissolution rate of CCB. In this work chemical-physical characterization studies were performed to evaluate the ability of HP-beta-Cyd to include CCB. We showed that only an external interaction could exist between CCB and HP-beta-Cyd that positively influences the water solubility of the drug (0.12 mg/ml at 25 degrees C for CCB-HP-beta-CyD system and 4.12 x 10(-3) mg/ml at 25 degrees C for free CCB). In vitro percutaneous experiments were performed using samples in solution and in suspension containing different Cyd concentrations. Both HP-beta-Cyd and DM-beta-Cyd enhanced drug flux through SCE by means of an increase of dissolution rate of the drug as well as a direct action on the stratum corneum (SC). Histological analysis of treated SCE showed a protective effect of the two Cyds towards an invasive action shown by CCB on SC.

Synthesis, characterization and in vitro evaluation of dimethyl-beta-cyclodextrin-4-biphenylylacetic acid conjugate.

Biphenylylacetic acid (BPAA) was linked to the free hydroxyl group of 2,6-di-O-methyl-beta-Cyclodextrin (DM-beta-CyD) through an ester linkage to obtain the site specific release of the drug to the colon. The conjugate at 1:1 mole ratio was separated from the reaction mixture by semipreparative reverse-phase HPLC and characterized by 1H-NMR, 13C-NMR, IR spectroscopy, mass spectrometry and elemental analysis. Chemico-physical characteristics, such as water solubility and dissolution rate, were evaluated comparatively to the BPAA-DM-beta-CyD inclusion complex. Hydrolysis rates were investigated in media simulating gastro-intestinal fluids and at pH 7.4 in the presence of porcine liver esterase. A rapid release of the drug was observed at acid pH value. In all cases a first order kinetic was observed, characterized by t1/2 value of 1.19, 19 and 4 h for chemical hydrolysis at pH 1.1, at pH 7.4 and enzymatic hydrolysis, respectively. In vitro permeation studies through caco-2 cells confirmed the ability of DM-beta-CyD to increase the absorption of included BPAA. A slow permeation was observed for the drug conjugate to DM-beta-CyD due to the slow release of BPAA.

Biomembrane model interaction and percutaneous absorption of papaverine through rat skin: effects of cyclodextrins as penetration enhancers.

The effects of different concentrations of beta-cyclodextrin (beta-CyD), hydroxypropyl-beta-cyclodextrin (HP-beta-CyD) and 2,6-di-O-methyl-beta-cyclodextrin (DM-beta-CyD) on percutaneous absorption of papaverine hydrochloride (PAP) were investigated. Abdominal rat skin mounted in Franz cells was used for in vitro experiments. To evaluate CyD interaction with a bilayer structure model, dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and DPPC-Chol (8:2 mole ratio) vesicles were used. CyD vesicle interaction was evaluated by differential scanning calorimetry. Permeation through rat skin and calorimetric experiments demonstrated that at low concentrations DM-beta-CyD shows higher enhancer activity as a possible result of a perturbing action on the skin by a complexation of its lipid components, but at higher concentrations HP-beta-CyD is the most effective. By considering that HP-beta-CyD presents a very moderate destabilizing action on the skin, we conclude that a 10% aqueous solution of this macrocycle appears to be the most suitable transdermal absorption enhancer for PAP.

In vitro and in vivo evaluation of caffeic and ferulic acids as topical photoprotective agents.

Topically-applied antioxidant drugs represent a successful strategy for protecting the skin against UV-mediated oxidative damage. However, they can afford to the skin a satisfactory photoprotection only if able to permeate through the stratum corneum and thus to reach deeper cutaneous layers. Caffeic and ferulic acids, dissolved in saturated aqueous solutions at pH 3 or 7.2, have been tested for their capability to permeate through excised human skin mounted in Franz cells. At both pH values, ferulic and, at a lower degree, caffeic acids appeared able to permeate through the stratum corneum. The known higher lipophilicity of ferulic acid may explain the fact that it permeates through the stratum corneum better than caffeic acid. However, vehicle pH values proved to have no influence on biophenol skin permeation profile; this observed lack of pH effect may reflect the drug higher concentration attainable in saturated solutions at high pH. On the basis of the findings obtained in these in vitro experiments, we designed the schedule of a series of in vivo experiments, carried out to evaluate the ability of caffeic and ferulic acids to reduce, in healthy human volunteers, UVB-induced skin erythema, monitored by means of reflectance spectrophotometry. Caffeic and ferulic acids, dissolved in saturated aqueous solution pH 7.2, proved to afford a significant protection to the skin against UVB-induced erythema. To conclude, we have confirmed, by means of in vitro and in vivo experiments, that caffeic and ferulic acids may be successfully employed as topical protective agents against UV radiation-induced skin damage; however their skin absorption is not influenced by the pH of the formulation.