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Effects of selective and nonselective PGE2 receptor agonists on cervical tensile strength and collagen organization and microstructure in the pregnant rat at term.

OBJECTIVE: The purpose of this study was to determine which of the 4 PGE2 receptors (EP1-EP4) is involved in cervical ripening in the rat, and to correlate its activity with changes in tensile strength and collagen microstructure. STUDY DESIGN: We assessed tensile strength after administration of selective and nonselective PGE2 receptor agonists. Quantification of collagen organization and microstructure was accomplished with polarized light microscopy and transmission electron microscopy. RESULTS: Selective agonists for EP1-3 did not produce significant differences when compared with each other or control animals. Significant differences in tensile strength, proportion of organized collagen, and microstructure were found between treatment and control animals with the nonselective receptor agonist (PGE2). This was taken as an indirect measure of EP4 activity. CONCLUSION: Changes in cervical collagen organization and microstructure are quantifiable and correlate with changes in tensile strength. These data implicate EP4 as the PGE2 receptor involved in producing these changes in the rat cervix.

Animals↗

Mifepristone-induced cervical ripening: structural, biomechanical, and molecular events.

OBJECTIVE: The purpose of this study was to assess the structural, biomechanical, and biochemical effects of mifepristone-induced progesterone withdrawal on the rat cervix to identify possible mechanisms by which mifepristone incites cervical ripening. STUDY DESIGN: After the administration of mifepristone, cervical tensile strength was determined by the cervical creep method. With polarized light microscopy and transmission electron microscopy, collagen organization and microstructure were quantified. Matrix metalloproteinase expression was assessed by Western Blot and Real-time reverse transcriptase-polymerase chain reaction. RESULTS: Mifepristone induced a decrease in cervical tensile strength at mid gestation that was associated with a decrease in collagen organization. Additionally, mifepristone led to collagen fragmentation with a significant decrease in fibril length and diameter, although fibril bundling remained unaffected. Matrix metalloproteinase-2 expression increased after the administration of mifepristone. CONCLUSION: Mifepristone-induced cervical ripening is associated with collagen degradation, and the collagenase activity of matrix metalloproteinase-2 may play a role in this process.

Animals↗

Synthesis and aqueous phase behavior of 1-glyceryl monooleyl ether.

Synthesis of 1-glyceryl monooleyl ether (GME) has been accomplished yielding material of high purity (99.6%). The aqueous phase behavior of synthesized lipid has been investigated by using polarized microscopy and small angle X-ray diffraction. As a result, a partial temperature-composition phase diagram has been constructed. GME forms a reversed micellar solution and reversed hexagonal liquid crystalline phase at low and high hydration, respectively. The hexagonal phase coexists with excess water and is stable up to about 63 degrees C. These findings make GME an interesting alternative to glycerol monoesters in various fields of applications.

Fatty Alcohols↗

Sealing smooth enamel surfaces with a newly devised adhesive patch: a radiochemical in vitro analysis.

OBJECTIVES: To assess the enamel-protective potential of a newly devised adhesive patch when used as a smooth surface sealant. METHODS: Eighty enamel discs were prepared from bovine lower central incisors and then irradiated. Twenty specimens were treated with one of three sealing options: enamel bond in a two-step application, the prototype of an adhesive patch, or a flowable resin. Unsealed enamel served as positive controls. Loss of apatite was determined using a radiochemical liquid scintillation method after immersion of the samples in saliva or lactic acid (n=10 per treatment group) each for up to 21 days, during which this experimental and control enamel surfaces were exposed to ten toothbrush strokes per day. Quantitative liquid scintillation data were verified by polarized light microscopy. RESULTS: With lactic acid exposure, a double layer of enamel bonding showed better enamel protection than untreated controls, but was significantly less effective than the adhesive patch or filled composite (P<0.05). No significant differences were noted between the latter two treatments. SIGNIFICANCE: It was concluded that the adhesive patch under investigation merits further studies to assess its potential as an inter-proximal sealant.

Acrylates↗

Surface characterization of precious alloys treated with thione metal primers.

OBJECTIVES: To characterize the effect of two thione metal primers with phosphate groups on the surface morphology and composition of two noble prosthodontic alloys. METHODS: Cast specimens from Argen 81(Au-Pd) and Argipal (Hi-Pd) alloys which were ground, polished and ultrasonicated in water, were divided in two groups (2 x 3) and treated with single layers of Alloy Primer (AP) and Metal Primer II (MP) primers respectively. The treated alloy surfaces were washed off with acetone and then examined by polarized light microscopy (PLM), reflection FTIR microspectroscopy (FTIRM) and X-ray photoelectron spectroscopy (XPS). RESULTS: After AP treatment, PLM revealed a crystalline phase (VBATDT) dispersed in an amorphous phase (MDP plus soluble VBATDT) on both the alloys tested. MP demonstrated a fibrial arrangement with the most dense structure found on the Hi-Pd alloy. FTIRM failed to clearly resolve the presence of SH peaks on alloy surfaces. Moreover, NH and PS peaks were identified denoting the presence of original thione tautomers. In both primers, phosphates were detected in a dissociative state (-PO(3)(2-)). FTIR molecular mapping confirmed separation of VBATDT from MDP and MEPS from residual MMA. XPS showed that on alloy surfaces approximately 50% of sulphur was in the sulphide state, the rest being organic sulphur. AP showed higher sulphide percentage than MP on both alloys and higher sulphide percentage on the Au-Pd alloy (p<0.05). CLINICAL SIGNIFICANCE: Phase separation of the primer components on alloy surfaces may adversely affect their clinical performance. Sulphide formation on alloy surfaces was confirmed only by XPS under ultra-high vacuum and not by environmental techniques like FTIR; this poses serious questions on the chemical bonding capacity of these primers with the noble alloys tested under environmental conditions.

Crystallization↗

Further characterization of theobroma oil-beeswax admixtures as lipid matrices for improved drug delivery systems.

There is an increasing interest in lipid based drug delivery systems due to factors such as better characterization of lipidic excipients and formulation versatility and the choice of different drug delivery systems. It is important to know the thermal characteristics, crystal habit, texture, and appearance of a new lipid matrix when determining its suitability for use in certain pharmaceutical application. It is line with this that this research was embarked upon to characterize mixtures of beeswax and theobroma oil with a view to applying their admixtures in drug delivery systems such as solid lipid nanoparticles and nanostructured lipid carriers. Admixtures of theobroma oil and beeswax were prepared to contain 25% w/w, 50% w/w, and 75% w/w of theobroma oil. The admixtures were analyzed by differential scanning calorimetry (DSC), small angle X-ray diffraction (SAXD), wide angle X-ray diffraction (WAXD), and isothermal heat conduction microcalorimetry (IMC). The melting behavior and microstructures of the lipid admixtures were monitored by polarized light microscopy (PLM). Transmission electron microscopy (TEM) was used to study the internal structures of the lipid bases. DSC traces indicated that the higher melting peaks were roughly constant for the different admixtures, but lower melting peaks significantly increased (p < 0.05). The admixture containing 25% w/w of theobroma oil possessed highest crystallinity index of 95.6%. WAXD studies indicated different reflections for the different lipid matrices. However, new interferences were detected for all the lipid matrix admixtures between 2theta = 22.0 degrees and 2theta = 25.0 degrees. The lipid matrices containing 50% w/w and 25% w/w of theobroma oil showed absence of the weak reflection characteristic of pure theobroma oil, while there was disappearance of the strong intensity reflection of beeswax in all the lipid matrix admixtures at all stages of the study. PLM micrographs revealed differences with regard to the thermal and optical behaviors depending on the composition of the matrix. The lipid matrix consisting of 75% w/w of theobroma oil showed a spherulite texture after 4 weeks of isothermal storage. Crystallization exotherms of lipid matrices containing 50% w/w and 25% w/w of theobroma oil showed change in modification after 30 min with the latter having a greater time-dependent crystallization. Generally, low non-integral Avrami exponents and growth rate constants were obtained for all the lipid matrices, with the admixture containing 25% w/w theobroma oil having the lowest Avrami exponent and growth rate constant. Based on the results obtained, admixtures containing 50% w/w and 75% w/w of theobroma oil could be applied in the formulation of solid lipid nanoparticles and nanostructured lipid carriers as these lipid matrices possessed crystal characteristics that favour such drug delivery systems.

Cacao↗

Phenytoin can accelerate the healing process after experimental myocardial infarction?

BACKGROUND: Over-degradation and/or inadequate accumulation of extracellular matrix after myocardial infarction (MI) may lead to adverse ventricular remodeling, even ventricular aneurysm or rupture. Phenytoin can increase gingival overgrowth by stimulating the proliferation of connective tissue, which implies a novel way to hasten the healing process after MI. METHODS: Experimental MI was induced by permanent coronary ligation. Surviving rats after MI were randomly divided into phenytoin, captopril, phenytoin plus captopril, operation control and sham operation group. Picrosirius red staining plus polarized microscopy was used for collagen analysis. Left ventricular passive pressure-volume relationship was determined ex vivo. The effects of phenytoin concentration gradient (0, 1.25, 2.5, 5.0, 10.0, and 20.0 microg/mL) on transforming growth factor-beta1 (TGF-beta1) mRNA and protein expression by neonatal rat cardiac fibroblast were determined using semi-quantitative RT-PCR and ELISA, respectively. Peritoneal macrophage was incubated with same gradient of phenytoin concentration. Then the supernatant was harvested to stimulate another 6 groups of cardiac fibroblast, to investigate possible role mediated by macrophage. RESULTS: Phenytoin treatment could promote type I collagen cross-linking level and ratio of type I/III collagen in the infarcted region and had no obvious side effect on interstitial collagen volume fraction, subtype ratio and distribution in non-infarcted region. Phenytoin-treated hearts exhibited attenuation of global ventricular dilation. Phenytoin alone had no direct effects on rat cardiac fibroblast proliferation and collagen production in vitro, but phenytoin-stimulated macrophage could exert a positive influence on cardiac fibroblast TGF-beta1 mRNA and protein production, which exhibited a dose-dependent manner. CONCLUSIONS: Phenytoin can accelerate the healing process in the infarcted region and has no obviously detrimental influence on collagen accumulation in non-infarcted region, which implies a potential benefit to patients undergoing early post-infarction ventricular remodeling process.

Animals↗

Hemozoin formation in Echinostoma trivolvis rediae.

Rediae of the trematode Echinostoma trivolvis, from naturally infected Helisoma trivolvis snails, form a black pigment while inside the snail host. Here we examine the black pigment to show that the insolubility characteristics in detergent and weak base solution are identical to Plasmodium falciparum hemozoin. Laser desorption mass spectrometry of the purified pigment demonstrates the presence of heme. Examination of purified pigment under polarized light microscopy illuminates ordered birefringent crystals. Field emission in lens scanning electron microscopy reveals irregular ovoid crystals of 200-300 nm in diameter. The purified pigment crystals seeded extension of monomeric heme onto the crystal which by Fourier Transform Infrared analysis is beta-hematin. Rediae of a second echinostome parasite, Echinostoma caproni, from experimentally infected Biomphalaria glabrata, do not produce measurable or recoverable heme crystals. These observations are consistent with heme crystal formation by a hematophagous parasite within a non-vertebrate intermediate host.

Animals↗

Isotropic medium chain mono-diglyceride/oil/water formulations for solubilization of lipophilic and hydrophilic drugs.

The aim of the study was to investigate isotropic mono- and diglyceride (MCMDG)/oil/water systems as vehicles for combinations of hydrophilic and lipophilic drugs. For two-component systems, MCMDG was mixed with various masses of water. For MCMDG/oily vehicles/water systems, mixtures were prepared by mixing oil and MCMDG prior to the addition of the appropriate masses of water. The isotropic region was examined by visual inspection and confirmed using polarized light microscopy. Viscosities of the systems were determined. Solubilities of hydrophilic (levamisole HCl) and lipopohilic (abamectin) drugs were determined in the isotropic formulations by HPLC analysis. The isotropic regions in the two-component and three-component systems had water contents of up to 18% at 25 degrees C. The isotropic formulations exhibited Newtonion flow. The viscosity of formulations having the same percentage of water increased with increasing ratio of MCMDG to oil in three-component systems. The solubilities of the levamisole HCl and abamectin were higher in the isotropic MCMDG/sesame oil/water formulations than in equivalent MCMDG/water formulations. In some formulations, the solubility of levamisole HCl was higher in the absence of abamectin than in combination with abamectin. Isotropic MCMDG/oil/water systems were obtained without the use of co-surfactants. Increasing water content in the system did not proportionally increase the solubility of hydrophilic drug. Solubilization of hydrophilic drug was affected by lipophilic drug in the presence or absence of SO and lipophilic drug solubility was affected by hydrophilic drug in the absence of SO. These systems are suitable vehicles to deliver both hydrophilic and lipophilic drugs and could be of interest for pharmaceutical formulations.

Benzoates↗

In vitro drug release mechanism and drug loading studies of cubic phase gels.

Glyceryl monooleate/water cubic phase systems were investigated as drug delivery systems, using salicylic acid as a model drug. The liquid crystalline phases formed by the glyceryl monooleate (GMO)/water systems were characterized by polarizing microscopy. In vitro drug release studies were performed and the influences of initial water content, swelling and drug loading on the drug release properties were evaluated. Water uptake followed second-order swelling kinetics. In vitro release profiles showed Fickian diffusion control and were independent on the initial water content and drug loading, suggesting GMO cubic phase gels suitability for use as drug delivery system.

Drug Carriers↗

Use of surfactants as plasticizers in preparing solid dispersions of poorly soluble API: selection of polymer-surfactant combinations using solubility parameters and testing the processability.

Formation of solid dispersions as a means to enhance the dissolution rate of poorly soluble Active pharmaceutical ingredients (APIs) typically employs hydrophilic polymer systems and surfactants. While the utility of the surfactant systems in solubilization is well known, the secondary effects of the same on processing and subsequent physical stability of the solid dispersions needs to be studied further. Physical blends of the poorly soluble API and hydrophilic polymers such as PVP-K30, Plasdone-S630, HPMC-E5, HPMCAS, and Eudragit L100 with mass ratio 1:1 were prepared. The surfactants tested in this study included Tween-80, Docusate sodium, Myrj-52, Pluronic-F68 and SLS. Thermal analysis of the API-polymer-surfactant blends suggested that the surfactants caused solvation/plasticization, manifesting in reduction of (i) the melting (T(m)) of API (ii) T(g) of the polymers and (iii) the combined T(g) of the solid dispersion formed from quench cooling. Explanation of these effects of surfactants is attempted based on their physical state (at the temperature of interest), HLB values and similarity of their solubility parameter values with respect to drug-polymer systems. Furthermore, extruded matrices containing different API-polymer (PVP-K30, Plasdone-S630, and HPMC-E5) mixtures prepared with and without surfactants, were produced by feeding the powder blend through a hot-melt extruder. The melt viscosity of the polymer blends was assessed by torque rheometry using a Haake Rheomix. The physicochemical properties of the extruded API-polymer-surfactant were characterized by differential scanning calorimetry, X-ray diffraction, Raman spectroscopy, and polarized microscopy. The results demonstrated that the glass transition temperature of the carrier polymers decreased as direct result of the surfactants in the extrudate, due to an increase in the chain mobility of polymers. A decrease in the melt viscosity was seen due to a plasticization of the polymer. The drug release profiles of the extruded solid dispersions containing intra granular surfactants were found to fit the dispersions with extra granularly added surfactants.

Calorimetry, Differential Scanning↗

Thermotropic behavior of asymmetric chain length catanionic surfactants: the influence of the polar head group.

Catanionic surfactants formed by the pairing of two ionic amphiphilic chains of opposite charge are now recognized as an important class of amphiphiles. Many aspects of their phase behavior have yet to be explored. In this work, two homologous series of catanionic surfactants were synthesized, based on the cationic headgroups trimethylammonium and pyridinium. Within each series, the headgroup and chain length of the cationic counterpart remains constant while for the anionic counterpart the headgroup is varied, while its alkyl chain length is also kept constant. Thus, one can directly monitor the influence of headgroup chemistry on the thermal behavior of these compounds. Differential scanning calorimetry (DSC) and polarizing light microscopy show that these compounds bear a rich and often complex thermotropic behavior, with the headgroup chemistry in some instances having a rather dramatic influence on phase behavior. Several liquid crystalline phases appear between the solid crystalline phase and the isotropic liquid phase. A qualitative correlation between the observed thermotropic behavior and the chemical nature of headgroup is presented.

Calorimetry, Differential Scanning↗

Iron dissolution in aqueous AOT solution.

The effects of sodium bis(2-ethylhexyl) sulfosuccinate (AOT) on the electrochemical behavior of iron were studied by potentiodynamic and potentiostatic techniques and open-circuit potential measurements. Experiments were made in both neutral and alkaline AOT solutions (pH 7 and pH 12). It was found that AOT-assisted dissolution is initiated on a passivated iron surface and that the oxidation current leads to the formation of a gel-like film on the electrode surface. This dissolution process was investigated as a function of pH, potential, and electrode rotation rate and the corrosion products were characterized by polarizing microscopy, SEM/EDX, and IR spectroscopy. The gel-like material is a mixed NaAOT-Fe(AOT)3 lamellar mesophase and a structure for this mesophase is proposed.

Journal Article↗

A novel cubic phase of medium chain lipid origin for the delivery of poorly water soluble drugs.

The existence of a novel cubic liquid crystalline phase is described within the pseudo-ternary system comprising lauric acid, monolaurin, and simulated endogenous intestinal fluid (SEIF). This phase behaviour has been characterized using cross-polarizing light microscopy (CPLM), and the structure of the cubic phase identified by small angle X-ray scattering (SAXS). The presence of the cubic phase was found to be temperature sensitive within the 20-37 degrees C range making it putative material for in situ gelation purposes. The cubic phase was shown to have a high capacity to solubilise a model poorly water-soluble drug, cinnarizine, and initial in vitro release data highlight the potential of this phase to provide sustained release. Absorption of cinnarizine from the cubic phase was studied in an unconscious rat model via duodenal administration and blood sampling via the carotid artery. The rate of absorption was significantly reduced when compared to a simple suspension formulation, a likely combination of retarded erosion of the cubic phase together with hindered drug release from the cubic matrix. The results of this study suggest that this cubic phase may potentially be of benefit in the delivery of poorly water-soluble compounds due to its high loading capacity and potential for sustained release. The ability to manipulate this system using temperature may warrant further interest in delivery applications via other routes of administration.

Animals↗

Synovial fluid analysis.

AsA prompt and accurate diagnosis of a painful, swollen joint is imperative, primarily in the case of a septic joint, as delayed therapy may result in progression of disease or permanent loss of function. Procurement and analysis of synovial fluid (SF) are paramount in helping the clinician to determine a patient's clinical condition and further course of treatment. Measurement of white blood cell (WBC) counts, crystal analysis by polarized microscopy, and microbiologic studies including Gram stain and culture are the SF parameters that are collectively most important in the ultimate determination by a clinician of the presence or absence of an infectious or inflammatory joint. It is important for the clinician to understand and recognize the limitations of various SF parameters to minimize under-treating patients with potentially serious joint pathology.

Arthritis↗

Synthesis and characterization of mesoporous silica films encapsulating titanium dioxide particles; Photodegradation of 2,4-dichlorophenol.

Degussa P-25 TiO2 encapsulated in a SBA-15 mesoporous thin film silica matrix prepared using a block copolymer templating method. The film structure was characterized using small angle X-ray diffraction (XRD), variable angle spectroscopic ellipsometry (VASE), and polarized light microscopy. The photoactivity of the films was assessed using HPLC and UV-vis spectroscopy to study the photodegradation of 2,4-dichlorophenol, an industrial pollutant, in water under 350 nm irradiation. The unoptimized film was found to be approximately half as photoactive as an equivalent dispersion of TiO2.

Chlorophenols↗

Variation in modern human enamel formation times.

Most of what we know about the timing of human enamel formation comes from radiographic studies on children of known age. Here, we present new longitudinal data derived from a histological analysis of tooth enamel. Two samples, one from southern Africa and one from northern Europe, contained all anterior and molar tooth types. Two further samples contained only one tooth type: canines from a medieval Danish sample and third molars from a modern North American sample. Data were collected on 326 molars and 352 anterior teeth. Each tooth was sectioned and prepared for polarized light microscopy. We used daily enamel cross striations to determine cuspal enamel formation time, recorded the periodicity of long-period striae in the lateral enamel, and used this value to calculate enamel formation times for each decile of crown length. We present data that reveal some of the processes whereby differences in enamel formation times arise between our samples. Mean cuspal enamel formation times were similar in southern African and northern European anterior teeth, but differed in certain molar cusps. All the southern African anterior teeth completed enamel formation earlier. The greatest difference in mean chronological age at enamel completion was 5.2 vs. 6.2 years of age in lower canines. However, enamel completion times in the molar teeth showed few differences between the samples, with mean times for the longest forming cusps all falling between 3.0 years and 3.45 years. Our data suggest fewer differences between samples and smaller ranges of variation than in many radiographic studies and present a more realistic picture of worldwide variation in enamel formation times.

Amelogenesis↗

T(2) relaxation time mapping reveals age- and species-related diversity of collagen network architecture in articular cartilage.

OBJECTIVE: The magnetic resonance imaging (MRI) parameter T(2) relaxation time has been shown to be sensitive to the collagen network architecture of articular cartilage. The aim of the study was to investigate the agreement of T(2) relaxation time mapping and polarized light microscopy (PLM) for the determination of histological properties (i.e., zone and fibril organization) of articular cartilage. METHODS: T(2) relaxation time was determined at 9.4 T field strength in healthy adult human, juvenile bovine and juvenile porcine patellar cartilage, and related to collagen anisotropy and fibril angle as measured by quantitative PLM. RESULTS: Both T(2) and PLM revealed a mutually consistent but varying number of collagen-associated laminae (3, 3-5 or 3-7 laminae in human, porcine and bovine cartilage, respectively). Up to 44% of the depth-wise variation in T(2) was accounted for by the changing anisotropy of collagen fibrils, confirming that T(2) contrast of articular cartilage is strongly affected by the collagen fibril anisotropy. A good correspondence was observed between the thickness of T(2)-laminae and collagenous zones as determined from PLM anisotropy measurements (r=0.91, r=0.95 and r=0.91 for human, bovine and porcine specimens, respectively). CONCLUSIONS: According to the present results, T(2) mapping is capable of detecting histological differences in cartilage collagen architecture among species, likely to be strongly related to the differences in maturation of the tissue. This diversity in the MRI appearance of healthy articular cartilage should also be recognized when using juvenile animal tissue as a model for mature human cartilage in experimental studies.

Adult↗