Formulation and Characterization of Magnetic Polyglutaraldehyde Nanoparticles as Carriers for Poly-l-Lysine-Methotrexate

Abstract

Using the statistically optimised method, submicron magnetic polyglutaraldehyde nanoparticles (Fe-PGNP) with free surface carboxylic groups have been synthesized. A model anticancer agent methotrexate (MTX) has been chemically bonded onto the surface of these particles using poly-l-Lysine (PL) as a spacer. The drug release characteristics of the final delivery device, i.e. [Fe-PGNP]-PL-MTX, has been elucidated at 37°C in a medium containing a proteolytic enzyme. Results demonstrate that using particles containing about 8% w/w of Fe₃O₄, and PL-MTX conjugate constituting 256 μg MTX per mg of PL, almost 50% of the conjugate can be covalently linked onto the carrier surface. Release studies failed to demonstrate the presence of free drug. However it appears that MTX-oligopeptides are released from the carrier as a result of the enzymatic hydrolysis of biodegradable bonds. It is suggested that [Fe-PGNP]-PL-MTX may be useful in the intracellular active targeting of bonded drug(s).     

纳米TiO2表面接枝甲基丙烯酸甲酯的聚合反应

利用表面接枝的方法,在纳米TiO2上接枝偶联剂-γMPS(3-(三甲氧硅基)丙基异丁烯酸)形成"杂化"单体,通过加入引发剂引发甲基丙烯酸甲酯(MMA)在粒子表面发生自由基聚合,形成PMMA/γ-MPS/TiO2纳米复合粒子。通过IR、元素分析、TGA等方法表征了PMMA/-γMPS/TiO2纳米复合粒子的结构,定量研究了TiO2/-γMPS单体接枝率和转化率,PMMA/-γMPS/TiO2纳米复合粒子的接枝率及其相应的影响因素。结果表明:PMMA以化学键的形式连接在纳米TiO2粒子表面并具有较高的接枝率。     

Cancer‐Cell‐Specific Induction of Apoptosis Using Mesoporous Silica Nanoparticles as Drug‐Delivery Vectors

Targeted delivery of the chemotherapeutic agent methotrexate (MTX) to cancer cells using poly(ethyleneimine)‐functionalized mesoporous silica particles as drug‐delivery vectors is reported. Due to its high affinity for folate receptors, the expression of which is elevated in cancer cells, MTX serves as both a targeting ligand and a cytotoxic agent. Enhanced cancer‐cell apoptosis (programmed cell death) relative to free MTX is thus observed at particle concentrations where nonspecific MTX‐induced apoptosis is not observed in the nontargeted healthy cell line, while corresponding amounts of free drug affect both cell lines equally. The particles remain compartmentalized in endo‐/lysosomes during the time of observation (up to 72 h), while the drug is released from the particle only upon cell entry, thereby inducing selective apoptosis in the target cells. As MTX is mainly attached to the particle surface, an additional advantage is that the presented carrier design allows for adsorption (loading) of additional drugs into the pore network for therapies based on a combination of drugs.     

Methotrexate-immobilized poly(ethylene glycol) magnetic nanoparticles for MR imaging and drug delivery

We report the development of a biostable methotrexate-immobilized iron oxide nanoparticle drug carrier that may potentially be used for real-time monitoring of drug delivery through magnetic resonance imaging. Methotrexate (MTX) was immobilized on the nanoparticle surface via a poly(ethylene glycol) self-assembled monolayer (PEG SAM). The cytotoxicity of the nanoparticle-drug conjugate (NP-PEG-MTX) to target cells was studied with 9L glioma cells. Cellular uptake experiments showed that the uptake of NP-PEG-MTX conjugates by glioma cells was considerably higher than that of control nanoparticles. Magnetic resonance imaging in 9L cells cultured with NP-PEG-MTX of various concentrations showed significant contrast enhancement. NP-PEG-MTX demonstrated higher cytotoxicity in 9L cells to free MTX in vitro. Leucovorin, an MTX antidote, was used to rescue the cells that had been exposed to NP-PEG-MTX or free MTX, and the experiment verified the biocompatibility of NP-PEG-MTX conjugates and the MTX on NP-PEG-MTX conjugates to be the true source of the cytotoxicity to the target cells. TEM results showed that NP-PEG-MTX conjugates were internalized into the 9L cellular cytoplasm and retained its crystal structure therein for up to 144 h, as identified by electron diffraction. This prolonged particle retention may allow physicians to image tumor cells exposed to the NP-PEG-MTX conjugate over an extended therapeutic time course.     

Influence of Magnesium Stearate on the Homogeneity of a Prednisone-Granule Ordered Mix

The use of ordered systems has been advocated in the formulation of microdose delivery systems to improve and maintain drug homogeneity during mixing. This study considered the effect of a lubricant such as magnesium stearate on the degree of homogeneity and stability of a preformed prednisone-granule ordered mix. Micronized prednisone was mixed with starch-lactose granules to produce an ordered mix of satisfactory homogeneity. Magnesium stearate in concentrations above and below the theoretical surface saturation of the granule caused negligible change in the degree of homogeneity. Sieve analysis of the mix and subsequent analysis of size fractions for prednisone allowed the prednisone distribution within the mix to be determined. Prednisone was found to be associated with the granules in all the mixes; the magnesium stearate did not compete for the surface adsorption sites and did not dislodge the drug from the granule surface, during mixing and mild demixing conditions. However, a decrease in surface adsorbed prednisone occurred in all mixes (with and without magnesium stearate) under more severe segregating conditions.

Recent research in drug homogeneity studies in microdose tablets has highlighted serious problems in dosage variation¹. Drugs are frequently micronized to improve their release from the solid dosage form. Micronization produces drug particles which are extremely cohesive and interactive. In practice, the adequate mixing of micronized powders with other excipients may be difficult to achieve since this cohesiveness produces aggregation of drug particles and interaction of the drug with the mixer surfaces. In recent years some research effort has been applied to using the interactive nature of drug particles to improve the homogeneity of mixes^2,3. Controlled adsorption of a micronized drug particle onto a carrier particle to produce an “ordered unit” has been shown to minimize segregation within the mix^4,5. Some of the factors affecting the degree of homogeneity of “ordered mixes” have been studied^6,7. However, little research has been conducted on the influence of other excipients on the homogeneity and stability of preformed ordered mixes. A cautionary note on the use of magnesium stearate in ordered mixtures indicated that the lubricant may displace salicylic acid from a sucrose carrier under conditions of segregation⁸. The purpose of this study, therefore, was to evaluate the influence of a third component such as magnesium stearate on the degree of homogeneity and stability of a preformed prednisone-starch lactose granule ordered system during a mixing process.     

Development and Optimization of a Methotrexate Topical Formulation

A topical methotrexate (MTX) formulation that would achieve optimal drug buildup in the epidermis and diminish potential systemic toxicity could be of great utility in the treatment of a variety of hyperproliferative skin disorders. In an attempt to develop an optimized MTX topical formulation containing pharmaceutically acceptable excipients, a 2³ factorial design was used to investigate the effects of a fatty alcohol, propylene glycol, and ethanol on the in vitro skin permeation and uptake of MTX. In vitro skin permeation studies were performed using excised human epidermis mounted in flow-through diffusion cells. The results of steady-state flux and skin uptake of MTX from these formulations ranged from 0.035 to 0.315 μg/cm²/hr and 1.146 to 7.929 μg/cm², respectively. Response surface analysis was used to determine the optimum formulation in terms of skin permeation and uptake of MTX.

An optimized cream formulation was developed and compared to a gel formulation containing 3% Azone in hairless mice to evaluate the uptake of MTX in the treated and untreated skin sites as well as the distribution of MTX in the blood and liver following topical application. The results of the in vivo study demonstrated the localization of MTX at the treated site for both formulations without significant uptake of MTX in the distant untreated epidermis and dermis. The levels of MTX in the blood and liver following topical application of the optimized cream were significantly less than those of the gel formulation with 3% Azone.     

Optical nonlinearities of Fe2O3 nanoparticles investigated by Z-scan technique

The single beam Z-scan technique has been used to measure the third-order nonlinear refractive index, γ, the two-photon absorption coefficient, ß, and the variation of refractive index per unit photoexcited carrier density σ_r, and the free carrier absorption cross section σ_ab for quantum-confined Fe₂O₃ nanoparticles coated by a layer of organic molecular in toluene and bare particles in hydrosol. For coated Fe₂O₃ nanoparticles, the ratio γ/ß is enhanced by a factor of five compared to the bare sample. Their figures of merit related to linear and two-photon absorption for ultrafast all-optical devices application were determined.     

Thickness dependence of structural and ferroelectric properties of sol-gel Pb(Zr0.56Ti0.44)0.90(Mg1/3Nb2/3)0.10O3 films

Sol-gel Pb(Zr_0.56Ti_0.44)_0.90(Mg_1/3Nb_2/3)_0.10O₃ (PZT-PMN) films were prepared onto the Ti/Pt/Ti bottom electrode by multilayer spin coating. The film thickness ranged from 0.22 to 0.88 μm. The Pt top electrodes were deposited on the PZT-PMN films by DC sputtering. The structural and ferroelectric properties of PZT-PMN films were investigated as a function of film thickness by X-ray diffraction (XRD), scanning electron microscopy (SEM), cross-sectional transmission electron microscopy (XTEM), and by measuring the relative permittivity. The film retains the tetragonal perovskite structure with the [111] and [100] preferred orientations perpendicular to the film surface independent of film thickness. The [100] texture increases with increasing film thickness although the [111] texture is always predominant. The film consists of columnar grains. The average grain size is nearly independent of film thickness. The surface layer containing fine grains about 30 nm in diameter is induced on the top of the film. As the film thickness exceeds 0.44 μm, the number of the fine grains decreases remarkably. The crystalline interface layer about 10 nm thick is formed between the film and the bottom electrode. This interface layer is composed of Pt, Pb, Zr, Ti and O, while it is rich in Ti and deficient in Pb and O as compared with the inside of the film. The measured relative permittivity of the film increases with increasing film thickness, following the low permittivity interface model. On the basis of this model, the relative permittivity is estimated to be 3200 for the intrinsic PZT-PMN film, 750 for the surface layer and 50 for the interface layer.     

Crystal engineering of lactose using electrospray technology: carrier for pulmonary drug delivery

Context: Dry powder inhalers (DPIs) consisting of a powder mixture containing coarse carrier particles (generally lactose) and micronized drug particles are used for lung drug delivery. The effective drug delivery to the lungs depends on size and shape of carrier particles. Thus, various methods have been proposed for engineering lactose particles to enhance drug delivery to lungs.

Objective: The objective of current work was to assess suitability of electrospray technology toward crystal engineering of lactose. Further, utility of the prepared lactose particles as a carrier in DPI was evaluated.

Materials and methods: Saturated lactose solutions were electrosprayed to obtain electrosprayed lactose (EL) particles. The polymorphic form of EL was determined using Fourier transform infrared spectroscopy, powder X-ray diffractometry, and differential scanning calorimetry. In addition, morphological, surface textural, and flow properties of EL were determined using scanning electron microscopy and Carr’s index, respectively. The aerosolization properties of EL were determined using twin-stage impinger and compared with commercial lactose particles [Respitose^® (SV003, Goch, Germany)] used in DPI formulations.

Results and discussion: EL was found to contain both isomers (α and β) of lactose having flow properties comparable to Respitose^® (SV003). In addition, the aerosolization properties of EL were found to be significantly improved when compared to Respitose^® (SV003) which could be attributed to morphological (high elongation ratio) and surface characteristic (smooth surface) alterations induced by electrospray technology.

Conclusion: Electrospray technology can serve as an alternative technique for continuous manufacturing of engineered lactose particles which can be used as a carrier in DPI formulations.     

Nucleation of Ge quantum dots on the C-alloyed Si(001) surface

Carbon pre-deposition onto the bare Si(001) surface has been shown to alter the (2×1) surface structure by formation of c(4×4) reconstructed domains containing a high C-concentration. Here we studied by ultra-high vacuum scanning tunneling microscopy the effect of this restructured surface on the initial stages of Ge nucleation by molecular beam epitaxy. Ge is found to form three-dimensional (3D) islands already at sub-monolayer coverage, resulting in a Volmer–Weber growth mode. Strain effects repel Ge adatoms from the C-rich domains, leading to enhanced Ge island formation on the C-free surface regions in between the c(4×4) areas. At a low growth temperature of 350°C, very small three-dimensional islands (3–5 nm in diameter, height 3–4 ML) with a density of nearly 1×10¹² cm⁻² are obtained for only 0.5 ML of Ge. At higher substrate temperatures of approximately 500°C this three-dimensional growth mode is less pronounced, but still evident. The initially nucleated three-dimensional islands define the positions of the larger quantum dots at higher Ge coverage, that exhibit enhanced photoluminescence (PL) properties.     

Structural aspects of self-assembly of thienoviologen molecular conductors on gold substrates

The adsorption of various monomethylated π-extended viologen compounds onto self-assembled layers of octadecylmercaptan (ODM) on gold was investigated with secondary ion mass spectroscopy (SIMS) and atomic force microscopy (AFM). SIMS results indicated that the monomethylated viologen 5-(4-pyridyl)-5′-[4-(N-methyl)pyridinio]-2,2′-bithiophene iodide (PT2) adsorbed much more strongly onto the ODM self-assembled layers at 50 μM adsorption concentration in ethanol, while very similar compounds were much less effective in their adsorption even at much higher adsorption concentrations. The formation of multilayers of PT2 could be observed by AFM. Some structural features of the viologens, (dipole moment and polarizability), were calculated with semi-empirical quantum chemical calculations (MOPAC/AM1). One compound, PF2, having a slightly larger molecular length, dipole moment and polarizability than PT2, did not assemble noticeably onto the surface due to its larger solubility. The large difference in efficiency of adsorption can be related to a combination of length and absence of hydrophilic groups at the free pyridine side of the monomethylated viologens.     

Solid-Phase Syntheses of Two Deacetyl-Thymosin β4 Analogues with Substitution at Position 12 and Their Effects on Impaired Blastogenic Response of T Lymphocytes of Uremic Patients

Two deacetyl-thymosin β₄ analogues containing Phe(4Br) or D-Phe(4Br) as position 12 were synthesized by the manual solid-phase method, and their immunological effects on the impaired blastogenic response of phytohemagglutinin-stimulated T lymphocytes of uremic patients with infectious diseases were studied. Bromination of the p-position of Phe¹² resulted in a marked restorative effect on the impaired blastogenic response of T lympocytes compared with that of our synthetic deacetyl-thymosin β₄ The synthetic [Phe(4Br)¹²]deacetyl-thymsin β₄ was approximately equal in potency to our synthetic [Phe(4F)¹²]deacetyl-thymosin β₄ in uremic patients, but the other analogue, [D-Phe(4Br)¹²]deacetyl-thymosin β₄, had no effect.     

AN INVESTIGATION ON THE OPTICAL PROPERTIES OF CARBON BLACK, FULLERITE, AND OTHER CARBONACEOUS MATERIALS IN RELATION TO THE SPECTRUM OF INTERSTELLAR EXTINCTION OF LIGHT

In this work, the optical properties in the UV-VIS of three carbon blacks having specific surface area from 145 to 7 m²/g and produced with furnace or thermal processes have been studied. The results have been compared to the optical properties (in the same spectral range) of C₆₀ fullerene photopolymer, to fullerite i.e., the carbon soot containing fullerenes and to a sample of carbonaceous matter containing carbyne. The scope of the work was to verify if any of the carbonaceous matter studied was able to match the interstellar extinction spectrum which shows a “bump” at 217.5 nm and which was originally attributed to interstellar graphitic particles. None of the materials studied has shown a peak at 217.5 nm. All the carbonaceous materials studied show maxima of absorption from 252 to 267 nm with the exclusion of C₆₀ photopolymer which has a completely peculiar and different spectrum from all other materials with three maxima at 271, 389 and 510 nm. All the carbon materials studied do not match the 217 nm peak, hence, cannot be considered the carrier of the interstellar “bump” but may be present in some circumstellar shells of late type stars, where the peaks at 240-250 nm have been recorded and attributed to a more ordered and partially graphitized carbonaceous matter. In the discussion, the matrix effect, the particle size, and the clumping of the material studied have been considered as cause of the shift of the peak to longer wavelength.

The carbon black samples have also been extracted with pentane or ethanol and the polycyclic aromatic hydrocarbons (PAH) identified have been discussed in the frame of recent works and in relation to the role that they play in the interstellar medium as potential carriers of the diffuse interstellar bands (DIBS).     

Temperature-dependent behavior of Langmuir monolayers of an amphiphilic spiropyran

The effect of subphase temperature on behaviors and morphologies of an amphiphilic spiropyran, 1′,3′-dihydro-3′,3′-dimethyl-6-nitro-1′-octadecyl-8-docosanoyloxymethylspiro[2H-1-benzopyran-2,2′[2H]indol] onto pure water was investigated by measurements of surface pressure–area isotherms and Brewster angle microscopy, respectively. Both depend strongly on the subphase temperature. The isotherms at 30 °C show a distinct phase transition at 7.5 mN m⁻¹, while no such phase transition is observed at 7 °C. The morphology changes drastically accompanying the phase transition at 7.5 mN m⁻¹, at which many large circular domains are formed. Furthermore, we have demonstrated that the morphology changes reversibly accompanying appreciable hysteresis in the isotherms upon continuous compression and expansion cycles.     

Development of drug adsorbates onto soluble inorganic silicate glass surface: example with acetaminophen

A ternary melt-derived inorganic glass system (Igl) of composition corresponding to 62SiO₂, 35Na₂O, 3Al₂O₃ (wt.%) has been formulated and studied as a drug carrier. The [Al₂O₃/Na₂O] ratio is less than one and the aluminium ion is a network former that retards the glass dissolution. The processing conditions lead to a brittle, easily grinding, amorphous product. The Igl structure was proven by IR-spectroscopy, energy-dispersive spectrometry, X-ray diffraction, scanning electron microscopy. A very important fact established is that the Igl corrosion (dissolution) is pH-dependent. Inorganic glass system was transformed into model acetaminophen (APH) adsorbate (APH/Igla 1:1(w/w)) with mild experimental conditions and evaluated as a drug carrier. No interactions between Igl and APH during the processing were proven. Besides, APH settles onto the glass surface as crystalline phase. A lower extent of corrosion, apparent solubility and delayed in vitro APH release from the adsorbate in water and artificial gastric juice in comparison to the samples untreated drug and APH/Iglm physical mixture were established. It is hypothesized that the glass decomposition products, formed into contact with a solvent, initiate interactions with APH at the glass/solution interface. Similar behaviour of the Igl and its drug adsorbates could be expected in gastro-intestinal tract.     

Effect of nerodilol and carvone on in vitro permeation of nicorandil across rat epidermal membrane

The objective of the study was to investigate the effect of nerodilol and carvone on the in vitro transdermal delivery of nicorandil so as to fabricate a membrane-moderated transdermal therapeutic system. The in vitro permeation studies were carried across the rat epidermal membrane from the hydroxypropyl methylcellulose (HPMC) gels (prepared with 70:30 v/v ethanol-water) containing selected concentrations of a terpene such as nerodilol (0% w/w, 4% w/w, 8% w/w, 10% w/w, or 12% w/w) or carvone (0% w/w, 4% w/w, 8% w/w, 12% w/w, or 16% w/w). The amount of nicorandil permeated (Q₂₄) from HPMC gel drug reservoir without a terpene was 3424.6 ± 51.4 μg/cm², and the corresponding flux of the drug was 145.5 ± 2.2 μg/cm²· h. The flux of nicorandil increased with an increase in terpene concentration in HPMC gel. It was increased ranging from 254.9 ± 3.1 to 375.7 ± 3.2 μg/cm²·h or 207.6 ± 4.7 to 356.7 ± 15.3 μg/cm²· h from HPMC gels containing nerodilol (4% w/w to 12% w/w) or carvone (4% w/w to 16% w/w), respectively. Nerodilol increased the flux of nicorandil by about 2.62-folds whereas carvone increased the flux of the drug by about 2.49-folds across the rat epidermal membrane. The results of the Fourier Transform Infrared (FT-IR) study indicated that the enhanced in vitro transdermal delivery of nicorandil might be due to the partial extraction of stratum corneum lipids by nerodilol or carvone. It was concluded that the terpenes, nerodilol and carvone, produced a marked penetration enhancing effect on the transdermal delivery of nicorandil that could be used in the fabrication of membrane-moderated transdermal therapeutic systems.     

乳糖酰化壳聚糖基自组装纳米载药体系的体外研究

制备乳糖酰化壳聚糖基自组装纳米微粒,用作抗肿瘤药物-甲氨喋呤(MTX)的载体,为肝靶向药物栽体系统的研究提供体外参考数椐.合成N-乳糖酰化-N-胆甾醇琥珀酰基壳聚糖(CLCS),并通过红外(IR)及核磁共振氢谱(1H NMR)对其进行结构确证.采用自组装的方法制备CLCS纳米微粒,并通过透射电镜及动态激光散射法对纳米微...     

Cisplatin-loaded porous Si microparticles capped by electroless deposition of platinum

The loading and release of the anti-cancer drug platinum cis-dichlorodiamine (cisplatin) from mesoporous silicon (pSi) microparticles is studied. The pSi microparticles are modified with 1-dodecene or with 1,12-undecylenic acid by hydrosilylation, and each modified pSi material acts as a reducing agent, forming a deposit of Pt on its surface that nucleates further deposition, capping the mesoporous structure and trapping free (unreduced) cisplatin within. Slow oxidation and hydrolytic dissolution of the Si/SiO(2) matrix in buffer solution or in culture medium leads to the release of drugs from the microparticles. The drug-loaded particles show significantly greater toxicity toward human ovarian cancer cells (in vitro), relative to an equivalent quantity of free cisplatin. This result is consistent with the mechanism of drug release, which generates locally high concentrations of the drug in the vicinity of the degrading particles. Control assays with pSi particles loaded in a similar manner with the therapeutically inactive trans isomer of the platinum drug, and with pSi particles containing no drug, result in low cellular toxicity. A hydrophobic pro-drug, cis,trans,cis-[Pt(NH(3))(2)(O(2)C(CH(2))(8)CH(3))(2)Cl(2)], is loaded into the pSi films from chloroform without concomitant reduction of the pSi carrier.     

碳化硼颗粒级配对硅反应结合碳化硼复合材料结构与性能的影响

本研究探讨了碳化硼原料颗粒尺寸对反应结合碳化硼复合材料相组成、结构与性能的影响。研究结果表明:颗粒级配可以使粉体堆积更加密实, 有效提高压制坯体的体积密度, 最终降低复合材料中游离Si的含量; 加入粗颗粒可减缓B₄C与Si的反应, 减少SiC相的生成; 当原料中粒径为3.5、14、28、45 μm的B₄C粉体按质量比为1.5 : 4 : 1.5 : 3配比时, 所制备的复合材料维氏硬度、抗弯强度、断裂韧性和体积密度分别为(29±5) GPa、(320±32) MPa、(3.9±0.2) MPa·m^1/2和2.51 g/cm³。在制备复合材料过程中减缓B₄C与Si反应速度、减少游离Si的含量和缩小Si区域尺寸是其性能升高的主要原因。     

pH- and thermo-sensitive MTX-loaded magnetic nanocomposites: synthesis,characterization, and in vitro studies on A549 lung cancer cell and MR imaging

In the current study, we proposed a facile method for fabrication of multifunctional pH- and thermo-sensitive magnetic nanocomposites (MNCs) as a theranostic agent for using in targeted drug delivery and magnetic resonance imaging (MRI). To this end, we decorated Fe₃O₄ magnetic nanoparticles (MNPs) with N,N-dimethylaminoethyl methacrylate (DMAEMA) and N-isopropylacrylamide (NIPAAm), best known for their pH- and thermo-sensitive properties, respectively. We also conjugated mesoporous silica nanoparticles (MSNs) to polymer matrix acting as drug container to enhance the drug encapsulation efficacy. Methotroxate (MTX) as a model drug was successfully loaded in MNCs (M-MNCs) via surface adsorption onto MSNs and electrostatic interaction between drug and carrier. The pH- and temperature-triggered release of MTX was concluded through the evaluation of in vitro release at both physiological and simulated tumor tissue conditions. Based on in vitro cytotoxicity assay results, M-MNCs significantly revealed higher antitumor activity compared to free MTX. In vitro MR susceptibility experiment showed that M-MNCs relatively possessed high transverse relaxivity (r₂) of about 0.15?mM^?1·ms^?1 and a linear relationship between the transverse relaxation rate (R₂) and the Fe concentration in the M-MNCs was also demonstrated. Therefore, the designed MNCs can potentially become smart drug carrier, while they also can be promising MRI negative contrast agent.