A simple and robust high-performance liquid chromatography coupled to a diode-array detector method for the analysis of genistein in mouse tissues

A simple liquid-liquid extraction procedure and quantification by high-performance liquid chromatography (HPLC) method coupled to a diode-array detector (DAD) of genistein (GEN) was developed in various mouse biological matrices. 7-ethoxycoumarin was used as internal standard (IS) and peaks were optimally separated using a Kinetex C18 column (2.6 µm, 150 mm X 2.10 mm I.D.) at 40 ºC with an isocratic elution of mobile phase with sodium dihydrogen phosphate 0.01M in water at pH 2.5 and methanol (55:45, v/v), at a flow rate of 0.25 mL/min. The injection volume was 10 µL. In all cases, the range of GEN recovery was higher than 61%. The low limit of quantification (LLOQ) was 25 ng/mL. The linearity of the calibration curves was satisfactory in all cases as shown by correlation coefficients >0.996. The within-day and between-day precisions were <15% and the accuracy ranged in all cases between 90.14 and 106.05%. This method was successfully applied to quantify GEN in liver, spleen, kidney and plasma after intravenous administration of a single dose (30 mg/Kg) in female BALB/C mice

Cytotoxicity of nanoscaled metal–organic frameworks

A series of fourteen porous Metal–Organic Frameworks (MOFs) with different compositions (Fe, Zn, and Zr; carboxylates or imidazolates) and structures have been successfully synthesised at the nanoscale and fully characterised by XRPD, FTIR, TGA, N2 porosimetry, TEM, DLS and z-potential. Their toxicological assessment was performed using two different cell lines: human epithelial cells from foetal cervical carcinoma (HeLa) and murine macrophage cell line (J774). It appears that MOF nanoparticles (NPs) exhibit low cytotoxicity, comparable to those of other commercialised nanoparticulate systems, the less toxic being the Fe carboxylate and the more toxic being the zinc imidazolate NPs. The cytotoxicity values, higher in J774 cells than in HeLa cells, are mainly function of their composition and cell internalisation capacity. Finally, cell uptake of one of the most relevant Fe-MOF-NPs for drug vectorisation has been investigated by confocal microscopy studies, and indicates a faster kinetics of cell penetration within J774 compared to HeLa cells

Chitosan-coated mesoporous MIL-100(Fe) nanoparticles as improved bio-compatible oral nanocarriers

Nanometric biocompatible Metal-Organic Frameworks (nanoMOFs) are promising candidates for drug delivery. Up to now, most studies have targeted the intravenous route, related to pain and severe complications; whereas nanoMOFs for oral administration, a commonly used non-invasive and simpler route, remains however unexplored. We propose here the biofriendly preparation of a suitable oral nanocarrier based on the benchmarked biocompatible mesoporous iron(III) trimesate nanoparticles coated with the bioadhesive polysaccharide chitosan (CS). This method does not hamper the textural/ structural properties and the sorption/release abilities of the nanoMOFs upon surface engineering. The interaction between the CS and the nanoparticles has been characterized through a combination of high resolution soft X-ray absorption and computing simulation, while the positive impact of the coating on the colloidal and chemical stability under oral simulated conditions is here demonstrated. Finally, the intestinal barrier bypass capability and biocompatibility of CS-coated nanoMOF have been assessed in vitro, leading to an increased intestinal permeability with respect to the noncoated material, maintaining an optimal biocompatibility. In conclusion, the preservation of the interesting physicochemical features of the CS-coated nanoMOF and their adapted colloidal stability and progressive biodegradation, together with their improved intestinal barrier bypass, make these nanoparticles a promising oral nanocarrier

Metal-Organic Frameworks for Drug Delivery Applications

In this work, a new type of particles denoted as MOFs or Metal-Organic Frameworks, have been studied as a new drug carriers. First, they were synthesised at the nanoscale (NPs) using, when possible, biofriendly methods. Their cytotoxicity, as well as that from their constitutive linkers, was evaluated by the MTT test in murine macrophage (J774) and in cervix carcinoma (HeLa) cell lines, observing: (i) a low cytotoxicity of MOFs, comparable with other described particulated systems, (ii) a strong influence of the composition (toxicity order: Fe<Zr<Zn; hydrophilic<hydrophobic linkers), (iii) a higher cytotoxicity in J774 than HeLa, due to their higher phagocytosis activity and (iv) MIL-100(Fe)_NPs was an excellent candidate for bioapplications (IC50=0.7 mg¿mL-1). MIL-100(Fe)_NPs are rapidly cell-uptaken, being immediately internalised in J774 cells. Next, the non-toxic bioflavonoid genistein (GEN) with antitumoral properties was successfully encapsulated in porous Fe or Zr carboxylate MOFs, achieving GEN payloads (160-340 g GEN¿mg formulation-1), higher than other existing formulations and dependent on their composition and topology. Furthermore, GEN was progressively released under simulated physiological conditions from 2 days to several weeks, being appropriate as long release drug delivery systems (DDS). Finally, the pharmacokinetics and bioavailability evaluation of MIL-100(Fe)_NPs¿ formulation was carried out in comparison with the free drug after its oral unique administration (30 mg¿Kg-1) to mice, depicting (i) higher and longer plasmatic levels, (ii) an increase in the relative and (iii) a better oral bioavailability and a mean residence time. Furthermore, higher and more prolonged drug levels were detected in organs, suggesting that the MOF worked as a drug shelter, protecting GEN from metabolisation. Finally, a new biologically active MOF (denoted BioMIL-5), based on the antibacterial and dermatologically active azelaic acid (AzA) and Zn, was successfully synthesised by a totally biofriendly route. Both active components BioMIL-5 were slowly released to the media, either pure water or bacteria medium, upon its structural degradation. Its antibacterial activity was evidenced in S. aureus and in S. epidermidis, observing an interesting additive effect with however high minimal inhibitory (MIC) and minimal bactericidal concentrations (MBC), in agreement with the isolated components. Finally, BioMIL-5 exhibited a lengthened bactericidal (4.3 mg¿mL-1) and bacteriostatic (0.9 and 1.7 mg¿mL-1) in contact with S. epidermidis for one week

Metal-Organic Frameworks for Drug Delivery Applications

In this work, a new type of particles denoted as MOFs or Metal-Organic Frameworks, have been studied as a new drug carriers. First, they were synthesised at the nanoscale (NPs) using, when possible, biofriendly methods. Their cytotoxicity, as well as that from their constitutive linkers, was evaluated by the MTT test in murine macrophage (J774) and in cervix carcinoma (HeLa) cell lines, observing: (i) a low cytotoxicity of MOFs, comparable with other described particulated systems, (ii) a strong influence of the composition (toxicity order: Fe<Zr<Zn; hydrophilic<hydrophobic linkers), (iii) a higher cytotoxicity in J774 than HeLa, due to their higher phagocytosis activity and (iv) MIL-100(Fe)_NPs was an excellent candidate for bioapplications (IC50=0.7 mg¿mL-1). MIL-100(Fe)_NPs are rapidly cell-uptaken, being immediately internalised in J774 cells. Next, the non-toxic bioflavonoid genistein (GEN) with antitumoral properties was successfully encapsulated in porous Fe or Zr carboxylate MOFs, achieving GEN payloads (160-340 g GEN¿mg formulation-1), higher than other existing formulations and dependent on their composition and topology. Furthermore, GEN was progressively released under simulated physiological conditions from 2 days to several weeks, being appropriate as long release drug delivery systems (DDS). Finally, the pharmacokinetics and bioavailability evaluation of MIL-100(Fe)_NPs¿ formulation was carried out in comparison with the free drug after its oral unique administration (30 mg¿Kg-1) to mice, depicting (i) higher and longer plasmatic levels, (ii) an increase in the relative and (iii) a better oral bioavailability and a mean residence time. Furthermore, higher and more prolonged drug levels were detected in organs, suggesting that the MOF worked as a drug shelter, protecting GEN from metabolisation. Finally, a new biologically active MOF (denoted BioMIL-5), based on the antibacterial and dermatologically active azelaic acid (AzA) and Zn, was successfully synthesised by a totally biofriendly route. Both active components BioMIL-5 were slowly released to the media, either pure water or bacteria medium, upon its structural degradation. Its antibacterial activity was evidenced in S. aureus and in S. epidermidis, observing an interesting additive effect with however high minimal inhibitory (MIC) and minimal bactericidal concentrations (MBC), in agreement with the isolated components. Finally, BioMIL-5 exhibited a lengthened bactericidal (4.3 mg¿mL-1) and bacteriostatic (0.9 and 1.7 mg¿mL-1) in contact with S. epidermidis for one week

A simple and robust high-performance liquid chromatography coupled to a diode-array detector method for the analysis of genistein in mouse tissues

A simple liquid-liquid extraction procedure and quantification by high-performance liquid chromatography (HPLC) method coupled to a diode-array detector (DAD) of genistein (GEN) was developed in various mouse biological matrices. 7-ethoxycoumarin was used as internal standard (IS) and peaks were optimally separated using a Kinetex C18 column (2.6 µm, 150 mm X 2.10 mm I.D.) at 40 ºC with an isocratic elution of mobile phase with sodium dihydrogen phosphate 0.01M in water at pH 2.5 and methanol (55:45, v/v), at a flow rate of 0.25 mL/min. The injection volume was 10 µL. In all cases, the range of GEN recovery was higher than 61%. The low limit of quantification (LLOQ) was 25 ng/mL. The linearity of the calibration curves was satisfactory in all cases as shown by correlation coefficients >0.996. The within-day and between-day precisions were <15% and the accuracy ranged in all cases between 90.14 and 106.05%. This method was successfully applied to quantify GEN in liver, spleen, kidney and plasma after intravenous administration of a single dose (30 mg/Kg) in female BALB/C mice

Cytotoxicity of nanoscaled metal–organic frameworks

A series of fourteen porous Metal–Organic Frameworks (MOFs) with different compositions (Fe, Zn, and Zr; carboxylates or imidazolates) and structures have been successfully synthesised at the nanoscale and fully characterised by XRPD, FTIR, TGA, N2 porosimetry, TEM, DLS and z-potential. Their toxicological assessment was performed using two different cell lines: human epithelial cells from foetal cervical carcinoma (HeLa) and murine macrophage cell line (J774). It appears that MOF nanoparticles (NPs) exhibit low cytotoxicity, comparable to those of other commercialised nanoparticulate systems, the less toxic being the Fe carboxylate and the more toxic being the zinc imidazolate NPs. The cytotoxicity values, higher in J774 cells than in HeLa cells, are mainly function of their composition and cell internalisation capacity. Finally, cell uptake of one of the most relevant Fe-MOF-NPs for drug vectorisation has been investigated by confocal microscopy studies, and indicates a faster kinetics of cell penetration within J774 compared to HeLa cells