Curcumin-loaded albumin submicron particles with potential as a cancer therapy: an in vitro study.

Curcumin (CUR), a polyphenolic compound, shows promising biological properties, particularly antioxidant activity. However, its medical applications are limited due to its low water solubility, bioavailability, and pH-instability. CUR-loaded albumin microparticles (CUR-HSA-MPs) of submicron size in the range of 800 to 900 nm and a zeta potential of -15 mV were prepared. The CUR loading efficiency was up to 65%. A maximum release of 37% of the encapsulated CUR was observed within 6 h when the CUR-HSA-MPs were dispersed in 50% ethanol in PBS at pH 7, while in RPMI 1640 medium the release was 7%. This demonstrates a sustainable release. The in vitro cytotoxicity of CUR-HSA-MPs showed promising anticancer potential against human hepatocellular carcinoma (Huh-7) and human breast adenocarcinoma (MCF-7) cell lines, although this effect was less pronounced in human dermal fibroblasts (HDFB) and human cholangiocyte (MMN) cell lines. Confocal microscopy was used to confirm the uptake of CUR-HSA-MPs by cancer cells. Our studies revealed that HSA-MPs are potentially promising vehicles for increasing the solubility and bioavailability of CUR.

Optimization of Image Quality and Organ Absorbed Dose for Pediatric Chest X-Ray Examination: In-House Developed Chest Phantom Study.

Purpose: This study aimed to identify proper exposure techniques to maintain optimal diagnostic image quality with minimum radiation dose for anteroposterior chest X-ray projection in pediatric patients. Methods: Briefly, an in-house developed pediatric chest phantom was constructed. Next, nanodot OSLDs were used for organ absorbed dose measurement and placed in the lung area, and the phantom was exposed to various exposure techniques (ranging from 50 to 70 kVp with 1.6, 2, and 2.5 mAs). After that, the phantom was used to assess image quality parameters, including SNR and CNR. Two radiologists assessed the subjective image quality using a visual grading analysis (VGA) technique. Finally, the figure of merit (FOM) was analyzed. Results: The developed phantom was constructed successfully and could be useful for dose measurement and image quality assessment. The absorbed dose varied from 0.009 to 0.031 mGy for the range of exposure techniques used. SNR and CNR showed a gradually increasing trend, while kVp and mAs values were increased. The highest kVp (70 kVp) produced the highest SNR and CNR, exhibiting a significant difference compared with 50 and 60 kVp (P < 0.05). The overall VGA score was 3.2 ± 0.3, and the low kVp technique demonstrated better image quality compared with the reference image. Conclusion: The optimized exposure technique was identified as 60 kV and 2.5 mAs, indicating the highest FOM score. This work revealed practicable techniques that could be implemented into clinical practice for performing pediatric chest radiography.

Fabrication and Characterization of Human Serum Albumin Particles Loaded with Non-Sericin Extract Obtained from Silk Cocoon as a Carrier System for Hydrophobic Substances.

Non-sericin (NS) extract was produced from the ethanolic extract of Bombyx mori silk cocoons. This extract is composed of both carotenoids and flavonoids. Many of these compounds are composed of substances of poor aqueous solubility. Thus, this study focused on the development of a carrier system created from biocompatible and biodegradable materials to improve the biological activity of NS extracts. Accordingly, NS was incorporated into human serum albumin template particles with MnCO3 (NS-HSA MPs) by loading NS into the preformed HAS-MnCO3 microparticles using the coprecipitation crosslinking dissolution technique (CCD-technique). After crosslinking and template dissolution steps, the NS loaded HSA particles are negatively charged, have a size ranging from 0.8 to 0.9 µm, and are peanut shaped. The degree of encapsulation efficiency ranged from 7% to 57% depending on the initial NS concentration and the steps of adsorption. In addition, NS-HSA MPs were taken up by human lung adenocarcinoma (A549 cell) for 24 h. The promotion of cellular uptake was evaluated by flow cytometry and the results produced 99% fluorescent stained cells. Moreover, the results from CLSM and 3D fluorescence imaging confirmed particle localization in the cells. Interestingly, NS-HSA MPs could not induce inflammation through nitric oxide production from macrophage RAW264.7 cells. This is the first study involving the loading of non-sericin extracts into HSA MPs by CCD technique to enhance the bioavailability and biological effects of NS. Therefore, HSA MPs could be utilized as a carrier system for hydrophobic substances targeting cells with albumin receptors.

Doxorubicin-Loaded Human Serum Albumin Submicron Particles: Preparation, Characterization and In Vitro Cellular Uptake.

Doxorubicin (DOX) is an effective anthracycline antibiotic drug which is commonly used in a broad range cancer therapy. However, due to dose depending side effects and toxicity to non-cancerous tissues, its clinical applications are restricted. To overcome these limitations, human serum albumin (HSA) has been investigated as a biocompatible drug delivery vehicle. In this study, human serum albumin submicron particles (HSA-MPs) were fabricated by using the Co-precipitation-Crosslinking-Dissolution technique (CCD technique) and DOX was loaded into the protein particles by absorption. DOX-HSA-MPs showed uniform peanut-like shape, submicron size and negative zeta-potential (-13 mV). The DOX entrapment efficiency was 25% of the initial amount. The in vitro release in phosphate buffered saline pH 7.4 was less than 1% within 5 h. In contrast, up to 40% of the entrapped DOX was released in presence of a protein digesting enzyme mixture (Pronase®) within the same time. In addition, in vitro cytotoxicity and cellular uptake of DOX-HSA-MPs were evaluated using the lung carcinoma cell line A549. The results demonstrated that DOX-HSA-MPs reduced the cell metabolic activities after 72 h. Interestingly, DOX-HSA-MPs were taken up by A549 cells up to 98% and localized in the cell lysosomal compartment. This study suggests that DOX-HSA-MPs which was fabricated by CCD technique is seen as a promising biopolymer particle as well as a viable alternative for drug delivery application to use for cancer therapy.

Albumin Submicron Particles with Entrapped Riboflavin-Fabrication and Characterization.

Although riboflavin (RF) belongs to the water-soluble vitamins of group B, its solubility is low. Therefore, the application of micro-formulations may help to overcome this limiting factor for the delivery of RF. In this study we immobilized RF in newly developed albumin submicron particles prepared using the Co-precipitation Crosslinking Dissolution technique (CCD-technique) of manganese chloride and sodium carbonate in the presence of human serum albumin (HSA) and RF. The resulting RF containing HSA particles (RF-HSA-MPs) showed a narrow size distribution in the range of 0.9 to 1 µm, uniform peanut-like morphology, and a zeta-potential of -15 mV. In vitro release studies represented biphasic release profiles of RF in a phosphate buffered saline (PBS) pH 7.4 and a cell culture medium (RPMI) 1640 medium over a prolonged period. Hemolysis, platelet activation, and phagocytosis assays revealed a good hemocompatibility of RF-HSA-MPs.

In-vitro haemocompatibility of dextran-protein submicron particles.

Blood compatibility is a key requirement to fulfil for intravenous administration of drug and oxygen carrier system. Recently, we published the fabrication of oxidised-dextran (Odex)-crosslinked protein particles by one-pot formulation. In the current study we investigate the haemocompatibility of these Odex - particles including albumin particles (Odex-APs) and haemoglobin particles (Odex-HbMPs). Odex-APs and Odex-HbMPs have a submicron size ranged 800-1000 nm with peanut-like shape and a negative surface charge. In vitro haemocompatibility assays included haemolysis test, indirect phagocytosis test and platelet activation test in human blood. Odex-APs and Odex-HbMPs did not provoke any undesirable effects on the blood cells. Firstly, the ratio of haemolysis after contacted with Odex-crosslinked protein particles were less than 5% and therefore the particles may be considered non-haemolytic. Secondly, the incubation of leukocyte with Odex-APs/HbMPs did not influence the phagocytosis of leukocyte. We conclude that our particles are not recognized by monocytes or granulocytes. Finally, exposure of Odex-APs/HbMPs to platelets did not cause an activation of platelets. Additionally, Odex-HbMP/AP did not enhance or attenuate agonist-induced platelet activation. We conclude that Odex-crosslinked protein particles exhibit a very good haemocompatibility and represent highly promising carriers for drugs or oxygen.

Antioxidative protection of haemoglobin microparticles (HbMPs) by PolyDopamine.

Clinically applicable haemoglobin-based oxygen carriers (HBOCs) should neither induce immunological nor toxic reactions. Additionally, Hb should be protected against oxidation. In the absence of protective enzymes (superoxide dismutase (SOD) and catalase (CAT)) Hb is oxidized to MetHb and thus losing its function of oxygen delivery. Alternatively, polydopamine (PD), a scavenger of free radicals, could be used for Hb protection against oxidation Therefore, we synthetized HbMPs modified with PD. The content of functional haemoglobin in these PD-HbMPs was twice higher than that in the control HbMPs due to the protective antioxidant effect of PD. In addition, the PD-HbMPs exhibited a high scavenging activity of free radicals including H2O2 and excellent biocompatibility. In contrast to monomeric dopamine, which has been shown to produce toxic effects on neurons due to formation of H2O2, hydroxyl radicals and superoxide during the process of auto-oxidation, PD-HbMPs are not neurotoxic. Consequently, the results presented here suggest a great potential of PD-HbMPs as HBOCs.

Improved oxygen storage capacity of haemoglobin submicron particles by one-pot formulation.

The coprecipitation-cross-linking-dissolution (CCD) technique for protein submicron particle fabrication was improved by omitting one preparation step using the macromolecular cross-linker, periodate-oxidized dextran (Odex, M.W. of 40 and 70 kDa). The coprecipitation and cross-linking of haemoglobin (Hb) were combined in one single step since the cross-linker is incorporated into the inorganic template, MnCO3, together with the protein. After removal of the MnCO3 templates by EDTA, the amount of entrapped Hb was 60 to 70% of the initial amount. This technique provides deformable Hb submicron particles (HbMP) with narrow size distribution between 800 and 1000 nm, uniform morphology and negative zeta-potential. More than 40% of Hb in the particles was able to carry oxygen over a storage period of 90 days. The results suggest that our new protein submicron particle fabrication technique minimizes the fabrication time and is very efficient and cost-effective.