Figure 2
FTIR spectra. Shows the Fourier Transform Infra Red Spectra of (a) BSA nanoparticles (b) ATV and (c) ATV-BSA nanoparticles in order of their arrangement from top to bottom. doi:10.1371/journal.pone.0086317.g002
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Currently, the discovery of effective chemotherapeutic agents poses a major challenge to the field of cancer biology. The present study focuses on enhancing the therapeutic and anti cancer properties of atorvastatin calcium loaded BSA (ATV-BSA) nanoparticles in vitro.
BSA-ATV nanoparticles were prepared using desolvation technique. The process para...
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... as synthesized ATV-BSA nanoparticles were highly stable in both water and cell media with an average hydrodynamic size in the range of 97–125 nm and polydispersity index between 0.29– 0.40. Interestingly the ATV-BSA nanoparticles showed an increased hydrodynamic diameter (361 nm) comparatively higher than bare nanoparticles thus enumerating the entrapment of the drug. The stability of the nanoparticles can be attributed to the significantly higher zeta potential. Negatively charged particles contribute to the high stability of the colloidal solution. Coulombic repulsive forces between the particles prevent them from agglomerating in the colloid state thus maintaining the stability of the system [19]. The zeta potential value of the ATV-BSA nanoparticles was 2 36.3 mV which is high thus contributing for a stable system. The surface morphology of the particles was determined by the Scanning Electron Microscopy (SEM). The images of the BSA nanoparticles and ATV-BSA nanoparticles revealed a spherical morphology of the particles as shown in Fig. 1. Majority of the particles showed uniform size distribution without any crystal precipitation. FTIR analysis helped in understanding the chemical modifications that occurred to the nanoparticles following drug interaction. Fig. 2 shows the FTIR spectra of BSA nanoparticles, ATV-BSA nanoparticles and ATV. The characteristic peaks of atorvastatin were recorded at 3364.21 cm 2 1 and 1649.81 cm 2 1 indicating aromatic N-H stretching and C = O stretching respectively, whereas the characteristic bands present in protein nanoparticles are Amide I at 1650 cm 2 1 , Amide II band at 1530 cm 2 1 and Amide III region at 1230 cm 2 1 . The ATV-BSA nanoparticle spectra were marked by the broadening of the N-H peaks indicating the possible interaction of the aromatic residues (tryptophan, tyrosine) of BSA with the drug. Further in order to confirm the nature of interaction of ATV to BSA and to corroborate the results obtained in FTIR spectroscopy, fluorescence spectroscopy was carried out. It also gives a clear picture on the internal environment around the vicinity of the fluorophore. Upon excitation at 293 nm, the tryptophan residues in BSA displayed a strong decrease in the intensity in the spectra of ATV-BSA nanoparticles in comparison with the bare nanoparticles (Fig. 3a). This decrease in the intensity could be due to fluorescence quenching attributed to the changes in the microenvironment of the tryptophan residues suggesting interac- tion of ATV and BSA. The quenching process also signifies the tertiary structural changes in BSA [20]. The hydrophobic binding pocket in the sub domain IIA would have undergone conformational changes with the possible interaction of BSA and ATV or overexposure of the residue to water [21]. The same quenching effects were noticed in Fig. 3b when the ATV-BSA nanoparticles were excited at 280 nm. This excitation wavelength corresponded to the tryptophan and tyrosine residues of the test sample. Hence the results obtained from FTIR and Fluorescence spectroscopy clearly indicate the major involvement of aromatic residues in the drug carrier interactions. We investigated the in vitro release profile of ATV from the nanoparticles at pre defined time intervals at pH 7.4. A biphasic drug release pattern was seen with the initial burst release for 2 hours followed by a controlled release of the drug. The plateau region seen after few hours corresponded to the sustained release of the drug making it an effective carrier. As shown in Fig. 4, about 85% of the drug was released in 72 hrs at specified intervals. The cross linking of the BSA nano carriers using glutaraldehyde was the major reason for the enhanced stability of the particles in PBS. Our results revealed that slow release of the drug in the media makes it an ideal carrier for drug delivery. Various combinations of the drug with the nanoparticle were taken and synthesis was done using the desolvation method. The supernatant after centrifugation was analysed to determine the amount of free drug. The encapsulation efficiency was found using the formula The drug: BSA combination with the best encapsulation efficiency was taken for further studies. As shown in Table 1, 10:20 ratio of drug: BSA gave the best encapsulation efficiency of 94 6 0.32%. The results of the haemolysis assay can be analysed from Figure 5. Three different concentrations of ATV alone, BSA nanoparticles, ATV-BSA nanoparticles were incubated with the blood samples and haemocompatibility effects were tested. Our results revealed that all the above tested concentrations neither exhibited haemolytic activity nor thrombus formation making it suitable for circulation in the blood. No adverse reactions could be observed between the serum proteins and the surface of the nanocarrier. Distilled water which was used as a positive control exhibited 100% haemolysis marked by complete lysis of the Red Blood Cells (RBC’s) as shown in Fig. 5. Whereas saline which was used as the negative control, bare drug and nanoformulation did not show any haemolysis or toxicity to the RBC’s making it a clinically promising formulation. Our results were consistent with results obtained by Wang et al, 2010 which discusses the haemocompatibility of docetaxel - loaded albumin nanoparticles [22]. Determination of ROS content. The cytotoxicity effects might have occurred through the induction of oxidative stress and apoptosis with possible involvement of overproduction of reactive oxygen species (ROS). In this regard, the ability of ATV-BSA nanoparticles to induce intra cellular oxidant production in MiaPaCa-2 cell lines was assessed by measuring DCF fluorescence as an indicator of ROS generation (Fig. 6). Various concentrations (10, 25, 50, 75, 100 m g/mL) of ATV-BSA and ATV were used for the study. The bare drug exhibited a concentration dependent production of ROS, but the results were not significant when compared to the ATV-BSA nanoparticles. Interestingly, 10, 25, 50 m g/mL of ATV-BSA did not induce significant generation of ROS. Whereas as the concentration of the nanoparticles increased further to 75 and 100 m g/mL a remarkable ROS production is seen. Our results revealed that though ROS production is not the ...
Citations
... Due to its remarkable structural similarity to human serum albumin (HSA), bovine serum albumin (BSA) has been extensively studied as a model protein in a wide range of scientific fields and therefore, determining BSA concentration in the biomedical, pharmaceutical, and food sectors is essential [54]. BSA is used in the growth of cells in culture [55], in vitro anticancer therapy using atorvastatin calciumloaded BSA (ATV-BSA) nanoparticles [56], so it is vital to measure BSA concentration. BSA can be extracted with 100% extraction efficiencies up to high protein concentration by aqueous biphasic systems of phosphonium-and ammonium-based ionic liquids [57]. ...
Numerous studies have used the Drude model to optimize silver-based Metal-Insulator-Metal plasmonic sensors for some limited sensing applications; however, Lab-on-a-chip sensing for cell protein concentration measurement still needs to be accomplished. Furthermore, little study has been conducted on sensing for the salinity measurement of seawater, Bovine serum albumin concentration measurement, human tissue categorization, mitigating the oxidation issue of these sensors, and nanofabrication challenges. In this article, a Metal-Insulator-Metal based oxidation free material (gold modeled with Drude-Lorentz model) built round-edged hexagonal plasmonic refractive index sensor with nanorods embedded both in the straight waveguide and resonator has been investigated using the Finite element method (FEM) for all of the above sensing applications. This sensor achieves 19.05 nm/g/100mL and 1476.6 nm/ppm sensitivity for cell protein concentration and salinity measurement. The sensor’s max sensitivity and Sensing Resolution are 9231.7 nm/RIU and 1.083∗10−7. This sensor has also been employed as an electromagnetic interference-free temperature sensor using toluene, achieving 5.16 nm/°C sensitivity. Thus, this label-free and low-footprint sensor can be employed to study the salinity of seawater, which is linked with global warming and the earth’s hydrologic cycle, study protein concentration for nanomedicine applications, and Lab-on-chip sensing as an oxidation-free alternative to silver-based sensors.
... The peak at 1656 cm −1 is attributed to the carbonyl C=O stretching of the amide I band, the most sensitive probe for detecting changes in the protein secondary structures. The amide II band located at 1542 cm −1 can be assigned to the N-H bending vibrations and the amide III band located at 1248 cm −1 is due to C-N and N-H in-plane stretching vibration [29,[67][68][69][70][71]]. ...
... The peak at 1656 cm −1 is attributed to the carbonyl C=O stretching of the amide I band, the most sensitive probe for detecting changes in the protein secondary structures. The amide II band located at 1542 cm −1 can be assigned to the N-H bending vibrations and the amide III band located at 1248 cm −1 is due to C-N and N-H in-plane stretching vibration [29,[67][68][69][70][71]. Comparing the FTIR spectra of ZnS-BSA with that of pure BSA, the changes in the intensity and positions in the characteristic IR bands can be noticed. ...
In this paper, ZnS nanoparticles were bioconjugated with bovine serum albumin and prepared in a form of nanosuspension using a wet circulation grinding. The stable nanosuspension with monomodal particle size distribution (d50 = 137 nm) and negative zeta potential (−18.3 mV) was obtained. The sorption kinetics and isotherm were determined. Interactions between ZnS and albumin were studied using the fluorescence techniques. The quenching mechanism, describing both static and dynamic interactions, was investigated. Various parameters were calculated, including the quenching rate constant, binding constant, stoichiometry of the binding process, and accessibility of fluorophore to the quencher. It has been found that tryptophan, in comparison to tyrosine, can be closer to the binding site established by analyzing the synchronous fluorescence spectra. The cellular mechanism in multiple myeloma cells treated with nanosuspension was evaluated by fluorescence assays for quantification of apoptosis, assessment of mitochondrial membrane potential and evaluation of cell cycle changes. The preliminary results confirm that the nontoxic nature of ZnS nanoparticles is potentially applicable in drug delivery systems. Additionally, slight changes in the secondary structure of albumin, accompanied by a decrease in α-helix content, were investigated using the FTIR method after analyzing the deconvoluted Amide I band spectra of ZnS nanoparticles conjugated with albumin. Thermogravimetric analysis and long-term stability studies were also performed to obtain a complete picture about the studied system.
... Several studies have used BSA as a nanocarrier for bioactive compounds by the desolvation method because it has high selectivity, great bioavailability, and potent therapeutic properties [20,26]. In this work, ethanol and glutaraldehyde were used as desolvation and crosslinking agents, respectively, in the desolvation method [21]. ...
Hepatocellular carcinoma (HCC) can be produced from aflatoxin B1 (AFB1) administration. Although berberine (BER) acts as an anticancer agent and can counteract the AFB1 effect, it has low bioavailability. Nanotechnology can overcome this problem. This research aimed to synthesize berberine nanoparticles (NPs) and then estimate their therapeutic effect compared to that of berberine against aflatoxin-induced hepatotoxicity. The desolvation method was used to prepare BER-NPs. Aflatoxicosis was induced by 5 consecutive intraperitoneal injections (IP) of 200 µg/kg/day AFB dissolved in dimethylsulfoxide (DMSO). After the induction period, two treatments were performed: the first with 100 mg/kg BER and the second with 10 mg/kg BER-NPs. Liver, kidney, and diabetic profiles were estimated by using standardized methods. Hepatic oxidative stress, inflammatory, cancer cell proliferation, and invasion markers were used by ELISA and qPCR techniques. The TEM image shows that both BSA NPs and BER-BSA NPs had spherical, regular, and uniform shapes. The BER encapsulation efficiency % was 78.5. The formed-BER-BSA NPs showed a loading capacity % of 7.71 and the synthesis yield % of 92.6. AFB1 increases pro-oxidant markers, decreases antioxidant systems, stimulates inflammatory enzymes, inhibits anti-inflammatory markers, decreases tumor suppressor enzymes, increases oncogenes, increases glycolytic activity, prevents cell death, and promotes cell growth. Most of the biochemical markers and hepatic architecture were normalized in the BER-BSA NP-treated group but not in the BER-treated group. Altogether, the obtained data proved that treatment with BER-NPs was more efficient than treatment with berberine against aflatoxicoses induced in rats.
... The pH of the albumin solution (XS3) significantly affected both the PS and the EE%. A smaller vesicular size was observed as the pH increased from 8 to 9: this could be attributed to the extension of the BSA backbone with loose unordered parts, by increasing the pH, allowing charged side chains to be accessible [62], which in turn increases the surface charge and reduces the particle attraction and agglomeration, leading to a reduction in the vesicular size [63]. This finding was in accordance with [37]. ...
Purpose
Albumin nanoparticles are promising carriers for therapeutic agents, owing to their biocompatibility, safety, and versatility in fabrication. The formulation of albumin nanoparticles is highly affected by many product and process variables, resulting in a great variation in these nanoparticles. The aim of this work was to formulate and optimize albumin nanoparticles loaded with silymarin, as a model drug with low bioavailability, for the treatment of hepatocellular carcinoma, using quality by design (QbD) approach.
Methods
A thorough risk assessment for albumin nanoparticles formulation was developed and a complete quality product profile was defined using the QbD approach. A D-optimal design was used to optimize the amount of albumin and drug, which significantly affected the particle size (PS) and the entrapment efficiency (EE%), which was further tested on hepatocellular carcinoma.
Results
A design space was constructed, with an optimized formula showing a PS of 135 nm, a polydispersity index (PDI) of 0.09, an EE% of 88%, and a zeta potential of − 12.5 mV. The optimized formula (O 1 ) with spherical particles, showed an extended-release rate as compared to free silymarin. Moreover, a pronounced anti-proliferation activity of O 1 was observed on human hepatocellular carcinoma cell line HepG2 than the free drug. The flow cytometric analysis of the cell cycle showed a significant suppression of the S-phase after treating the HepG2 cell with O 1 , but not with free silymarin.
Conclusion
Thus, a detailed QbD study has been conducted, with deep product and process understanding, and resulted in a successful formulation of silymarin albumin nanoparticles for the suppression of hepatocellular carcinoma.
... 21 The use of methanolethanol as desolvation solution and dropwise addition of this solution greatly affect the size and solubility of BSA nanoparticles. 70 Aliquot of 12 mL of methanolethanol solution recorded to be sufficient volume for the successful preparation of a small BSA NPs. The methanol-ethanol addition at a speed range of 1 mL min À1 produced nanoparticles with a size around 100 nm. ...
The study planned to estimate biological parameters linked to rheumatoid arthritis (RA) patients, detecting the influence of MTX and biotherapy treatments on these parameters and synthesizing methotrexate bovine serum albumin nanoparticles linked to folate (FA-MTX-BSA NPs) to reduce the overwhelming expression of inflammatory cytokines. Inflammatory parameters showed significant increases in newly diagnosed and MTX-receiving groups while no changes were observed in the biotherapy-maintained group. MTX-loaded BSA nanoparticles were fabricated by the desolvation method and further linked to activated folic acid to obtain FA-MTX-BSA NPs. FA-MTX-BSA NPs were successfully characterized within the nanoscale range using different screening techniques. FA-MTX-BSA NPs showed an in vitro release in a sustained manner. The potential of MTX, MTX-BSA NPs, and FA-MTX-BSA NPs in inducing cytokine level reduction was detected. Significant decreases in interleukin-1 beta (IL-1b), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-a) levels were obtained in cultures treated with FA-MTX-BSA NPs compared to the untreated culture in a dose-dependent pattern. Furthermore, FA-MTX-BSA NPs comparing with MTX and MTX-BSA NPs exhibited a significant advanced effect in decreasing cytokines levels. Accordingly, the conjunction of BSA NPs and MTX linked to folate potentially reduced cytokines manifestation in RA.
... Bovine serum albumin, a nano-carrier, can encapsulate several bioactive components and deliver different drugs and natural products with high selectivity, high bioavailability, and potent therapeutic properties 23,26,27 . In the current study, BBR-BSA NPs were synthesized by the desolvation method, using ethanol as a desolvating agent and glutaraldehyde as the crosslinking agent. ...
Berberine (BBR) is an isoquinoline alkaloid with several clinical therapeutic applications. Its low water solubility, absorption, and cellular bioavailability diminish BBR's therapeutic efficacy. In this study, BBR was encapsulated into bovine serum albumin nanoparticles (BSA NPs) core to reduce BBR limitations and enhance its clinical therapeutic properties. Several physicochemical characterization tools, such as Dynamic Light Scattering and Ultraviolet–Visible spectroscopic measurements, field emission transmission electron microscopy surface morphology, Fourier transforms infrared spectroscopy, thermal stability analysis, and releasing studies, were used to evaluate the BBR-BSA NPs. Compared to BBR, BBR-BSA nanoparticles demonstrated superior free radical scavenging and antioxidant capacities, anti-hemolytic and anticoagulant efficacies, and antimicrobial activities, as demonstrated by the findings of the in vitro studies. Furthermore, a stressed pancreatic rat model was induced using a high-fat, high-sucrose diet plus carbon tetrachloride injection. The in vivo results revealed that BBR-BSA NPs substantially restored peripheral glucose metabolism and insulin sensitivity. Oral administration of BBR-BSA NPs also improved pancreatic β-cells homeostasis, upregulated pancreatic antioxidant mechanisms, inhibited oxidants generation, and attenuated oxidative injury in the stressed pancreatic tissues. In conclusion, our in vitro and in vivo results confirmed that BBR-BSA NPs demonstrated more potent antioxidant properties and restored pancreatic homeostasis compared to BBR.
... Hypercholesterolemia, comorbidity of obesity, has been identified as an independent risk factor for breast cancer [6,7]. Statins, the 3-hydroxy-3methylglutaryl (HMG) -CoA reductase inhibitors, beyond their cardiovascular effects, they have been reported to have possible benefits as anticancer, and inhibition of cancer progression [8,9]. Besides, a potential role for statins as a radiosensitizer for aggressive breast cancer has been suggested [10]. ...
... Apart from their cardiovascular effects, statins have demonstrated significant although heterogeneous, anti-tumor activities in preventing breast cancer progression [9,42]. Our data showed that SIM increased significantly oxidative stress markers, LDH release, caspase 3 activity, Bax/ BCL2 ratio, and increased glucose uptake. ...
... Similarly, Sanchez et al. [46] reported that statin-induced growth inhibition of breast cancer due to apoptotic and necrotic cell death by an increase in the formation of superoxide and oxidative stress in cytotoxic activity [47]. SIM reduces not only serum cholesterol levels but also mevalonate synthesis, a precursor of several major products, regulating the cell cycle, and signal transduction involved in cell proliferation, differentiation, and apoptosis [9,42]. Moreover, a significant increase in LDH release in C2C12 cells exposed to SIM was reported [48]. ...
Tamoxifen (TAM) is a nonsteroidal antiestrogen drug, used in the prevention and treatment of all stages of hormone-responsive breast cancer. Simvastatin (SIM), a lipid-lowering agent, has been shown to inhibit cancer cell growth. The study aimed at investigating the impact of using SIM with TAM in estrogen receptor-positive (ER+) breast cancer cell line, T47D, as well as in mice-bearing Ehrlich solid tumor. The cell line was treated with different concentrations of TAM or/and SIM for 72 h. The effects of treatment on cytotoxicity, oxidative stress markers, apoptosis, angiogenesis, and metastasis were investigated. Our results showed that the combination treatment decreased the oxidative stress markers, glucose uptake, VEGF, and MMP 2 &9 in the cell line compared to TAM-treated cells. Drug interaction of TAM and SIM was synergistic in T47D by increasing the apoptotic makers Bax/BCL-2 ratio and caspase 3 activity. Additionally, in vivo, the combination regimen resulted in a non-significant decrease in the tumor volume compared to TAM treated group. Moreover, the combined treatment decreased the protein expression of TNF-α, NF-kB compared to control. In conclusion, our results suggest that SIM may serve as a promising treatment with TAM for improving the efficacy against estrogen receptor-positive (ER+) breast cancer.
... This result confirmed that BG@BSA NPs are highly stable. The electrostatic repulsive force of the negatively charged surface of NPs attributed to the higher stability of the colloidal solution [24]. The particle size analysis of BG@BSA NPs is shown in Fig. 6b. ...
A new method for targeting lung infections is of great interest using biodegradable nanoparticles. In this study, bergenin-loaded BSA NPs were developed against lung injury. Briefly, bergenin-loaded bovine serum albumin nanoparticles (BG@BSA NPs) were synthesized and characterized. HPLC recorded the major peak of bergenin. UV–Vis spectra had an absorbance at 376 nm. XRD revealed the presence of crystalline particles. FTIR confirmed the occurrence of functionalized molecules in the synthesized NPs. The particles were highly stable with a net negative charge of − 24.2. The morphology of NPs was determined by SEM and TEM. The mean particle size was 124.26 nm. The production of NO by NR8383 cells was decreased by BG@BSA NPs. Also, in mice, lipopolysaccharide-mediated acute lung inflammation was induced. BG@BSA NPs reduced macrophages and neutrophils in BALF and remarkably enhanced wet weight-to-dry weight (W/D) ratios and myeloperoxidase (MPO) activity. Further, BG@BSA NPs inhibited the production of inflammatory cells as well as tumor necrosis factor. The histopathological studies revealed that the damage and neutrophil infiltration were greatly inhibited by BG@BSA NPs. This indicates that BG@BSA NPs may be used to treat lung infections. Therefore, this study has given new insight into producing an active drug for the treatment of lung-associated diseases in the future.
... This result con rmed that BG@BSA NPs are highly stable. The electrostatic repulsive force of the negatively charged surface of NPs attributed to the higher stability of the colloidal solution [24]. The particle size analysis of BG@BSA NPs was shown in Fig. 6b. ...
A new method for targeting lung infections is of great interest using biodegradable nanoparticles. In this study, bergenin loaded BSA NPs were developed against lung injury. Briefly, BG@BSA NPS were synthesized and characterized. HPLC recorded the major peak of bergenin. UV-vis spectra had an absorbance at 376nm. XRD revealed the presence of crystalline particles. FTIR confirmed the occurrence of functionalized molecules in the synthesized NPS. The particles were highly stable with a net negative charge of -24.2. The morphology of NPS were determined by SEM and TEM. The mean particle size was 124.26nm. The production of NO by NR8383 cells was decreased by BG@BSA NPs. Also, in mice, lipopolysaccharide mediated acute lung inflammation was induced. BG@BSA NPs reduced macrophages and neutrophils in BALF and remarkably enhanced wet-to-dry weight (W/D) ratios and myeloperoxidase (MPO) activity. Further, BG@BSA NPs inhibited the production of inflammatory cells as well as tumour necrosis factor. The histopathological studies revealed that the damage and neutrophil infiltration was greatly inhibited by BG@BSA NPs. This indicates that BG@BSA NPs may be used to treat lung infections Therefore, this study has given new insight into producing an active drug for the treatment of lung associated diseases in the future.
... The enhanced activity for nanoparticle formulation was attributed to the improved cellular uptake of ATV-BSA. 144 5-Fluorouracil (5-FU) is an analogue of the pyrimidine, and it is a chemotherapeutic anticancer agent that belongs to the family of the antimetabolites. 145 It exerts its anticancer effects through inhibition of thymidylate synthase, followed by incorporation of its metabolites into RNA and DNA, and it was widely used, in particular, for colorectal cancer. ...
Albumin is an appealing carrier in nanomedicine because of its unique features. First, it is the most abundant protein in plasma, endowing high biocompatibility, biodegradability, nonimmunogenicity, and safety for its clinical application. Second, albumin chemical structure and conformation allows interaction with many different drugs, potentially protecting them from elimination and metabolism in vivo, thus improving their pharmacokinetic properties. Finally, albumin can interact with receptors overexpressed in many diseased tissues and cells, providing a unique feature for active targeting of the disease site without the addition of specific ligands to the nanocarrier. For this reason, albumin, characterized by an extended serum half-life of around 19 days, has the potential of promoting half-life extension and targeted delivery of drugs. Therefore, this article focuses on the importance of albumin as a nanodrug delivery carrier for hydrophobic drugs, taking advantage of the passive as well as active targeting potential of this nanocarrier. Particular attention is paid to the breakthrough NAB-Technology, with emphasis on the advantages of Nab-Paclitaxel (Abraxane), compared to the solvent-based formulations of Paclitaxel, i.e., CrEL-paclitaxel (Taxol) in a clinical setting. Finally, the role of albumin in carrying anticancer compounds is depicted, with a particular focus on the albumin-based formulations that are currently undergoing clinical trials. The article sheds light on the power of an endogenous substance, such as albumin, as a drug delivery system, signifies the importance of the drug vehicle in drug performance in the biological systems, and highlights the possible future trends in the use of this drug delivery system.
