Effects of Sonication Processing on the Behavior of the Synthesis Human Serum Albumin-SPIONs Loaded PLGA Nanoparticles

This paper reports the most prominent contributions in the field of biodegradable polymeric nanoparticles from poly (lactic-co-glycolic acid) (PLGA) used as a protein/drug delivery. We use a combination of Human Serum Albumin (HSA)-superparamagnetic iron oxide nanoparticles (SPIONs) loaded PLGA nanoparticles. To obtain protein stabilization, the optimization of each step of synthesis nanoparticle is required. One of the most common problems in encapsulating protein to PLGA nanoparticles is the presence of several challenges as a problem of instability. We explained how the effect of the various sonication processing on the synthesis HSA-SPIONs loaded PLGA nanoparticles would be one of the crucial parameters for stability.


Introduction
Nanoparticles are dense and spherical structures range from 100 nm -200 nm in size and are made from natural or synthetic polymers. Various medications can be delivered using nanoparticles, such as hydrophilic small drug, hydrophobic small drug, vaccines, and biological macromolecules. Nanoparticles also allow the administration of specific organs or cells or controlled drug delivery.
In connections with the safety of the polymers used for encapsulation, Poly (lactic-co-glycolic acid) (PLGA) is one of the most successfully used biodegradable polymers, because its hydrolysis leads to metabolite monomers, lactic acid How to cite this paper: Vidawati, S., Barbosa, S., Taboada, P. and Mosquera, V. The development of nanotechnology is explained in medical sciences, e.g.
SPIONs (Superparamagnetic Iron Oxide Nanoparticles). SPIONs appear with significant potential application in Magnetic Resonance Imaging (MRI), drug delivery, magnetic hyperthermia, tissue repair, detoxification of biological fluids, and in cell separation, etc. The development of nanoparticles for the delivery of contrast agents has emerged in recent years because of the possibility of producing multifunctional nanoparticles that can specifically target tumors [1]. PLGA is used to formulate nanoparticles that encapsulate superparamagnetic iron oxide for MRI. This system enhances the imaging effect along with increasing the half-life of nanoparticles in the bloodstream, thereby reducing side effects [2].
This encapsulation of these therapeutic proteins in PLGA nanoparticles has emerged as a promising alternative to overcome all these problems as well as to contribute with certain additional benefits. Combining proteins into the polymer matrix provides protection against enzymatic and hydrolytic degradation in vivo, maintains their integrity and activity, can increase their bioavailability and in some cases can target therapeutic protein to the target area. Biodegradable nanoparticles production contains stable therapeutic proteins, mostly in terms of technical barriers. The precise assessment of the stability and quantifying of protein encapsulation remains difficult for major tasks and barrier prior to analysis [3] [4] [5] [6]. To enable protein stabilization, the optimization of each step of nanoparticles production is required. One of the most common techniques for encapsulating proteins into PLGA nanoparticles presents several challenges as a matter of instability [7]. Often protein instability is closely related to the presence of water or interfaces during particle preparation and some new techniques. Proteins from therapeutic should be studied on a case-by-case basis, so as to bring to the stage of future processing and stress factors that damage them.
To address this problem, many studies have focused on optimizing the formulation process in order to improve protein stability during the processing of procedures. The purpose of this study is investigated the effect of sonication processing on the behavior of the encapsulated PLGA nanoparticles for protein/drug delivery. For this study, we used HSA as a protein model. We use encapsuled PLGA loaded combinations of SPIONs and HSA. These nanoparticles are characterized for their physicochemical properties.
The importance of mixing kinetics solution containing hydrophobic and non-solvent compounds is highlighted. They require accurate processing parameters of possible sonication of fabrication for applications, nanoparticulate delivery systems. Sonication is used in a variety of physical, chemical and biological processes. Sonication is a highly effective processing method for generation and the application of nanosize, homogenizing, emulsifying, and dispersing for physical processes. Sonication with high-powered pulses is used to increase the dispersion of nanoparticles in the preparation of nanofluids. High-intensity sonication is used for the processing of liquids such as mixing, emulsifying, dispersing and de-agglomeration, or milling. When liquids are sonicated with high intensity, the sound waves that spread to the liquid media produce high-pressure (compression) and low-pressure (rarefaction) cycles, with rates depending on the frequency. Variations of the sonication intensity probe are studied to determine its effect on the characteristics of nanoparticles, such as average agglomerate size, polydispersity of the solution, and surface charge. The processing conditions have an important effect on the morphology, particle size and the formation of a stable nanoparticles phase.

Synthesis of HSA-SPIONs-PLGA Nanoparticles
Preparation of the polymer is encapsulated PLGA nanoparticles containing a combination of SPIONs and HSA prepared by using the multiple emulsion sol-

Characterization of Nanoparticles
In this study, all of nanoparticles were characterized using TEM image, Zeta Potential, and UV-Vis spectroscopy measurements.
The Transmission electron microscopy (TEM) is used for described particle

SPIONs
In this study, oleic acid-stabilized SPIONs were obtained with a co-precipitation method [3]. Without the coating, SPIONs tend to be aggregated, they are also hydrophobic and, when injected into the bloodstream, are coated by plasma proteins (called opsonization). The hydrophilic coating may prevent or significantly reduse opsonization, and through electrostatic interactions or steric hindrance, it decreases the aggregation of the SPIONs.
TEM image of the size and morphology of SPIONs is shown in Figure 1.  The Zeta potential of SPIONs in this study around +45 mV, the zeta potential representing a surface charge of the particles in a colloidal suspension, is one of the most important factors defining their stability, a tendency to aggregate (thus defines them effective size), as well as their ability to bind serum proteins. In spite of the fact that most of them are charged negatively, the more positive the charge of a SPION is, the stronger its ability to bind serum proteins [8]. The ultrasonic emulsification has been studied for decades and has recently garnered increased interest [10] [11]. The study compares the ultrasonic emulsification with the dispersing rotors [11] [12] finding ultrasound to be competitive or even superior in terms of droplet size and energetic efficiency. Oil-in-water emulsions, the system is also found useful for biodegradable nanoparticles preparations using the solvent extraction/evaporation method. The sonication power is controlled by the transfer oscillation amplitude. To measure the power consumed for emulsification, the power intake of a high frequency generator was recorded using a standard household power monitor. For 100%, 80% and 60% of the maximum amplitude, the power intake is of 32 W, 25 W and 17 W, respectively. The actual power assessment transferred to the emulsion is usually done by measuring the heat taken by the emulsion, which for current ultrasonic flow-through cell will be difficult to do with reasonable accuracy. However, it makes sense to assume that the power consumption by the generator should be comparable to that delivered to the emulsion [13] [14]. PLGA nanoparticles, forming emulsion in ultrasonic flow-cell via rapid, oil-in-water emulsions, the particle sizes is increased with less sonication power, although the difference is less pronounced than observed for the oil emulsions. By increasing the concentration of the polymer solution, and hence its viscosity, larger particles are produced. Obviously, viscosity is not the only physicochemical. Set the emulsification parameter, as it has been noted for the oil-in-water emulsion. Factors such as the tension of the interface and surfactants conformity are equally important, especially with regard to droplet coalescence.

HSA-SPIONs Loaded PLGA Nanoparticles
The size and morphology of HSA-SPIONs loaded PLGA nanoparticles of this experiment is characterized by TEM. TEM image are used to obtain important information about the main size and morphology of nanoparticles. TEM is an important technique whose unique ability to probing the internal structure of individual nanoparticles.
The results TEM images of HSA-SPIONs loaded PLGA nanoparticles are displayed interesting phenomenon information in the synthesis of HSA-SPIONs loaded PLGA nanoparticles processing with the sonication variation parameters. Figure 2 to number 5 has provided information about TEM image 0.05 g/ml HSA-SPIONs loaded PLGA nanoparticles which multple parameters of sonication processing combinations for mixture the PLGA + SPIONs + HSA solutions and in vitro release of HSA from HSA-SPIONs loaded PLGA nanoparticles. In this study, HSA-SPIONs loaded PLGA nanoparticles had a zeta potential between −4.2 mV until −10.15 mV.   and all result of this study according to Gopferich et al. [14] about the release of protein. From biodegradable nanoparticles in Figure 4(a), HSA release around 5.5% to 120 hours (see Figure 4(b)). The release profile is conducted, HSA release about 15% to 70 hours (see Figure   5(b)).

Conclusion
Summarizing our data, we argue that the sonication step is to be an important parameter on the synthesis HSA-SPIONs loaded PLGA nanoparticles. The