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Another type of silica nanoparticles that has been studied in the present work is aminopropyl functionalized silica nanoparticles AMNPs , which were prepared via cocondensation sol-gel process of tetraethoxysilane TEOS and 3-aminopropyl triethoxysilane. They are also considered as promising drug carriers [ 16 , 17 ]. The choice of these types of silica nanoparticles is not random.

Silica Nanoparticles as Drug Delivery System for Immunomodulator GMDP - ASME

According to the literature data [ 18 — 20 ], amorphous silica and cationic silica nanoparticles are accepted as having low cytotoxicity and genotoxicity. However, the toxicity of silica nanoparticles is affected by many factors: surface chemistry, porosity, particle size, concentration, time of incubation and mode of administration into human organism [ 18 — 21 ]. A measure of cytotoxicity of each material must be clearly tested because the extent to which un- and functionalized mesoporous silica are toxic to mammalian cells has not yet been fully explored.

Various techniques are applied for drug immobilization, including adsorption, covalent attachment, and entrapment in polymers [ 22 ]. Drug adsorption on nanoparticles due to noncovalent interactions is easy to perform and widely used for drug loading. It is an attractive way to bind, deliver, and release actual drug without needing any triggers [ 23 ]. Sodium chloride high purity and double distilled water were used for preparation of isotonic solution.

The unmodified silica was synthesized via HCl-catalyzed sol-gel procedure of TEOS in the presence of pore forming agent sucrose as described elsewhere [ 24 ]. But in our procedure of the synthesis the silica precursor was not prehydrolyzed. After removal of the pore forming agent by water extraction and drying the mesoporous, unmodified silica was obtained.

Silica Nanoparticles as Drug Delivery System for Immunomodulator GMDP

The powders were milled and pressed into discs with KBr. The FTIR spectra of the sucrose and fructose containing materials are also recorded. The synthesized silica powders were investigated by SAXS. Theory of the method is presented in [ 26 , 27 ]. The n sil value is reported in [ 26 ]; the n m value of isotonic solution was measured and found to be 1.

The average radius of the particles was calculated as. The Q values can be calculated according to the data tabulated in [ 27 ]. The number concentration of the particles N per m 3 was calculated from 2. Peritoneal fluid of 40 women with endometriosis 32 with mild endometriosis stage II of disease according to the classification of American Society for Reproductive Medicine and 8 with severe endometriosis III-IV stages of disease who underwent laparoscopic examination for pelvic pain or infertility was used as the material.

Informed consent was given from each woman participating in our study, according to local Ethic Committee protocol. All patients ranged in age from 20 to 40 years and were not taking any hormone therapy at least 3 months prior to collection of the samples. Samples of peritoneal fluid were aspirated into the sterile tubes from the Douglas pouch during laparoscopic surgery and immediately used for the investigation. The standard procedure of isolation of the peritoneal mononuclear cells MNC by centrifugation in density gradient of Ficoll-Urografin d-1, was performed. Cells, which were incubated in RPMI medium in the absence of the nanoparticles at the same conditions, were used as control.

After incubation the viability of macrophages was estimated using the trypan blue dye exclusion assay, and the membrane expression of some functional molecules HLA-DR, CD11b, CD95, CD36, CD was defined using monoclonal antibodies by flow cytometry method. Production of the reactive oxygen species by macrophages after its incubation with the silica nanoparticles was estimated using the test of spontaneous NBTsp and zymosan stimulated NBTst reduction of nitrotetrazolium blue.

Immobilization of the labeled IgG onto the silica nanoparticles was carried out from isotonic solution. The level of the peritoneal macrophages reacted with the silica nanoparticles was estimated by flow cytometry as the amount of fluorescence-bright cells. The parameters of cells incubated only in the presence of culture media RPMI were used as the control.

One hundred thousand cells per tube were used for immunofluorescence staining. The data were presented as the percentage of stained cells in the macrophage population. Total RNA was isolated from pure fraction of peritoneal macrophages using the standard acid guanidinium thiocyanate-phenol-chloroform method. Sequences of corresponding genes cloned were used as controls.

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All variables were checked for normal distribution with the Kolmogorov-Smirnov test. Student's t -test was used to compare results between groups with normal distribution. Mann-Whitney test was used to compare results between groups with nonnormal distribution. Figure 1 shows FTIR spectra of the synthesized silica materials. Thus, the FTIR spectra indicate that the synthesized silica particles have different surface functional groups.

So, the prepared materials are amorphous. As has been mentioned above, amorphous silica has low toxic effects upon living organisms [ 18 — 20 ]. The influence of hour incubation of peritoneal macrophages with different types of silica nanoparticles upon functional activity of macrophages. No significant changes in the expression of scavenger receptors CD and CD36 molecules and spontaneous and zymozan-stimulated NBT-activity of the macrophages were observed after incubation of the macrophages with the studied silica nanoparticles.

So, the synthesized silica nanoparticles are nontoxic in relation to the peritoneal macrophages. The data characterizing the intensity of interaction between the peritoneal macrophages and studied silica nanoparticles are presented in Table 2. Characteristics of the intensity of interaction between peritoneal macrophages and different silica nanoparticles after 1-hour and hour incubation.

P : The level of statistically significant differences between different time of incubation and between different types of silica nanoparticles, P 1 is given in relation to the data after 1-hour incubation; P 2 is given in relation to the data for AMNPs. Thus, the cellular uptake of UMNPs by macrophages is intensive and time dependent. As the nanoparticles of these two types have the same size but different surface properties, it is likely that the last factor plays an important role.

However the interpretation of the shift of amide II band is doubtful because of overlapping of this band and the N—H deformation band of the aminopropyl groups in the region. The shifts testify about adsorption of GMDP on the silica materials via hydrogen bonding [ 33 ]. The obtained results are presented in Table 3.

Thus, GMDP stimulates the expression of the scavenger receptors by the peritoneal macrophages of women with endometriosis. In this case, similar to the experiments with scavenger receptors, the immobilization of GMDP upon UMNPs led to the decrease of the immunostimulatory action of the drug. This leads to the loss of the immunomodulatory effect of GMDP. This suppressor factor plays the important role in the regulation of the macrophages activation via PRRs molecules, preventing the undesirable hyperactivation of phagocytes [ 34 ].

To restore the balance between the pro- and anti-inflammatory signals, entering in the macrophages after its stimulation by GMDP, the high level of suppressor activity is kept in the cells.


It has been shown that macrophages produce a huge amount of different biologically active molecules, which are involved in the realization of different stages of phagocytosis [ 36 ]. In our study we estimated the effect of GMDP on the synthesis of proteolytic enzymes relevant to the family of matrix metalloproteinases MMPs. It is known that MMPs are proteolytic enzymes involved in extracellular matrix and basement membranes degradation.

MMPs activity is negatively regulated by its specific inhibitors - TIMPs tissue inhibitors of matrix metalloproteinases [ 37 ]. It has been shown that MMPs play an active role in the development of many pathological conditions, including tissue destruction, cancer invasion and metastasis, angiogenesis, and apoptosis [ 37 ].

Supposedly, MMPs may be actively involved in pathogenic mechanisms of endometriosis development. Recent studies have demonstrated that the content of MMP-1, MMP-2, and MMP-7 in the peritoneal fluid of women, with endometriosis is increased in comparison to that of healthy women, and the level of their inhibitors TIMPs , on the contrary, is reduced [ 38 ].

Our earlier investigations have showed the imbalance of MMPs and their inhibitors synthesis in the endometrial tissue of women with endometriosis as well as during the development of experimental endometriosis in rats [ 39 ]. However, according to the literature data, production of MMP-9 by macrophages of women with endometriosis is suppressed. Expression and secretion of matrix metalloproteinase MMP-9 by the macrophages serve to degrade the extracellular matrix of cells that are designated for phagocytosis [ 40 ]. Journals and Conferences. Learn More.

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Adsorption capacities of mesoporous silica materials having various surface functional groups hydroxyl, phenyl, mercaptopropyl, aminopropyl at pH values of 4. It was … More. Composites of cardiovascular drug molsidomine with silica materials unmodified and mercaptopropyl modified were prepared by 2 methods, adsorption and sol-gel technology. Pythagora: Roma; Dzierzbicka K, Wardowska A, Trzonkowski P: Recent developments in the synthesis and biological activity of muramylpeptides.

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