Activation of Neutrophils by Mucin–Vaterite Microparticles
Nano- and microparticles enter the body through the respiratory airways and the digestive system, or form as biominerals in the gall bladder, salivary glands, urinary bladder, kidney, or diabetic pancreas. Calcium, magnesium, and phosphate ions can precipitate from biological fluids in the presence...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/18/10579 |
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| author | Elena Mikhalchik Liliya Yu. Basyreva Sergey A. Gusev Oleg M. Panasenko Dmitry V. Klinov Nikolay A. Barinov Olga V. Morozova Alexander P. Moscalets Liliya N. Maltseva Lyubov Yu. Filatova Evgeniy A. Pronkin Julia A. Bespyatykh Nadezhda G. Balabushevich |
| author_facet | Elena Mikhalchik Liliya Yu. Basyreva Sergey A. Gusev Oleg M. Panasenko Dmitry V. Klinov Nikolay A. Barinov Olga V. Morozova Alexander P. Moscalets Liliya N. Maltseva Lyubov Yu. Filatova Evgeniy A. Pronkin Julia A. Bespyatykh Nadezhda G. Balabushevich |
| author_sort | Elena Mikhalchik |
| collection | DOAJ |
| description | Nano- and microparticles enter the body through the respiratory airways and the digestive system, or form as biominerals in the gall bladder, salivary glands, urinary bladder, kidney, or diabetic pancreas. Calcium, magnesium, and phosphate ions can precipitate from biological fluids in the presence of mucin as hybrid nanoparticles. Calcium carbonate nanocrystallites also trap mucin and are assembled into hybrid microparticles. Both mucin and calcium carbonate polymorphs (calcite, aragonite, and vaterite) are known to be components of such biominerals as gallstones which provoke inflammatory reactions. Our study was aimed at evaluation of neutrophil activation by hybrid vaterite–mucin microparticles (CCM). Vaterite microparticles (CC) and CCM were prepared under standard conditions. The diameter of CC and CCM was 3.3 ± 0.8 µm and 5.8 ± 0.7 µm, with ƺ-potentials of −1 ± 1 mV and −7 ± 1 mV, respectively. CC microparticles injured less than 2% of erythrocytes in 2 h at 1.5 mg mL<sup>−1</sup>, and no hemolysis was detected with CCM; this let us exclude direct damage of cellular membranes by microparticles. Activation of neutrophils was analyzed by luminol- and lucigenin-dependent chemiluminescence (Lum-CL and Luc-CL), by cytokine gene expression (IL-6, IL-8, IL-10) and release (IL-1β, IL-6, IL-8, IL-10, TNF-α), and by light microscopy of stained smears. There was a 10-fold and higher increase in the amplitude of Lum-CL and Luc-CL after stimulation of neutrophils with CCM relative to CC. Adsorption of mucin onto prefabricated CC microparticles also contributed to activation of neutrophil CL, unlike mucin adsorption onto yeast cell walls (zymosan); adsorbed mucin partially suppressed zymosan-stimulated production of oxidants by neutrophils. Preliminary treatment of CCM with 0.1–10 mM NaOCl decreased subsequent activation of Lum-CL and Luc-CL of neutrophils depending on the used NaOCl concentration, presumably because of the surface mucin oxidation. Based on the results of ELISA, incubation of neutrophils with CCM downregulated IL-6 production but upregulated that of IL-8. IL-6 and IL-8 gene expression in neutrophils was not affected by CC or CCM according to RT<sup>2</sup>-PCR data, which means that post-translational regulation was involved. Light microscopy revealed adhesion of CC and CCM microparticles onto the neutrophils; CCM increased neutrophil aggregation with a tendency to form neutrophil extracellular traps (NETs). We came to the conclusion that the main features of neutrophil reaction to mucin–vaterite hybrid microparticles are increased oxidant production, cell aggregation, and NET-like structure formation, but without significant cytokine release (except for IL-8). This effect of mucin is not anion-specific since particles of powdered kidney stone (mainly calcium oxalate) in the present study or calcium phosphate nanowires in our previous report also activated Lum-CL and Luc-CL response of neutrophils after mucin sorption. |
| first_indexed | 2024-03-09T23:45:41Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T23:45:41Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-50da3da0de0f4dfca23f9f64a65559812023-11-23T16:44:11ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-09-0123181057910.3390/ijms231810579Activation of Neutrophils by Mucin–Vaterite MicroparticlesElena Mikhalchik0Liliya Yu. Basyreva1Sergey A. Gusev2Oleg M. Panasenko3Dmitry V. Klinov4Nikolay A. Barinov5Olga V. Morozova6Alexander P. Moscalets7Liliya N. Maltseva8Lyubov Yu. Filatova9Evgeniy A. Pronkin10Julia A. Bespyatykh11Nadezhda G. Balabushevich12Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, RussiaFederal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, RussiaFederal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, RussiaFederal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, RussiaFederal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, RussiaFederal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, RussiaFederal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, RussiaFederal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, RussiaFaculty of Chemistry, Lomonosov Moscow State University, 119991 Moscow, RussiaFaculty of Chemistry, Lomonosov Moscow State University, 119991 Moscow, RussiaFederal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, RussiaFederal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, RussiaFaculty of Chemistry, Lomonosov Moscow State University, 119991 Moscow, RussiaNano- and microparticles enter the body through the respiratory airways and the digestive system, or form as biominerals in the gall bladder, salivary glands, urinary bladder, kidney, or diabetic pancreas. Calcium, magnesium, and phosphate ions can precipitate from biological fluids in the presence of mucin as hybrid nanoparticles. Calcium carbonate nanocrystallites also trap mucin and are assembled into hybrid microparticles. Both mucin and calcium carbonate polymorphs (calcite, aragonite, and vaterite) are known to be components of such biominerals as gallstones which provoke inflammatory reactions. Our study was aimed at evaluation of neutrophil activation by hybrid vaterite–mucin microparticles (CCM). Vaterite microparticles (CC) and CCM were prepared under standard conditions. The diameter of CC and CCM was 3.3 ± 0.8 µm and 5.8 ± 0.7 µm, with ƺ-potentials of −1 ± 1 mV and −7 ± 1 mV, respectively. CC microparticles injured less than 2% of erythrocytes in 2 h at 1.5 mg mL<sup>−1</sup>, and no hemolysis was detected with CCM; this let us exclude direct damage of cellular membranes by microparticles. Activation of neutrophils was analyzed by luminol- and lucigenin-dependent chemiluminescence (Lum-CL and Luc-CL), by cytokine gene expression (IL-6, IL-8, IL-10) and release (IL-1β, IL-6, IL-8, IL-10, TNF-α), and by light microscopy of stained smears. There was a 10-fold and higher increase in the amplitude of Lum-CL and Luc-CL after stimulation of neutrophils with CCM relative to CC. Adsorption of mucin onto prefabricated CC microparticles also contributed to activation of neutrophil CL, unlike mucin adsorption onto yeast cell walls (zymosan); adsorbed mucin partially suppressed zymosan-stimulated production of oxidants by neutrophils. Preliminary treatment of CCM with 0.1–10 mM NaOCl decreased subsequent activation of Lum-CL and Luc-CL of neutrophils depending on the used NaOCl concentration, presumably because of the surface mucin oxidation. Based on the results of ELISA, incubation of neutrophils with CCM downregulated IL-6 production but upregulated that of IL-8. IL-6 and IL-8 gene expression in neutrophils was not affected by CC or CCM according to RT<sup>2</sup>-PCR data, which means that post-translational regulation was involved. Light microscopy revealed adhesion of CC and CCM microparticles onto the neutrophils; CCM increased neutrophil aggregation with a tendency to form neutrophil extracellular traps (NETs). We came to the conclusion that the main features of neutrophil reaction to mucin–vaterite hybrid microparticles are increased oxidant production, cell aggregation, and NET-like structure formation, but without significant cytokine release (except for IL-8). This effect of mucin is not anion-specific since particles of powdered kidney stone (mainly calcium oxalate) in the present study or calcium phosphate nanowires in our previous report also activated Lum-CL and Luc-CL response of neutrophils after mucin sorption.https://www.mdpi.com/1422-0067/23/18/10579mucinvateritemicroparticleschemiluminescencecytokinesneutrophils |
| spellingShingle | Elena Mikhalchik Liliya Yu. Basyreva Sergey A. Gusev Oleg M. Panasenko Dmitry V. Klinov Nikolay A. Barinov Olga V. Morozova Alexander P. Moscalets Liliya N. Maltseva Lyubov Yu. Filatova Evgeniy A. Pronkin Julia A. Bespyatykh Nadezhda G. Balabushevich Activation of Neutrophils by Mucin–Vaterite Microparticles International Journal of Molecular Sciences mucin vaterite microparticles chemiluminescence cytokines neutrophils |
| title | Activation of Neutrophils by Mucin–Vaterite Microparticles |
| title_full | Activation of Neutrophils by Mucin–Vaterite Microparticles |
| title_fullStr | Activation of Neutrophils by Mucin–Vaterite Microparticles |
| title_full_unstemmed | Activation of Neutrophils by Mucin–Vaterite Microparticles |
| title_short | Activation of Neutrophils by Mucin–Vaterite Microparticles |
| title_sort | activation of neutrophils by mucin vaterite microparticles |
| topic | mucin vaterite microparticles chemiluminescence cytokines neutrophils |
| url | https://www.mdpi.com/1422-0067/23/18/10579 |
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