In vitro testing of genotoxic and apoptotic potential of two synthesized nanomaterials by DNA fragmentation assay
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Nanomaterials represent a relatively new and developing class of materials that are widely used because of their unique mechanical and physiochemical properties. Due to the particles’ size and the ability to easily enter the cells and interact with different cellular components, nanoparticles can exhibit toxic effects in biological systems, so it is necessary to conduct a thorough in vitro biocompatibility testing. Our aim was to examine the in vitro genotoxic and apoptotic potential of two different synthesized nanomaterials intended for bone tissue engineering and regenerative applications: biphasic calcium phosphate coated with poly- D,L-lactide-co-glycolide (CP/PLGA) and cobalt-substituted hydroxyapatite (CoHAp), by DNA fragmentation assay. NB4 cells (acute promyelocytic leukemia cell line) were incubated with different concentrations of CP/PLGA and CoHAp nanomaterials for 24 hours. Cells cultured in standard medium, without nanomaterials, were used as negative control and cells tr...eated with hydrogen peroxide were used as positive control. After incubation with materials the genomic DNA was extracted from the cells and applied on horizontal agarose gel electrophoresis. The results of DNA fragmentation assay were observed on gel documentation system. Higher examined concentrations of CP/PLGA caused DNA fragmentation at some extent with DNA laddering at approximately 180 bp which is characteristic for later stages of apoptosis, while DNA laddering was not seen in the case of CoHAp at the same concentrations. Although both materials are in the form of nanoparticles, a different effect in apoptotic DNA fragmentation was observed which suggests that the chemical composition of nanomaterials notably affects the genotoxicity and apoptosis potential in addition to the particle size.
Keywords:
DNA fragmentation assay / nanomaterials / genotoxic potential / apoptotic potential / poly(D,L-lactide-co-glycolide) / cobalt-substituted hydroxyapatiteSource:
Program and the Book of Abstracts / Seventeenth Young Researchers' Conference Materials Sciences and Engineering, December 5-7, 2018, Belgrade, Serbia, 2018, 69-69Publisher:
- Belgrade : Institute of Technical Sciences of SASA
Funding / projects:
- Virtual human osteoarticular system and its application in preclinical and clinical practice (RS-41017)
Institution/Community
Институт техничких наука САНУ / Institute of Technical Sciences of SASATY - CONF AU - Stojanović, Sanja AU - Bajić, Lazar AU - Cvetković, Vladimir AU - Ignjatović, Nenad AU - Najman, Stevo PY - 2018 UR - https://dais.sanu.ac.rs/123456789/4723 AB - Nanomaterials represent a relatively new and developing class of materials that are widely used because of their unique mechanical and physiochemical properties. Due to the particles’ size and the ability to easily enter the cells and interact with different cellular components, nanoparticles can exhibit toxic effects in biological systems, so it is necessary to conduct a thorough in vitro biocompatibility testing. Our aim was to examine the in vitro genotoxic and apoptotic potential of two different synthesized nanomaterials intended for bone tissue engineering and regenerative applications: biphasic calcium phosphate coated with poly- D,L-lactide-co-glycolide (CP/PLGA) and cobalt-substituted hydroxyapatite (CoHAp), by DNA fragmentation assay. NB4 cells (acute promyelocytic leukemia cell line) were incubated with different concentrations of CP/PLGA and CoHAp nanomaterials for 24 hours. Cells cultured in standard medium, without nanomaterials, were used as negative control and cells treated with hydrogen peroxide were used as positive control. After incubation with materials the genomic DNA was extracted from the cells and applied on horizontal agarose gel electrophoresis. The results of DNA fragmentation assay were observed on gel documentation system. Higher examined concentrations of CP/PLGA caused DNA fragmentation at some extent with DNA laddering at approximately 180 bp which is characteristic for later stages of apoptosis, while DNA laddering was not seen in the case of CoHAp at the same concentrations. Although both materials are in the form of nanoparticles, a different effect in apoptotic DNA fragmentation was observed which suggests that the chemical composition of nanomaterials notably affects the genotoxicity and apoptosis potential in addition to the particle size. PB - Belgrade : Institute of Technical Sciences of SASA C3 - Program and the Book of Abstracts / Seventeenth Young Researchers' Conference Materials Sciences and Engineering, December 5-7, 2018, Belgrade, Serbia T1 - In vitro testing of genotoxic and apoptotic potential of two synthesized nanomaterials by DNA fragmentation assay SP - 69 EP - 69 UR - https://hdl.handle.net/21.15107/rcub_dais_4723 ER -
@conference{ author = "Stojanović, Sanja and Bajić, Lazar and Cvetković, Vladimir and Ignjatović, Nenad and Najman, Stevo", year = "2018", abstract = "Nanomaterials represent a relatively new and developing class of materials that are widely used because of their unique mechanical and physiochemical properties. Due to the particles’ size and the ability to easily enter the cells and interact with different cellular components, nanoparticles can exhibit toxic effects in biological systems, so it is necessary to conduct a thorough in vitro biocompatibility testing. Our aim was to examine the in vitro genotoxic and apoptotic potential of two different synthesized nanomaterials intended for bone tissue engineering and regenerative applications: biphasic calcium phosphate coated with poly- D,L-lactide-co-glycolide (CP/PLGA) and cobalt-substituted hydroxyapatite (CoHAp), by DNA fragmentation assay. NB4 cells (acute promyelocytic leukemia cell line) were incubated with different concentrations of CP/PLGA and CoHAp nanomaterials for 24 hours. Cells cultured in standard medium, without nanomaterials, were used as negative control and cells treated with hydrogen peroxide were used as positive control. After incubation with materials the genomic DNA was extracted from the cells and applied on horizontal agarose gel electrophoresis. The results of DNA fragmentation assay were observed on gel documentation system. Higher examined concentrations of CP/PLGA caused DNA fragmentation at some extent with DNA laddering at approximately 180 bp which is characteristic for later stages of apoptosis, while DNA laddering was not seen in the case of CoHAp at the same concentrations. Although both materials are in the form of nanoparticles, a different effect in apoptotic DNA fragmentation was observed which suggests that the chemical composition of nanomaterials notably affects the genotoxicity and apoptosis potential in addition to the particle size.", publisher = "Belgrade : Institute of Technical Sciences of SASA", journal = "Program and the Book of Abstracts / Seventeenth Young Researchers' Conference Materials Sciences and Engineering, December 5-7, 2018, Belgrade, Serbia", title = "In vitro testing of genotoxic and apoptotic potential of two synthesized nanomaterials by DNA fragmentation assay", pages = "69-69", url = "https://hdl.handle.net/21.15107/rcub_dais_4723" }
Stojanović, S., Bajić, L., Cvetković, V., Ignjatović, N.,& Najman, S.. (2018). In vitro testing of genotoxic and apoptotic potential of two synthesized nanomaterials by DNA fragmentation assay. in Program and the Book of Abstracts / Seventeenth Young Researchers' Conference Materials Sciences and Engineering, December 5-7, 2018, Belgrade, Serbia Belgrade : Institute of Technical Sciences of SASA., 69-69. https://hdl.handle.net/21.15107/rcub_dais_4723
Stojanović S, Bajić L, Cvetković V, Ignjatović N, Najman S. In vitro testing of genotoxic and apoptotic potential of two synthesized nanomaterials by DNA fragmentation assay. in Program and the Book of Abstracts / Seventeenth Young Researchers' Conference Materials Sciences and Engineering, December 5-7, 2018, Belgrade, Serbia. 2018;:69-69. https://hdl.handle.net/21.15107/rcub_dais_4723 .
Stojanović, Sanja, Bajić, Lazar, Cvetković, Vladimir, Ignjatović, Nenad, Najman, Stevo, "In vitro testing of genotoxic and apoptotic potential of two synthesized nanomaterials by DNA fragmentation assay" in Program and the Book of Abstracts / Seventeenth Young Researchers' Conference Materials Sciences and Engineering, December 5-7, 2018, Belgrade, Serbia (2018):69-69, https://hdl.handle.net/21.15107/rcub_dais_4723 .