16 research outputs found
ΠΠΎΠ·Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ ΡΠΈΡΠΏΠ»Π°ΡΠΈΠ½Π° Π½Π° ΠΌΡΠ»ΡΡΠΈΠΏΠΎΡΠ΅Π½ΡΠ½ΡΠ΅ ΠΌΠ΅Π·Π΅Π½Ρ ΠΈΠΌΠ½ΡΠ΅ ΡΡΡΠΎΠΌΠ°Π»ΡΠ½ΡΠ΅ ΠΊΠ»Π΅ΡΠΊΠΈ ΠΆΠΈΡΠΎΠ²ΠΎΠΉ ΡΠΊΠ°Π½ΠΈ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° ΠΏΡΠΈ ΡΠ°Π·Π»ΠΈΡΠ½ΠΎΠΌ ΡΡΠΎΠ²Π½Π΅ ΠΊΠΈΡΠ»ΠΎΡΠΎΠ΄Π°
Objective: To evaluate the damaging effects of cisplatin on MMSCs from adipose tissue in a phase of active proliferation and the state of the monolayer, which was exposed at standard (20%) and reduced to 1% and 5% level of oxygen.Methods: The incubation MMSC with cisplatin was performed on cultures of 2 passage in a state in monolayer and cultures in the active growth phase. Profile surface markers of MMSC determined by flow cytometry. MMSCs viability after incubation with cisplatin was detected by the number of apoptotic and necrotic cells using ANNEXIN V-FITC - PI Kit (Immunotech, France). Standard culture conditions (~ 20% O2) created in a CO2 incubator (Sanyo, Japan), 5% O2 created using multigas incubator (Sanyo, Japan), 1% O2 - using an airtight chamber (Stemcell Technologies, USA).Results: Incubation of monolayer MMSC with cisplatin at a concentration of 10 ug/ml for 72 hours leads to death of half of the cells in culture under 20% O2, 5% O2 and 1% O2. Cisplatin increased the fracture of PI+-cell, and PI+/Ann+-cells under all culture conditions. The short-term exposure with cisplatin (24 and 48 hours) did not cause the damaging effect. Effects of cisplatin on the MMSC in the growth phase for 48 hours led to accumulation of Ann+-cells and PI+/Ann +-cells under all culture conditions. However least damaging effect of cisplatin was observed in culture under hypoxic conditions (1% O2).Conclusion: These data suggest that monolayer MMSCs are dying primarily through necrosis, whereas MMSC in the growth phase in response to cisplatin treatment are dying by apoptosis, regardless the oxygen tension.Π¦Π΅Π»Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ: ΠΎΡΠ΅Π½ΠΈΡΡ ΠΏΠΎΠ²ΡΠ΅ΠΆΠ΄Π°ΡΡΠ΅Π΅ Π²ΠΎΠ·Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ ΡΠΈΡΠΏΠ»Π°ΡΠΈΠ½Π° Π½Π° ΠΠΠ‘Π ΠΆΠΈΡΠΎΠ²ΠΎΠΉ ΡΠΊΠ°Π½ΠΈ, Π½Π°Ρ
ΠΎΠ΄ΡΡΠΈΠ΅ΡΡ Π² ΡΠ°Π·Π΅ Π°ΠΊΡΠΈΠ²Π½ΠΎΠΉ ΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠΈ ΠΈ Π² ΡΠΎΡΡΠΎΡΠ½ΠΈΠΈ ΠΌΠΎΠ½ΠΎΡΠ»ΠΎΡ, ΠΏΡΠΈ ΡΡΠ°Π½Π΄Π°ΡΡΠ½ΠΎΠΌ (20%) ΠΈ ΡΠ½ΠΈΠΆΠ΅Π½Π½ΠΎΠΌ Π΄ΠΎ 1% ΠΈ 5% ΡΡΠΎΠ²Π½Π΅ ΠΊΠΈΡΠ»ΠΎΡΠΎΠ΄Π° Π² ΡΡΠ΅Π΄Π΅ ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΠΎΠ²Π°Π½ΠΈΡ.ΠΠ΅ΡΠΎΠ΄Ρ: ΠΡΠ΅Π½ΠΊΡ Π²Π»ΠΈΡΠ½ΠΈΡ ΡΠΈΡΠΏΠ»Π°ΡΠΈΠ½Π° Π½Π° ΠΠΠ‘Π ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ Π½Π° ΠΊΡΠ»ΡΡΡΡΠ°Ρ
2 ΠΏΠ°ΡΡΠ°ΠΆΠ° Π² ΡΠΎΡΡΠΎΡΠ½ΠΈΠΈ ΠΌΠΎΠ½ΠΎΡΠ»ΠΎΡ ΠΈ Π² ΠΊΡΠ»ΡΡΡΡΠ°Ρ
Π² Π°ΠΊΡΠΈΠ²Π½ΠΎΠΉ ΡΠ°Π·Π΅ ΡΠΎΡΡΠ°. ΠΡΠΎΡΠΈΠ»Ρ ΠΏΠΎΠ²Π΅ΡΡ
Π½ΠΎΡΡΠ½ΡΡ
ΠΌΠ°ΡΠΊΠ΅ΡΠΎΠ² ΠΠΠ‘Π ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΠΏΡΠΎΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΠΎΡΠ»ΡΠΎΡΠΈΠΌΠ΅ΡΡΠΈΠΈ. ΠΠΈΠ·Π½Π΅ΡΠΏΠΎΡΠΎΠ±Π½ΠΎΡΡΡ ΠΠΠ‘Π ΠΏΠΎΡΠ»Π΅ ΠΈΠ½ΠΊΡΠ±Π°ΡΠΈΠΈ Ρ ΡΠΈΡΠΏΠ»Π°ΡΠΈΠ½ΠΎΠΌ ΠΎΡΠ΅Π½ΠΈΠ²Π°Π»ΠΈ ΠΏΠΎ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Ρ Π°ΠΏΠΎΠΏΡΠΎΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΈ Π½Π΅ΠΊΡΠΎΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΊΠ»Π΅ΡΠΎΠΊ Ρ ΠΏΠΎΠΌΠΎΡΡΡ Π½Π°Π±ΠΎΡΠ° ANNEXIN V-FITC β PI (Immunotech, Π€ΡΠ°Π½ΡΠΈΡ). Π‘ΡΠ°Π½Π΄Π°ΡΡΠ½ΡΠ΅ ΡΡΠ»ΠΎΠ²ΠΈΡ ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΠΎΠ²Π°Π½ΠΈΡ (~20% Π2) ΡΠΎΠ·Π΄Π°Π²Π°Π»ΠΈ Π² Π‘Π2-ΠΈΠ½ΠΊΡΠ±Π°ΡΠΎΡΠ΅ (Sanyo, Π―ΠΏΠΎΠ½ΠΈΡ), 5% Π2 ΡΠΎΠ·Π΄Π°Π²Π°Π»ΠΈ ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΡ ΠΌΡΠ»ΡΡΠΈΠ³Π°Π·ΠΎΠ²ΡΠΉ ΠΈΠ½ΠΊΡΠ±Π°ΡΠΎΡ (Sanyo, Π―ΠΏΠΎΠ½ΠΈΡ), 1% Π2 β ΠΈΡΠΏΠΎΠ»ΡΠ·ΡΡ Π³Π΅ΡΠΌΠ΅ΡΠΈΡΠ½ΡΡ ΠΊΠ°ΠΌΠ΅ΡΡ (Stemcell Technologies, Π‘Π¨Π).Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ: ΠΠ½ΠΊΡΠ±Π°ΡΠΈΡ ΠΠΠ‘Π Π² ΡΠΎΡΡΠΎΡΠ½ΠΈΠΈ ΠΌΠΎΠ½ΠΎΡΠ»ΠΎΡ Ρ ΡΠΈΡΠΏΠ»Π°ΡΠΈΠ½ΠΎΠΌ Π² ΠΊΠΎΠ½ΡΠ΅Π½ΡΡΠ°ΡΠΈΠΈ 10 ΠΌΠΊΠ³/ΠΌΠ» Π² ΡΠ΅ΡΠ΅Π½ΠΈΠ΅ 72 ΡΠ°ΡΠΎΠ² Π²ΡΠ·ΡΠ²Π°Π»Π° Π³ΠΈΠ±Π΅Π»Ρ ΠΏΡΠ°ΠΊΡΠΈΡΠ΅ΡΠΊΠΈ ΠΏΠΎΠ»ΠΎΠ²ΠΈΠ½Ρ ΠΊΠ»Π΅ΡΠΎΠΊ Π² ΠΊΡΠ»ΡΡΡΡΠ΅ ΠΊΠ°ΠΊ ΠΏΡΠΈ 20% Π2, ΡΠ°ΠΊ ΠΈ ΠΏΡΠΈ 5% ΠΈ 1% Π2 Π² ΡΡΠ΅Π΄Π΅. ΠΠΎ Π²ΡΠ΅Ρ
ΡΡΠ»ΠΎΠ²ΠΈΡΡ
ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΠ²Π΅Π»ΠΈΡΠΈΠ²Π°Π»Π°ΡΡ Π΄ΠΎΠ»Ρ PI+-ΠΊΠ»Π΅ΡΠΎΠΊ ΠΈ PI+/Ann+-ΠΊΠ»Π΅ΡΠΎΠΊ. ΠΠΎΠ»Π΅Π΅ ΠΊΡΠ°ΡΠΊΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½ΡΠ΅ ΡΠΊΡΠΏΠΎΠ·ΠΈΡΠΈΠΈ Ρ ΡΠΈΡΠΏΠ»Π°ΡΠΈΠ½ΠΎΠΌ (24 ΠΈ 48 ΡΠ°ΡΠΎΠ²) Π½Π΅ ΠΎΠΊΠ°Π·ΡΠ²Π°Π»ΠΈ Π²ΡΡΠ°ΠΆΠ΅Π½Π½ΠΎΠ³ΠΎ ΠΏΠΎΠ²ΡΠ΅ΠΆΠ΄Π°ΡΡΠ΅Π³ΠΎ ΡΡΡΠ΅ΠΊΡΠ°. ΠΠΎΠ·Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ ΡΠΈΡΠΏΠ»Π°ΡΠΈΠ½Π° Π½Π° ΠΠΠ‘Π Π² Π°ΠΊΡΠΈΠ²Π½ΠΎΠΉ ΡΠ°Π·Π΅ ΡΠΎΡΡΠ° Π² ΡΠ΅ΡΠ΅Π½ΠΈΠ΅ 48 ΡΠ°ΡΠΎΠ² ΡΠΎΠΏΡΠΎΠ²ΠΎΠΆΠ΄Π°Π»ΠΎΡΡ Π½Π°ΠΊΠΎΠΏΠ»Π΅Π½ΠΈΠ΅ΠΌ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π° Ann+-ΠΊΠ»Π΅ΡΠΎΠΊ ΠΈ PI+/Ann+-ΠΊΠ»Π΅ΡΠΎΠΊ. ΠΡΠΈ ΡΡΠΎΠΌ Π½Π°ΠΈΠΌΠ΅Π½ΡΡΠ΅Π΅ ΠΏΠΎΠ²ΡΠ΅ΠΆΠ΄Π°ΡΡΠ΅Π΅ Π²ΠΎΠ·Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ ΡΠΈΡΠΏΠ»Π°ΡΠΈΠ½ ΠΎΠΊΠ°Π·Π°Π» Π½Π° ΠΊΠ»Π΅ΡΠΊΠΈ, ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΡΠ΅ΠΌΡΠ΅ Π² ΡΡΠ»ΠΎΠ²ΠΈΡΡ
Π³ΠΈΠΏΠΎΠΊΡΠΈΠΈ (1% Π2).ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅: ΠΠΎΠ»ΡΡΠ΅Π½Π½ΡΠ΅ Π΄Π°Π½Π½ΡΠ΅ ΡΠ²ΠΈΠ΄Π΅ΡΠ΅Π»ΡΡΡΠ²ΡΡΡ, ΡΡΠΎ ΠΠΠ‘Π Π² ΡΠΎΡΡΠΎΡΠ½ΠΈΠΈ ΠΌΠΎΠ½ΠΎΡΠ»ΠΎΡ ΠΏΠΎΠ³ΠΈΠ±Π°ΡΡ ΠΏΡΠ΅ΠΈΠΌΡΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎ ΠΏΡΡΠ΅ΠΌ Π½Π΅ΠΊΡΠΎΠ·Π°, ΡΠΎΠ³Π΄Π° ΠΊΠ°ΠΊ Π΄Π»Ρ ΠΊΡΠ»ΡΡΡΡ ΠΠΠ‘Π Π² ΡΠ°Π·Π΅ ΡΠΎΡΡΠ° Π² ΠΎΡΠ²Π΅Ρ Π½Π° Π²ΠΎΠ·Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ Π±ΡΠ»ΠΎ Ρ
Π°ΡΠ°ΠΊΡΠ΅ΡΠ½ΠΎ Π½Π°ΠΊΠΎΠΏΠ»Π΅Π½ΠΈΠ΅ ΠΊΠ»Π΅ΡΠΎΠΊ, ΠΏΠΎΠ³ΠΈΠ±Π°ΡΡΠΈΡ
ΠΏΡΡΠ΅ΠΌ Π°ΠΏΠΎΠΏΡΠΎΠ·Π°, Π²Π½Π΅ Π·Π°Π²ΠΈΡΠΈΠΌΠΎΡΡΠΈ ΠΎΡ ΡΡΠΎΠ²Π½Ρ ΡΠΎΠ΄Π΅ΡΠΆΠ°Π½ΠΈΡ ΠΊΠΈΡΠ»ΠΎΡΠΎΠ΄Π° Π² ΡΡΠ΅Π΄Π΅
Osteogenic Potential Reduction in Mesenchymal Stem Cells under Prolonged Simulated Microgravity
Increasing the duration of orbital space flights up to 6β12 months and planning interplanetary missions actualizes the need for a better understanding of the mechanisms of osteopenia caused by microgravity. Investigation of mesenchymal stem cells (MSCs) that support the tissue homeostasis under microgravity conditions allows a deeper insight into the processes underlying bone loss. The purpose of this study was to investigate the osteogenic potential of MSCs under prolonged simulated microgravity by clinorotation. Using the method of mineralized matrix detection, it has been found that MSCs osteogenic potential decreased after long-term clinorotation. The investigation of major osteogenic gene expression has showed decreased transΒcriptional activity in RUNX2, ALPL-1, Col-1, but increased expression of PPARΞ³. One of the reasons for the decreased osteogenic potential of MSCs may be an increased level of reactive oxygen species (ROS) after 30 days of clinorotation. ROS may affect cellular signaling cascades, such as Wnt, Hedgehog and FOXO pathways, thereby leading to a shift of the differentiation potential to adipogenesis
BONE LOSS RECOVERY IN MICE FOLLOWING MICROGRAVITY WITH CONCURRENT BONE-COMPARTMENT-SPECIFIC OSTEOCYTE CHARACTERISTICS
Space missions provide the opportunity to investigate the influence of gravity on the dynamic remodelling processes in bone. Mice were examined following space flight and subsequent recovery to determine the effects on bone compartment-specific microstructure and composition. The resulting bone loss following microgravity recovered only in trabecular bone, while in cortical bone the tissue mineral density was restored after only one week on Earth. Detection of TRAP-positive bone surface cells in the trabecular compartment indicated increased resorption following space flight. In cortical bone, a persistent reduced viability of osteocytes suggested an impaired sensitivity to mechanical stresses. A compartment-dependent structural recovery from microgravity-induced bone loss was shown, with a direct osteocytic contribution to persistent low bone volume in the cortical region even after a recovery period. Trabecular recovery was not accompanied by changes in osteocyte characteristics. These post-space-flight findings will contribute to the understanding of compositional changes that compromise bone quality caused by unloading, immobilisation, or disuse