Bone marrow stromal cells (BMSCs CD45‐/CD44+/CD73+/CD90+) isolated from osteoporotic mice SAM/P6 as a novel model for osteoporosis investigation

Sikora, Mateusz; Śmieszek, Agnieszka; Marycz, Krzysztof

doi:10.1111/jcmm.16667

Cited by 17 publications

(16 citation statements)

References 91 publications

(179 reference statements)

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“…In order to induce the osteogenic differentiation of BMSCs, the cultures were propagated in the osteogenic medium prepared as described previously [ 20 , 25 ]: MEM-α medium (Minimum Essential Medium Eagle—Alpha Modification) was supplemented with ascorbic acid (50 µg/mL), β glycerol phosphate disodium salt hydrate (10 nM) and 15% addition of FBS. The osteogenesis was performed for 10 days, and the media were changed twice a week.…”

Section: Methodsmentioning

confidence: 99%

“…The immunocytochemical staining technique was used to visualise the cells’ ultrastructure and protein accumulation, as described previously [ 8 ] and accordingly to manufacturers’ instructions. Mitochondrial network was stained using Mito Red dye (Sigma-Aldrich, Munich, Germany) at the concentration of 1:1000 in CGM (complete growth medium) for 30 min at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

“…The miRNA and mRNA transcripts levels were analysed using RT-qPCR (reverse transcription quantitative polymerase chain reaction) technique as described previously in detail [ 8 , 28 ]. The cells were homogenised using 1 mL of Extrazol® (Blirt DNA, Gdańsk, Poland).…”

Section: Methodsmentioning

confidence: 99%

“…Recently, bone marrow-derived mesenchymal stem/stromal cells (BMSCs) have been proposed to have clinical potential for osteoporosis-related bone fractures regeneration due to their unique physiological properties [ 7 , 8 ]. Numerous studies showed that BMSCs compared to the other stem cells populations exhibit a higher ability to stimulate and enhance bone regeneration due to their paracrine activity [ 9 , 10 ].…”

Section: Introductionmentioning

confidence: 99%

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MiR-21-5p regulates the dynamic of mitochondria network and rejuvenates the senile phenotype of bone marrow stromal cells (BMSCs) isolated from osteoporotic SAM/P6 mice

et al. 2023

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Background Progression of senile osteoporosis is associated with deteriorated regenerative potential of bone marrow-derived mesenchymal stem/stromal cells (BMSCs). According to the recent results, the senescent phenotype of osteoporotic cells strongly correlates with impaired regulation of mitochondria dynamics. Moreover, due to the ageing of population and growing osteoporosis incidence, more efficient methods concerning BMSCs rejuvenation are intensely investigated. Recently, miR-21-5p was reported to play a vital role in bone turnover, but its therapeutic mechanisms in progenitor cells delivered from senile osteoporotic patients remain unclear. Therefore, the goal of this paper was to investigate for the first time the regenerative potential of miR-21-5p in the process of mitochondrial network regulation and stemness restoration using the unique model of BMSCs isolated from senile osteoporotic SAM/P6 mice model. Methods BMSCs were isolated from healthy BALB/c and osteoporotic SAM/P6 mice. We analysed the impact of miR-21-5p on the expression of crucial markers related to cells’ viability, mitochondria reconstruction and autophagy progression. Further, we established the expression of markers vital for bone homeostasis, as well as defined the composition of extracellular matrix in osteogenic cultures. The regenerative potential of miR-21 in vivo was also investigated using a critical-size cranial defect model by computed microtomography and SEM–EDX imaging. Results MiR-21 upregulation improved cells’ viability and drove mitochondria dynamics in osteoporotic BMSCs evidenced by the intensification of fission processes. Simultaneously, miR-21 enhanced the osteogenic differentiation of BMSCs evidenced by increased expression of Runx-2 but downregulated Trap, as well as improved calcification of extracellular matrix. Importantly, the analyses using the critical-size cranial defect model indicated on a greater ratio of newly formed tissue after miR-21 application, as well as upregulated content of calcium and phosphorus within the defect site. Conclusions Our results demonstrate that miR-21-5p regulates the fission and fusion processes of mitochondria and facilitates the stemness restoration of senile osteoporotic BMSCs. At the same time, it enhances the expression of RUNX-2, while reduces TRAP accumulation in the cells with deteriorated phenotype. Therefore, miR-21-5p may bring a novel molecular strategy for senile osteoporosis diagnostics and treatment.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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MiR-21-5p regulates the dynamic of mitochondria network and rejuvenates the senile phenotype of bone marrow stromal cells (BMSCs) isolated from osteoporotic SAM/P6 mice

et al. 2023

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“…Immunocytochemical (ICC) staining was performed using previously described protocol s [24,31]. Here, Eca EPCs cultures were performed in 24-well dishes covered by glass cover slides.…”

Section: Immunocytochemical Detectionmentioning

confidence: 99%

Obesity affects mitochondrial metabolism and proliferative potential of equine endometrial progenitor cells

Śmieszek

Marcinkowska

Pielok

et al. 2021

Preprint

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The study aimed to investigate the influence of obesity on cellular features of equine endometrial progenitor cells (Eca EPCs), including viability, proliferation capacity, mitochondrial metabolism, and oxidative homeostasis. Eca EPCs derived from non-obese (Non-OB) and obese (OB) mares were characterised by cellular phenotype and multipotency.Obesity-induced changes in the activity of Eca EPCs include the decline of their proliferative activity, clonogenic potential, mitochondrial metabolism and enhanced oxidative stress. Eca EPCs isolated from obese mares were characterised by an increased occurrence of early apoptosis, loss of mitochondrial dynamics, and senescence-associated phenotype. Attenuated metabolism of Eca EPCs OB was related to increased expression of pro-apoptotic markers (CASP9, BAX, P53, P21), enhanced expression of OPN, PI3K and AKT, simultaneously with decreased signalling stabilising cellular homeostasis (including mitofusin, SIRT1, FOXP3).Obesity alters functional features and self-renewal potential of endometrial progenitor cells. The impaired cytophysiology of progenitor cells from obese endometrium predict lower regenerative capacity if used as autologous transplants.

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G/ β‐ TCP composite scaffold material promotes osteogenic differentiation of bone marrow mesenchymal stem cells

Su,

Liao,

Yuan

et al. 2023

J Biomed Mater Res

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To explore self‐made graphene/β Graphene (G)/β‐ tricalcium phosphate, G/β‐ The effect of TCP composite scaffold material on osteogenic differentiation of BMSC. Preparation of G/β‐ TCP composite material was used to investigate the effect of composite material on bone marrow mesenchymal stem cell ossification/β‐ TCP material was used to treat primary BMSCs of rats. Cell morphology changes were observed under scanning electron microscopy, cell cycle and proliferation were detected by flow cytometry, and gene expression of chondrogenic genes Fibronectin, collagen I, collagen II, ICAM, and VCAM was detected by q‐PCR. In addition, using osteogenic induction medium and G/β‐ TCP composite materials were co treated with BMSCs, and ALP and alizarin red staining were used to observe the effect of the materials on osteogenic differentiation. q‐PCR was used to detect the gene expression of osteogenic related genes Runx2, OCN, and OPN. G/ β‐ After the TCP composite was co cultured with BMSC, the proportion of G0/G1 phase of BMSC cells was significantly increased, the cell proliferation ability was enhanced, and the gene expression of fibronectin, collagen I, collagen II, ICAM, and VCAM were significantly increased. The ALP staining results indicate that BMSC in G/β‐ After treatment with TCP composite material, significant enhancement of osteogenic ability was observed at 7,14 and 21 days. In addition, BMSC in G/β‐ A significant increase in calcium deposition was observed at 7,14 and 21 days after treatment with TCP composite materials. The effect of different time points on the expression of osteogenic related genes varies. At 7 and 14 days, the expression of RUNX2 was significantly reduced compared to the control, but significantly increased at 21 days; OCN significantly increased on the 21st day; OPN significantly increased at 14 days. G/β‐ TCP materials significantly promote the osteogenic differentiation of BMSCs.

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Bone marrow stromal cells (BMSCs CD45^‐/CD44⁺/CD73⁺/CD90⁺) isolated from osteoporotic mice SAM/P6 as a novel model for osteoporosis investigation

Abstract: This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Cited by 17 publications

References 91 publications

MiR-21-5p regulates the dynamic of mitochondria network and rejuvenates the senile phenotype of bone marrow stromal cells (BMSCs) isolated from osteoporotic SAM/P6 mice

MiR-21-5p regulates the dynamic of mitochondria network and rejuvenates the senile phenotype of bone marrow stromal cells (BMSCs) isolated from osteoporotic SAM/P6 mice

Obesity affects mitochondrial metabolism and proliferative potential of equine endometrial progenitor cells

G/ β‐ TCP composite scaffold material promotes osteogenic differentiation of bone marrow mesenchymal stem cells

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