Hypothermia reduces VEGF-165 expression, but not osteogenic differentiation of human adipose stem cells under hypoxia

Leegwater, Nick C.; Bakker, Astrid D.; Hogervorst, Jolanda M. A.; Nolte, Peter A.; Klein‐Nulend, Jenneke

doi:10.1371/journal.pone.0171492

Cited by 8 publications

(9 citation statements)

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“…During fracture healing, hypoxia re-establishes oxygen supply by promoting angiogenesis via VEGF (Leegwater et al, 2017;Schipani, Maes, Carmeliet, & Semenza, 2009), which is a key component of bone repair (Farre-Guasch et al, 2018;Wang et al, 2011). Our implicating that IL4Rα/mTOR is the major signaling pathway contributing to the osteopenic phenotype in Fbn1 +/− mice (Chen et al, 2015).…”

Section: Hasc Proliferationmentioning

confidence: 77%

“…During fracture healing, hypoxia re‐establishes oxygen supply by promoting angiogenesis via VEGF (Leegwater et al, ; Schipani, Maes, Carmeliet, & Semenza, ), which is a key component of bone repair (Farre‐Guasch et al, ; Wang et al, ). Our findings also showed that hypoxia strongly enhanced VEGF expression in hASCs by ~4.0‐fold at Day 2 and 7 (Figures a,b–a,b).…”

Section: Discussionmentioning

confidence: 99%

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IL‐6 counteracts the inhibitory effect of IL‐4 on osteogenic differentiation of human adipose stem cells

Bastidas‐Coral

Hogervorst

Forouzanfar

et al. 2019

Journal Cellular Physiology

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Fracture repair is characterized by cytokine production and hypoxia. To better predict cytokine modulation of mesenchymal stem cell (MSC)‐aided bone healing, we investigated whether interleukin 4 (IL‐4), IL‐6, and their combination, affect osteogenic differentiation, vascular endothelial growth factor (VEGF) production, and/or mammalian target of rapamycin complex 1 (mTORC1) activation by MSCs under normoxia or hypoxia. Human adipose stem cells (hASCs) were cultured with IL‐4, IL‐6, or their combination for 3 days under normoxia (20% O 2) or hypoxia (1% O 2), followed by 11 days without cytokines under normoxia or hypoxia. Hypoxia did not alter IL‐4 or IL‐6‐modulated gene or protein expression by hASCs. IL‐4 alone decreased runt‐related transcription factor 2 (RUNX2) and collagen type 1 (COL1) gene expression, alkaline phosphatase (ALP) activity, and VEGF protein production by hASCs under normoxia and hypoxia, and decreased mineralization of hASCs under hypoxia. In contrast, IL‐6 increased mineralization of hASCs under normoxia, and enhanced RUNX2 gene expression under normoxia and hypoxia. Neither IL‐4 nor IL‐6 affected phosphorylation of the mTORC1 effector protein P70S6K. IL‐4 combined with IL‐6 diminished the inhibitory effect of IL‐4 on ALP activity, bone nodule formation, and VEGF production, and decreased RUNX2 and COL1 expression, similar to IL‐4 alone, under normoxia and hypoxia. In conclusion, IL‐4 alone, but not in combination with IL‐6, inhibits osteogenic differentiation and angiogenic stimulation potential of hASCs under normoxia and hypoxia, likely through pathways other than mTORC1. These results indicate that cytokines may differentially affect bone healing and regeneration when applied in isolation or in combination.

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Section: Hasc Proliferationmentioning

confidence: 77%

Section: Discussionmentioning

confidence: 99%

IL‐6 counteracts the inhibitory effect of IL‐4 on osteogenic differentiation of human adipose stem cells

Bastidas‐Coral

Hogervorst

Forouzanfar

et al. 2019

Journal Cellular Physiology

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“…Mild hypothermia alleviates hypoxic ischemic damage and increases tolerance to hypoxia via small ubiquitin-like modifier proteins of marrow-derived MSCs 34 , 35 . Although hypothermia reduces the gene expression of vascular endothelial growth factor, it does not affect the differentiation or apoptosis of MSCs exposed to hypoxia 36 . It can be seen that cryo-temperature pretreatment can enable MSCs to realize their potential under harsh conditions.…”

Section: Discussionmentioning

confidence: 95%

Cryo-Temperature Pretreatment Increases the Pro-Angiogenic Capacity of Three-Dimensional Mesenchymal Stem Cells via the PI3K-AKT Pathway

Zhu

Zhuang

et al. 2022

Cell Transplant

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To increase the potential and effectiveness of three-dimensional (3D) mesenchymal stem cells (MSCs) for clinical applications, this study explored the effects of short cryo-temperature pretreatment on MSC function. Adipose-derived MSCs (A-MSCs) were cultured via the ordinary monolayer method and 3D hanging drop spheroid method. When the cells adhered to the wall or formed a spheroid, they were subjected to hypothermic stress at 4°C for 1 h and then divided into three recovery periods at 37°C, specifically 0, 12, and 24 h. The control group was not subjected to any treatment throughout the study. Monolayer and 3D spheroid A-MSCs were analyzed via RNA sequencing after hypothermic stress at 4°C for 1 h. Subsequently, each group of cells was collected and subjected to phenotype identification via flow cytometry, and mRNA expression was detected via reverse transcription–quantitative polymerase chain reaction analysis. Western blot analysis was performed to analyze the PI3K-AKT signaling pathway in A-MSCs. The effects of A-MSCs on angiogenesis in vivo were examined using a chick chorioallantoic membrane assay. Transwell assays were performed to determine whether the culture supernatant from each group could induce the chemotaxis of human umbilical vein endothelial cells (HUVECs). Three-dimensional spheroid culture did not change the phenotype of A-MSCs. The expression of fibroblast growth factors, hepatocyte growth factors, and other angiogenesis-related factors in A-MSCs was upregulated. A-MSCs subjected to hypothermic stress promoted angiogenesis under both monolayer and 3D spheroid cultures. Moreover, the chemotaxis of HUVECs to the 3D spheroid culture supernatant increased substantially. Short cryo-temperature pretreatment could stimulate 3D spheroid A-MSCs and activate the PI3K-AKT pathway. This approach has the advantages of promoting angiogenesis and maintaining cell viability.

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“…It might also have implications for bone healing, as osteoblast activity is severely impaired and osteoclast activity promoted in murine calvarial cells subjected to 34 °C [13]. We previously determined that hypothermia reduces vascular endothelial growth factor 165 ( VEGF-165 ) protein expression under hypoxia, but we did not find the osteogenic differentiation of human adipose stem cells to be impaired [19]. Since the CFCT treatments in our study were applied intermittently for 30 min, the temperature drop is short-lived.…”

Section: Discussionmentioning

confidence: 99%

Continuous-flow cryocompression therapy penetrates to bone level in hip fracture patients in a numerical simulation

et al. 2019

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Background The aim of this study was to define deep tissue temperature during cryotherapy in postoperative hip fracture patients, by using measured skin temperature as input parameter for a simple numerical model. Second, an association was investigated between pain and tissue temperature distribution, to assess cryotherapy-induced analgesia of soft tissue-derived pain. Methods Data from 35 participants in an ongoing trial was used. In three subjects who consented on optional measurements, skin temperature was measured in 3 days during and after cryotherapy. A simple numerical model was developed to calculate tissue temperature distribution during cryotherapy. Results Inter and intrasubject skin temperature displayed high variation: trochanter 11–27 °C, mid-femur 11–24 °C, distal femur 10–16 °C. Predicted temperatures decreased to 20 °C at 1 cm, 26 °C at 2 cm, and 30 °C at 3 cm tissue depth. Smallest soft tissue layer was measured at the trochanter; 42% had less than 30 mm and 21% had less than 20 mm. Numeric rating scale pain varied (mean = 2.14; SD = 1.92), and no association was found between pain and decrease in temperature ( r = 0.064; p = 0.204). Conclusions Cryotherapy was predicted to reduce temperature up to 3 cm; in cachectic patients, this reaches the bone, where it might have implications for bone tissue healing when treated for a prolonged period of time. Cryotherapy-induced analgesia is likely to originate from skin analgesia rather than analgesia of muscle or bone-derived pain.

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Hypothermia reduces VEGF-165 expression, but not osteogenic differentiation of human adipose stem cells under hypoxia

Cited by 8 publications

References 50 publications

IL‐6 counteracts the inhibitory effect of IL‐4 on osteogenic differentiation of human adipose stem cells

IL‐6 counteracts the inhibitory effect of IL‐4 on osteogenic differentiation of human adipose stem cells

Cryo-Temperature Pretreatment Increases the Pro-Angiogenic Capacity of Three-Dimensional Mesenchymal Stem Cells via the PI3K-AKT Pathway

Continuous-flow cryocompression therapy penetrates to bone level in hip fracture patients in a numerical simulation

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