Progressive liver fibrosis occurs in response to chronic liver injury and, if uncontrolled, can lead to cirrhosis, cancer and end-stage liver disease. For these reasons, there is increasing interest in finding new anti-fibrotic treatments. Extracellular vesicles (EVs) are lipid bilayer membrane-bound vesicles involved in cell-to-cell communication both in physiological and pathophysiological processes, as inflammation and fibrosis. Mesenchymal stromal cell (MSC)-derived EVs were reported to exert anti-inflammatory and anti-fibrotic functions. Moreover, previous studies have demonstrated that human liver stem cells (HLSCs), a hepatic multipotent population of adult stem cells with MSC-like phenotype, release EVs with pro-regenerative activity. The aim of this work is to set up an in vitro model of hepatic fibrosis using a human hepatic stellate cell line (LX-2) and then to study the potential anti-fibrotic effects of stem cell-EVs on LX-2. The stimulation of LX-2 with 10 ng/mL of TGF-β1 increases the expression of the pro-fibrotic markers α-SMA, COL1α1, TGF-B1 and matrix metalloproteinase 2 (MMP2). There is also an increase of pro-collagen I α1 released in LX-2 cell supernatant. The incubation of activated LX-2 with stem cell-EVs shows a dose-dependent downregulation of α-SMA. However, only in the presence of HLSC-EVs a slight decrease in the expression of pro-fibrotic genes COL1A1, TGF-β1 and MMP2 has been observed. In conclusion, this study demonstrates the TGF-β1-dependent in vitro activation of LX-2. Moreover, this study shows that bone marrow (BM)-MSC-derived EVs can partially revert the LX-2 activated phenotype, while HLSC-EVs have a more potent in vitro anti-fibrotic effect on the same cells.
Progressive liver fibrosis occurs in response to chronic liver injury and, if uncontrolled, can lead to cirrhosis, cancer and end-stage liver disease. For these reasons, there is increasing interest in finding new anti-fibrotic treatments. Extracellular vesicles (EVs) are lipid bilayer membrane-bound vesicles involved in cell-to-cell communication both in physiological and pathophysiological processes, as inflammation and fibrosis. Mesenchymal stromal cell (MSC)-derived EVs were reported to exert anti-inflammatory and anti-fibrotic functions. Moreover, previous studies have demonstrated that human liver stem cells (HLSCs), a hepatic multipotent population of adult stem cells with MSC-like phenotype, release EVs with pro-regenerative activity. The aim of this work is to set up an in vitro model of hepatic fibrosis using a human hepatic stellate cell line (LX-2) and then to study the potential anti-fibrotic effects of stem cell-EVs on LX-2. The stimulation of LX-2 with 10 ng/mL of TGF-β1 increases the expression of the pro-fibrotic markers α-SMA, COL1α1, TGF-B1 and matrix metalloproteinase 2 (MMP2). There is also an increase of pro-collagen I α1 released in LX-2 cell supernatant. The incubation of activated LX-2 with stem cell-EVs shows a dose-dependent downregulation of α-SMA. However, only in the presence of HLSC-EVs a slight decrease in the expression of pro-fibrotic genes COL1A1, TGF-β1 and MMP2 has been observed. In conclusion, this study demonstrates the TGF-β1-dependent in vitro activation of LX-2. Moreover, this study shows that bone marrow (BM)-MSC-derived EVs can partially revert the LX-2 activated phenotype, while HLSC-EVs have a more potent in vitro anti-fibrotic effect on the same cells.
Study of the effects of stem cell derived EVs on an in vitro model of hepatic fibrosis
CECCOTTI, ELENA
2019/2020
Abstract
Progressive liver fibrosis occurs in response to chronic liver injury and, if uncontrolled, can lead to cirrhosis, cancer and end-stage liver disease. For these reasons, there is increasing interest in finding new anti-fibrotic treatments. Extracellular vesicles (EVs) are lipid bilayer membrane-bound vesicles involved in cell-to-cell communication both in physiological and pathophysiological processes, as inflammation and fibrosis. Mesenchymal stromal cell (MSC)-derived EVs were reported to exert anti-inflammatory and anti-fibrotic functions. Moreover, previous studies have demonstrated that human liver stem cells (HLSCs), a hepatic multipotent population of adult stem cells with MSC-like phenotype, release EVs with pro-regenerative activity. The aim of this work is to set up an in vitro model of hepatic fibrosis using a human hepatic stellate cell line (LX-2) and then to study the potential anti-fibrotic effects of stem cell-EVs on LX-2. The stimulation of LX-2 with 10 ng/mL of TGF-β1 increases the expression of the pro-fibrotic markers α-SMA, COL1α1, TGF-B1 and matrix metalloproteinase 2 (MMP2). There is also an increase of pro-collagen I α1 released in LX-2 cell supernatant. The incubation of activated LX-2 with stem cell-EVs shows a dose-dependent downregulation of α-SMA. However, only in the presence of HLSC-EVs a slight decrease in the expression of pro-fibrotic genes COL1A1, TGF-β1 and MMP2 has been observed. In conclusion, this study demonstrates the TGF-β1-dependent in vitro activation of LX-2. Moreover, this study shows that bone marrow (BM)-MSC-derived EVs can partially revert the LX-2 activated phenotype, while HLSC-EVs have a more potent in vitro anti-fibrotic effect on the same cells.File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.14240/3634