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Hypoxia-preconditioned mesenchymal stem cells attenuate bleomycin-induced pulmonary fibrosis.

Lan YW, Choo KB, Chen CM, Hung TH, Chen YB, Hsieh CH, Kuo HP, Chong KY - Stem Cell Res Ther (2015)

Bottom Line: The pulmonary respiratory functions significantly improved for up to 18 days of hypoxia-preconditioned MSC treatment.Histopathologic examination observed a significant amelioration of the lung fibrosis.Our data further demonstrated that upregulation of hepatocyte growth factor possibly played an important role in mediating the therapeutic effects of transplanted hypoxia-preconditioned MSCs.

View Article: PubMed Central - PubMed

Affiliation: Division of Biotechnology, Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, Republic of China. bublelanwilliam@gmail.com.

ABSTRACT

Introduction: Idiopathic pulmonary fibrosis is a progressive diffuse parenchymal lung disorder of unknown etiology. Mesenchymal stem cell (MSC)-based therapy is a novel approach with great therapeutic potential for the treatment of lung diseases. Despite demonstration of MSC grafting, the populations of engrafted MSCs have been shown to decrease dramatically 24 hours post-transplantation due to exposure to harsh microenvironments. Hypoxia is known to induce expression of cytoprotective genes and also secretion of anti-inflammatory, anti-apoptotic and anti-fibrotic factors. Hypoxic preconditioning is thought to enhance the therapeutic potency and duration of survival of engrafted MSCs. In this work, we aimed to prolong the duration of survival of engrafted MSCs and to enhance the effectiveness of idiopathic pulmonary fibrosis transplantation therapy by the use of hypoxia-preconditioned MSCs.

Methods: Hypoxic preconditioning was achieved in MSCs under an optimal hypoxic environment. The expression levels of cytoprotective factors and their biological effects on damaged alveolar epithelial cells or transforming growth factor-beta 1-treated fibroblast cells were studied in co-culture experiments in vitro. Furthermore, hypoxia-preconditioned MSCs (HP-MSCs) were intratracheally instilled into bleomycin-induced pulmonary fibrosis mice at day 3, and lung functions, cellular, molecular and pathological changes were assessed at 7 and 21 days after bleomycin administration.

Results: The expression of genes for pro-survival, anti-apoptotic, anti-oxidant and growth factors was upregulated in MSCs under hypoxic conditions. In transforming growth factor-beta 1-treated MRC-5 fibroblast cells, hypoxia-preconditioned MSCs attenuated extracellular matrix production through paracrine effects. The pulmonary respiratory functions significantly improved for up to 18 days of hypoxia-preconditioned MSC treatment. Expression of inflammatory factors and fibrotic factor were all downregulated in the lung tissues of the hypoxia-preconditioned MSC-treated mice. Histopathologic examination observed a significant amelioration of the lung fibrosis. Several LacZ-labeled MSCs were observed within the lungs in the hypoxia-preconditioned MSC treatment groups at day 21, but no signals were detected in the normoxic MSC group. Our data further demonstrated that upregulation of hepatocyte growth factor possibly played an important role in mediating the therapeutic effects of transplanted hypoxia-preconditioned MSCs.

Conclusion: Transplantation of hypoxia-preconditioned MSCs exerted better therapeutic effects in bleomycin-induced pulmonary fibrotic mice and enhanced the survival rate of engrafted MSCs, partially due to the upregulation of hepatocyte growth factor.

No MeSH data available.


Related in: MedlinePlus

Hypoxia-preconditioned mesenchymal stem cell transplantation improves long-term attenuation of bleomycin-induced airway constriction and lung edema. (A) Pulmonary functions were evaluated as enhanced respiratory pause (Penh) values in animals that received normoxia-preconditioned mesenchymal stem cells (NP-MSCs), hypoxia-preconditioned mesenchymal stem cells (HP-MSCs) or phosphate-buffered saline (PBS) for 18 days following bleomycin (BLM) administration on day 3. Whole-body plethysmography was employed and Penh was used as a noninvasive index of airway dysfunction. (B) After sacrificing the mice on day 21, the lung wet-to-dry weight ratio was determined to assess pulmonary edema. Each dot represents an individual mouse with the mean shown for n ≥ 5 per group. *P < 0.05, **P < 0.01. Ctrl, control.
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Fig4: Hypoxia-preconditioned mesenchymal stem cell transplantation improves long-term attenuation of bleomycin-induced airway constriction and lung edema. (A) Pulmonary functions were evaluated as enhanced respiratory pause (Penh) values in animals that received normoxia-preconditioned mesenchymal stem cells (NP-MSCs), hypoxia-preconditioned mesenchymal stem cells (HP-MSCs) or phosphate-buffered saline (PBS) for 18 days following bleomycin (BLM) administration on day 3. Whole-body plethysmography was employed and Penh was used as a noninvasive index of airway dysfunction. (B) After sacrificing the mice on day 21, the lung wet-to-dry weight ratio was determined to assess pulmonary edema. Each dot represents an individual mouse with the mean shown for n ≥ 5 per group. *P < 0.05, **P < 0.01. Ctrl, control.

Mentions: To evaluate the therapeutic efficacy of HP-MSCs in the BLM-induced pulmonary fibrosis mouse model, WBP was used to monitor lung functions of the treated mice. The respiratory parameter, in particular Penh, was measured as a noninvasive index of BLM-induced airway dysfunction [44]. In the experiments, the Penh values showed a two-fold increment at day 21 after BLM treatment compared with the PBS control group (0.61 ± 0.06 versus 1.16 ± 0.33, P < 0.05; Figure 4A). In addition, the BLM HP-MSC group had the lowest Penh value which was approximately the same as the PBS control; however, the BLM NP-MSC group showed no significant improvements in lung function in the fibrosis animal model (0.70 ± 0.07 in BLM HP-MSCs versus 1.19 ± 0.34 (BLM NP-MSCs) and 1.16 ± 0.33 (BLM control); Figure 4A). In addition, compared with the PBS control, the mice treated with BLM for 21 days showed a significant increment in the lung wet-to-dry ratio, indicating the degree of pulmonary edema caused by an inflammatory response in the lung. Likewise, the BLM HP-MSC group showed a lower wet-to-dry ratio than the BLM NP-MSC and BLM control groups at day 18 after stem cells transplantation (0.32 ± 0.05 in BLM HP-MSC group versus 0.43 ± 0.05 (BLM NP-MSC) and 0.42 ± 0.10 (BLM control); Figure 4B) also indicating that transplantation of HP-MSCs significantly reduced lung fluid content caused by BLM treatment. Taken together, our results indicated that transplanted HP-MSCs showed better therapeutic effects and improved lung functions in mice with BLM-induced pulmonary fibrosis.Figure 4


Hypoxia-preconditioned mesenchymal stem cells attenuate bleomycin-induced pulmonary fibrosis.

Lan YW, Choo KB, Chen CM, Hung TH, Chen YB, Hsieh CH, Kuo HP, Chong KY - Stem Cell Res Ther (2015)

Hypoxia-preconditioned mesenchymal stem cell transplantation improves long-term attenuation of bleomycin-induced airway constriction and lung edema. (A) Pulmonary functions were evaluated as enhanced respiratory pause (Penh) values in animals that received normoxia-preconditioned mesenchymal stem cells (NP-MSCs), hypoxia-preconditioned mesenchymal stem cells (HP-MSCs) or phosphate-buffered saline (PBS) for 18 days following bleomycin (BLM) administration on day 3. Whole-body plethysmography was employed and Penh was used as a noninvasive index of airway dysfunction. (B) After sacrificing the mice on day 21, the lung wet-to-dry weight ratio was determined to assess pulmonary edema. Each dot represents an individual mouse with the mean shown for n ≥ 5 per group. *P < 0.05, **P < 0.01. Ctrl, control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4487587&req=5

Fig4: Hypoxia-preconditioned mesenchymal stem cell transplantation improves long-term attenuation of bleomycin-induced airway constriction and lung edema. (A) Pulmonary functions were evaluated as enhanced respiratory pause (Penh) values in animals that received normoxia-preconditioned mesenchymal stem cells (NP-MSCs), hypoxia-preconditioned mesenchymal stem cells (HP-MSCs) or phosphate-buffered saline (PBS) for 18 days following bleomycin (BLM) administration on day 3. Whole-body plethysmography was employed and Penh was used as a noninvasive index of airway dysfunction. (B) After sacrificing the mice on day 21, the lung wet-to-dry weight ratio was determined to assess pulmonary edema. Each dot represents an individual mouse with the mean shown for n ≥ 5 per group. *P < 0.05, **P < 0.01. Ctrl, control.
Mentions: To evaluate the therapeutic efficacy of HP-MSCs in the BLM-induced pulmonary fibrosis mouse model, WBP was used to monitor lung functions of the treated mice. The respiratory parameter, in particular Penh, was measured as a noninvasive index of BLM-induced airway dysfunction [44]. In the experiments, the Penh values showed a two-fold increment at day 21 after BLM treatment compared with the PBS control group (0.61 ± 0.06 versus 1.16 ± 0.33, P < 0.05; Figure 4A). In addition, the BLM HP-MSC group had the lowest Penh value which was approximately the same as the PBS control; however, the BLM NP-MSC group showed no significant improvements in lung function in the fibrosis animal model (0.70 ± 0.07 in BLM HP-MSCs versus 1.19 ± 0.34 (BLM NP-MSCs) and 1.16 ± 0.33 (BLM control); Figure 4A). In addition, compared with the PBS control, the mice treated with BLM for 21 days showed a significant increment in the lung wet-to-dry ratio, indicating the degree of pulmonary edema caused by an inflammatory response in the lung. Likewise, the BLM HP-MSC group showed a lower wet-to-dry ratio than the BLM NP-MSC and BLM control groups at day 18 after stem cells transplantation (0.32 ± 0.05 in BLM HP-MSC group versus 0.43 ± 0.05 (BLM NP-MSC) and 0.42 ± 0.10 (BLM control); Figure 4B) also indicating that transplantation of HP-MSCs significantly reduced lung fluid content caused by BLM treatment. Taken together, our results indicated that transplanted HP-MSCs showed better therapeutic effects and improved lung functions in mice with BLM-induced pulmonary fibrosis.Figure 4

Bottom Line: The pulmonary respiratory functions significantly improved for up to 18 days of hypoxia-preconditioned MSC treatment.Histopathologic examination observed a significant amelioration of the lung fibrosis.Our data further demonstrated that upregulation of hepatocyte growth factor possibly played an important role in mediating the therapeutic effects of transplanted hypoxia-preconditioned MSCs.

View Article: PubMed Central - PubMed

Affiliation: Division of Biotechnology, Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan, Republic of China. bublelanwilliam@gmail.com.

ABSTRACT

Introduction: Idiopathic pulmonary fibrosis is a progressive diffuse parenchymal lung disorder of unknown etiology. Mesenchymal stem cell (MSC)-based therapy is a novel approach with great therapeutic potential for the treatment of lung diseases. Despite demonstration of MSC grafting, the populations of engrafted MSCs have been shown to decrease dramatically 24 hours post-transplantation due to exposure to harsh microenvironments. Hypoxia is known to induce expression of cytoprotective genes and also secretion of anti-inflammatory, anti-apoptotic and anti-fibrotic factors. Hypoxic preconditioning is thought to enhance the therapeutic potency and duration of survival of engrafted MSCs. In this work, we aimed to prolong the duration of survival of engrafted MSCs and to enhance the effectiveness of idiopathic pulmonary fibrosis transplantation therapy by the use of hypoxia-preconditioned MSCs.

Methods: Hypoxic preconditioning was achieved in MSCs under an optimal hypoxic environment. The expression levels of cytoprotective factors and their biological effects on damaged alveolar epithelial cells or transforming growth factor-beta 1-treated fibroblast cells were studied in co-culture experiments in vitro. Furthermore, hypoxia-preconditioned MSCs (HP-MSCs) were intratracheally instilled into bleomycin-induced pulmonary fibrosis mice at day 3, and lung functions, cellular, molecular and pathological changes were assessed at 7 and 21 days after bleomycin administration.

Results: The expression of genes for pro-survival, anti-apoptotic, anti-oxidant and growth factors was upregulated in MSCs under hypoxic conditions. In transforming growth factor-beta 1-treated MRC-5 fibroblast cells, hypoxia-preconditioned MSCs attenuated extracellular matrix production through paracrine effects. The pulmonary respiratory functions significantly improved for up to 18 days of hypoxia-preconditioned MSC treatment. Expression of inflammatory factors and fibrotic factor were all downregulated in the lung tissues of the hypoxia-preconditioned MSC-treated mice. Histopathologic examination observed a significant amelioration of the lung fibrosis. Several LacZ-labeled MSCs were observed within the lungs in the hypoxia-preconditioned MSC treatment groups at day 21, but no signals were detected in the normoxic MSC group. Our data further demonstrated that upregulation of hepatocyte growth factor possibly played an important role in mediating the therapeutic effects of transplanted hypoxia-preconditioned MSCs.

Conclusion: Transplantation of hypoxia-preconditioned MSCs exerted better therapeutic effects in bleomycin-induced pulmonary fibrotic mice and enhanced the survival rate of engrafted MSCs, partially due to the upregulation of hepatocyte growth factor.

No MeSH data available.


Related in: MedlinePlus