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The Therapeutic Effects of Optimal Dose of Mesenchymal Stem Cells in a Murine Model of an Elastase Induced-Emphysema.

Kim YS, Kim JY, Huh JW, Lee SW, Choi SJ, Oh YM - Tuberc Respir Dis (Seoul) (2015)

Bottom Line: However, the optimal dose of mesenchymal stem cells (MSCs) for the most effective therapy has not yet been determined.But activity of matrix metalloproteinase 9 increased by emphysematous lung was decreased by intravenously injected MSCs.This is the first study on the optimal dose of MSCs as a therapeutic candidate.

View Article: PubMed Central - PubMed

Affiliation: Asan Institute for Life Sciences, Seoul, Korea. ; University of Ulsan College of Medicine, Seoul, Korea.

ABSTRACT

Background: Chronic obstructive pulmonary disease is characterized by emphysema, chronic bronchitis, and small airway remodeling. The alveolar destruction associated with emphysema cannot be repaired by current clinical practices. Stem cell therapy has been successfully used in animal models of cigarette smoke- and elastase-induced emphysema. However, the optimal dose of mesenchymal stem cells (MSCs) for the most effective therapy has not yet been determined. It is vital to determine the optimal dose of MSCs for clinical application in emphysema cases.

Methods: In the present study, we evaluated the therapeutic effects of various doses of MSCs on elastase-induced emphysema in mice. When 3 different doses of MSCs were intravenously injected into mice treated with elastase, only 5×10(4) MSCs showed a significant effect on the emphysematous mouse lung. We also identified action mechanisms of MSCs based on apoptosis, lung regeneration, and protease/antiprotease imbalance.

Results: The MSCs were not related with caspase-3/7 dependent apoptosis. But activity of matrix metalloproteinase 9 increased by emphysematous lung was decreased by intravenously injected MSCs. Vascular endothelial growth factor were also increased in lung from MSC injected mice, as compared to un-injected mice.

Conclusion: This is the first study on the optimal dose of MSCs as a therapeutic candidate. This data may provide important basic data for determining dosage in clinical application of MSCs in emphysema patients.

No MeSH data available.


Related in: MedlinePlus

The effects on protease and anti-protease in elastase induced emphysema with or without mesenchymal stem cells (MSCs). C57BL/6J mice were intratracheally applied with 0.4 U of elastase on day 0 and then intravenously injected with MSCs on day 7. Lung tissues were collected on day 14 (n=6-9 per group). (A) Matrix metalloproteinase (MMP) 2, MMP9, and MMP12 mRNA expression were measured with quantitative polymerase chain reaction (PCR) using SYBR Green and normalized by β-actin expression and then displayed with ratio to control group. Values are presented as the mean±SEM. *p<0.05, as compared with control group. (B) Activity of MMP2 and 9 were measured with gelatin zymography. (C) The mRNA expression of SLPI (left) and tissue inhibitor of metalloproteinases-1 (TIMP1) (right) were measured with quantitative PCR using SYBR Green and normalized by β-actin expression and then displayed with ratio to control group. PBS: phosphate buffered saline; Ela: elastase only; Ela+5×104: elastase+5×104 MSC.
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Figure 3: The effects on protease and anti-protease in elastase induced emphysema with or without mesenchymal stem cells (MSCs). C57BL/6J mice were intratracheally applied with 0.4 U of elastase on day 0 and then intravenously injected with MSCs on day 7. Lung tissues were collected on day 14 (n=6-9 per group). (A) Matrix metalloproteinase (MMP) 2, MMP9, and MMP12 mRNA expression were measured with quantitative polymerase chain reaction (PCR) using SYBR Green and normalized by β-actin expression and then displayed with ratio to control group. Values are presented as the mean±SEM. *p<0.05, as compared with control group. (B) Activity of MMP2 and 9 were measured with gelatin zymography. (C) The mRNA expression of SLPI (left) and tissue inhibitor of metalloproteinases-1 (TIMP1) (right) were measured with quantitative PCR using SYBR Green and normalized by β-actin expression and then displayed with ratio to control group. PBS: phosphate buffered saline; Ela: elastase only; Ela+5×104: elastase+5×104 MSC.

Mentions: To identify action mechanisms of MSCs in this model of elastase-induced emphysema, we obtained protein and mRNA in lung tissue from three of the mouse groups (control, elastase-only, and elastase- and 5×104 MSC-treated groups). The therapeutic mechanisms are divided in apoptosis, protease/antiprotease and regeneration by growth factor. First, we test caspase 3/7 dependent apoptosis. Caspase 3/7 activity to detect cellular apoptosis is not significantly different among the three groups (Figure 2). Second, we performed an evaluation based on protease/antiprotease imbalance in the same three groups. The mRNA expression of MMP2, MMP9, and MMP12 were measured using quantitative PCR, and data were presented as a ratio of the control group (Figure 3A). In the mouse group treated with elastase only, mRNA expression of MMP12 was higher than in the control group. The mouse group treated with both elastase and MSCs may be also higher than in the control group but there is not statistical difference. Next, we performed gelatin zymography to observe the activity of MMP2 and MMP9 (Figure 3B). The activity of MMP2 and MMP9 were induced in elastase only treated mouse group compared to the control group. In elastase and MSC treated mouse group, the MMP9 activity was decreased compared with the elastase only treated mouse group. The mRNA expression of SLPI and TIMP1 (representative antiprotease) were measured by quantitative PCR, and data were displayed as a ratio of the control group (Figure 3C). SLPI and TIMP1 expression in lung is not significantly different among the three groups. HGF, FGF2, and VEGF production related to the lung regeneration was also measured in lung homogenate using ELISA. HGF and FGF2 were not significantly different among the three groups (Figure 4A, B). The VEGF in lung tissue were decreased in elastase only treated mouse group compared to control mouse group and were recovered in elastase and MSCs treated mouse group (Figure 4C).


The Therapeutic Effects of Optimal Dose of Mesenchymal Stem Cells in a Murine Model of an Elastase Induced-Emphysema.

Kim YS, Kim JY, Huh JW, Lee SW, Choi SJ, Oh YM - Tuberc Respir Dis (Seoul) (2015)

The effects on protease and anti-protease in elastase induced emphysema with or without mesenchymal stem cells (MSCs). C57BL/6J mice were intratracheally applied with 0.4 U of elastase on day 0 and then intravenously injected with MSCs on day 7. Lung tissues were collected on day 14 (n=6-9 per group). (A) Matrix metalloproteinase (MMP) 2, MMP9, and MMP12 mRNA expression were measured with quantitative polymerase chain reaction (PCR) using SYBR Green and normalized by β-actin expression and then displayed with ratio to control group. Values are presented as the mean±SEM. *p<0.05, as compared with control group. (B) Activity of MMP2 and 9 were measured with gelatin zymography. (C) The mRNA expression of SLPI (left) and tissue inhibitor of metalloproteinases-1 (TIMP1) (right) were measured with quantitative PCR using SYBR Green and normalized by β-actin expression and then displayed with ratio to control group. PBS: phosphate buffered saline; Ela: elastase only; Ela+5×104: elastase+5×104 MSC.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4499592&req=5

Figure 3: The effects on protease and anti-protease in elastase induced emphysema with or without mesenchymal stem cells (MSCs). C57BL/6J mice were intratracheally applied with 0.4 U of elastase on day 0 and then intravenously injected with MSCs on day 7. Lung tissues were collected on day 14 (n=6-9 per group). (A) Matrix metalloproteinase (MMP) 2, MMP9, and MMP12 mRNA expression were measured with quantitative polymerase chain reaction (PCR) using SYBR Green and normalized by β-actin expression and then displayed with ratio to control group. Values are presented as the mean±SEM. *p<0.05, as compared with control group. (B) Activity of MMP2 and 9 were measured with gelatin zymography. (C) The mRNA expression of SLPI (left) and tissue inhibitor of metalloproteinases-1 (TIMP1) (right) were measured with quantitative PCR using SYBR Green and normalized by β-actin expression and then displayed with ratio to control group. PBS: phosphate buffered saline; Ela: elastase only; Ela+5×104: elastase+5×104 MSC.
Mentions: To identify action mechanisms of MSCs in this model of elastase-induced emphysema, we obtained protein and mRNA in lung tissue from three of the mouse groups (control, elastase-only, and elastase- and 5×104 MSC-treated groups). The therapeutic mechanisms are divided in apoptosis, protease/antiprotease and regeneration by growth factor. First, we test caspase 3/7 dependent apoptosis. Caspase 3/7 activity to detect cellular apoptosis is not significantly different among the three groups (Figure 2). Second, we performed an evaluation based on protease/antiprotease imbalance in the same three groups. The mRNA expression of MMP2, MMP9, and MMP12 were measured using quantitative PCR, and data were presented as a ratio of the control group (Figure 3A). In the mouse group treated with elastase only, mRNA expression of MMP12 was higher than in the control group. The mouse group treated with both elastase and MSCs may be also higher than in the control group but there is not statistical difference. Next, we performed gelatin zymography to observe the activity of MMP2 and MMP9 (Figure 3B). The activity of MMP2 and MMP9 were induced in elastase only treated mouse group compared to the control group. In elastase and MSC treated mouse group, the MMP9 activity was decreased compared with the elastase only treated mouse group. The mRNA expression of SLPI and TIMP1 (representative antiprotease) were measured by quantitative PCR, and data were displayed as a ratio of the control group (Figure 3C). SLPI and TIMP1 expression in lung is not significantly different among the three groups. HGF, FGF2, and VEGF production related to the lung regeneration was also measured in lung homogenate using ELISA. HGF and FGF2 were not significantly different among the three groups (Figure 4A, B). The VEGF in lung tissue were decreased in elastase only treated mouse group compared to control mouse group and were recovered in elastase and MSCs treated mouse group (Figure 4C).

Bottom Line: However, the optimal dose of mesenchymal stem cells (MSCs) for the most effective therapy has not yet been determined.But activity of matrix metalloproteinase 9 increased by emphysematous lung was decreased by intravenously injected MSCs.This is the first study on the optimal dose of MSCs as a therapeutic candidate.

View Article: PubMed Central - PubMed

Affiliation: Asan Institute for Life Sciences, Seoul, Korea. ; University of Ulsan College of Medicine, Seoul, Korea.

ABSTRACT

Background: Chronic obstructive pulmonary disease is characterized by emphysema, chronic bronchitis, and small airway remodeling. The alveolar destruction associated with emphysema cannot be repaired by current clinical practices. Stem cell therapy has been successfully used in animal models of cigarette smoke- and elastase-induced emphysema. However, the optimal dose of mesenchymal stem cells (MSCs) for the most effective therapy has not yet been determined. It is vital to determine the optimal dose of MSCs for clinical application in emphysema cases.

Methods: In the present study, we evaluated the therapeutic effects of various doses of MSCs on elastase-induced emphysema in mice. When 3 different doses of MSCs were intravenously injected into mice treated with elastase, only 5×10(4) MSCs showed a significant effect on the emphysematous mouse lung. We also identified action mechanisms of MSCs based on apoptosis, lung regeneration, and protease/antiprotease imbalance.

Results: The MSCs were not related with caspase-3/7 dependent apoptosis. But activity of matrix metalloproteinase 9 increased by emphysematous lung was decreased by intravenously injected MSCs. Vascular endothelial growth factor were also increased in lung from MSC injected mice, as compared to un-injected mice.

Conclusion: This is the first study on the optimal dose of MSCs as a therapeutic candidate. This data may provide important basic data for determining dosage in clinical application of MSCs in emphysema patients.

No MeSH data available.


Related in: MedlinePlus