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Adipose Mesenchymal Stem Cell Secretome Modulated in Hypoxia for Remodeling of Radiation-Induced Salivary Gland Damage.

An HY, Shin HS, Choi JS, Kim HJ, Lim JY, Kim YM - PLoS ONE (2015)

Bottom Line: Repeated systemic infusion with the hAdMSC secretome resulted in improved salivation capacity and increased levels of salivary proteins, including amylase and EGF, relative to the PBS group.The hAdMSC secretome strongly induced proliferation of HPEC and led to a significant decrease in cell death in vivo and in vitro.Moreover, the anti-apoptotic effects of the hAdMSC secretome were found to be promoted after hypoxia-preconditioning relative to normoxia-cultured hAdMSC secretome.

View Article: PubMed Central - PubMed

Affiliation: Department of Otorhinolaryngology-Head and Neck Surgery, Inha University College of Medicine, Incheon, Republic of Korea; Translational Research Center, Inha University College of Medicine, Incheon, Republic of Korea.

ABSTRACT

Background and purpose: This study was conducted to determine whether a secretome from mesenchymal stem cells (MSC) modulated by hypoxic conditions to contain therapeutic factors contributes to salivary gland (SG) tissue remodeling and has the potential to improve irradiation (IR)-induced salivary hypofunction in a mouse model.

Materials and methods: Human adipose mesenchymal stem cells (hAdMSC) were isolated, expanded, and exposed to hypoxic conditions (O2 < 5%). The hypoxia-conditioned medium was then filtered to a high molecular weight fraction and prepared as a hAdMSC secretome. The hAdMSC secretome was subsequently infused into the tail vein of C3H mice immediately after local IR once a day for seven consecutive days. The control group received equal volume (500 μL) of vehicle (PBS) only. SG function and structural tissue remodeling by the hAdMSC secretome were investigated. Human parotid epithelial cells (HPEC) were obtained, expanded in vitro, and then irradiated and treated with either the hypoxia-conditioned medium or a normoxic control medium. Cell proliferation and IR-induced cell death were examined to determine the mechanism by which the hAdMSC secretome exerted its effects.

Results: The conditioned hAdMSC secretome contained high levels of GM-CSF, VEGF, IL-6, and IGF-1. Repeated systemic infusion with the hAdMSC secretome resulted in improved salivation capacity and increased levels of salivary proteins, including amylase and EGF, relative to the PBS group. The microscopic structural integrity of SG was maintained and salivary epithelial (AQP-5), endothelial (CD31), myoepithelial (α-SMA) and SG progenitor cells (c-Kit) were successfully protected from radiation damage and remodeled. The hAdMSC secretome strongly induced proliferation of HPEC and led to a significant decrease in cell death in vivo and in vitro. Moreover, the anti-apoptotic effects of the hAdMSC secretome were found to be promoted after hypoxia-preconditioning relative to normoxia-cultured hAdMSC secretome.

Conclusion: These results show that the hAdMSC secretome from hypoxic-conditioned medium may provide radioprotection and tissue remodeling via release of paracrine mediators.

No MeSH data available.


Related in: MedlinePlus

Recovery of salivary hypofunction by hAdMSC secretome infusion.(A) Salivary flow rate (SFR) was calculated at pre-IR and 4 and 16 weeks post-IR. The changes in SFR after IR were expressed by the ratio of post-IR SFR to pre-IR SFR (Mean ± SEM). (B) Time to salivation (lag time, LT) was measured and the ratios of post-IR LT to pre-IR LT were presented. (C) Western blotting of amylase in saliva at 2 and 4 weeks post-IR. (D) The salivary amylase activity was examined by the Assay Kit and fold changes in activity level are presented. (E) EGF contents were measured at each time point and the average contents are presented. Data are presented as the mean ± SEM. Two-way ANOVA, Bonferroni post hoc test (A and B), one-way ANOVA, Tukey’s pot hoc test (D and E). *, compared to CON; #, compared to IR + PBS. *P <0.05, **P < .01, ***P <0.001, #P < 0.05, and ##P < 0.01. CON, normal control group (n = 5–7); IR + PBS, PBS-treated group (n = 5–6); IR + SEC, secretome-treated group (n = 5–6).
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pone.0141862.g004: Recovery of salivary hypofunction by hAdMSC secretome infusion.(A) Salivary flow rate (SFR) was calculated at pre-IR and 4 and 16 weeks post-IR. The changes in SFR after IR were expressed by the ratio of post-IR SFR to pre-IR SFR (Mean ± SEM). (B) Time to salivation (lag time, LT) was measured and the ratios of post-IR LT to pre-IR LT were presented. (C) Western blotting of amylase in saliva at 2 and 4 weeks post-IR. (D) The salivary amylase activity was examined by the Assay Kit and fold changes in activity level are presented. (E) EGF contents were measured at each time point and the average contents are presented. Data are presented as the mean ± SEM. Two-way ANOVA, Bonferroni post hoc test (A and B), one-way ANOVA, Tukey’s pot hoc test (D and E). *, compared to CON; #, compared to IR + PBS. *P <0.05, **P < .01, ***P <0.001, #P < 0.05, and ##P < 0.01. CON, normal control group (n = 5–7); IR + PBS, PBS-treated group (n = 5–6); IR + SEC, secretome-treated group (n = 5–6).

Mentions: The amount of salivation and lag time were measured pre-IR and 4 and 16 weeks post-IR (Fig 4A and 4B). The changes in SFR and lag time after IR were calculated by the post-IR to pre-IR ratio. IR significantly decreased the ratio of SFR and increased the ratio of the lag time at 16 weeks after IR (Fig 4A, P < 0.001 and P < 0.01, respectively). hAdMSC infusion significantly improved the ratio of SFR at 16 weeks after IR (Fig 4A, P < 0.01) and the lag time at 4 weeks after IR relative to the PBS group (Fig 4B, P < 0.05).


Adipose Mesenchymal Stem Cell Secretome Modulated in Hypoxia for Remodeling of Radiation-Induced Salivary Gland Damage.

An HY, Shin HS, Choi JS, Kim HJ, Lim JY, Kim YM - PLoS ONE (2015)

Recovery of salivary hypofunction by hAdMSC secretome infusion.(A) Salivary flow rate (SFR) was calculated at pre-IR and 4 and 16 weeks post-IR. The changes in SFR after IR were expressed by the ratio of post-IR SFR to pre-IR SFR (Mean ± SEM). (B) Time to salivation (lag time, LT) was measured and the ratios of post-IR LT to pre-IR LT were presented. (C) Western blotting of amylase in saliva at 2 and 4 weeks post-IR. (D) The salivary amylase activity was examined by the Assay Kit and fold changes in activity level are presented. (E) EGF contents were measured at each time point and the average contents are presented. Data are presented as the mean ± SEM. Two-way ANOVA, Bonferroni post hoc test (A and B), one-way ANOVA, Tukey’s pot hoc test (D and E). *, compared to CON; #, compared to IR + PBS. *P <0.05, **P < .01, ***P <0.001, #P < 0.05, and ##P < 0.01. CON, normal control group (n = 5–7); IR + PBS, PBS-treated group (n = 5–6); IR + SEC, secretome-treated group (n = 5–6).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4631328&req=5

pone.0141862.g004: Recovery of salivary hypofunction by hAdMSC secretome infusion.(A) Salivary flow rate (SFR) was calculated at pre-IR and 4 and 16 weeks post-IR. The changes in SFR after IR were expressed by the ratio of post-IR SFR to pre-IR SFR (Mean ± SEM). (B) Time to salivation (lag time, LT) was measured and the ratios of post-IR LT to pre-IR LT were presented. (C) Western blotting of amylase in saliva at 2 and 4 weeks post-IR. (D) The salivary amylase activity was examined by the Assay Kit and fold changes in activity level are presented. (E) EGF contents were measured at each time point and the average contents are presented. Data are presented as the mean ± SEM. Two-way ANOVA, Bonferroni post hoc test (A and B), one-way ANOVA, Tukey’s pot hoc test (D and E). *, compared to CON; #, compared to IR + PBS. *P <0.05, **P < .01, ***P <0.001, #P < 0.05, and ##P < 0.01. CON, normal control group (n = 5–7); IR + PBS, PBS-treated group (n = 5–6); IR + SEC, secretome-treated group (n = 5–6).
Mentions: The amount of salivation and lag time were measured pre-IR and 4 and 16 weeks post-IR (Fig 4A and 4B). The changes in SFR and lag time after IR were calculated by the post-IR to pre-IR ratio. IR significantly decreased the ratio of SFR and increased the ratio of the lag time at 16 weeks after IR (Fig 4A, P < 0.001 and P < 0.01, respectively). hAdMSC infusion significantly improved the ratio of SFR at 16 weeks after IR (Fig 4A, P < 0.01) and the lag time at 4 weeks after IR relative to the PBS group (Fig 4B, P < 0.05).

Bottom Line: Repeated systemic infusion with the hAdMSC secretome resulted in improved salivation capacity and increased levels of salivary proteins, including amylase and EGF, relative to the PBS group.The hAdMSC secretome strongly induced proliferation of HPEC and led to a significant decrease in cell death in vivo and in vitro.Moreover, the anti-apoptotic effects of the hAdMSC secretome were found to be promoted after hypoxia-preconditioning relative to normoxia-cultured hAdMSC secretome.

View Article: PubMed Central - PubMed

Affiliation: Department of Otorhinolaryngology-Head and Neck Surgery, Inha University College of Medicine, Incheon, Republic of Korea; Translational Research Center, Inha University College of Medicine, Incheon, Republic of Korea.

ABSTRACT

Background and purpose: This study was conducted to determine whether a secretome from mesenchymal stem cells (MSC) modulated by hypoxic conditions to contain therapeutic factors contributes to salivary gland (SG) tissue remodeling and has the potential to improve irradiation (IR)-induced salivary hypofunction in a mouse model.

Materials and methods: Human adipose mesenchymal stem cells (hAdMSC) were isolated, expanded, and exposed to hypoxic conditions (O2 < 5%). The hypoxia-conditioned medium was then filtered to a high molecular weight fraction and prepared as a hAdMSC secretome. The hAdMSC secretome was subsequently infused into the tail vein of C3H mice immediately after local IR once a day for seven consecutive days. The control group received equal volume (500 μL) of vehicle (PBS) only. SG function and structural tissue remodeling by the hAdMSC secretome were investigated. Human parotid epithelial cells (HPEC) were obtained, expanded in vitro, and then irradiated and treated with either the hypoxia-conditioned medium or a normoxic control medium. Cell proliferation and IR-induced cell death were examined to determine the mechanism by which the hAdMSC secretome exerted its effects.

Results: The conditioned hAdMSC secretome contained high levels of GM-CSF, VEGF, IL-6, and IGF-1. Repeated systemic infusion with the hAdMSC secretome resulted in improved salivation capacity and increased levels of salivary proteins, including amylase and EGF, relative to the PBS group. The microscopic structural integrity of SG was maintained and salivary epithelial (AQP-5), endothelial (CD31), myoepithelial (α-SMA) and SG progenitor cells (c-Kit) were successfully protected from radiation damage and remodeled. The hAdMSC secretome strongly induced proliferation of HPEC and led to a significant decrease in cell death in vivo and in vitro. Moreover, the anti-apoptotic effects of the hAdMSC secretome were found to be promoted after hypoxia-preconditioning relative to normoxia-cultured hAdMSC secretome.

Conclusion: These results show that the hAdMSC secretome from hypoxic-conditioned medium may provide radioprotection and tissue remodeling via release of paracrine mediators.

No MeSH data available.


Related in: MedlinePlus