Adipose Mesenchymal Stem Cell Secretome Modulated in Hypoxia for Remodeling of Radiation-Induced Salivary Gland Damage
Adipose Mesenchymal Stem Cell Secretome Modulated in Hypoxia for Remodeling of Radiation-Induced Salivary Gland Damage
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DOI：
10.1371/journal.pone.0141862
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发表时间：
2015-11-03
期刊：
影响因子：
3.7
通讯作者：
Kim, Young-Mo
中科院分区：
文献类型：
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作者：
An, Hye-Young;Shin, Hyun-Soo;Kim, Young-Mo
背景和目的本研究旨在确定经低氧条件调节以包含治疗因子的来自间充质干细胞（MSC）的分泌组是否有助于唾液腺（SG）组织重塑并具有改善小鼠模型中辐射（IR）诱导的唾液功能减退的潜力。并暴露于缺氧条件（O-2 < 5%）。然后将缺氧条件培养基过滤成高分子量部分并制备为hAdMSC分泌蛋白组。随后在局部IR后立即将hAdMSC分泌蛋白组输注到C3 H小鼠的尾静脉中，每天一次，连续七天。对照组仅接受等体积（500 μ L）的溶媒（PBS）。研究了hAdMSC分泌组的SG功能和结构组织重塑。人腮腺上皮细胞（HPEC），获得，在体外扩增，然后照射和处理的缺氧条件培养基或常氧对照培养基。细胞增殖和IR诱导的细胞死亡进行了检查，以确定hAdMSC分泌组发挥其effects.ResultsThe条件hAdMSC分泌组含有高水平的GM-CSF，VEGF，IL-6，和IGF-1的机制。与PBS组相比，反复全身输注hAdMSC分泌蛋白质组导致唾液分泌能力改善和唾液蛋白质水平增加，包括淀粉酶和EGF。SG的显微结构完整性得以保持，唾液上皮细胞（AQP-5）、内皮细胞（CD 31）、肌上皮细胞（α-SMA）和SG祖细胞（c-Kit）成功地受到保护，免受辐射损伤并重塑。hAdMSC分泌组强烈诱导HPEC的增殖，并导致体内和体外细胞死亡的显着减少。此外，hAdMSC分泌组的抗凋亡作用被发现后，促进缺氧预处理相对于normoxia培养的hAdMSC secretome.ConclusionThese结果表明，hAdMSC分泌组从缺氧条件培养基可能提供辐射保护和组织重塑通过释放旁分泌介质。
Background and PurposeThis 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 MethodsHuman adipose mesenchymal stem cells (hAdMSC) were isolated, expanded, and exposed to hypoxic conditions (O-2 < 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 mu L) 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.ResultsThe 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 (alpha-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.ConclusionThese results show that the hAdMSC secretome from hypoxic-conditioned medium may provide radioprotection and tissue remodeling via release of paracrine mediators.