Influences of donor and host age on human muscle-derived stem cell-mediated bone regeneration.
Influences of donor and host age on human muscle-derived stem cell-mediated bone regeneration.
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DOI：
10.1186/s13287-018-1066-z
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发表时间：
2018-11-21
影响因子：
7.5
通讯作者：
Huard J
中科院分区：
文献类型：
--
作者：
Gao X;Lu A;Tang Y;Schneppendahl J;Liebowitz AB;Scibetta AC;Morris ER;Cheng H;Huard C;Amra S;Wang B;Hall MA;Lowe WR;Huard J
人肌源性干细胞（hMDSC）已被证明在用慢病毒骨形态发生蛋白2（LBMP 2）转导时能有效地再生骨。然而，hMDSCs的年龄和动物宿主是否影响hMDSCs的骨再生能力及其机制尚不清楚，这促使了目前的研究。我们分离了三个性别匹配的年轻和老年骨骼肌干细胞群体，并在Lenti-BMP 2/绿色荧光蛋白（GFP）转导前后使用团块培养测试细胞年龄对体外成骨分化的影响。我们进一步研究了hMDSC和动物宿主的年龄对hMDSC介导的骨再生的影响，在一个临界大小的颅骨骨缺损模型在体内。采用微电脑断层扫描（CT）、组织学和免疫组化评价体外成骨分化和矿化以及体内骨再生。采用Western印迹、定量聚合酶链反应（PCR）和氧化应激试验检测年龄对hMDSC细胞存活和成骨相关基因的影响。采用酶联免疫吸附试验（ELISA）测定血清胰岛素样生长因子1（IGF 1）和核因子-κ B受体激活因子配体（RANKL）。我们发现LBMP 2/GFP转导显著增强了体外hMDSC的成骨分化，与供体年龄无关。我们还发现老年小鼠与年轻小鼠LBMP 2/GFP转导的hMDSC一样有效地再生年轻和老年小鼠的功能性骨。这些发现与较低的磷酸化p38 MAPK表达和相似的细胞存活基因和成骨相关基因的表达水平在老年hMDSC相对于年轻的hMDSC。老年细胞表现出相当的抗氧化应激。然而，年轻和年老的供体细胞在年老的宿主中再生的骨比年轻的宿主少。老年宿主的骨再生受损与较高的骨重建相关，因为RANKL血清水平较高，IGF-1水平较低。当用LBMP 2/GFP转导hMDSC时，hMDSC介导的骨再生不受供体年龄的损害，但宿主的年龄对hMDSC介导的骨再生产生不利影响。无论供体和宿主年龄如何，hMDSCs都能形成功能性骨，这表明hMDSCs是一种有前途的骨再生细胞资源。本文的在线版本（10.1186/s13287-018-1066-z）包含补充材料，可供授权用户使用。
Human muscle-derived stem cells (hMDSCs) have been shown to regenerate bone efficiently when they were transduced with Lenti-viral bone morphogenetic protein 2 (LBMP2). However, whether the age of hMDSCs and the animal host affect the bone regeneration capacity of hMDSCs and mechanism are unknown which prompted the current study. We isolated three gender-matched young and old populations of skeletal muscle stem cells, and tested the influence of cells’ age on in vitro osteogenic differentiation using pellet culture before and after Lenti-BMP2/green fluorescent protein (GFP) transduction. We further investigated effects of the age of hMDSCs and animal host on hMDSC-mediated bone regeneration in a critical-size calvarial bone defect model in vivo. Micro-computer tomography (CT), histology, and immunohistochemistry were used to evaluate osteogenic differentiation and mineralization in vitro and bone regeneration in vivo. Western blot, quantitative polymerase chain reaction (PCR), and oxidative stress assay were performed to detect the effects of age of hMDSCs on cell survival and osteogenic-related genes. Serum insulin-like growth factor 1 (IGF1) and receptor activator of nuclear factor-kappa B ligand (RANKL) were measured with an enzyme-linked immunosorbent assay (ELISA). We found LBMP2/GFP transduction significantly enhanced osteogenic differentiation of hMDSCs in vitro, regardless of donor age. We also found old were as efficient as young LBMP2/GFP-transduced hMDSCs for regenerating functional bone in young and old mice. These findings correlated with lower phosphorylated p38MAPK expression and similar expression levels of cell survival genes and osteogenic-related genes in old hMDSCs relative to young hMDSCs. Old cells exhibited equivalent resistance to oxidative stress. However, both young and old donor cells regenerated less bone in old than young hosts. Impaired bone regeneration in older hosts was associated with high bone remodeling due to higher serum levels of RANKL and lower level of IGF-1. hMDSC-mediated bone regeneration was not impaired by donor age when hMDSCs were transduced with LBMP2/GFP, but the age of the host adversely affected hMDSC-mediated bone regeneration. Regardless of donor and host age, hMDSCs formed functional bone, suggesting a promising cell resource for bone regeneration. The online version of this article (10.1186/s13287-018-1066-z) contains supplementary material, which is available to authorized users.
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影响因子：
10.5
作者：
Salama R;Sadaie M;Hoare M;Narita M
通讯作者：
Narita M
影响因子：
7.4
作者：
Choudhery MS;Badowski M;Muise A;Pierce J;Harris DT
通讯作者：
Harris DT
影响因子：
3.7
作者：
Beane OS;Fonseca VC;Cooper LL;Koren G;Darling EM
通讯作者：
Darling EM
影响因子：
14
作者：
Jing, Wei;Smith, Andrew A.;Helms, Jill A.
通讯作者：
Helms, Jill A.
影响因子：
14
作者：
Liu, Hen-Yu;Huang, Chiung-Fang;Deng, Win-Ping
通讯作者：
Deng, Win-Ping