1. 四川大学华西广安医院烧伤整形科,四川,广安,638001
2. 陆军军医大学第一附属医院整形美容外科,重庆,400038
Published:2019,
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REN Zhangxia, LI Fan, YANG Huijia, et al. Study on keloid tumor growth affected by osteoblast-specific factor 2 interacting with mesenchymal stem cells. [J]. null (9):695(2019)
目的 探讨骨母细胞特异性因子2(Periostin)调控 瘢痕疙瘩间充质干细胞(KMLSCs)对体外及裸鼠体内瘢痕疙瘩样瘤体生长的影响。方法 从瘢痕疙瘩中分离培养KMLSCs,设置实验Periostin干扰组、空载体组和未转染组,根据干预目的构建Periostin慢病毒载体并转染对应组细胞。利用Western Blot技术体外检测各组细胞Periostin、α平滑肌肌动蛋白(α-SMA)和血管内皮生长因子(VEGF)的表达情况;制备丝素蛋白-壳聚糖支架复合体,与细胞共培养后,扫描电镜观察支架的结构、KMLSCs的生长情况。建立裸鼠皮下瘢痕疙瘩样瘤体移植模型,移植8周后取出瘤体,测量3组肿瘤体积,通过荧光共聚焦技术检测各组组织中Periostin、α-SMA、VEGF以及Western Blot技术评估Erk的表达情况。结果 体外培养的3组KMLSCs中Periostin在空载体组和未转染组有表达,而干扰组则表达微弱;同时3组细胞均不表达αSMA和VEGF。制备的支架复合体呈多孔结构,KMLSCs在复合体支架中生长良好;各分组瘤体移植模型在体内裸鼠皮下成活良好。干扰组中Periostin表达下调后,瘤体体积显著减小,同时α-SMA、VEGF、Erk表达减弱,差异有统计学意义(P<0.05),并呈现一致性。结论 Periostin可能通过调控KMLSCs的功能,参与了模型中瘢痕疙瘩样肿块的生长。
Objective To investigate the effect of osteoblastspecific factor 2 (OSF2
Periostin) on keloid mesenchymal stem cells(KMLSCs)growth in vitro through nude mice model. Methods KMLSCs was isolated from keloid.The experiments were divided into three groups:the Periostin group
the blankvector group and the nontransfected group. The Periostin lentiviral vector were constructed and infected into the corresponding KMLSCs. The expression of Periostin
alpha smooth muscle actin (α-SMA)
and vascular endothelial growth factor(VEGF) were determined in vitro by Western blotting.Chitosansilk fibroin porpous scaffolds were made through freeze drying.The KMLSCs were cocultured with the scaffolds
and the cellscaffolds and the growth of cells were detected by scanning electron microscopy.The cellscaffold complexes were subcutaneously implanted into the nude mices.After 8 weeeks the mices were killed to measure the volume of the keloid.The expression of Periostin
αSMA
and VEGF in keloid were detected by immunofluorescent staining
and the expression of Erk was detected by Western blotting. Results In the blankvector group and the nontransfected group
the Periostin were expressed normally in KMLSCs
but it was Weakly expressed in the Periostin group.And the αSMA and VEGF were not expressed in KMLSCs cultured in vitro.The cells grew well on composite scaffolds.Tumor transplantation models of different groups survived well subcutaneously in vivo in nude mice.However
when the expression of Periostin were downregulated
the volume of keloid was statistically smaller(P<0.05)
and the expression of αSMA
VEGF and Erk siginificantly attenuated(P<0.05). Conclusion Periostin can regulate the functions of KMLSCs in vivo and promote the growth of keloid in nude mice.
骨母细胞特异性因子2间质干细胞瘢痕疙瘩α-SMAVEGF
osteoblastspecific factor 2(OSF2Periostin)mesenchymal stem cellkeloidαSMAvascular endothelial growth factor
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