PRF及所含三因子对大鼠脂肪干细胞迁移的影响

上海口腔医学 ›› 2015, Vol. 24 ›› Issue (6): 667-673.

PRF及所含三因子对大鼠脂肪干细胞迁移的影响

高洁¹，王明国²，杨帅²，李秀梅³，杨世茂²，李雪⁴

1.解放军济南军区第401医院口腔科，山东青岛266071；
2.济南市中心医院口腔科，山东济南250013；
3.济南市人民政府机关门诊部口腔科，山东济南250099；
4.潍坊医学院口腔医院，山东潍坊261053

收稿日期:2015-05-19 出版日期:2015-12-25 发布日期:2016-01-04
通讯作者: 王明国，Tel:0531-85695171，E-mail：wmgsh@163.com E-mail:gaojierizhao@126.com
作者简介:高洁1987-，女，硕士，住院医师
基金资助:
Supported by Science and Technology Development Program of Shandong Province (2014GSF11804).

Effects of PRF and released three growth factors on migration of rat adipose tissue-derived stem cells

GAO Jie¹, WANG Ming-guo², YANG Shuai², LI Xiu-mei³, YANG Shi-mao², LI Xue⁴.

1.Department of Stomatology, the 401st Hospital of Jinan Military Area, PLA.Qingdao 266071;
2.Department of Stomatology, Jinan Central Hospital. Jinan 250013;
3.Department of Stomatology, Jinan Municipal People’s Government. Jinan 250099;
4.Hospital of Stomatology, Weifang Medical University. Weifang 261053, Shandong Province, China

Received:2015-05-19 Online:2015-12-25 Published:2016-01-04
Contact: 山东省科技发展项目（2014GSF11804） E-mail:gaojierizhao@126.com

摘要/Abstract

摘要： 目的研究富血小板纤维蛋白(platelet-rich fibrin,PRF)及所含三因子TGF-β1、PDGF-AB、 VEGF对培养的大鼠脂肪干细胞(adipose tissue-derived stem cell,ADSCs)迁移的影响，并初步探讨其机制。方法无菌切除SD大鼠腹股沟处的白色脂肪组织，采用酶消化法原代培养SD大鼠ADSCs，多向诱导分化法鉴定ADSCs，一次离心法提取制备PRF膜。采用划痕实验和Transwell实验检测细胞的迁移能力，实时荧光定量PCR分析迁移相关因子MT1-MMP和MMP-2 mRNA 的表达情况。采用SPSS13.0软件包对数据进行统计学分析。结果 Transwell实验显示，PRF组的迁移细胞数显著高于阴性对照组（P＜0.05）和抑制剂组（P＜0.05）；不同浓度的TGFβ1、PDGF-AB、VEGF组的组内迁移细胞数的差异显著（P﹤0.05）。经PCR检测，PRF组细胞基因MMP2和MT1-MMP较对照组的表达显著上调（P＜0.05），TGF-β1、 PDGF-AB和 VEGF的细胞基因MMP2和MT1-MMP较对照组表达均上调，组间差异显著（P＜0.05）。结论 PRF促进了ADSCs的迁移，PRF所分泌的三因子均可不同程度地促进ADSCs的迁移，并呈一定的量-效关系，其迁移能力的增加可能与MT1-MMP和MMP-2 mRNA的上调密切相关。

关键词: 富血小板纤维蛋白, 脂肪干细胞, 细胞迁移, 转化生长因子, 血小板源性生长因子AB, 血管内皮细胞生长因子

Abstract: PURPOSE To analyze the effects of PRF and released three growth factors on migration of rat adipose tissue-derived stem cells and to investigate the mechanism of migration. METHODS The inguinal adipose tissue of rat was excised at aseptic condition to obtain primary ADSCs by enzyme digestion. Multi-directional differentiation was used to identify the ADSCs. PRF membrane was acquired through one time centrifuge. The cell migration was examined by Transwell assay and wound healing assay. The mRNA expression of MMP2 and MT1-MMP was tested by real-time PCR. Statistical analysis was performed using SPSS 13.0 software package. RESULTS Cell migration test showed that the migration of rat ADSCs in PRF group were significantly higher than those in the negative group（P＜0.05） and inhibitor group（P＜0.05）. The ADSCs migration effects in three growth factors group at different concentrations showed significant difference（P＜0.05）. Real-time PCR showed that gene expressions of MMP2 and MT1-MMP were significantly higher in PRF group than control group (P＜0.05). PCR showed that gene expressions of MMP2 and MT1-MMP were significantly higher in three growth factors group than control group (P＜0.05). CONCLUTIONS: PRF and three growth factors consistently enhanced the migration of rat ADSCs in a dose-response manner. The migration increase of rat ADSCs may be associated with the up-regulation of MMP2 and MT1-MMP gene expression.

Key words: PRF, ADSCs, Migration, TGF-β1, PDGF-AB, VEGF

中图分类号:

R782

引用本文

高洁，王明国，杨帅，李秀梅，杨世茂，李雪. PRF及所含三因子对大鼠脂肪干细胞迁移的影响[J]. 上海口腔医学, 2015, 24(6): 667-673.

GAO Jie, WANG Ming-guo, YANG Shuai, LI Xiu-mei, YANG Shi-mao, LI Xue.. Effects of PRF and released three growth factors on migration of rat adipose tissue-derived stem cells[J]. Shanghai Journal of Stomatology, 2015, 24(6): 667-673.

使用本文

/ 推荐

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://shkqyx.magtechjournal.com/CN/

https://shkqyx.magtechjournal.com/CN/Y2015/V24/I6/667

参考文献

[1] Zuk PA, Zhu M, Ashjian P, et al. Human adipose tissue is a source of multipotent stem cells [J]. Mol Biol Cell, 2002, 13(12): 4279-4295.
[2] Choukroun J, Adda F, Schoeffler C, et al. Une opportunitéen paro-implantologie: Le PRF[J]. Implantodontie, 2001, 42: 55-62.
[3] Dohan Ehrenfest DM, de Peppo GM, Doglioli P, et al. Slow release of growth facto-rs and thrombospondin-1 in Choukroun’s platelet-rich fibrin (PRF): a gold standard t-o achieve for all surgical platelet concentrates tehnologies [J]. Growth Factors, 2009,27(1): 63-69.
[4] Lee JW, Kim SG, Kim JY, et al. Restoration of a peri-implant defect by platelet-richfibrin [J]. Oral Surg Oral Med Oral Pathol Oral Radiol, 2012, 113(4): 459-463.
[5] Toffler M, Toscano N, Holtzclaw D. Osteotome-mediated sinus floor elevation using only platelet-rich fibrin: an early report on 110 patients [J]. Implant Dent, 2010, 19(5): 447-456.
[6] Peck MT, Marnewick J, Stephen L. Alveolar ridge preservation using leukocyte and platelet-rich fibrin: a report of a case [J].Case Rep Dent, 2011, 2011: 345048.
[7] Chen FM, Wu LA, Zhang M, et al. Homing of endogenous stem/progenitor cells for in situ tissue regeneration: Promises, strategies, and translational perspectives [J]. Biomaterials, 2011, 32(12): 3189-3209.
[8] 杨世茂,王明国, 李静. 富血小板纤维蛋白与富血小板血浆体外释放生长因子的比较及其对脂肪干细胞增殖分化的影响 [J]. 华西口腔医学杂志, 2012, 30(6): 641-644.
[9] Ji B, Sheng L, Chen G, et al. The combination use of platelet-rich fibrin and treated dentin matrix for tooth root regeneration by cell homing [J]. Tissue Eng Part A, 2015,21(1-2): 26-34.
[10] 范文娟, 杨明, 张晨, 等. 富血小板纤维蛋白对人牙龈成纤维细胞增殖、迁移和分泌Ι型胶原的影响 [J]. 中华口腔医学杂志, 2013, 48(2): 72-76.
[11] Chang IC, Tsai CH, Chang YC. Platelet-rich fibrin modulates the expression of extr-acellular signal-regulated protein kinase and osteoprotegerin in human osteoblasts [J]. J Biomed Mater Res A, 2010, 95(1): 327-332.
[12] Tang J, Wang J, Kong X, et al. Vascular endothelial growth factor promotes cardiac stem cell migration via the PI3K/Akt pathway [J]. Exp Cell Res, 2009, 315(20): 3521-3531.
[13] Ries C, Egea V, Karow M, et al. MMP-2, MT1-MMP, and TIMP-2 are essential for the invasive capacity of human mesenchymal stem cells: differential regulation by inflammatory cytokines [J]. Blood, 2007, 109(9): 4055-4063.
[14] Cho A, Reidy MA. Matrix metalloproteinase-9 is necessary for the regulation of smooth muscle cell replication and migration after arterial injury [J]. Circ Res, 2002,91(9): 845-851.
[15] Matter CM, Rozenberg I, Jaschko A, et al. Effects of tacrolimus or sirolimus on proliferation of vascular smooth muscle and endothelial cells [J]. J Cardiovasc Pharmacol, 2006, 48(6): 286-292.
[16] Kamath AT, Rochat AF, Valenti MP, et al. Adult-like anti-mycobacterial T cell and in vivo dendritic cell responses following neonatal immunization with Ag85B-ESAT-6 in the IC31 adjuvant [J]. PloS One, 2008, 3(11): e3683.

相关文章 15

[1] 白月辉, 刘阳, 崔玉兰, 姜杉杉, 商庆龙, 赵琛. 慢性睡眠剥夺对大鼠髁突软骨的影响[J]. 上海口腔医学, 2022, 31(2): 148-155.

[2] 徐方芳, 蒋备战. DKK1对脂多糖感染人牙髓细胞生物学行为的影响[J]. 上海口腔医学, 2021, 30(4): 355-359.

[3] 朱陈元, 徐玲, 于卫强. 小檗碱通过JNK信号通路调控大鼠脂肪干细胞成骨向分化[J]. 上海口腔医学, 2021, 30(3): 258-262.

[4] 黄稍稍, 谭荣才, 邝晓岚. Bio-Oss骨粉联合富血小板纤维蛋白在牙槽骨缺损种植引导骨再生后的骨量变化[J]. 上海口腔医学, 2020, 29(4): 427-430.

[5] 吴峥嵘, 左园林, 李朝晖. 自体牙本质颗粒结合富血小板纤维蛋白膜治疗93例下颌第一磨牙根分叉病变效果评价[J]. 上海口腔医学, 2020, 29(2): 213-216.

[6] 谢慧, 解永富, 刘勤, 商玲燕, 陈敏箴. 注射型富血小板纤维蛋白在上颌窦外提升术中的应用效果[J]. 上海口腔医学, 2019, 28(1): 71-75.

[7] 郭延伟, 杨世茂. TGF-β3转染脂肪干细胞复合OGP-HA-ChS支架对兔髁突受损软骨的修复作用[J]. 上海口腔医学, 2018, 27(6): 567-573.

[8] 沈敏华, 黄远亮, 厉祯, 张迎娣, 何燕萍, 王磊. 富血小板纤维蛋白治疗Ⅱ度根分叉病变的效果评价[J]. 上海口腔医学, 2018, 27(5): 508-512.

[9] 越杨, 陈卓, 谢冰, 姚海亮, 张娴, 闫利辉, 张志华. 正畸牙移动中牙周组织VEGF的表达及在骨改建中的作用机制探讨[J]. 上海口腔医学, 2018, 27(1): 18-21.

[10] 王贵玲, 于雅琼, 郭佳杰, 仇丽鸿. 转化生长因子β3对脂多糖诱导的成骨细胞中IL-6表达的影响[J]. 上海口腔医学, 2017, 26(1): 21-25.

[11] 浦光瑞, 刘法昱, 王博. 牛蒡苷元对人口腔鳞癌裸鼠移植瘤生长及VEGF蛋白表达的影响[J]. 上海口腔医学, 2015, 24(4): 400-403.

[12] 张疆弢, 梅梅, 江策, 丰雷, 黄瑾, 王家佳. rhPDGF-BB与rhTGF-β₁联合应用对大鼠正畸牙破骨细胞 FAK蛋白表达的影响[J]. 上海口腔医学, 2015, 24(4): 423-427.

[13] 何璇,韦维,陈文霞. 富血小板纤维蛋白凝胶三维结构及其对人牙髓细胞体外增殖的影响[J]. 上海口腔医学, 2015, 24(3): 263-268.

[14] 张疆弢,梅梅,李平,张跃蓉,江策,丰雷,黄瑾,王家佳. 丹参对大鼠正畸牙牙周组织TGF-β₁表达的影响[J]. 上海口腔医学, 2015, 24(3): 288-293.

[15] 孙晓琳,周延民,赵静辉,郑玲,杨婷婷. 富血小板纤维蛋白对体外培养的成骨细胞生物学特性的影响[J]. 上海口腔医学, 2015, 24(1): 61-64.

编辑推荐

Metrics

阅读次数

全文

摘要

本文评价

摘要

参考文献

相关文章

编辑推荐

Metrics

本文评价

回顶部