Chondrogenesis of adipose stem cells in a porous PLGA scaffold impregnated with plasmid DNA containing SOX trio (SOX-5,-6 and-9) genes
Chondrogenesis of adipose stem cells in a porous PLGA scaffold impregnated with plasmid DNA containing SOX trio (SOX-5,-6 and-9) genes
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DOI：
10.1016/j.biomaterials.2011.02.054
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发表时间：
2011-07-01
期刊：
影响因子：
14
通讯作者：
Lee, Jin H.
中科院分区：
文献类型：
--
作者：
Im, Gun-Ii;Kim, Hye-Joung;Lee, Jin H.
我们开发了一种软骨形成支架系统，其中将含有SOX三重（SOX-5、-6和-9）基因的质粒DNA（pDNA）掺入PLGA支架中并缓慢释放以抑制接种在支架中的脂肪干细胞（ASC）。本研究的目的是测试该系统在体外和体内诱导ASCs软骨分化的功效。通过沉淀/颗粒沥滤法制备pDNA/PEI-PEG复合物掺入的PLGA/Pluronic F127多孔支架。将以下五种pDNA掺入支架中：1）没有插入基因的pECFP-C1载体（对照组）; 2）SOX-5质粒; 3）SOX-6质粒; 4）SOX-9质粒;和5）三分之一剂量的每种质粒（SOX-5、-6和-9）。将ASC接种在掺入pDNA的PLGA支架上，并在软骨形成培养基中培养21天。也从兔子中分离出ASC，将其接种在掺入了pDNA的PLGA支架中，然后植入在髌骨沟上产生的骨软骨缺损中。植入后8周处死家兔，进行大体和显微镜分析。转染细胞的百分比在第14天最高，约70%。21天后，与每个基因结合的PLGA支架显示出相应基因和蛋白质的表达显著增加。糖胺聚糖（GAG）测定和番红-O染色显示SOX trio pDNA掺入支架中蛋白聚糖的产生增加。COL 2A 1基因和蛋白在SOX trio pDNA掺入的支架中比对照中显著增加，而COL 10A 1蛋白表达降低。体内研究的大体和组织学结果显示，ASCs/SOX三重pDNA掺入PLGA支架中的软骨再生增强。(C)2011爱思唯尔有限公司保留所有权利。
We developed a chondrogenic scaffold system in which plasm id DNA (pDNA) containing SOX trio (SOX-5, -6, and -9) genes was incorporated into a PLGA scaffold and slowly released to transfect adipose stem cells (ASCs) seeded in the scaffold. The purpose of this study was to test the in vitro and in vivo efficacy of the system to induce chondrogenic differentiation of ASCs. The pDNA/PEI-PEG complex-incorporated PLGA/Pluronic F127 porous scaffolds were fabricated by a precipitation/particulate leaching method. The following five kinds of pDNA were incorporated into the scaffolds: 1)pECFP-C1 vector without an interposed gene (control group); 2) SOX-5 plasmids; 3) SOX-6 plasmids; 4) SOX-9 plasmids; and 5) one-third doses of each plasmid (SOX-5, -6, and -9). ASCs were seeded on pDNA-incorporated PLGA scaffolds and cultured in chondrogenic media for 21 days. ASCs were also isolated from rabbits, seeded in pDNA-incorporated PLGA scaffolds, and then implanted in the osteochondral defect created on the patellar groove. The rabbits were sacrificed and analyzed grossly and microscopically 8 weeks after implantation. The percentage of transfected cells was highest on day 14, around 70%. After 21 days, PLGA scaffolds incorporated with each gene showed markedly increased expression of the corresponding gene and protein. Glycosaminoglycan (GAG) assay and Safranin-O staining showed an increased proteoglycan production in SOX trio pDNA-incorporated scaffolds. The COL2A1 gene and protein were notably increased in SOX trio pDNA-incorporated scaffolds than in the control, while COL10A1 protein expression decreased. Gross and histological findings from the in vivo study showed enhanced cartilage regeneration in ASCs/SOX trio pDNA-incorporated PLGA scaffolds. (C) 2011 Elsevier Ltd. All rights reserved.