Effects of phosphorylatable short peptide-conjugated chitosan-mediated IL-1Ra and igf-1 gene transfer on articular cartilage defects in rabbits.
Effects of phosphorylatable short peptide-conjugated chitosan-mediated IL-1Ra and igf-1 gene transfer on articular cartilage defects in rabbits.
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DOI：
10.1371/journal.pone.0112284
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Song W
中科院分区：
文献类型：
--
作者：
Zhao R;Peng X;Li Q;Song W
此前，我们报道了利用可磷酸化短肽连接的壳聚糖(PSP-CS)来提高质粒DNA-壳聚糖(PDNA/CS)的转染率。在本研究中，我们观察了PSP-CS介导的白介素1受体拮抗剂蛋白(IL-1ra)联合胰岛素样生长因子-1(IGF-1)对兔软骨细胞和兔软骨缺损模型的影响。构建pBudCE4.1-IL-1Ra+IGF-1、pBudCE4.1-IL-1Ra和pBudCE4.1-IGF-1，并与PSP-CS结合形成PDNA/PSP-CS复合体。将这些复合体导入兔原代软骨细胞或注射到关节腔内。治疗7周后，处死所有兔并进行分析。在细胞培养上清液和滑液中均检测到高水平的IL-1ra和IGF-1表达。在体外，转基因复合体可显著促进软骨细胞的增殖，促进糖胺聚糖和II型胶原的合成，抑制软骨细胞的凋亡和一氧化氮的合成。在体内，外源基因导致关节滑液中II型胶原合成增加，NO和GAG浓度降低；组织学研究显示，PDNA/PSP-CS治疗导致不同程度的透明样软骨修复和Mankin评分下降。在体外和体内，两个基因的共同表达比单独一个基因产生更大的影响。结果表明，PSP-CS是一种很好的用于软骨缺损基因治疗的候选细胞，IGF-1和IL-1ra的共同表达对软骨缺损具有良好的生物学效应。
Previously, we reported an improvement in the transfection efficiency of the plasmid DNA-chitosan (pDNA/CS) complex by the utilization of phosphorylatable short peptide-conjugated chitosan (pSP-CS). In this study, we investigated the effects of pSP-CS-mediated gene transfection of interleukin-1 receptor antagonist protein (IL-1Ra) combined with insulin-like growth factor-1 (IGF-1) in rabbit chondrocytes and in a rabbit model of cartilage defects. pBudCE4.1-IL-1Ra+igf-1, pBudCE4.1-IL-1Ra and pBudCE4.1-igf-1 were constructed and combined with pSP-CS to form pDNA/pSP-CS complexes. These complexes were transfected into rabbit primary chondrocytes or injected into the joint cavity. Seven weeks after treatment, all rabbits were sacrificed and analyzed. High levels of IL-1Ra and igf-1 expression were detected both in the cell culture supernatant and in the synovial fluid. In vitro, the transgenic complexes caused significant proliferation of chondrocytes, promotion of glycosaminoglycan (GAG) and collagen II synthesis, and inhibition of chondrocyte apoptosis and nitric oxide (NO) synthesis. In vivo, the exogenous genes resulted in increased collagen II synthesis and reduced NO and GAG concentrations in the synovial fluid; histological studies revealed that pDNA/pSP-CS treatment resulted in varying degrees of hyaline-like cartilage repair and Mankin score decrease. The co-expression of both genes produced greater effects than each single gene alone both in vitro and in vivo. The results suggest that pSP-CS is a good candidate for use in gene therapy for the treatment of cartilage defects and that igf-1 and IL-1Ra co-expression produces promising biologic effects on cartilage defects.
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在软骨组织损伤和兔子修复中选择合适的参考基因，以归一化实时PCR。
影响因子：
5.6
作者：
Peng XX;Zhao RL;Song W;Chu HR;Li M;Song SY;Li GZ;Liang DC
通讯作者：
Liang DC
影响因子：
2.3
作者：
Madry, Henning;Orth, Patrick;Cucchiarini, Magali
通讯作者：
Cucchiarini, Magali
影响因子：
3.1
作者：
Madry H;Kaul G;Zurakowski D;Vunjak-Novakovic G;Cucchiarini M
通讯作者：
Cucchiarini M
DOI：
10.1302/0301-620x.89b5.18343
发表时间：
2007-05-01
影响因子：
--
作者：
Goodrich, L. R.;Hidaka, C.;Nixon, A. J.
通讯作者：
Nixon, A. J.
DOI：
10.1088/1748-6041/9/2/025015
发表时间：
2014-04
期刊：
Biomedical materials (Bristol, England)
影响因子：
--
作者：
Clark A;Milbrandt TA;Hilt JZ;Puleo DA
通讯作者：
Puleo DA