Release of Bioactive Adeno-Associated Virus from Fibrin Scaffolds: Effects of Fibrin Glue Concentrations
Release of Bioactive Adeno-Associated Virus from Fibrin Scaffolds: Effects of Fibrin Glue Concentrations
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DOI：
10.1089/ten.tea.2010.0586
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发表时间：
2011-08-01
影响因子：
4.1
通讯作者：
Chu, Constance R.
中科院分区：
文献类型：
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作者：
Lee, Hannah H.;Haleem, Amgad M.;Chu, Constance R.
纤维蛋白胶 (FG) 用于各种临床应用和实验室，用于局部持续释放对组织工程潜在重要的因子。然而，不同纤维蛋白原浓度对生物活性腺相关病毒 (AAV) 的 FG 支架递送的影响尚未确定。这项研究的目的是检验 FG 浓度改变 AAV 释放曲线，从而影响 AAV 生物利用度的假设。使用 HEK-293 细胞测量从 FG 释放的 AAV-GFP 的基因转移效率。使用水貂肺细胞测定法并通过测量人间充质干细胞 (hMSC) 中软骨特异性基因表达的诱导来评估从 FG 释放的 AAV 转化生长因子-β1 (TGF-β(1)) 的生物活性。未稀释的 FG 具有较长的凝血时间、较小的孔径、较粗的纤维和较慢的溶解速率，导致 AAV 的释放减少。 25% 和 50% FG 的 AAV 释放和基因转移效率高于 75% 和 100% FG 的 AAV 释放和基因转移效率。与未稀释 FG 的 hMSC 相比，稀释 FG 转导的 hMSC 释放 AAV-TGF-β(1)，导致生物活性 TGF-β(1) 浓度更高，软骨特异性基因表达上调更大。这项研究显示，从稀释的 FG 中释放的生物活性 AAV-TGF-β(1) 的释放、转导效率和对 hMSC 的软骨形成作用有所改善，为优化这种临床可用的治疗性基因传递支架（在软骨再生和其他组织工程应用中）提供了重要的信息。
Fibrin glue (FG) is used in a variety of clinical applications and in the laboratory for localized and sustained release of factors potentially important for tissue engineering. However, the effect of different fibrinogen concentrations on FG scaffold delivery of bioactive adeno-associated viruses (AAVs) has not been established. This study was performed to test the hypothesis that FG concentration alters AAV release profiles, which affect AAV bioavailability. Gene transfer efficiency of AAV-GFP released from FG was measured using HEK-293 cells. Bioactivity of AAV transforming growth factor-beta1 (TGF-beta(1)) released from FG was assessed using the mink lung cell assay, and by measuring induction of cartilage-specific gene expression in human mesenchymal stem cells (hMSCs). Nondiluted FG had longer clotting times, smaller pore sizes, thicker fibers, and slower dissolution rate, resulting in reduced release of AAV. AAV release and gene transfer efficiency was higher with 25% and 50% FG than with the 75% and 100% FG. AAV-TGF-beta(1) released from dilute-FG transduced hMSCs, resulting in higher concentrations of bioactive TGF-beta(1) and greater upregulation of cartilage-specific gene expression compared with hMSC from undiluted FG. This study, showing improved release, transduction efficiency, and chondrogenic effect on hMSC of bioactive AAV-TGF-beta(1) released from diluted FG, provides information important to optimization of this clinically available scaffold for therapeutic gene delivery, both in cartilage regeneration and for other tissue engineering applications.