Chemically modified RNA activated matrices enhance bone regeneration.
Chemically modified RNA activated matrices enhance bone regeneration.
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DOI：
10.1016/j.jconrel.2015.09.050
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发表时间：
2015-11-28
期刊：
影响因子：
--
通讯作者：
Salem AK
中科院分区：
文献类型：
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作者：
Elangovan S;Khorsand B;Do AV;Hong L;Dewerth A;Kormann M;Ross RD;Sumner DR;Allamargot C;Salem AK
迫切需要改进的治疗剂以实现可预测的骨再生。使用非病毒载体的基因治疗是安全和有效的靶细胞的，是一种有前途的方法，以克服蛋白质递送的生长因子的缺点。在这里，我们调查的转染效率，细胞毒性，成骨潜能和体内骨再生能力的化学修饰的核糖核酸（cmRNA）（编码BMP-2）与聚乙烯亚胺（PEI）复合，并与PEI复合与传统的质粒DNA（编码BMP-2）进行比较。以胺（N）与磷酸盐（P）的比率为10制造聚合复合物，并使用人骨髓基质细胞（BMSC）表征转染效率。通过测定骨特异性基因、骨钙素和碱性磷酸酶的表达以及通过检测骨基质沉积来验证用这些复合物处理的BMSCs的成骨潜力。使用大鼠颅骨骨缺损模型，结果表明，与PEI-质粒DNA（BMP-2）激活的基质相比，PEI-cmRNA（编码BMP-2）激活的基质促进了显著增强的骨再生。我们的概念验证研究表明，装载有携带编码成骨蛋白的cmRNA的非病毒载体的支架可能是刺激骨再生的有力工具，具有显著的临床转化潜力。
There exists a dire need for improved therapeutics to achieve predictable bone regeneration. Gene therapy using non-viral vectors that are safe and efficient at transfecting target cells is a promising approach to overcoming the drawbacks of protein delivery of growth factors. Here, we investigated the transfection efficiency, cytotoxicity, osteogenic potential and in vivo bone regenerative capacity of chemically modified ribonucleic acid (cmRNA) (encoding BMP-2) complexed with polyethylenimine (PEI) and made comparisons with PEI complexed with conventional plasmid DNA (encoding BMP-2). The polyplexes were fabricated at an amine (N) to phosphate (P) ratio of 10 and characterized for transfection efficiency using human bone marrow stromal cells (BMSCs). The osteogenic potential of BMSCs treated with these polyplexes was validated by determining the expression of bone-specific genes, osteocalcin and alkaline phosphatase as well as through the detection of bone matrix deposition. Using a calvarial bone defect model in rats it was shown that PEI-cmRNA (encoding BMP-2)-activated matrices promoted significantly enhanced bone regeneration compared to PEI-plasmid DNA (BMP-2)-activated matrices. Our proof of concept study suggests that scaffolds loaded with non-viral vectors harboring cmRNA encoding osteogenic proteins may be a powerful tool for stimulating bone regeneration with significant potential for clinical translation.