Research Department of General Surgery, Division of Surgery and Interventional Science, Faculty of Medical Sciences, University College London, London, UK.
Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Centre, Austrian Cluster for Tissue Regeneration, European Institute of Excellence on Tissue Engineering and Regenerative Medicine Research (Expertissues EEIG) Vienna Branch, Vienna, Austria.
J Tissue Eng Regen Med. 2018 Jan;12(1):e250-e260. doi: 10.1002/term.2406. Epub 2017 Jun 1.
Gene-activated matrix (GAM)-based therapeutics for tissue regeneration are limited by efficacy, the lack of spatiotemporal control and availability of target cells, all of which impact negatively on their translation to the clinic. Here, an advanced ultrasound-responsive GAM is described containing target cells that facilitates matrix-assisted sonoporation (MAS) to induce osteogenic differentiation. Ultrasound-responsive GAMs consisting of fibrin/collagen hybrid-matrices containing microbubbles, bone morphogenetic protein BMP2/7 coexpression plasmids together with C2C12 cells were treated with ultrasound either in vitro or following parenteral intramuscular implantation in vivo. Using direct measurement for alkaline phosphatase activity, von Kossa staining and immunohistochemical analysis for osteocalcin expression, MAS-stimulated osteogenic differentiation was confirmed in the GAMs in vitro 7 days after treatment with ultrasound. At day 30 post-treatment with ultrasound, ectopic osteogenic differentiation was confirmed in vivo using X-ray microcomputed tomography and histological analysis. Osteogenic differentiation was indicated by the presence of ectopic bone structures in all animals treated with MAS. In addition, bone volumes in this group were statistically greater than those in the control groups. This novel approach of incorporating a MAS capability into GAMs could be exploited to facilitate ex vivo gene transfer with subsequent surgical implantation or alternatively provide a minimally invasive means of stimulating in situ transgene delivery for osteoinductive gene-based therapies. Copyright © 2017 John Wiley & Sons, Ltd.
基于基因激活基质（GAM）的组织再生治疗方法受到疗效、缺乏时空控制和靶细胞可用性的限制，所有这些都对其向临床转化产生负面影响。本文描述了一种先进的超声响应 GAM，其中包含靶细胞，可促进基质辅助声孔（MAS）诱导成骨分化。体外或经肌肉内注射体内植入后，用超声处理含有微泡的纤维蛋白/胶原杂化基质、骨形态发生蛋白 BMP2/7 共表达质粒和 C2C12 细胞的超声响应 GAMs。通过直接测量碱性磷酸酶活性、Von Kossa 染色和骨钙素表达的免疫组织化学分析，在超声处理后 7 天，体外 GAMs 中证实了 MAS 刺激的成骨分化。在超声处理后 30 天，通过 X 射线微计算机断层扫描和组织学分析在体内证实了异位成骨分化。MAS 治疗的所有动物均存在异位骨结构，表明成骨分化。此外，该组的骨体积明显大于对照组。将 MAS 能力纳入 GAMs 的这种新方法可用于促进基因转移，随后进行手术植入，或者提供一种微创刺激原位转基因传递的方法，用于成骨诱导基因治疗。版权所有 © 2017 约翰威立父子公司
