In situ bone tissue engineering via ultrasound-mediated gene delivery to endogenous progenitor cells in mini-pigs.
In situ bone tissue engineering via ultrasound-mediated gene delivery to endogenous progenitor cells in mini-pigs.
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DOI：
10.1126/scitranslmed.aal3128
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发表时间：
2017-05-17
影响因子：
17.1
通讯作者：
Gazit D
中科院分区：
文献类型：
--
作者：
Bez M;Sheyn D;Tawackoli W;Avalos P;Shapiro G;Giaconi JC;Da X;David SB;Gavrity J;Awad HA;Bae HW;Ley EJ;Kremen TJ;Gazit Z;Ferrara KW;Pelled G;Gazit D
每年使用自体移植物或同种异体移植物进行超过200万次骨移植手术。然而，这两种选择都有缺点，并且仍然存在明显的医学需求，需要开发用于大面积骨丢失和骨折不愈合的新疗法。我们假设，局部超声介导的，微泡增强的治疗基因传递到内源性干细胞将诱导有效的骨再生和骨折修复。为了验证这一假设，我们通过手术在尤卡坦小型猪的胫骨中制造了一个临界尺寸的骨折，这是一种临床相关的大型动物模型。将胶原支架植入骨折处，以促进内源性间充质干/祖细胞（MSC）招募到骨折部位。两周后，经皮超声介导的报告基因成功转染骨折部位40%的细胞，流式细胞仪显示80%的转染细胞表达MSC标记。人骨形态发生蛋白-6（BMP-6）质粒DNA使用超声在相同的动物模型中递送，导致BMP-6的瞬时表达和分泌定位于骨折区域。显微计算机断层扫描和生物力学分析表明，超声介导的BMP-6基因递送导致治疗后6周所有动物的完全放射学和功能性骨折愈合，而对照组动物的骨不连明显。总的来说，这些研究结果表明，超声介导的基因传递到内源性间充质祖细胞可以有效地治疗大型动物的不愈合骨折，从而解决了一个主要的骨科未满足的需求，并提供了新的可能性，为临床翻译。
More than 2 million bone-grafting procedures are performed each year using autografts or allografts. However, both options carry disadvantages, and there remains a clear medical need for the development of new therapies for massive bone loss and fracture nonunions. We hypothesized that localized ultrasound-mediated, microbubble-enhanced therapeutic gene delivery to endogenous stem cells would induce efficient bone regeneration and fracture repair. To test this hypothesis, we surgically created a critical-sized bone fracture in the tibiae of Yucatán mini-pigs, a clinically relevant large animal model. A collagen scaffold was implanted in the fracture to facilitate recruitment of endogenous mesenchymal stem/progenitor cells (MSCs) into the fracture site. Two weeks later, transcutaneous ultrasound-mediated reporter gene delivery successfully transfected 40% of cells at the fracture site, and flow cytometry showed that 80% of the transfected cells expressed MSC markers. Human bone morphogenetic protein-6 (BMP-6) plasmid DNA was delivered using ultrasound in the same animal model, leading to transient expression and secretion of BMP-6 localized to the fracture area. Micro–computed tomography and biomechanical analyses showed that ultrasound-mediated BMP-6 gene delivery led to complete radiographic and functional fracture healing in all animals 6 weeks after treatment, whereas nonunion was evident in control animals. Collectively, these findings demonstrate that ultrasound-mediated gene delivery to endogenous mesenchy-mal progenitor cells can effectively treat nonhealing bone fractures in large animals, thereby addressing a major orthopedic unmet need and offering new possibilities for clinical translation.
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影响因子：
5.1
作者：
Betz, O. B.;Betz, V. M.;Evans, C. H.
通讯作者：
Evans, C. H.
影响因子：
82.9
作者：
Bonadio, J;Smiley, E;Goldstein, S
通讯作者：
Goldstein, S
影响因子：
2.6
作者：
Flierl MA;Smith WR;Mauffrey C;Irgit K;Williams AE;Ross E;Peacher G;Hak DJ;Stahel PF
通讯作者：
Stahel PF
影响因子：
12.4
作者：
Kimelman-Bleich, Nadav;Pelled, Gadi;Gazit, Dan
通讯作者：
Gazit, Dan
影响因子：
12.4
作者：
Endoh, M;Koibuchi, N;Kaneda, Y
通讯作者：
Kaneda, Y