A light and electron microscopic study of ectopic tendon and ligament formation induced by bone morphogenetic protein-13 adenoviral gene therapy
A light and electron microscopic study of ectopic tendon and ligament formation induced by bone morphogenetic protein-13 adenoviral gene therapy
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DOI：
10.3171/jns.2001.95.2.0298
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发表时间：
2001-08-01
影响因子：
4.1
通讯作者：
Kallmes, DF
中科院分区：
文献类型：
--
作者：
Helm, GA;Li, JZ;Kallmes, DF
对象。骨形态发生蛋白（BMPs）参与许多组织的生长和发育，但其在骨骼发育中的作用及其诱导异位和正位成骨的独特能力引起了最大的兴趣。BMP-13基因的表达主要局限于发育过程中软骨内骨形成区域的肥大软骨细胞，以及成年人成熟的关节软骨。此外，在啮齿类动物皮下或肌肉注射时，将BMP-13应用于胶原载体可诱导新肌腱/新寡韧带形成。本研究的目的是确定组织学和超微结构。肌内注射第一代BMP-13腺病毒载体后发生的变化。将3.75 X 10（10）个斑块形成单位的腺病毒(Ad)-BMP-13或Ad- β -半乳糖苷酶注射至胸裸大鼠大腿肌肉组织，在注射后2天至100天的不同时间点对该区域进行光镜和电镜检查。早在注射Ad-BMP-13后2天，就观察到祖细胞在转导的肌纤维之间浸润。这些细胞随后增殖、分化并分泌大量胶原细胞外基质。注射后100天，处理后的组织显示出新肌腱/新寡韧带的组织学和超微结构外观，与周围肌肉有明显的界限。在处理过的组织内也可见小的骨和纤维软骨灶。一项短期的溴脱氧尿苷研究也表明，早在注射后48小时，Ad-BMP-13注射部位的间充质细胞就会快速增殖。在所有时间点，对照ad - β -gal注射部位只含有正常肌肉，没有炎症或间充质细胞增殖的证据。本研究结果表明，在未来，BMP-13基因的使用可能对肌腱和韧带撕裂和撕脱伤的愈合具有治疗作用。
Object. Bone morphogenetic proteins (BMPs) are involved in the growth and development of many tissues, but it is their role in skeletal development and their unique ability to induce ectopic and orthotopic osteogenesis that have attracted the greatest interest. Expression of the BMP-13 gene is predominantly localized to hypertrophic chondrocytes in regions of endochondral bone formation during development, as well as in mature articular cartilage in the adult. In addition, the application of BMP-13 on a collagen carrier induces neotendon/neoligament formation when delivered subcutaneously or intramuscularly in rodents. The aim of the present study was to determine the histological and ultrastructural. changes that occur after the intramuscular injection of a first-generation BMP-13 adenoviral vector.Methods. Athymic nude rats were injected with 3.75 X 10(10) plaque-forming units of adenovirus (Ad)-BMP-13 or Ad-beta -galactosidase in the thigh musculature, and the region was examined using light and electron microscopy at various time points between 2 days and 100 days postinjection. As early as 2 days after injection of Ad-BMP-13, progenitor cells were observed infiltrating between the transduced muscle fibers. These cells subsequently proliferated, differentiated, and secreted large amounts of collagenous extracellular matrix. By 100 days postinjection, the treated tissue displayed the histological and ultrastructural appearance of neotendon/neoligament, which was clearly demarcated from the surrounding muscle. Small foci of bone and fibrocartilage were also seen within the treated tissue. A short-term bromodeoxyuridine study also demonstrated rapid mesenchymal cell proliferation at the Ad-BMP-13 injection site as early as 48 hours postinjection. At all time points, the control AD-beta -gal injection sites were found to contain only normal muscle, without evidence of inflammation or mesenchymal cell proliferation.Conclusions. The results of this study indicate that in the future the use of the BMP-13 gene may have therapeutic utility for the healing of tendon and ligament tears and avulsion injuries.