The Angiopoietin-1 Variant COMP-Ang1 Enhances BMP2-Induced Bone Regeneration with Recruiting Pericytes in Critical Sized Calvarial Defects.
The Angiopoietin-1 Variant COMP-Ang1 Enhances BMP2-Induced Bone Regeneration with Recruiting Pericytes in Critical Sized Calvarial Defects.
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DOI：
10.1371/journal.pone.0140502
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Koh JT
中科院分区：
文献类型：
--
作者：
Choi H;Jeong BC;Hur SW;Kim JW;Lee KB;Koh JT
颅面骨缺损是临床常见的骨缺损，其重建需要血管生成和成骨之间的协调耦合。在这项研究中，我们探讨了软骨寡聚基质蛋白促血管生成素1（COMP-Ang 1），促血管生成素1的合成和可溶性变体，对骨形态发生蛋白2（BMP 2）诱导的颅骨再生，招募和血管周细胞的成骨分化的影响。在C57 BL/6小鼠中产生临界尺寸的颅骨缺损，并使用可吸收的胶原海绵将COMP-Ang 1和/或BMP 2蛋白递送到缺损中。3周后，使用显微计算机断层扫描和组织学检查评估骨再生。使用抗NG 2和抗CD 31抗体的免疫荧光染色检查周细胞向缺损中的募集。采用Boyden小室法、钙化结节染色法、RT-PCR和Western blot法检测周细胞的体外募集和成骨分化。COMP-Ang 1和BMP 2的联合给药协同增强骨修复，沿着增加的CD 31（内皮细胞标记物）和NG 2（周细胞的特异性标记物）阳性细胞群。在体外培养的周细胞一致地表明，周细胞浸润到膜孔的Boyden室更增强的组合处理。此外，该组合进一步增加成骨细胞特异性基因表达，包括骨唾液蛋白（BSP），骨钙素（OCN）和osterix（OSX），Smad/1/5/8的磷酸化和矿化结节形成。COMP-Ang 1可以增强BMP 2诱导的颅骨再生，增加周细胞募集。联合递送蛋白可能是修复颅骨损伤的治疗策略。
Craniofacial bone defects are observed in a variety of clinical situations, and their reconstructions require coordinated coupling between angiogenesis and osteogenesis. In this study, we explored the effects of cartilage oligomeric matrix protein-angiopoietin 1 (COMP-Ang1), a synthetic and soluble variant of angiopoietin 1, on bone morphogenetic protein 2 (BMP2)-induced cranial bone regeneration, and recruitment and osteogenic differentiation of perivascular pericytes. A critical-size calvarial defect was created in the C57BL/6 mouse and COMP-Ang1 and/or BMP2 proteins were delivered into the defects with absorbable collagen sponges. After 3 weeks, bone regeneration was evaluated using micro-computed tomography and histologic examination. Pericyte recruitment into the defects was examined using immunofluorescence staining with anti-NG2 and anti-CD31 antibodies. In vitro recruitment and osteoblastic differentiation of pericyte cells were assessed with Boyden chamber assay, staining of calcified nodules, RT-PCR and Western blot analyses. Combined administration of COMP-Ang1 and BMP2 synergistically enhanced bone repair along with the increased population of CD31 (an endothelial cell marker) and NG2 (a specific marker of pericyte) positive cells. In vitro cultures of pericytes consistently showed that pericyte infiltration into the membrane pore of Boyden chamber was more enhanced by the combination treatment. In addition, the combination further increased the osteoblast-specific gene expression, including bone sialoprotein (BSP), osteocalcin (OCN) and osterix (OSX), phosphorylation of Smad/1/5/8, and mineralized nodule formation. COMP-Ang1 can enhance BMP2-induced cranial bone regeneration with increased pericyte recruitment. Combined delivery of the proteins might be a therapeutic strategy to repair cranial bone damage.