Combination therapy with BMP-2 and a systemic RANKL inhibitor enhances bone healing in a mouse critical-sized femoral defect.
Combination therapy with BMP-2 and a systemic RANKL inhibitor enhances bone healing in a mouse critical-sized femoral defect.
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DOI：
10.1016/j.bone.2015.12.052
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发表时间：
2016-03
期刊：
影响因子：
4.1
通讯作者：
Lieberman JR
中科院分区：
文献类型：
--
作者：
Bougioukli S;Jain A;Sugiyama O;Tinsley BA;Tang AH;Tan MH;Adams DJ;Kostenuik PJ;Lieberman JR
重组人BMP-2 （rhBMP-2）是一种有效的骨诱导剂，不仅与骨形成有关，而且与破骨细胞发生和骨吸收有关。骨保护素（Osteoprotegerin， OPG）是一种阻断破骨细胞分化和功能的RANKL抑制剂。我们假设局部BMP-2（重组蛋白或基因治疗的产物）联合全身OPG-Fc在促进骨修复方面比单独BMP-2更有效。为了验证这一假设，我们使用了小鼠临界大小的股骨缺损模型。用rhBMP-2 （I组）、rhBMP-2和全身OPG （II组）、rhBMP-2和延迟给药OPG （III组）、用含有BMP-2基因的慢病毒载体（LV-BMP-2； IV组）、LV-BMP-2和全身OPG （V组）、单独载体（VI组）和单独给药OPG （VII组）转导小鼠BM细胞。BMP-2（单独或联合OPG）治疗的所有骨缺损均愈合，而单独使用载体或单独使用OPG治疗的动物骨形成最小。显微ct分析显示，rhBMP-2 + OPG组和LV-BMP-2 + OPG组骨体积（BV）显著高于单独rhBMP-2组（p < 0.01）和单独LV-BMP-2组（p < 0.001）。在组织形态学测量中也观察到类似的结果，与rhBMP-2 + OPG缺损相比，rhBMP-2单独缺损的骨面积（B.Ar）显著降低（p < 0.005），而与所有BMP-2 + OPG处理组相比，lm -BMP-2缺损的B.Ar显著降低（p≤0.01）。TRAP染色显示，早在术后7天，未接受OPG （rhBMP-2、LV-BMP-2和海绵单独）的组就出现了主要的破骨细胞反应。总之，我们证明，与BMP-2单独相比，局部递送BMP-2（重组蛋白或基因治疗）联合全身给药OPG可改善小鼠临界大小骨缺损的骨愈合。这些数据表明破骨细胞可以减少BMP-2的愈合反应，因此抑制RANKL可能会增强BMP-2的疗效。
Recombinant human BMP-2 (rhBMP-2) is a potent osteoinductive agent, but has been associated not only with bone formation, but also osteoclastogenesis and bone resorption. Osteoprotegerin (OPG) is a RANKL inhibitor that blocks differentiation and function of osteoclasts. We hypothesized that the combination of local BMP-2 (recombinant protein or a product of gene therapy) plus systemic OPG-Fc is more effective than BMP-2 alone in promoting bone repair. To test this hypothesis we used a mouse critical-sized femoral defect model. Col2.3eGFP (osteoblastic marker) male mice were treated with rhBMP-2 (group I), rhBMP-2 and systemic OPG (group II), rhBMP-2 and delayed administration of OPG (group III), mouse BM cells transduced with a lentiviral vector containing the BMP-2 gene (LV-BMP-2; group IV), LV-BMP-2 and systemic OPG (group V), a carrier alone (group VI) and administration of OPG alone (group VII). All bone defects treated with BMP-2 (alone or combined with OPG) healed, whereas minimal bone formation was noted in animals treated with the carrier alone or OPG alone. MicroCT analysis showed that bone volume (BV) in rhBMP-2 + OPG and LV-BMP-2 + OPG groups was significantly higher compared to rhBMP-2 alone (p < 0.01) and LV-BMP-2 alone (p < 0.001). Similar results were observed in histomorphometry, with rhBMP-2 alone defects exhibiting significantly lower bone area (B.Ar) compared to rhBMP-2 + OPG defects (p < 0.005) and LV-BMP-2 defects having a significantly lower B.Ar compared to all BMP-2 + OPG treated groups (p ≤ 0.01). TRAP staining demonstrated a major osteoclast response in the groups that did not receive OPG (rhBMP-2, LV-BMP-2 and sponge alone) beginning as early as 7 days post-operatively. In conclusion, we demonstrated that locally delivered BMP-2 (recombinant protein or gene therapy) in combination with systemically administered OPG improved bone healing compared to BMP-2 alone in a mouse critical-sized bone defect. These data indicate that osteoclasts can diminish healing responses to BMP-2 and that RANKL inhibition may thus accentuate BMP-2 efficacy.