Effect of Freeze-Dried Allograft Bone With Human Basic Fibroblast Growth Factor Containing a Collagen-Binding Domain From Clostridium histolyticum Collagenase on Bone Formation After Lumbar Posterolateral Fusion Surgery in Rats
Effect of Freeze-Dried Allograft Bone With Human Basic Fibroblast Growth Factor Containing a Collagen-Binding Domain From Clostridium histolyticum Collagenase on Bone Formation After Lumbar Posterolateral Fusion Surgery in Rats
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DOI：
10.1097/brs.0000000000002074
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发表时间：
2017-09-01
期刊：
影响因子：
3
通讯作者：
Takaso, Masashi
中科院分区：
文献类型：
--
作者：
Inoue, Gen;Uchida, Kentaro;Takaso, Masashi
研究设计。一项实验研究。客观。目的：评价碱性成纤维细胞生长因子(BFGF)与多囊肾病域(PKD)和胶原结合域(CBD)融合的冻干骨移植在促进大鼠腰椎后外侧融合中的作用。有报道表明，碱性成纤维细胞生长因子是一种有效的生长因子，具有促进骨再生的潜能，但由于其从宿主部位扩散到体液中，其效率迅速下降。在先前的研究中，我们开发了一种含有C组裂性胶原酶的PKD和CBD的碱性成纤维细胞生长因子融合蛋白(bFGF-PKD-CBD)，在较低浓度下应用于大鼠股骨表面可显著促进骨形成。这种新蛋白在大鼠腰椎后外侧融合模型中加速骨融合的潜力还有待研究。双侧L4-L5后外侧融合，每侧使用150 mg FDBA粉末。雄性SD大鼠20只，体重200~250g/只，随机分为两组：FDBA+磷酸盐缓冲液对照组(对照组)和0.58nmolbFGF-PKD-CBD组(bFGF-PKD-CBD组)，每组10只。通过X线片、显微计算机断层扫描和组织学(苏木精-伊红和von Kossa染色)评价bFGF-PKD-CBD的效果。BFGF-PKD-CBD组后外侧损伤区移植骨体积及椎板表面和脊髓突表面新骨形成体积均显著高于对照组。组织学上，bFGF-PKD-CBD组新生骨形成及周围软骨细胞和成纤维细胞明显增多。FDBA复合bFGF-PKD-CBD有望成为一种促进脊柱融合的材料，而基于FDBA的bFGF-PKD-CBD定位递送系统可能为促进脊柱融合提供新的治疗途径。
Study Design. An experimental study.Objective. To evaluate the effectiveness of freeze-dried bone allograft (FDBA) with basic fibroblast growth factor (bFGF) fused with the polycystic kidney disease domain (PKD) and the collagen-binding domain (CBD) of Clostridium histolyticum collagenase, for the acceleration of lumbar posterolateral fusion in rats.Summary of Background Data. Reports indicate bFGF is an effective growth factor with osteogenic potential for promoting bone regeneration, although its efficiency decreases rapidly following its diffusion in body fluid from the host site. We developed a bFGF fusion protein containing the PKD and the CBD of C histolyticum collagenase (bFGF-PKD-CBD), which markedly enhanced bone formation at a relatively low concentration when applied to the surface of rat femurs in a previous study. The potential of this novel protein to accelerate bone fusion in a rat model of lumbar posterolateral fusion has yet to be investigated.Methods. Bilateral L4-L5 posterolateral fusions were performed, using 150 mg of FDBA powder per side. A total of 20 male Sprague-Dawley rats weighing 200 to 250 g/each were divided into two groups of 10 rats: FDBA was incubated with either phosphate-buffered saline (control group) or 0.58 nmol bFGF-PKD-CBD (bFGF-PKD-CBD group) before fusion surgery. The effect of bFGF-PKD-CBD was estimated using radiographs, microcomputed tomography, and histology (hematoxylin-eosin and von Kossa staining).Results. Both grafted bone volume in the posterolateral lesion and the volume of new bone formation on the surface of laminae and spinal processes were significantly higher in the bFGF-PKD-CBD group than in the control group. Histologically, new bone formation and surrounding chondrocytes and fibroblasts were prominent in the bFGF-PKD-CBD group.Conclusion. FDBA infused with bFGF-PKD-CBD may be a promising material for accelerating spinal fusion, and the FDBA-based delivery system for localizing bFGF-PKD-CBD may offer novel therapeutic approaches to augment spinal fusion.