Evaluation of Osteogenic Cell Differentiation in Response to Bone Morphogenetic Protein or Demineralized Bone Matrix in a Critical Sized Defect Model Using GFP Reporter Mice
Evaluation of Osteogenic Cell Differentiation in Response to Bone Morphogenetic Protein or Demineralized Bone Matrix in a Critical Sized Defect Model Using GFP Reporter Mice
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DOI：
10.1002/jor.22657
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发表时间：
2014-09-01
影响因子：
2.8
通讯作者：
Lieberman, Jay R.
中科院分区：
文献类型：
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作者：
Alaee, Farhang;Hong, Seung-Hyun;Lieberman, Jay R.
我们评价了骨祖细胞对rhBMP-2和DBM在转基因小鼠临界大小缺损中的反应。这些小鼠表达Col3.6GFP托帕兹（一种前成骨细胞标记物）、Col2.3GFP祖母绿（一种成骨细胞标记物）和α-平滑肌肌动蛋白（α-SMA-Cherry，一种周细胞/肌成纤维细胞标记物）。我们使用X光片、冷冻和常规组织学分析评估了不同时间点的缺损愈合情况。使用新的计算机辅助算法定量对应于新骨形成区域的感兴趣区域中的GFP信号。rhBMP-2治疗的所有缺损均愈合。相比之下，DBM组（27/30）和对照组（28/30）中的大多数缺损没有愈合。前成骨细胞的定量显示在第7天在Col3.6GFP托帕兹小鼠中的最大反应（% GFP +细胞/ TV）（7.2% +/-6.0，p < 0.05，与第14、21、28和56天相比）。在第14天（8.04% +/-5.0）和第21天（8.31% +/-4.32）观察到Col2.3GFP细胞的最大应答，p < 0.05。相比之下，DBM和对照组在所有时间点均显示出有限的成骨反应。总之，我们证明了BMP和DBM诱导了截然不同的成骨反应，这将影响它们作为骨移植替代品的临床应用。(C)2014骨科研究学会。由威利期刊公司出版
We evaluated the osteoprogenitor response to rhBMP-2 and DBM in a transgenic mouse critical sized defect. The mice expressed Col3.6GFPtopaz (a pre-osteoblastic marker), Col2.3GFPemerald (an osteoblastic marker) and a-smooth muscle actin (alpha-SMA-Cherry, a pericyte/myofibroblast marker). We assessed defect healing at various time points using radiographs, frozen, and conventional histologic analyses. GFP signal in regions of interest corresponding to the areas of new bone formation was quantified using a novel computer assisted algorithm. All defects treated with rhBMP-2 healed. In contrast, the majority of the defects in the DBM (27/30) and control (28/30) groups did not heal. Quantitation of pre-osteoblasts demonstrated a maximal response (% GFP + cells/ TV) in the Col3.6GFPtopaz mice at day 7 (7.2% +/- 6.0, p < 0.05 compared to days 14, 21, 28, and 56). The maximal response of the Col2.3GFP cells was seen at days 14 (8.04% +/- 5.0) and 21 (8.31% +/- 4.32), p < 0.05. In contrast, DBM and control groups showed a limited osteogenic response at all time points. In conclusion, we demonstrated that the BMP and DBM induce vastly different osteogenic responses which should influence their clinical application as bone graft substitutes. (C) 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.