Effect of concentrated growth factor fibrin membrane during distraction osteogenesis of the rabbit mandible

doi:10.3969/j.issn.2095-4344.2017.08.005

Abstract

Abstract:

BACKGROUND: Concentrated growth factor (CGF) fibrin membrane is involved in metabolism regulation, which has been applied in bone remodeling. Previous Studies have found that the cytoplasts of osteoblasts, interstitial cells and osteocytes in distraction osteogenesis are positive for osteoprotegerin and receptor activator nuclear factor kappa B ligand (RANKL).
OBJECTIVE: To explore the mechanism of bone orthopedic effect and the promotion effect of CGF fibrin membrane on the distraction osteogenesis of the rabbit mandible.
METHODS: Twenty-four white rabbits were randomly selected. Distraction osteogenesis was performed unilaterally in the rabbit left mandible (control group); CGF fibrin membrane was fixed on the inner surface of distractor, and the distraction gap was then completely wrapped prior to distraction osteogenesis in the right mandible (experimental group). At 1, 7, 14 and 28 days of consolidation period, the rabbits were sacrificed and the bilateral mandibles were taken for hematoxylin-eosin staining, western blot assay and immunohistochemical staining to detect the expression of RANKL and osteoprotegerin in the newborn bones; the osteogenesis in the distraction gap was observed and compared between two groups.
RESULTS AND CONCLUSION: The osteoprotegerin positive cells and positive area percentage in the experimental group were significantly higher than those in the control group at 1, 7 and 14 days after distraction osteogenesis (P < 0.01). The RANKL positive rate and positive area percentage in the experimental group were significantly higher than those in the control group at 1 and 14 days after distraction osteogenesis (P < 0.05 or P < 0.01). Western blot assay showed a higher ratio of RANKL/osteoprotegerin in the control group than the experimental group (P < 0.01). Our findings indicate that CGF fibrin membrane can promote osteogenesis and mineralization in the distraction gap of the rabbit mandible, which is beneficial for distraction osteogenesis in the mandible.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Palete-Rich-Plasma, RANK Ligand, Osteoprotegerin, Tissue Engineeirng

CLC Number:

R318

Tian Lu-ming1, Xin Zeng-xi1, Ma Yun-sheng2. Effect of concentrated growth factor fibrin membrane during distraction osteogenesis of the rabbit mandible[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(8): 1172-1177.

Figures/Tables 2

2.1 实验动物数量分析实验选用白兔24只，实验过程无脱失，全部进入结果分析。 2.2 术后一般情况及标本观察实验动物全部均存活，精神饮食尚好，体质量略有下降，两侧下颌骨均存在偏颌现象，日常活动正常，牵张器无松动、断裂和变形的(图1，2)进入结果分析，处死后可见牵张侧下颌骨延长。 2.3 骨牵引区组织学观察见图3。对照组：稳定期第1天，牵引区内新生血管和细胞较实验组少；第7天，牵引区内胶原纤维排列紊乱，但部分胶原纤维束可见沿牵引方向排列(图3B)；第14天，对照组间隙的胶原纤维束密集，牵引中心区骨小梁纤细，形态幼稚，其表面也可观察到大量成骨细胞(图3D)。第28天，对照组牵引区中心区域出现细小骨小梁，沿着骨切开方向排列；牵张间隙内新骨组织沿牵引方向生长，成骨方式主要是膜内成骨，在牵引器松动的标本早期仍可观察到散在的软骨岛。实验组：稳定期第1天，见牵引区间隙内为无序排列的纤维结缔组织，结缔组织间可见新生血管，炎性细胞，大量原发性间充质细胞，成骨细胞和成纤维细胞；第7天，见牵引区间隙内充满沿牵张方向排列的胶原纤维，胶原纤维内出现由间充质细胞分化而来的成纤维细胞，成骨细胞及其前体细胞，并可观察到骨基质形成和矿化，牵引区主体为纤维结缔组织(图3A)；第14天，实验组骨切开边缘处及骨膜下有少量新生骨小梁组织(图3C)；第28天，实验组骨小梁变粗大、致密，相互连续，但排列不规则。 2.4 骨保护素、核因子κB受体活化因子配体的免疫组织化学染色结果牵引结束第1天，两组牵引骨痂处的骨髓衬里细胞骨保护素表达呈强阳性，核因子κB受体活化因子配体表达呈弱阳性(图4A，B)。第7天，两组牵引骨痂处的骨髓衬里细胞骨保护素表达呈阳性，核因子κB受体活化因子配体表达呈强阳性。第14天，两组牵引骨痂处的骨髓衬里细胞骨保护素表达阳性，核因子κB受体活化因子配体表达呈强阳性(图4C，D)。第28天，两组牵引骨痂处的骨髓衬里细胞骨保护素、核因子κB受体活化因子配体表达呈弱阳性。骨保护素、核因子κB受体活化因子配体表达强度具有时间变化特点。与对照组比较，实验组在第7，14天骨保护素表达的阳性细胞率及阳性面积百分比升高(P < 0.05或P < 0.01)，以后逐渐下降；第1，14天核因子κB受体活化因子配体表达的阳性细胞率及阳性面积百分比升高(P < 0.05或P < 0.01)，以后逐渐下降；第28天骨保护素、核因子κB受体活化因子配体仅有微弱表达(表1-4)。 2.5 Western blot表达骨保护素和核因子κB受体活化因子配体的结果用quantity one软件分析计算灰度值，当在第7天时，对照组、实验组内的骨保护素和核因子κB受体活化因子配体表达水平均较强，骨保护素两组间比较差异有显著性意义(P < 0.05)，核因子κB受体活化因子配体两组在第1天时差异有显著性意义(P < 0.05)，到第7天时差异无显著性意义；核因子κB受体活化因子配体/骨保护素比值第7天较第1天时降低，对照组、实验组间比值差异有显著性意"

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