Divergent Resorbability and Effects on Osteoclast Formation of Commonly Used Bone Substitutes in a Human In Vitro-Assay
Divergent Resorbability and Effects on Osteoclast Formation of Commonly Used Bone Substitutes in a Human In Vitro-Assay
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DOI：
10.1371/journal.pone.0046757
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发表时间：
2012-10-10
期刊：
影响因子：
3.7
通讯作者：
Lange, Tobias
中科院分区：
文献类型：
--
作者：
Keller, Johannes;Brink, Silja;Lange, Tobias
生物活性骨替代材料是一种有价值的替代自体骨移植修复骨缺损的材料。然而，临床研究报告了许多常用生物材料的成功率不同。虽然成骨细胞传统上被认为是调节骨整合的关键角色，但越来越多的证据表明，骨吸收破骨细胞对应用生物材料的寿命至关重要。由于目前还没有关于生物材料吸收能力的标准化数据，我们采用了体外实验方法对十种常用的骨替代材料进行了比较。人外周血单个核细胞(PBMC)在牙本质芯片与生物材料或单独牙本质(对照组)共同存在的情况下分化为破骨细胞，时间为28天。在第0天和第14天通过光学显微镜监测破骨细胞的成熟情况，每周两次评估细胞外pH的材料依赖性变化。第28天用TRAP染色定量成熟破骨细胞，并测定其在牙本质(甲苯胺蓝染色)和生物材料(扫描电子显微镜、扫描电子显微镜)上的吸收活性。所分析的生物材料引起了特定的pH变化，这与破骨细胞多核(r=0.942；p=0.034)和生物材料上的活性(r=0.594；p=0.041)相关。骨膜导致pH值、破骨细胞核数和牙本质吸收显著降低，而Tutogen牛和Tutobone人显著增加了这三个参数。此外，天然生物材料比合成生物材料吸收更快，导致不同的相对吸收系数，这表明骨替代材料是否导致生物材料或周围骨的平衡吸收或优先吸收。综上所述，这项研究首次比较了广泛使用的生物材料对破骨细胞形成和吸收能力的影响，这一无偏见的方法现在可能有助于改进骨替代材料的临床前评估。
Bioactive bone substitute materials are a valuable alternative to autologous bone transplantations in the repair of skeletal defects. However, clinical studies have reported varying success rates for many commonly used biomaterials. While osteoblasts have traditionally been regarded as key players mediating osseointegration, increasing evidence suggests that bone-resorbing osteoclasts are of crucial importance for the longevity of applied biomaterials. As no standardized data on the resorbability of biomaterials exists, we applied an in vitro-assay to compare ten commonly used bone substitutes. Human peripheral blood mononuclear cells (PBMCs) were differentiated into osteoclasts in the co-presence of dentin chips and biomaterials or dentin alone (control) for a period of 28 days. Osteoclast maturation was monitored on day 0 and 14 by light microscopy, and material-dependent changes in extracellular pH were assessed twice weekly. Mature osteoclasts were quantified using TRAP stainings on day 28 and their resorptive activity was determined on dentin (toluidin blue staining) and biomaterials (scanning electron microscopy, SEM). The analyzed biomaterials caused specific changes in the pH, which were correlated with osteoclast multinuclearity (r = 0.942; p = 0.034) and activity on biomaterials (r = 0.594; p = 0.041). Perossal led to a significant reduction of pH, nuclei per osteoclast and dentin resorption, whereas Tutogen bovine and Tutobone human strikingly increased all three parameters. Furthermore, natural biomaterials were resorbed more rapidly than synthetic biomaterials leading to differential relative resorption coefficients, which indicate whether bone substitutes lead to a balanced resorption or preferential resorption of either the biomaterial or the surrounding bone. Taken together, this study for the first time compares the effects of widely used biomaterials on osteoclast formation and resorbability in an unbiased approach that may now aid in improving the preclinical evaluation of bone substitute materials.