In vitro characterization of an osteoinductive biphasic calcium phosphate in combination with recombinant BMP2.
In vitro characterization of an osteoinductive biphasic calcium phosphate in combination with recombinant BMP2.
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DOI：
10.1186/s12903-016-0263-3
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发表时间：
2016-08-02
期刊：
影响因子：
2.9
通讯作者：
Miron RJ
中科院分区：
文献类型：
--
作者：
Shuang Y;Yizhen L;Zhang Y;Fujioka-Kobayashi M;Sculean A;Miron RJ
用生长因子和骨移植材料修复牙槽骨缺损在现代牙科中发挥了举足轻重的作用。重组人骨形态发生蛋白-2 (rhBMP2) 是一种骨诱导生长因子，能够招募细胞并向成骨细胞谱系分化，已与各种生物材料结合使用，以进一步增强新骨形成。最近，一组新型双相磷酸钙（BCP）骨移植材料通过异位骨形成的迹象被证明具有骨诱导特性。本研究的目的是研究 rhBMP2 联合骨诱导 BCP 骨移植对成骨细胞行为的影响。将 MC3T3-E1 前成骨细胞接种于 1) 对照组织培养塑料、2) 10 mg 单独的 BCP、3) 100 ng rhBMP2 和 4) 100 ng rhBMP2+ 10 mg BCP 上，并通过 Transwell 室分析细胞招募、通过 MTS 测定进行增殖以及通过碱性磷酸酶 (ALP) 活性、茜素红染色和实时评估进行分化。 PCR 检测成骨细胞分化标记物，包括 Runx2、胶原蛋白 1、ALP 和骨钙素 (OCN)。 rhBMP2能够显着上调细胞募集，而添加BCP以及单独的BCP没有额外的能力来改善成骨细胞募集。 BCP 和 rhBMP2 都能在播种后 3 天和 5 天显着增加细胞增殖，并且当 rhBMP2 与 BCP 结合时，细胞数量进一步增加。此外，与单独接种 rhBMP2 或单独 BCP 的细胞相比，rhBMP2 与 BCP 的组合显着提高了接种后 7 天和 14 天的 ALP 活性、14 天的茜素红染色以及 Runx2、ALP 和骨钙素的 mRNA 水平。本研究的结果表明：1) BCP 骨颗粒的骨诱导潜力在体外成骨细胞分化方面与 rhBMP2 具有同等的骨促进作用，2) 与单独的 rhBMP2 或单独的 BCP 相比，BCP 颗粒与 rhBMP2 组合能够进一步增加体外成骨细胞的骨促进分化。未来需要进一步进行动物测试来研究这种组合方法对新骨形成的影响。
The repair of alveolar bone defects with growth factors and bone grafting materials has played a pivotal role in modern dentistry. Recombinant human bone morphogenetic protein-2 (rhBMP2), an osteoinductive growth factor capable of cell recruitment and differentiation towards the osteoblast lineage, has been utilized in combination with various biomaterials to further enhance new bone formation. Recently, a group of novel biphasic calcium phosphate (BCP) bone grafting materials have been demonstrated to possess osteoinductive properties by demonstrating signs of ectopic bone formation. The aim of the present study was to study the effects of rhBMP2 in combination with osteoinductive BCP bone grafts on osteoblast cell behaviour. MC3T3-E1 pre-osteoblasts were seeded on 1) control tissue culture plastic, 2) 10 mg of BCP alone, 3) 100 ng rhBMP2, and 4) 100 ng rhBMP2+ 10 mg of BCP and analyzed for cell recruitment via a Transwell chamber, proliferation via an MTS assay and differentiation as assessed by alkaline phosphatase (ALP) activity, alizarin red staining and real-time PCR for osteoblast differentiation markers including Runx2, collagen1, ALP, and osteocalcin (OCN). rhBMP2 was able to significantly upregulate cell recruitment whereas the addition of BCP as well as BCP alone had no additional ability to improve osteoblast recruitment. Both BCP and rhBMP2 were able to significantly increase cell proliferation at 3 and 5 days post seeding and cell number was further enhanced when rhBMP2 was combined with BCP. In addition, the combination of rhBMP2 with BCP significantly improved ALP activity at 7 and 14 days post seeding, alizarin red staining at 14 days, and mRNA levels of Runx2, ALP and osteocalcin when compared to cells seeded with rhBMP2 alone or BCP alone. The results from the present study demonstrate that 1) the osteoinductive potential of BCP bone particles is equally as osteopromotive as rhBMP2 on in vitro osteoblast differentiation and 2) BCP particles in combination with rhBMP2 is able to further increase the osteopromotive differentiation of osteoblasts in vitro when compared to either rhBMP2 alone or BCP alone. Future animal testing is further required to investigate this combination approach on new bone formation.