Galli Carlo, Macaluso Guido M, Guizzardi Stefano, Vescovini Rosanna, Passeri Mario, Passeri Giovanni
Unit of Periodontology, University of Parma, Parma, Italy.
J Periodontol. 2006 Jul;77(7):1223-8. doi: 10.1902/jop.2006.050304.
Bone regeneration techniques increasingly rely on the use of exogenous molecules able to enhance tissue formation in pathologic and traumatic defects. An enamel matrix derivative (EMD) has been largely used to promote tooth ligament regeneration within periodontal pockets. Recent evidence suggests that EMD may contribute to inducing osteoblast growth and differentiation. We investigated the effects of EMD on growth and osteogenic marker modulation in human mandibular osteoblasts.
We focused our attention on cell growth by 3-(4,5-dimethyl[thiazol-2-yl]-3,5-diphery)tetradium bromide (MTT) assay, cell differentiation, mineralized nodule formation, and, in particular, the expression of receptor activator of nuclear factor-kappa B ligand (RANKL), the main osteoclast differentiation factor, and its decoy receptor, osteoprotegerin (OPG), by enzyme-linked immunosorbent assay.
Cell growth was significantly increased by EMD. Similarly, a significantly higher quantity of OPG and a lower amount of RANKL were detectable in groups treated with 50 and 100 microg/ml at weeks 1, 2, and 3, and alkaline phosphatase activity and osteocalcin production were enhanced in cultures treated with 50 and 100 microg/ml at weeks 2 and 3. Mineralized nodules appeared bigger and more numerous in cultures treated with 50 and 100 microg/ml EMD.
EMD was able to enhance osteoblast cell growth and the expression of markers of osteoblastic phenotype and differentiation. EMD also seemed able to create a favorable osteogenic microenvironment by reducing RANKL release and enhancing osteoblastic OPG production.
骨再生技术越来越依赖于使用能够促进病理性和创伤性缺损组织形成的外源性分子。釉基质衍生物(EMD)已被广泛用于促进牙周袋内的牙周韧带再生。最近的证据表明,EMD可能有助于诱导成骨细胞的生长和分化。我们研究了EMD对人下颌骨成骨细胞生长和成骨标志物调节的影响。
我们通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法关注细胞生长、细胞分化、矿化结节形成,特别是通过酶联免疫吸附测定法检测核因子κB受体激活剂配体(RANKL)(主要的破骨细胞分化因子)及其诱饵受体骨保护素(OPG)的表达。
EMD显著促进细胞生长。同样,在第1、2和3周用50和100μg/ml处理的组中,可检测到显著更高量的OPG和更低量的RANKL,并且在第2和3周用50和100μg/ml处理的培养物中碱性磷酸酶活性和骨钙素产生增加。在用50和100μg/ml EMD处理的培养物中,矿化结节显得更大且更多。
EMD能够促进成骨细胞生长以及成骨细胞表型和分化标志物的表达。EMD似乎还能够通过减少RANKL释放和增强成骨细胞OPG产生来创造有利的成骨微环境。