Stem cell and regenerative medicine group, National Institute of Genetic Engineering and Biotechnology, Tehran, 14965/161, Iran; Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, 14115-175, Iran; Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, 14115-175, Iran.
Colloids Surf B Biointerfaces. 2019 Jan 1;173:662-671. doi: 10.1016/j.colsurfb.2018.10.035. Epub 2018 Oct 17.
A natural peptide motif in the first helix of osteocalcin (OCN) is used to promote nucleation and crystallization of hydroxyapatite (HA) in hard tissue. The capability of osteocalcin mimetic peptides to induce osteogenic activity of osteoblast cells leading to in-vitro mineralization is demonstrated. An osteocalcin-derived peptide consisting of thirteen amino acids is synthesized in both acidic (OSC) and amidic (OSN) forms and added into the human osteoblast-like cells (MG63) culture. The viability, proliferation, alkaline phosphatase activity, HA deposition and osteogenic gene expression by osteoblast cells are evaluated. It is revealed that the addition of 100 μg/ml of peptides enhances the proliferation rate and total protein content of osteoblast cells. Alkaline phosphatase activity is significantly higher in the presence of peptides which in turn stimulated RNA expression of collagen type I and osteopontin in a phosphate-dependent manner. Alizarin red staining and calcium content measurement show that mineral deposition is considerably increased. Ultrastructural characterization of MG63 cultures confirms the crystalline nature and chemical composition of HA mineral formation in the presence of peptides. It is confirmed that the osteocalcin-derived peptide, particularly in amidic form (OSN), is able to act as a bioactive inducer of mineralization process and hence accelerating bone tissue regeneration.
骨钙素（OCN）第一螺旋中的天然肽基序用于促进硬组织中羟磷灰石（HA）的成核和结晶。证明了骨钙素模拟肽能够诱导成骨细胞的成骨活性，从而导致体外矿化。合成了由十三个氨基酸组成的骨钙素衍生肽，其呈酸性（OSC）和酰胺（OSN）两种形式，并添加到人成骨样细胞（MG63）培养物中。评估了成骨细胞的活力、增殖、碱性磷酸酶活性、HA 沉积和成骨基因表达。结果表明，添加 100μg/ml 的肽可提高成骨细胞的增殖率和总蛋白含量。碱性磷酸酶活性在存在肽的情况下显著升高，这反过来又以磷酸盐依赖的方式刺激 I 型胶原和骨桥蛋白的 RNA 表达。茜素红染色和钙含量测量表明矿化沉积显著增加。MG63 培养物的超微结构特征证实了在肽存在下 HA 矿化形成的结晶性质和化学成分。证实骨钙素衍生肽，特别是酰胺形式（OSN），能够作为矿化过程的生物活性诱导剂，从而加速骨组织再生。
