Cytodifferentiation activity of synthetic human enamel sheath protein peptides
Cytodifferentiation activity of synthetic human enamel sheath protein peptides
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DOI：
10.1111/j.1600-0765.2010.01279.x
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发表时间：
2010-10-01
影响因子：
3.5
通讯作者：
Fukae, M.
中科院分区：
文献类型：
--
作者：
Kakegawa, A.;Oida, S.;Fukae, M.
背景与目的：釉鞘蛋白（enamelsheathprotein，ESP）参与牙齿发育过程中釉鞘的形成。17 kDa ESP是通过从鞘蛋白（成釉蛋白/釉蛋白）（猪釉基质蛋白之一）的N-末端侧蛋白水解加工的一步切割产物。牙釉质鞘蛋白在狗颊裂模型中表现出牙周膜和牙骨质再生活性，并促进培养的人牙周膜（HPDL）细胞的细胞分化。本研究的目的是确定具有细胞分化活性的人ESP的C-末端侧序列上的肽段对培养的HPDL cells.Material and Methods：在人ESP C-末端侧序列的基础上合成的肽被测试其增加碱性磷酸酶（ALP）和矿化活性的培养的HPDL cells的能力。骨钙素，骨桥蛋白和骨涎蛋白的表达测定通过半定量PCR，因此被确定为矿化组织分化的特异性指标。结果：从人ESP的多个合成肽增加ALP活性和刺激基质矿化在长期培养的HPDL细胞。半定量PCR显示骨钙素、骨桥蛋白和骨涎蛋白的表达相对于对照值增加。SDKPPKPELPGVDF具有最强的细胞分化活性在所有的合成肽tested.Conclusion：一个特定的肽序列来自于C-末端侧的人ESP促进HPDL细胞的细胞分化和矿化活性在细胞培养系统中。
Background and Objective:Enamel sheath protein (ESP) is involved in the construction of the enamel sheath during tooth development. The 17 kDa ESP is a one-step cleavage product processed by proteolysis from the N-terminal side of sheathlin (ameloblastin/amelin), one of the porcine enamel matrix proteins. Enamel sheath protein exhibits periodontal ligament and cementum regeneration activity in a buccal dehiscence model in dogs, and promotes the cytodifferentiation of cultured human periodontal ligament (HPDL) cells. The aim of this study was to determine the peptide segment on the C-terminal side sequence of the human ESP that possesses a cytodifferentiation activity on cultured HPDL cells.Material and Methods:The peptides synthesized on the basis of human ESP C-terminal side sequence were tested for their ability to increase the alkaline phosphatase (ALP) and mineralization activity of cultured HPDL cells. The expressions of osteocalcin, osteopontin and bone sialoprotein were measured by semi-quantitative PCR and therefore were determined to be specific indicators of mineralized tissue differentiation.Results:Multiple synthetic peptides from the human ESP increased the ALP activity and stimulated matrix mineralization in long-term cultures of HPDL cells. Semi-quantitative PCR demonstrated the osteocalcin, osteopontin and bone sialoprotein expressions to increase relative to the control values. The peptide SDKPPKPELPGVDF had the strongest cytodifferentiation activity among all the synthetic peptides tested.Conclusion:A specific peptide sequence derived from the C-terminal side of the human ESP promotes the cytodifferentiation and mineralization activity of HPDL cells in a cell culture system.