Isolation and characterization of apical papilla cells from root end of human third molar and their differentiation into cementoblast cells: an in vitro study.

IF 3.7 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS Biological Procedures Online Pub Date : 2023-01-23 DOI:10.1186/s12575-023-00190-6

Morvarid Ebadi, Amirfarhang Miresmaeili, Sarah Rajabi, Shahrokh Shojaei, Sareh Farhadi

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引用次数: 1

Abstract

Background: Periodontal regeneration, treatment of periodontal-related diseases and improving the function of implants are global therapeutic challenges. The differentiation of human stem cells from apical papilla into cementoblasts may provide a strategy for periodontitis treatment. This study aimed to evaluate the differentiation of primary human stem cells apical papilla (hSCAPs) to cementoblast cells.

Material and methods: SCAPs cells were isolated from human third molar and then incubated for 21 days in a differentiation microenvironment. Alkaline phosphatase (ALP) and Alizarin red S staining assays were performed to evaluate the calcium deposition and formation of hydroxyapatite in the cultured hSCAPs microenvironment. Real-time polymerase chain reaction (RT-PCR) assay was performed for cementum protein 1 (CEMP1), collagen type I (COL1), F-Spondin (SPON1), osteocalcin (OCN), and osteopontin (OPN) as specific markers of cementoblasts and their progenitors.

Results: ALP phosphatase activity in day 21 of treatment demonstrated a significant increase in ALP compared to the control. Alizarin red S staining assay showed that the differentiated hSCAPs offered a great amount of calcium deposition nodules compared to the control. The increased expression level of CEMP1, OCN, OPN, COL1 and Spon1 was observed in days 7, 14 and 21 compared to the control, while greatest expression level was observed in day 21.

Conclusion: In conclusion, the differentiation microenviroment is convenient and useful for promoting the differentiation of hSCAPs into cementoblast.

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人第三磨牙根端根尖乳头细胞的分离、鉴定及其向成胶细胞分化的体外研究。

背景:牙周再生、牙周相关疾病的治疗和提高种植体的功能是全球治疗的挑战。人根尖乳头干细胞向成胶细胞的分化可能为牙周炎的治疗提供一种策略。本研究旨在探讨人原代干细胞尖乳头(hSCAPs)向成水泥细胞的分化。材料和方法:从人第三磨牙中分离SCAPs细胞，在分化微环境中培养21 d。采用碱性磷酸酶(ALP)和茜素红S染色法观察培养hSCAPs微环境中钙沉积和羟基磷灰石的形成。采用实时聚合酶链反应(RT-PCR)检测骨水泥蛋白1 (CEMP1)、I型胶原蛋白(COL1)、F-Spondin (SPON1)、骨钙素(OCN)和骨桥蛋白(OPN)作为成骨水泥细胞及其祖细胞的特异性标志物。结果:与对照组相比，治疗第21天ALP磷酸酶活性明显升高。茜素红S染色结果表明，分化后的hSCAPs与对照相比有大量的钙沉积结节。与对照组相比，CEMP1、OCN、OPN、COL1和Spon1的表达量在第7、14和21天均有所增加，且在第21天表达量最高。结论:该分化微环境有利于促进hSCAPs向成水泥细胞分化。

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来源期刊

Biological Procedures Online 生物-生化研究方法

CiteScore

10.50

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： iological Procedures Online publishes articles that improve access to techniques and methods in the medical and biological sciences. We are also interested in short but important research discoveries, such as new animal disease models. Topics of interest include, but are not limited to: Reports of new research techniques and applications of existing techniques Technical analyses of research techniques and published reports Validity analyses of research methods and approaches to judging the validity of research reports Application of common research methods Reviews of existing techniques Novel/important product information Biological Procedures Online places emphasis on multidisciplinary approaches that integrate methodologies from medicine, biology, chemistry, imaging, engineering, bioinformatics, computer science, and systems analysis.