Periodontal Specific Differentiation of Dental Follicle Stem Cells into Osteoblast, Fibroblast, and Cementoblast
Periodontal Specific Differentiation of Dental Follicle Stem Cells into Osteoblast, Fibroblast, and Cementoblast
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DOI：
10.1089/ten.tec.2014.0603
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发表时间：
2015-10-01
影响因子：
3
通讯作者：
Jayakumar, R.
中科院分区：
文献类型：
--
作者：
Sowmya, S.;Chennazhi, K. P.;Jayakumar, R.
牙囊是牙囊干细胞（DFC）的来源，其具有向牙周谱系分化的潜能。因此，DFC在牙齿组织工程中具有价值。本研究的目的是评估生长因子类型和浓度对DFC分化为牙周特异性谱系的影响。从人牙囊中分离DFC，并表征其表达间充质标记物。细胞CD-73、CD-44和CD-90阳性; CD-33、CD-34和CD-45阴性。CD-29和CD-31的表达几乎可以忽略不计。细胞还表达牙周韧带和牙骨质标记物，如牙周韧带相关蛋白-1（PLAP-1）、成纤维细胞生长因子-2（FGF-2）和牙骨质蛋白-1（CEMP-1），然而，成骨细胞标记物的表达不存在。此外，在三种不同的诱导培养基中培养DFC以分析成骨细胞、成纤维细胞和成牙骨质细胞的分化。Runt相关转录因子2（RUNX-2）、碱性磷酸酶（ALP）活性、茜素染色、钙定量、1型胶原（Col-1）和骨桥蛋白（OPN）表达证实了DFC的成骨分化。在含有重组人FGF-2（rhFGF-2）的培养基中培养的DFC显示随着rhFGF-2浓度的增加PLAP-1、FGF-2和COL-1表达增强，从而证实牙周膜成纤维细胞分化。类似地，在含有重组人牙骨质蛋白-1（rhCEMP-1）的培养基中培养的DFC显示相对于rhCEMP-1增强的骨唾液酸蛋白-2（BSP-2）、CEMP-1和COL-1表达，这证实了成牙骨质细胞分化。与非诱导培养基中培养的DFCs相比，诱导培养基中培养的DFCs成骨细胞、成纤维细胞和成牙骨质细胞相关基因的表达增强。因此，通过定量分析证明了DFCs依赖生长因子分化为牙周特异性谱系。
The dental follicle is a source of dental follicle stem cells (DFCs), which have the potential to differentiate into the periodontal lineage. DFCs therefore are of value in dental tissue engineering. The purpose of this study was to evaluate the effect of growth factor type and concentration on DFC differentiation into periodontal specific lineages. DFCs were isolated from the human dental follicle and characterized for the expression of mesenchymal markers. The cells were positive for CD-73, CD-44, and CD-90; and negative for CD-33, CD-34, and CD-45. The expression of CD-29 and CD-31 was almost negligible. The cells also expressed periodontal ligament and cementum markers such as periodontal ligament-associated protein-1 (PLAP-1), fibroblast growth factor-2 (FGF-2), and cementum protein-1 (CEMP-1), however, the expression of osteoblast markers was absent. Further, the DFCs were cultured in three different induction medium to analyze the osteoblastic, fibroblastic, and cementoblastic differentiation. Runt-related transcription factor 2 (RUNX-2), alkaline phosphatase (ALP) activity, alizarin staining, calcium quantification, collagen type-1 (Col-1), and osteopontin (OPN) expression confirmed the osteoblastic differentiation of DFCs. DFCs cultured in recombinant human FGF-2 (rhFGF-2) containing medium showed enhanced PLAP-1, FGF-2, and COL-1 expression with increasing concentration of rhFGF-2 which thereby confirmed periodontal ligament fibroblastic differentiation. Similarly, DFCs cultured in recombinant human cementum protein-1 (rhCEMP-1) containing medium showed enhanced bone sialoprotein-2 (BSP-2), CEMP-1, and COL-1 expression with respect to rhCEMP-1 which confirmed cementoblastic differentiation. The expression of osteoblast, fibroblast, and cementoblast-related genes of DFCs cultured in induction medium was enhanced in comparison to DFCs cultured in noninduction medium. Thus, growth factor-dependent differentiation of DFCs into periodontal specific lineages was proved by quantitative analysis.