CAJ | 학술논문

背景：牙周膜干细胞是一类起源于牙组织的成体干细胞，具有良好的成骨分化能力，有望在骨组织工程中得到应用。 　　目的：观察成骨诱导液对牙周膜干细胞成骨分化能力及细胞早期凋亡的影响。 　　方法：从原代牙周膜组织中分离得到牙周膜干细胞，以1×104/cm2浓度铺板后开始诱导。利用1，10，100 nmol/L地塞米松、β-磷酸甘油钠、维生素C为成骨诱导剂，以碱性磷酸酶活性检测、茜素红矿化结节染色、荧光定量PCR等方法对细胞成骨情况进行鉴定，采用AnnexinV/PI双染法检测细胞凋亡情况。 　　结果与结论：地塞米松可有效诱导牙周膜干细胞成骨分化，可显著提高碱性磷酸酶活性，促进茜素红矿化结节形成，提高成骨相关基因骨粘连蛋白及Ⅰ型胶原表达。根据碱性磷酸酶活性和矿化结节实验结果，地塞米松的成骨诱导具有浓度梯度效应，其中100 nmol/L 地塞米松具有最佳成骨诱导能力。细胞凋亡结果提示，地塞米松诱导的成骨分化具有一定促凋亡作用，可诱导牙周膜细胞的早期凋亡。
배경：아주막간세포시일류기원우아조직적성체간세포，구유량호적성골분화능력，유망재골조직공정중득도응용。 　　목적：관찰성골유도액대아주막간세포성골분화능력급세포조기조망적영향。 　　방법：종원대아주막조직중분리득도아주막간세포，이1×104/cm2농도포판후개시유도。이용1，10，100 nmol/L지새미송、β-린산감유납、유생소C위성골유도제，이감성린산매활성검측、천소홍광화결절염색、형광정량PCR등방법대세포성골정황진행감정，채용AnnexinV/PI쌍염법검측세포조망정황。 　　결과여결론：지새미송가유효유도아주막간세포성골분화，가현저제고감성린산매활성，촉진천소홍광화결절형성，제고성골상관기인골점련단백급Ⅰ형효원표체。근거감성린산매활성화광화결절실험결과，지새미송적성골유도구유농도제도효응，기중100 nmol/L 지새미송구유최가성골유도능력。세포조망결과제시，지새미송유도적성골분화구유일정촉조망작용，가유도아주막세포적조기조망。
BACKGROUND:Periodontal ligment stem cells are adult stem cells derived from dental tissues, which possess the capability of osteogenic differentiation. They are promising“seed cells”in bone tissue engineering. OBJECTIVE:To investigate the effects of osteogenic induction medium on the osteogenic differentiation and early apoptosis of periodontal ligment stem cells. METHODS:Periodontal ligment stem cells were isolated from primary periodontal ligament tissues and seeded on the plate at a density of 1×104/cm2. Osteogenic induction medium containing 1, 10 and 100 nmol/L dexamethasone,β-glycerophosphate and ascorbic acid were used. Alkaline phosphatase activity, mineral deposition staining and real time PCR were performed to assess the osteogenic capacity. AnnexinV/PI staining was used to detected apoptosis. RESULTS AND CONCLUSION:Dexamethasone effectively promoted osteogenic differentiation of periodontal ligment stem cells, by up-regulating alkaline phosphatase activity, enhancing mineral deposition and increasing osteogenic related gene (osteonectin and type Ⅰ col agen) expression. Alkaline phosphatase activity and mineralized nodule detection showed that osteogenic effects of dexamethasone were gradient-dependent, and 100 nmol/L was the best concentration. Apoptosis detection results indicated that osteogenic differentiation of dexamethasone for periodontal ligment stem cells could also induce cellapoptosis to some extent.