CAJ | 학술논문

目的：研究人骨形态发生蛋白-2（bone morphogenetic protein，BMP-2）对人炎症牙髓组织来源的牙髓干细胞（dental pulp stem cells from inflamed pulps，DPSCs-IPs）体外骨向诱导分化的影响。方法：取第三代 DPSCs-IPs，按是否于培养基中加入 BMP-2分成：BMP-2＋DPSCs-IPs 组（实验组）和 DPSCs-IPs 组（对照组）。无成骨诱导条件培养1周后，对各组分泌的胶原基质行 Tri-chrome 染色，观察并比较实验组和对照组之间的骨向诱导分化情况；实时定量 RT-PCR 检测二者的Ⅰ型胶原蛋白（collagenⅠ，COL-Ⅰ）mRNA 表达情况。在成骨诱导条件下，培养3周后对各组分泌的钙化基质行 Von Kossa 染色，通过实时定量 RT-PCR 检测转录因子及成骨相关基因的表达。结果：无成骨诱导培养1周后，实验组 DPSCs-IPs 分泌的胶原基质 Trichrome 染色较对照组深， COL-ⅠmRNA 的表达水平显著上调（P ＜0．05），说明 BMP-2可诱导 DPSCs-IPs 沉积更多的胶原基质；成骨诱导培养3周后，相对于对照组，实验组 DPSCs-IPs 沉积了更多的钙化基质，Nanog、八聚体结合转录因子4（octamer-binding transcription factor 4，Oct4）、性别决定区因子（SRY-related high-mobility group box 2，Sox2）等转录因子表达升高，碱性磷酸酶（alkaline phosphatase，ALP）、骨钙素（osteocalcin，OCN）、骨涎蛋白（bone sialoprotein，BSP），牙骨质蛋白1（cementum protein 1，CEMP-1）、COL-Ⅰ等成骨相关基因表达水平显著上调（P ＜0．05）。结论：BMP-2在成骨诱导条件下可促进 DPSCs-IPs 进行体外骨向诱导分化。
목적：연구인골형태발생단백-2（bone morphogenetic protein，BMP-2）대인염증아수조직래원적아수간세포（dental pulp stem cells from inflamed pulps，DPSCs-IPs）체외골향유도분화적영향。방법：취제삼대 DPSCs-IPs，안시부우배양기중가입 BMP-2분성：BMP-2＋DPSCs-IPs 조（실험조）화 DPSCs-IPs 조（대조조）。무성골유도조건배양1주후，대각조분비적효원기질행 Tri-chrome 염색，관찰병비교실험조화대조조지간적골향유도분화정황；실시정량 RT-PCR 검측이자적Ⅰ형효원단백（collagenⅠ，COL-Ⅰ）mRNA 표체정황。재성골유도조건하，배양3주후대각조분비적개화기질행 Von Kossa 염색，통과실시정량 RT-PCR 검측전록인자급성골상관기인적표체。결과：무성골유도배양1주후，실험조 DPSCs-IPs 분비적효원기질 Trichrome 염색교대조조심， COL-ⅠmRNA 적표체수평현저상조（P ＜0．05），설명 BMP-2가유도 DPSCs-IPs 침적경다적효원기질；성골유도배양3주후，상대우대조조，실험조 DPSCs-IPs 침적료경다적개화기질，Nanog、팔취체결합전록인자4（octamer-binding transcription factor 4，Oct4）、성별결정구인자（SRY-related high-mobility group box 2，Sox2）등전록인자표체승고，감성린산매（alkaline phosphatase，ALP）、골개소（osteocalcin，OCN）、골연단백（bone sialoprotein，BSP），아골질단백1（cementum protein 1，CEMP-1）、COL-Ⅰ등성골상관기인표체수평현저상조（P ＜0．05）。결론：BMP-2재성골유도조건하가촉진 DPSCs-IPs 진행체외골향유도분화。
Objective:To investigate the influence of bone morphogenetic protein-2(BMP-2)on osteogenic differentiation of human inflammatory dental pulp stem cells(DPSCs-IPs)in vitro.Methods:Passage 3 cells dental pulp stem cells from inflamed teeth in medi-um with or without osteogenic matter.According to whether 100 μg /L BMP-2 was added they were divided into four groups:group 1 , DPSCs-IPs culturing with 100 μg /L BMP-2 and osteogenic medium containing;group 2,DPSCs-IPs culturing with 100 μg /L BMP-2 but without osteogenic medium containing;group 3,DPSCs-IPs culturing without 100 μg /L BMP-2 but with osteogenic medium contai-ning;group 4,DPSCs-IPs culturing without 100 μg /L BMP-2 and osteogenic medium containing.After one week of their culture in nonosteogenic condition,we used Trichrome to stain the secretion of collagen matrix,and compared the difference of osteogenic differ-entiation between them.Real-time RT-PCR was used to test the expression of COL-ⅠmRNA.Three weeks after their culture in osteo-genic condition,the cell differentiation was examined by von Kossa staining.Real-time RT-PCR was used to test nuclear transcription factor and osteogenesis related molecules gene expression.Results:One week later,Trichrome staining matrix secretion of DPSCs-IPs with BMP-2 group is more obvious than the control group in the nonosteogenic medium.Real-time RT-PCR tests showed that expression of COL-ⅠmRNA increased (P <0.05).The results suggested that BMP-2 could induce DPSCs-IPs deposit more collagen matrix;3 weeks later,more mineralized matrix secretion of DPSCs-IPs can be observed in the osteogenic medium.In addition,real-time RT-PCR tests showed that experimental group cells express more Nanog,Oct4 and SOX2.And the osteogenesis related molecules ALP,OCN, BSP,CEMP-1 and COL-Ⅰ expression increased significantly compared with the control group (P <0.05 ).Conclusions:BMP-2 can promote osteogenic differentiation of human inflammatory dental pulp stem cells in vitro in osteogenic condition.