牙周膜干细胞聚合体：一种潜在骨组织再生方法

Periodontal ligament stem cell pellet:a potential method for regenerating bone tissue

目的:探索牙周膜干细胞的成骨能力。方法:体外培养牙周膜干细胞(PDLSCs),经体外鉴定、扩增构建PDLSCs细胞膜片(CS)和细胞聚合体(CP),通过大体观察、HE和免疫组化染色、扫描电镜进行形态学检测,实时定量PCR检测CS和CP成骨相关基因的表达水平,并观察裸鼠体内成骨能力。结果:成功分离、培养并鉴定PDLSCs后,构建PDLSCs CS和CP。组织学观察发现CP内细胞外基质丰富;免疫组化染色发现这些细胞外基质中骨涎蛋白(BSP)、碱性磷酸酶(ALP)和I型胶原表达呈阳性;CP上BSP、ALP和Runx2基因表达水平明显高于CS;体内实验发现CP组大量骨样组织形成,而CS组可见纤维样骨组织形成。结论:PDLSCs细胞聚合体技术为修复牙槽骨缺损提供了一种新的方法。

AIM： To investigate the osteogenesis potential of periodontal ligament stem cell（PDLSC） pellet（CP）. METHODS： PDLSCs were in vitro cultured,identified and CP was constructed. CP was examined by HE and IHC staining,scanning electron microscope（SEM） and real-time PCR for the observation of relevant osteogenic gene expression. Cell sheet（CS） was used as the control. CS and CP were respectively mixed with ceramic bovine bone（CBB） and then transplanted into the back of nude mice for the observation of bone formation. RESULTS： CP showed a better elasticity and shape. CP contained much more extracellular matrix（ECM） than CS,and CP was positive for the expression of alkaline phosphatase（ALP）,bone sialoprotein（BSP） and collagen I（COL-I）. In addition,CP expressed higher levels of osteogenic genes than CS counterparts. In vivo study revealed that CP group showed lots of bone-like tissues in CBB,but CS group showed a lot of fiber-like tissues. CONCLUSION： PDLSCs CP may be a new strategy for regenerating bone tissue.