Davies O G, Cooper P R, Shelton R M, Smith A J, Scheven B A
School of Dentistry, University of Birmingham, St Chad's Queensway, Birmingham, B4 6NN, UK,
J Bone Miner Metab. 2015 Jul;33(4):371-82. doi: 10.1007/s00774-014-0601-y. Epub 2014 Jul 6.
Stem-cell-based therapies provide a biological basis for the regeneration of mineralised tissues. Stem cells isolated from adipose tissue (ADSCs), bone marrow (BMSCs) and dental pulp (DPSCs) have the capacity to form mineralised tissue. However, studies comparing the capacity of ADSCs with BMSCs and DPSCs for mineralised tissue engineering are lacking, and their ability to regenerate dental tissues has not been fully explored. Characterisation of the cells using fluorescence-activated cell sorting and semi-quantitative reverse transcription PCR for MSC markers indicated that they were immunophenotypically similar. Alizarin red (AR) staining and micro-computed tomography (µCT) analyses demonstrated that the osteogenic potential of DPSCs was significantly greater than that of BMSCs and ADSCs. Scanning electron microscopy and AR staining showed that the pattern of mineralisation in DPSC cultures differed from ADSCs and BMSCs, with DPSC cultures lacking defined mineralised nodules and instead forming a diffuse layer of low-density mineral. Dentine matrix components (DMCs) were used to promote dentinogenic differentiation. Their addition to cultures resulted in increased amounts of mineral deposited in all three cultures and significantly increased the density of mineral deposited in BMSC cultures, as determined by µCT analysis. Addition of DMCs also increased the relative gene expression levels of the dentinogenic markers dentine sialophosphoprotein and dentine matrix protein 1 in ADSC and BMSC cultures. In conclusion, DPSCs show the greatest potential to produce a comparatively high volume of mineralised matrix; however, both dentinogenesis and mineral volume was enhanced in ADSC and BMSC cultures by DMCs, suggesting that these cells show promise for regenerative dental therapies.
基于干细胞的疗法为矿化组织的再生提供了生物学基础。从脂肪组织(ADSCs)、骨髓(BMSCs)和牙髓(DPSCs)中分离出的干细胞具有形成矿化组织的能力。然而,缺乏比较ADSCs与BMSCs和DPSCs在矿化组织工程方面能力的研究,并且它们再生牙组织的能力尚未得到充分探索。使用荧光激活细胞分选和针对间充质干细胞标志物的半定量逆转录PCR对细胞进行表征表明,它们在免疫表型上相似。茜素红(AR)染色和显微计算机断层扫描(µCT)分析表明,DPSCs的成骨潜力明显大于BMSCs和ADSCs。扫描电子显微镜和AR染色显示,DPSC培养物中的矿化模式与ADSCs和BMSCs不同,DPSC培养物中缺乏明确的矿化结节,而是形成了一层低密度矿化的弥漫层。牙本质基质成分(DMCs)被用于促进牙本质生成分化。将它们添加到培养物中导致所有三种培养物中沉积的矿物质数量增加,并且通过µCT分析确定,显著增加了BMSC培养物中沉积矿物质的密度。添加DMCs还增加了ADSC和BMSC培养物中牙本质生成标志物牙本质涎磷蛋白和牙本质基质蛋白1的相对基因表达水平。总之,DPSCs显示出产生相对大量矿化基质的最大潜力;然而,DMCs增强了ADSC和BMSC培养物中的牙本质生成和矿化量,表明这些细胞在再生牙科治疗方面具有前景。