Viña-Almunia Jose, Mas-Bargues Cristina, Borras Consuelo, Gambini Juan, El Alami Marya, Sanz-Ros Jorge, Peñarrocha Miguel, Vina Jose
Int J Oral Maxillofac Implants. 2017 November/December;32(6):1251–1256. doi: 10.11607/jomi.5529. Epub 2017 Sep 22.
To analyze, in vitro, the influence of O₂ pressure on the adhesion, proliferation, and osteogenic differentiation of human dental pulp stem cells (DPSC) on β-tricalcium phosphate (β-TCP) scaffold.
DPSC, positive for the molecular markers CD133, Oct4, Nestin, Stro-1, and CD34, and negative for CD45, were isolated from extracted third molars. Experiments were started by seeding 200,000 cells on β-TCP cultured under 3% or 21% O₂ pressure. No osteogenic medium was used. Eight different cultures were performed at each time point under each O₂ pressure condition. Cell adhesion, proliferation, and differentiation over the biomaterial were evaluated at 7, 13, 18, and 23 days of culture. Cell adhesion was determined by light microscopy, proliferation by DNA quantification, and osteogenic differentiation by alkaline phosphatase (ALP) activity analysis.
DPSC adhered to β-TCP with both O₂ conditions. Cell proliferation was found from day 7 of culture. Higher values were recorded at 3% O₂ in each time point. Statistically significant differences were recorded at 23 days of culture (P = .033). ALP activity was not detectable at 7 days. There was, however, an increase in ALP activity over time in both groups. At 13, 18, and 23 days of culture, higher ALP activity was recorded under 3% O₂ pressure. Statistical differences were found at day 23 (P = .014).
DPSC display capacity of adhering to β-TCP under 3% or 21% O₂ pressure conditions. Cell proliferation on β-TCP phosphate is significantly higher at 3% than at 21% O₂ pressure, the most frequently used O₂ tension. β-TCP can itself promote osteogenic differentiation of DPSC and is enhanced under 3% O₂ compared with 21%.
在体外分析氧气压力对人牙髓干细胞（DPSC）在β-磷酸三钙（β-TCP）支架上的黏附、增殖和成骨分化的影响。
从拔除的第三磨牙中分离出分子标志物CD133、Oct4、巢蛋白、Stro-1和CD34呈阳性且CD45呈阴性的DPSC。实验通过将200,000个细胞接种到在3%或21%氧气压力下培养的β-TCP上开始。未使用成骨培养基。在每个氧气压力条件下的每个时间点进行8种不同的培养。在培养的第7、13、18和23天评估细胞在生物材料上的黏附、增殖和分化。通过光学显微镜确定细胞黏附，通过DNA定量确定增殖，通过碱性磷酸酶（ALP）活性分析确定成骨分化。
在两种氧气条件下DPSC均黏附于β-TCP。从培养第7天开始发现细胞增殖。在每个时间点，3%氧气条件下记录到更高的值。在培养23天时记录到统计学上的显著差异（P = 0.033）。在第7天未检测到ALP活性。然而，两组中ALP活性均随时间增加。在培养的第13、18和23天，3%氧气压力下记录到更高的ALP活性。在第23天发现统计学差异（P = 0.014）。
DPSC在3%或21%氧气压力条件下均表现出黏附于β-TCP的能力。在β-TCP磷酸盐上的细胞增殖在3%氧气压力下显著高于最常用的21%氧气张力。β-TCP本身可促进DPSC的成骨分化，与21%相比在3%氧气条件下增强。
