Low-intensity laser phototherapy enhances the proliferation of dental pulp stem cells under nutritional deficiency
Low-intensity laser phototherapy enhances the proliferation of dental pulp stem cells under nutritional deficiency
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DOI：
10.1590/1807-3107bor-2016.vol30.0080
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发表时间：
2016-01-01
影响因子：
2.5
通讯作者：
MARQUES, Marcia Martins
中科院分区：
文献类型：
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作者：
MOURA-NETTO, Cacio;FERREIRA, Leila Soares;MARQUES, Marcia Martins
未成熟恒牙的牙外伤可损伤牙髓血管，导致牙髓坏死和根尖发育停止。利用人脱落乳牙干细胞进行组织工程的研究已取得了可喜的成果。激光光疗法（LPT）能够影响这些细胞的增殖和分化，这将有助于促进组织工程。将SHED（第八代）接种到96孔培养板（103个细胞/孔）中，并在补充有15%确定的胎牛血清（FBS）的培养基中生长12小时。在确定适当的营养缺乏状态（5%FBS）后，将细胞分为四组：1）G1 - 15%FBS（阳性对照）; 2）G2 - 5%FBS（阴性对照）; 3）G3 - 5%FBS +LPT 3 J/cm（2）;和4）G4 - 5%FBS +LPT 5 J/cm（2）。对于LPT组，使用连续波InGaAlP二极管激光器（660 nm，光斑尺寸为0.028 cm（2），10 mW）以点状和接触模式进行两次激光照射，间隔6 h。在第二次激光照射后（0 h）和24、48和72 h后立即通过MTT还原试验评估细胞活力。我们发现，G3和G4与G2相比呈现出显著更高的细胞生长速率（p < 0.01）。此外，在整个实验中，G4表现出与G1相似的细胞生长速率（p > 0.05）。这些结果表明，在营养缺乏的情况下，5 J/cm（2）的LPT可以促进SHEDs的生长。因此，LPT可能是一个有价值的辅助治疗组织工程时，使用干细胞来源于乳牙牙髓。
Dental trauma in immature permanent teeth can damage pulp vascularization, which leads to necrosis and cessation of apexogenesis. Studies on tissue engineering using stem cells from human exfoliated deciduous teeth (SHEDs) have yielded promising results. Laser phototherapy (LPT) is able to influence the proliferation and differentiation of these cells, which could improve tissue engineering. SHEDs (eighth passage) were seeded into 96-well culture plates (103 cells/well) and were grown in culture medium supplemented with 15% defined fetal bovine serum (FBS) for 12 h. After determining the appropriate nutrition deficiency status (5% FBS), the cells were assigned into four groups: 1) G1 - 15% FBS (positive control); 2) G2 - 5% FBS (negative control); 3) G3 - 5% FBS+LPT 3 J/cm(2); and 4) G4 - 5% FBS+LPT 5 J/cm(2). For the LPT groups, two laser irradiations at 6 h intervals were performed using a continuous wave InGaAlP diode laser (660 nm, with a spot size of 0.028 cm(2), 10 mW) in punctual and contact mode. Cell viability was assessed via an MTT reduction assay immediately after the second laser irradiation (0 h) and 24, 48, and 72 h later. We found that G3 and G4 presented a significantly higher cell growth rate when compared with G2 (p < 0.01). Moreover, G4 exhibited a similar cell growth rate as G1 throughout the entire experiment (p > 0.05). These findings indicate that LPT with 5 J/cm(2) can enhance the growth of SHEDs during situations of nutritional deficiency. Therefore, LPT could be a valuable adjunct treatment in tissue engineering when using stem cells derived from the dental pulp of primary teeth.