不同光照能量LED红光照射牙周膜干细胞与根尖乳头干细胞的增殖
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摘要
背景:光生物调节是一种新型治疗方案,LED红光能否影响牙源干细胞增殖,是探索颌面部再生治疗新方案的关键。目的:比较不同光照能量LED红光对人牙周膜干细胞与人根尖乳头干细胞增殖的影响。方法:分离培养、流式鉴定获取人牙周膜干细胞和人根尖乳头干细胞。MTT法检测不同光照能量0,1,3,5 J/cm~2对两种牙源性干细胞增殖的影响。结果与结论:①非光照下根尖乳头干细胞增殖速度较牙周膜干细胞更快,1,3,5 J/cm~2 LED红光均能促进两种牙源性干细胞增殖;②LED红光能促进牙周膜干细胞缓慢增殖期和对数生长期(1-7d)增殖,其中5J/cm~2对早期影响最明显,后期1 J/cm~2增殖作用最强;③LED红光对根尖乳头干细胞对数生长后期(5-7 d)影响较明显,3 J/cm~2LED红光的增殖作用于第7天达到峰值;④根据人牙周膜干细胞和人根尖乳头干细胞对LED红光的时效依赖及量效依赖性,选定特定的光照能量及时间,可快速有效地促进细胞增殖。
BACKGROUND: Photobiomodulation is a new treatment option. Whether red light-emitting diodes can affect the proliferation and regeneration of dental stem cells is the key to exploring new treatments for maxillofacial regeneration OBJECTIVE: To compare the effects of red light-emitting diodes with different light energies on the proliferation of human periodontal ligament stem cells and apical papillary stem cells.METHODS: Periodontal ligament stem cells and apical papillary stem cells were isolated and cultured by flow cytometry. Proliferation of these cells was detected using MTT assay after exposure to red light-emitting diodes at different light energies(0, 1, 3, and 5 J/cm2).RESULTS AND CONCLUSION: Apical papillary stem cells proliferated faster than periodontal ligament stem cells under non-lighting environment. Red light emitting diodes at 1, 3, and 5 J/cm~2 promoted the proliferation of two kinds of odontogenic stem cells. The red light-emitting diode promoted the proliferation of periodontal ligament stem cells at proliferative and logarithmic growth phase(1-7 days). And 5 J/cm~2 red light emitting diode had the most obvious effect in early stage, and 1 J/cm~2 had the strongest effect in the late stage. The red light-emitting diode had a significant effect on the logarithmic growth(5-7 days) of apical papilla stem cells, and under 3 J/cm~2 red light-emitting diode the proliferation peaked on the 7 th day. Therefore, the red light-emitting diode can promote the proliferation of human periodontal ligament stem cells and apical papillary stem cells in dose-and time-dependent manner. Exposure to red light-emitting diode at proper energy and time can promote cell proliferation quickly and effectively.
BACKGROUND: Photobiomodulation is a new treatment option. Whether red light-emitting diodes can affect the proliferation and regeneration of dental stem cells is the key to exploring new treatments for maxillofacial regeneration OBJECTIVE: To compare the effects of red light-emitting diodes with different light energies on the proliferation of human periodontal ligament stem cells and apical papillary stem cells.METHODS: Periodontal ligament stem cells and apical papillary stem cells were isolated and cultured by flow cytometry. Proliferation of these cells was detected using MTT assay after exposure to red light-emitting diodes at different light energies(0, 1, 3, and 5 J/cm2).RESULTS AND CONCLUSION: Apical papillary stem cells proliferated faster than periodontal ligament stem cells under non-lighting environment. Red light emitting diodes at 1, 3, and 5 J/cm~2 promoted the proliferation of two kinds of odontogenic stem cells. The red light-emitting diode promoted the proliferation of periodontal ligament stem cells at proliferative and logarithmic growth phase(1-7 days). And 5 J/cm~2 red light emitting diode had the most obvious effect in early stage, and 1 J/cm~2 had the strongest effect in the late stage. The red light-emitting diode had a significant effect on the logarithmic growth(5-7 days) of apical papilla stem cells, and under 3 J/cm~2 red light-emitting diode the proliferation peaked on the 7 th day. Therefore, the red light-emitting diode can promote the proliferation of human periodontal ligament stem cells and apical papillary stem cells in dose-and time-dependent manner. Exposure to red light-emitting diode at proper energy and time can promote cell proliferation quickly and effectively.
引文
[1]童晓洁,兰泽栋,郑雅心,等.骨形成蛋白2和牙胚细胞联合作用诱导人牙周膜干细胞表达成牙骨质细胞的表型研究[J].中国美容医学, 2014, 23(16):1343-1348.
[2]Abe S, Yamaguchi S, Watanabe A, et al. Hard tissue regeneration capacity of apical pulp derived cells(APDCs)from human tooth with immature apex. Biochem Biophys Res Commun. 2008;371(1):90-93.
[3]Bakopoulou A, Leyhausen G, Volk J, et al. Comparative analysis of in vitro osteo/odontogenic differentiation potential of human dental pulp stem cells(DPSCs)and stem cells from the apical papilla(SCAP). Arch Oral Biol. 2011;56(7):709-721.
[4]Huang GT, Sonoyama W, Liu Y, et al. The hidden treasure in apical papilla:the potential role in pulp/dentin regeneration and bioroot engineering. J Endod. 2008;34(6):645-651.
[5]Merry GF, Munk MR, Dotson RS, et al. Photobiomodulation reduces drusen volume and improves visual acuity and contrast sensitivity in dry age-related macular degeneration. Acta Ophthalmol. 2017;95(4):e270-e277.
[6]项红升,李明,霍荣龄,等.LED应用于光疗的研究进展[J].北京生物医学工程, 2005,24(4):311-315.
[7]Peng F, Wu H, Zheng Y, et al. The effect of noncoherent red light irradiation on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells. Lasers Med Sci. 2012;27(3):645-653.
[8]Ding G, Niu J, Wei F. Current understanding of orofacial tissue derived mesenchymal stem cells:an immunological perspective. Histol Histopathol. 2015;30(3):255-265.
[9]Seo BM, Miura M, Gronthos S, et al. Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet. 2004; 364(9429):149-155.
[10] Sonoyama W, Liu Y, Fang D, et al. Mesenchymal stem cell-mediated functional tooth regeneration in swine. PLoS One.2006;1:e79.
[11] Sonoyama W, Liu Y, Yamaza T, et al. Characterization of the apical papilla and its residing stem cells from human immature permanent teeth:a pilot study. J Endod. 2008;34(2):166-171.
[12] Li Y, Zhang J, Xu Y, et al. Effects of 630 nm Red and 460 nm Blue Light Emitting Diode Irradiation on Healing of the Skin Wound in Japanese Big-ear White Rabbit. Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2017;39(3):301-306.
[13] Mamalis A, Jagdeo J. High-Fluence Light-Emitting Diode-Generated Red Light Modulates the Transforming Growth Factor-Beta Pathway in Human Skin Fibroblasts.Dermatol Surg. 2018;44(10):1317-1322.
[14] Parker S. Low-level laser use in dentistry. Br Dent J. 2007;202(3):131-138.
[15] Yang D, Yi W, Wang E, et al. Effects of light-emitting diode irradiation on the osteogenesis of human umbilical cord mesenchymal stem cells in vitro. Sci Rep. 2016;6:37370.
[16] Fekrazad R, Asefi S, Allahdadi M, et al. Effect of Photobiomodulation on Mesenchymal Stem Cells. Photomed Laser Surg. 2016;34(11):533-542.
[17] Pagin MT, de Oliveira FA, Oliveira RC, et al. Laser and light-emitting diode effects on pre-osteoblast growth and differentiation. Lasers Med Sci. 2014;29(1):55-59.
[18] Horvát-Karajz K, Balogh Z, Kovács V, et al. In vitro effect of carboplatin, cytarabine, paclitaxel, vincristine, and low-power laser irradiation on murine mesenchymal stem cells. Lasers Surg Med. 2009;41(6):463-469.
[19] Asai T, Suzuki H, Kitayama M, et al. The long-term effects of red light-emitting diode irradiation on the proliferation and differentiation of osteoblast-like MC3T3-E1 cells. Kobe J Med Sci. 2014;60(1):E12-18.
[20] Mamalis A, Jagdeo J. Light-emitting diode-generated red light inhibits keloid fibroblast proliferation. Dermatol Surg. 2015;41(1):35-39.
[21] Mamalis A, Garcha M, Jagdeo J. Light emitting diode-generated blue light modulates fibrosis characteristics:fibroblast proliferation, migration speed, and reactive oxygen species generation. Lasers Surg Med. 2015;47(2):210-215.
[22]赵璐,于莉,袁萍,等.根尖乳头干细胞与牙周膜干细胞的生物学行为比较[J].中国组织工程研究,2016,20(1):113-117.
[23] Wu HP, Persinger MA. Increased mobility and stem-cell proliferation rate in Dugesia tigrina induced by 880nm light emitting diode. J Photochem Photobiol B. 2011;102(2):156-160.
[24] Mandrillo PM, Fischetto G, Odorisio P, et al. Effects of light-emitting diode(led 640nm)on human gingival fibroblasts:a comparative in vitro study. Oral Implantol(Rome). 2017;10(2):151-161.
[2]Abe S, Yamaguchi S, Watanabe A, et al. Hard tissue regeneration capacity of apical pulp derived cells(APDCs)from human tooth with immature apex. Biochem Biophys Res Commun. 2008;371(1):90-93.
[3]Bakopoulou A, Leyhausen G, Volk J, et al. Comparative analysis of in vitro osteo/odontogenic differentiation potential of human dental pulp stem cells(DPSCs)and stem cells from the apical papilla(SCAP). Arch Oral Biol. 2011;56(7):709-721.
[4]Huang GT, Sonoyama W, Liu Y, et al. The hidden treasure in apical papilla:the potential role in pulp/dentin regeneration and bioroot engineering. J Endod. 2008;34(6):645-651.
[5]Merry GF, Munk MR, Dotson RS, et al. Photobiomodulation reduces drusen volume and improves visual acuity and contrast sensitivity in dry age-related macular degeneration. Acta Ophthalmol. 2017;95(4):e270-e277.
[6]项红升,李明,霍荣龄,等.LED应用于光疗的研究进展[J].北京生物医学工程, 2005,24(4):311-315.
[7]Peng F, Wu H, Zheng Y, et al. The effect of noncoherent red light irradiation on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells. Lasers Med Sci. 2012;27(3):645-653.
[8]Ding G, Niu J, Wei F. Current understanding of orofacial tissue derived mesenchymal stem cells:an immunological perspective. Histol Histopathol. 2015;30(3):255-265.
[9]Seo BM, Miura M, Gronthos S, et al. Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet. 2004; 364(9429):149-155.
[10] Sonoyama W, Liu Y, Fang D, et al. Mesenchymal stem cell-mediated functional tooth regeneration in swine. PLoS One.2006;1:e79.
[11] Sonoyama W, Liu Y, Yamaza T, et al. Characterization of the apical papilla and its residing stem cells from human immature permanent teeth:a pilot study. J Endod. 2008;34(2):166-171.
[12] Li Y, Zhang J, Xu Y, et al. Effects of 630 nm Red and 460 nm Blue Light Emitting Diode Irradiation on Healing of the Skin Wound in Japanese Big-ear White Rabbit. Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2017;39(3):301-306.
[13] Mamalis A, Jagdeo J. High-Fluence Light-Emitting Diode-Generated Red Light Modulates the Transforming Growth Factor-Beta Pathway in Human Skin Fibroblasts.Dermatol Surg. 2018;44(10):1317-1322.
[14] Parker S. Low-level laser use in dentistry. Br Dent J. 2007;202(3):131-138.
[15] Yang D, Yi W, Wang E, et al. Effects of light-emitting diode irradiation on the osteogenesis of human umbilical cord mesenchymal stem cells in vitro. Sci Rep. 2016;6:37370.
[16] Fekrazad R, Asefi S, Allahdadi M, et al. Effect of Photobiomodulation on Mesenchymal Stem Cells. Photomed Laser Surg. 2016;34(11):533-542.
[17] Pagin MT, de Oliveira FA, Oliveira RC, et al. Laser and light-emitting diode effects on pre-osteoblast growth and differentiation. Lasers Med Sci. 2014;29(1):55-59.
[18] Horvát-Karajz K, Balogh Z, Kovács V, et al. In vitro effect of carboplatin, cytarabine, paclitaxel, vincristine, and low-power laser irradiation on murine mesenchymal stem cells. Lasers Surg Med. 2009;41(6):463-469.
[19] Asai T, Suzuki H, Kitayama M, et al. The long-term effects of red light-emitting diode irradiation on the proliferation and differentiation of osteoblast-like MC3T3-E1 cells. Kobe J Med Sci. 2014;60(1):E12-18.
[20] Mamalis A, Jagdeo J. Light-emitting diode-generated red light inhibits keloid fibroblast proliferation. Dermatol Surg. 2015;41(1):35-39.
[21] Mamalis A, Garcha M, Jagdeo J. Light emitting diode-generated blue light modulates fibrosis characteristics:fibroblast proliferation, migration speed, and reactive oxygen species generation. Lasers Surg Med. 2015;47(2):210-215.
[22]赵璐,于莉,袁萍,等.根尖乳头干细胞与牙周膜干细胞的生物学行为比较[J].中国组织工程研究,2016,20(1):113-117.
[23] Wu HP, Persinger MA. Increased mobility and stem-cell proliferation rate in Dugesia tigrina induced by 880nm light emitting diode. J Photochem Photobiol B. 2011;102(2):156-160.
[24] Mandrillo PM, Fischetto G, Odorisio P, et al. Effects of light-emitting diode(led 640nm)on human gingival fibroblasts:a comparative in vitro study. Oral Implantol(Rome). 2017;10(2):151-161.