纳米珍珠粉对小鼠骨髓间充质干细胞活性影响研究
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摘要
目的探讨纳米珍珠粉(nano-pearl powder,NPP)对小鼠骨髓间充质干细胞活性的影响。方法研究于2016年8月至2017年3月在中南大学湘雅医学院附属海口医院实验中心进行。将NPP以100、300、500μg/mL的质量浓度与小鼠骨髓间充质干细胞共培养24 h(NPP组),用CCK8法测定吸光度值,计算细胞相对增殖率,评定其细胞毒性级别;用Annexin-FITC/PI法检测细胞凋亡率。两者综合评定NPP对小鼠骨髓间充质干细胞活性的影响,并与相同质量浓度纳米羟基磷灰石(nano-hydroxyapatite,NHA)(NHA组)进行对比。结果 NPP与NHA细胞毒性检测均合格,NPP在100μg/mL时细胞相对增殖率最高;NPP组细胞凋亡率均低于NHA组,在质量浓度为100和500μg/mL时的差异有统计学意义(P <0.01)。结论 NPP细胞毒性检测合格,其对小鼠骨髓间充质干细胞活性的影响优于NHA。
Objective To investigate the effect of nano-pearl powder(NPP)on cell viability of mouse bone marrow mesenchymal stem cells. Methods Mouse bone marrow mesenchymal stem cells(m BMMSCs)were co-cultured with NPP at concentration of 100,300,500 μg/mL for 24 h(NPP group). CCK8 test was carried out to evaluate the relative growth rate and cytotoxicity. The apoptosis rate of cells was measured by Annexin-FITC/PI method. The effect of NPP on mBMMSCs activity was analyzed by the above two approaches,and the results were compared with nano-hydroxyapatite(NHA group)at the same concentration. Results The cytotoxicity tests of NPP and NHA were qualified,and the relative proliferation rates of mBMMSCs in NPP groups were highest at 100 μg/mL. The apoptotic rates of mBMMSCs in NPP groups were all lower than those in NHA groups. The difference was statistically significant at 100 and 500 μg/mL(P <0.01). Conclusion The cytotoxicity tests of NPP are qualified,and its effect on the cell viability of mBMMSCs is better than that of NHA.
Objective To investigate the effect of nano-pearl powder(NPP)on cell viability of mouse bone marrow mesenchymal stem cells. Methods Mouse bone marrow mesenchymal stem cells(m BMMSCs)were co-cultured with NPP at concentration of 100,300,500 μg/mL for 24 h(NPP group). CCK8 test was carried out to evaluate the relative growth rate and cytotoxicity. The apoptosis rate of cells was measured by Annexin-FITC/PI method. The effect of NPP on mBMMSCs activity was analyzed by the above two approaches,and the results were compared with nano-hydroxyapatite(NHA group)at the same concentration. Results The cytotoxicity tests of NPP and NHA were qualified,and the relative proliferation rates of mBMMSCs in NPP groups were highest at 100 μg/mL. The apoptotic rates of mBMMSCs in NPP groups were all lower than those in NHA groups. The difference was statistically significant at 100 and 500 μg/mL(P <0.01). Conclusion The cytotoxicity tests of NPP are qualified,and its effect on the cell viability of mBMMSCs is better than that of NHA.
引文
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[23]贾健,李艳博,郭彩霞.纳米材料诱导细胞死亡的研究进展[J].中国药理学与毒理学杂志,2016,30(4):421-428.
[24] Horie M,Nishio K,Kato H,et al. Evaluation of cellular influences caused by calcium carbonate nanoparticles[J]. Chem Biol Interact,2014,210:64-76.
[25] Denu RA,Hematti P. Effects of oxidative stress on mesenchymal stem cell biology[J]. Oxid Med Cell Longev,2016,2016:2989076.
[26] Chaturvedi R,Singha PK,Dey S. Water soluble bioactives of nacre mediate antioxidant activity and osteoblast differentiation[J]. PloS One,2013,8(12):e84584.
[2]许艳慧,李四群,李志安.纳米羟基磷灰石复合材料的研究——纳米羟基磷灰石的细胞毒性[J].实用口腔医学杂志,2004,20(2):147-150.
[3]乔广艳,沈庆平,苏俭生. 3种临床常用烤瓷合金对小鼠成纤维细胞L929的毒性评价[J].上海口腔医学,2010,19(1):72-76.
[4] Tropel P,No?l D,Platet N,et al. Isolation and characterisation of mesenchymal stem cells from adult mouse bone marrow[J]. Exp Cell Res,2004,295(2):395-406.
[5] Bruder SP,Kurth AA,Shea M,et al. Bone regeneration by implantation of purified,culture-expanded human mesenchymal stem cells[J]. J Orthop Res,1998,16(2):155-162.
[6] Shafiee A,Seyedjafari E,Soleimani M,et al. A comparison between osteogenic differentiation of human unrestricted somatic stem cells and mesenchymal stem cells from bone marrow and adipose tissue[J]. Biotechnol Lett,2011,33(6):1257-1264.
[7] Sepulveda P,Bressiani AH,Bressiani JC,et al. In vivo evaluation of hydroxyapatite foams[J]. J Biomed Mater Res,2002,2(4):587-592.
[8] Suto M,Nemoto E,Kanaya S,et al. Nanohydroxyapatite increases BMP-2 expression via a p38 MAP kinase dependent pathway in periodontal ligament cells[J]. Arch Oral Biol,2013,58(8):1021-1028.
[9]潘泽民,刘开泰,杨磊,等.砷对细胞和生物作用的研究进展[J].环境与健康杂志,2003,20(3):184-186.
[10]张艳芳,金婵,马春旺,等.几种纳米材料细胞毒性效应的研究现状[J].第二军医大学学报,2010,31(11):1234-1238.
[11] Xu Z,Liu C,Wei J,et al. Effects of four types of hydroxyapatite nanoparticles with different nanocrystal morphologies and sizes on apoptosis in rat osteoblasts[J]. J Appl Toxicol,2012,32(6):429-435.
[12] Yuan Y,Liu C,Qian J,et al. Size-mediated cytotoxicity and apoptosis of hydroxyapatite nanoparticles in human hepatoma HepG2 cells[J]. Biomaterials,2010,31(4):730-740.
[13] Xu J,Xu P,Li Z,et al. Oxidative stress and apoptosis induced by hydroxyapatite nanoparticles in C6 cells[J]. J Biomed Mater Res A,2012,100(3):738-745.
[14]沈链链,钟义红,李世芬,等.珍珠粉的食用安全性毒理学评价[J].包头医学院学报,2013,29(5):21-24.
[15]吴家瑜,竹剑平.水溶性珍珠粉的毒理实验研究[J].中华中医药学刊,2009,27(2):397-399.
[16]毛秋华.纳米级淡水珍珠粉的制备及其生物相容性初步研究[D].长沙:中南大学,2014.
[17] Shen Y,Zhu J,Zhang H,et al. In vitro osteogenetic activity of pearl[J]. Biomaterials,2006,27(2):281-287.
[18] Xu M,Li Y,Suo H,et al. Fabricating a pearl/PLGA composite scaffold by the low-temperature deposition manufacturing technique for bone tissue engineering[J]. Biofabrication,2010,2(2):025002.
[19] Chandra J,Samali A,Orrenius S. Triggering and modulation of apoptosis by oxidative stress[J]. Free Radic Biol Med,2000,29(3-4):323-333.
[20] Yang SR,Rahman I,Trosko JE,et al. Oxidative stress-induced biomarkers for stem cell-based chemical screening[J]. Prev Med,2012,54(Suppl):S42-S49.
[21] Liu Z,Jiao Y,Wang Y,et al. Polysaccharides-based nanoparticles as drug deliverysystems[J]. Adv Drug Deliv Rev,2008,60(15):1650-1662.
[22] Yuan Y,Liu C,Qian J,et al. Size-mediated cytotoxicity and apoptosis of hydroxyapatite nanoparticles in human hepatoma HepG2 cells[J]. Biomaterials,2010,31(4):730-740.
[23]贾健,李艳博,郭彩霞.纳米材料诱导细胞死亡的研究进展[J].中国药理学与毒理学杂志,2016,30(4):421-428.
[24] Horie M,Nishio K,Kato H,et al. Evaluation of cellular influences caused by calcium carbonate nanoparticles[J]. Chem Biol Interact,2014,210:64-76.
[25] Denu RA,Hematti P. Effects of oxidative stress on mesenchymal stem cell biology[J]. Oxid Med Cell Longev,2016,2016:2989076.
[26] Chaturvedi R,Singha PK,Dey S. Water soluble bioactives of nacre mediate antioxidant activity and osteoblast differentiation[J]. PloS One,2013,8(12):e84584.