牙科烤瓷合金对金黄地鼠颊囊黏膜中凋亡相关蛋白表达的影响
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摘要
背景:有研究证实,牙科烤瓷合金具有不同程度的细胞毒性,可导致细胞凋亡,但以往关于牙科烤瓷合金引起细胞凋亡的研究多采用体外实验,无法很好地模拟口腔环境。目的:观察牙科烤瓷合金对金黄地鼠颊囊黏膜中凋亡相关蛋白表达的影响。方法:取60-70 d龄雄性金黄地鼠36只(北京维通利华实验动物技术有限公司提供),随机分6组,分别在颊囊黏膜表面缝合固定镍铬烤瓷合金、钴铬烤瓷合金、纯钛烤瓷合金、钯基烤瓷合金、金铂烤瓷合金及聚丙烯(阴性对照)。固定14,28 d,切取与试件接触的颊囊黏膜,Western Blot检测黏膜细胞中Caspase-3、Caspase-8、Caspase-9蛋白表达情况。结果与结论:(1)固定14,28 d时,金铂烤瓷合金组Caspase-3蛋白表达量与阴性对照组比较无明显差异(P>0.05),其余4种烤瓷合金组Caspase-3蛋白表达量高于阴性对照组(P <0.05);(2)固定14 d时,各烤瓷合金组Caspase-8蛋白表达量均高于阴性对照组(P<0.05);固定28d时,镍铬烤瓷合金组、钯基烤瓷合金组Caspase-8蛋白表达量高于阴性对照组(P<0.05),其余烤瓷合金组Caspase-3蛋白表达量与阴性对照组比较无明显差异(P> 0.05);(3)固定14 d时,各烤瓷合金组Caspase-9蛋白表达量均高于阴性对照组(P <0.05);固定28 d时,钴铬合金组Caspase-9蛋白表达量与阴性对照组比较无明显差异(P> 0.05),其余烤瓷合金组Caspase-9蛋白表达量均高于阴性对照组(P <0.05);(4)结果表明,5种牙科烤瓷合金均不同程度引起凋亡相关信号分子的表达增加。
BACKGROUND: Dental porcelain alloys hold different levels of cytotoxicity, which can lead to cell apoptosis. However, the in vitro researches on dental ceramic alloy causing apoptosis are mainly conducted, in which the oral environment cannot be simulated well. OBJECTIVE: To investigate the effects of dental ceramic alloys on apoptotic protein expression in the oral buccal mucosa of golden hamsters. METHODS: Thirty-six 60-70-day-old male golden hamsters provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. were randomly divided into six groups. Five kinds of dental ceramic alloys(nickel-chromium, cobalt chromium, CPTi, palladium based and gold-platinum alloys) and polypropylene(negative control) were fixed on the oral mucosa of golden hamsters in corresponding groups. The mucosae contacting with the specimens were removed at 14 and 28 days, respectively. The protein expression levels of Caspase-3, Caspase-8, and Caspase-9 in the buccal mucosa were detected by western blot assay. RESULTS AND CONCLUSION:(1) At 14 and 28 days of fixation, there was no significant difference in the Caspase-3 protein level between gold-platinum alloy group and negative control groups(P > 0.05), and the level of Caspase-3 protein in the remaining four groups was significantly higher than that in the negative control group(P < 0.05).(2) At 14 days, the Caspase-8 protein level in each group was significantly higher than that in the negative control group(P < 0.05). After 28 days of fixation, the protein level of Caspase-8 in the nickel-chromium and palladium group was higher than that in the negative control group(P < 0.05), while the protein level of Caspase-8 in the other groups was not significantly different from that in the negative control group(P > 0.05).(3) At 14 days, the Caspase-9 protein level in each group was higher than that in the negative control group(P < 0.05). At 28 days, there was no significant difference in the Caspase-9 protein level between cobalt-chromium and negative control groups(P > 0.05), while the Caspase-9 protein level in the remaining groups was higher than that in the negative control group(P < 0.05). These results indicate that the use of five dental alloys can upregulate the levels of apoptosis-related signal molecules to different degrees.
BACKGROUND: Dental porcelain alloys hold different levels of cytotoxicity, which can lead to cell apoptosis. However, the in vitro researches on dental ceramic alloy causing apoptosis are mainly conducted, in which the oral environment cannot be simulated well. OBJECTIVE: To investigate the effects of dental ceramic alloys on apoptotic protein expression in the oral buccal mucosa of golden hamsters. METHODS: Thirty-six 60-70-day-old male golden hamsters provided by Beijing Vital River Laboratory Animal Technology Co., Ltd. were randomly divided into six groups. Five kinds of dental ceramic alloys(nickel-chromium, cobalt chromium, CPTi, palladium based and gold-platinum alloys) and polypropylene(negative control) were fixed on the oral mucosa of golden hamsters in corresponding groups. The mucosae contacting with the specimens were removed at 14 and 28 days, respectively. The protein expression levels of Caspase-3, Caspase-8, and Caspase-9 in the buccal mucosa were detected by western blot assay. RESULTS AND CONCLUSION:(1) At 14 and 28 days of fixation, there was no significant difference in the Caspase-3 protein level between gold-platinum alloy group and negative control groups(P > 0.05), and the level of Caspase-3 protein in the remaining four groups was significantly higher than that in the negative control group(P < 0.05).(2) At 14 days, the Caspase-8 protein level in each group was significantly higher than that in the negative control group(P < 0.05). After 28 days of fixation, the protein level of Caspase-8 in the nickel-chromium and palladium group was higher than that in the negative control group(P < 0.05), while the protein level of Caspase-8 in the other groups was not significantly different from that in the negative control group(P > 0.05).(3) At 14 days, the Caspase-9 protein level in each group was higher than that in the negative control group(P < 0.05). At 28 days, there was no significant difference in the Caspase-9 protein level between cobalt-chromium and negative control groups(P > 0.05), while the Caspase-9 protein level in the remaining groups was higher than that in the negative control group(P < 0.05). These results indicate that the use of five dental alloys can upregulate the levels of apoptosis-related signal molecules to different degrees.
引文
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[6]Y-T 0127.13-2009,中华人民共和国医药行业标准,口腔医疗器械生物学评价:试验方法:口腔黏膜刺激试验[S].北京:中国标准出版社,2009.
[7]ISO 10993-12-2007.Biological evaluation of medical devices-Part 12:Sample preparation and reference materials,2007.
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[12]谢颖.线粒体损伤拮抗剂在Cr(VI)所致肝细胞凋亡中的保护作用[D].长沙:中南大学,2014.
[13]李秀梅战,德松.Caspase-3表达强度评价齿科合金生物相容性的实验研究[J].中国医学工程,2011,19(7):70-73.
[14]郭毅.纳米级氧化锆与钛合金颗粒诱导大鼠海马凋亡的对比性研究[D].徐州:徐州医学院,2013.
[15]万书健,苏俭生.3种铸造金属全冠戴用后对外周静脉血淋巴细胞凋亡的影响[J].口腔医学,2009,29(2):73-76.
[16]焦俊霞,高维娟.细胞凋亡的信号转导机制研究进展[J].中国老年学,2010,30(6):853-856.
[17]Nutt LK,Pataer A,Pahler J,et al.Bax and Bak promote apoptosis by modulating endoplasmic reticular and mitochondrial Ca2+stores.J Biol Chem.2002;277(11):9219-9225.
[18]Brenner C,Kroemer G.Apoptosis.Mitochondria--the death signal integrators.Science.2000;289(5482):1150-1151.
[19]Huang DC,Strasser A.BH3-Only proteins-essential initiators of apoptotic cell death.Cell.2000;103(6):839-842.
[20]吴科达,王远亮,潘君.生物材料诱导细胞凋亡的研究进展[J].生物医学工程学杂志,2005,22(2):413-419.
[21]陈书宝,张睿,李锐.不同齿科合金对成纤维细胞增殖凋亡的影响及机制研究[J].医学研究杂志,2018,47(1):103-107.
[2]林泓磊,江磊,张长源,等.反复熔铸对钴铬烤瓷合金细胞相容性的影响[J].上海口腔医学,2017,26(3):246-250
[3]Faccioni F,Franceschetti P,Cerpelloni M,et al.In vivo study on metal release from fixed orthodontic appliances and DNA damage in oral mucosa cells.Am J Orthod Dentofacial Orthop.2003;124(6):687-694.
[4]Cortizo MC,DeMele MF,et al.Metallic dental material biocompatibility in osteoblast like cells:correlation with metal in release.Bio Trace Elem Res.2004;100(2)151-168.
[5]Schedle A,Samorapoompichit P,Rausch-Fan XH,et al.Response of L-929 fibroblasts,human gingival fibroblasts,and human tissue mast cells to various metal cations.J Dent Res.1995;74(8):1513-1520.
[6]Y-T 0127.13-2009,中华人民共和国医药行业标准,口腔医疗器械生物学评价:试验方法:口腔黏膜刺激试验[S].北京:中国标准出版社,2009.
[7]ISO 10993-12-2007.Biological evaluation of medical devices-Part 12:Sample preparation and reference materials,2007.
[8]Wolf BB,Green DR.Suicidal tendencies:apoptotic cell death by caspase family proteinases.J Biol Chem.1999;274(29):20049-20052.
[9]Joseph EK,Levine JD.Caspasesignalling in neuropathic and inflammatory pain in the rat.Eur J Neurosci.2004;20(11):2896-2902.
[10]孟贺,李秀梅,徐艳丽,等.3种牙科金属材料对L929细胞的毒性及对凋亡相关基因与蛋白表达的影响[J].上海口腔医学,2013,22(1):30-34.
[11]孟贺,丁洁,李任,等.4种牙科金属材料对成纤维细胞L929凋亡相关基因及蛋白表达的影响[J].华西口腔医学杂志,2013,31(3):242-246.
[12]谢颖.线粒体损伤拮抗剂在Cr(VI)所致肝细胞凋亡中的保护作用[D].长沙:中南大学,2014.
[13]李秀梅战,德松.Caspase-3表达强度评价齿科合金生物相容性的实验研究[J].中国医学工程,2011,19(7):70-73.
[14]郭毅.纳米级氧化锆与钛合金颗粒诱导大鼠海马凋亡的对比性研究[D].徐州:徐州医学院,2013.
[15]万书健,苏俭生.3种铸造金属全冠戴用后对外周静脉血淋巴细胞凋亡的影响[J].口腔医学,2009,29(2):73-76.
[16]焦俊霞,高维娟.细胞凋亡的信号转导机制研究进展[J].中国老年学,2010,30(6):853-856.
[17]Nutt LK,Pataer A,Pahler J,et al.Bax and Bak promote apoptosis by modulating endoplasmic reticular and mitochondrial Ca2+stores.J Biol Chem.2002;277(11):9219-9225.
[18]Brenner C,Kroemer G.Apoptosis.Mitochondria--the death signal integrators.Science.2000;289(5482):1150-1151.
[19]Huang DC,Strasser A.BH3-Only proteins-essential initiators of apoptotic cell death.Cell.2000;103(6):839-842.
[20]吴科达,王远亮,潘君.生物材料诱导细胞凋亡的研究进展[J].生物医学工程学杂志,2005,22(2):413-419.
[21]陈书宝,张睿,李锐.不同齿科合金对成纤维细胞增殖凋亡的影响及机制研究[J].医学研究杂志,2018,47(1):103-107.