烤瓷合金对人牙龈成纤维细胞生物学行为的影响
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摘要
目的:探讨镍铬烤瓷合金、钴铬烤瓷合金和金铂烤瓷合金,以及镍铬烤瓷合金表面镀金对体外培养的人牙龈成纤维细胞(Human gingival fibroblast,HGF)分泌免疫细胞因子白细胞介素-6(Interleukin-6,IL-6)、生长与增殖活性以及附着生长能力等生物学行为的影响。从而评价其生物相容性,为临床烤瓷合金的选择提供参考。
方法:
1于临床取得正常牙龈组织,采用组织块贴壁法进行人牙龈成纤维细胞的原代培养,运用细胞免疫组织化学染色方法通过波形丝蛋白、角蛋白染色鉴定细胞来源。
2用不同烤瓷合金提浸液分别对人牙龈成纤维细胞进行培养,以未处理的DMEM培养液作为阴性对照。通过酶联免疫吸附试验(Enzyme-linked immunosorbent assay,ELISA)抗体加心法测定细胞培养上清液中IL-6的分泌量;通过四甲基偶氮唑盐微量酶反应比色法(Methylthiazoletrazolium,MTT)检测烤瓷合金提浸液对人牙龈成纤维细胞生长与增殖的影响。
3以盖玻片接种细胞作为阴性对照,在不同烤瓷合金表面分别接种人牙龈成纤维细胞。采用MTT法检测人牙龈成纤维细胞在不同烤瓷合金表面的附着生长与增殖活性;扫描电镜观察细胞形态与排列附着情况。
结果:
1细胞免疫组织化学染色,波形丝蛋白染色阳性、角蛋白染色阴性,证实细胞符合中胚层来源的成纤维细胞特点。
2不同烤瓷合金提浸液培养人牙龈成纤维细胞,ELISA结果显示:
6小时细胞培养上清液中IL-6的浓度均值分别为:对照组:133.75±12.50pg/ml,镍铬烤瓷合金组(A):180.00±10.36 pg/ml,钴铬烤瓷合金组(B):168.50±10.08 pg/ml,镍铬烤瓷合金镀金组(C):145.00±8.90 pg/ml,金铂烤瓷合金组(D):138.75±10.31 pg/ml。A、B组与对照组之间有显著性差异(P<0.05);C、D组与对照组之间无显著性差异(P>0.05);A组与B组、C组与D组比较,无显著性差异(P>0.05);A组与C组、A组与D组、B组与C组、B组与D组两两比较,有显著性差异(P<0.05)。
24小时对照组与A、B、C、D组细胞上清液中IL-6的浓度均值分别为:189.50±10.12pg/ml,228.75±8.54pg/ml,223.75±7.50pg/ml , 200.50±9.57pg/ml , 196.50±10.79pg/ml。A、B组与对照组之间有显著性差异(P<0.05);C、D组与对照组之间无显著性差异(P>0.05);A组与B组、C组与D组比较,无显著性差异(P>0.05);A组与C组、A组与D组、B组与C组、B组与D组两两比较,有显著性差异(P<0.05)。
3不同烤瓷合金提浸液培养人牙龈成纤维细胞,MTT比色实验结果显示:
与对照组相比,6小时镍铬烤瓷合金组(A),钴铬烤瓷合金组(B),镍铬烤瓷合金镀金组(C),金铂烤瓷合金组(D)细胞相对增殖率均值分别为:113.07±4.16%,110.50±4.55%,102.57±4.36%,99.00±3.17%。A、B组与对照组之间有显著性差异(P<0.05);C、D组与对照组之间无显著性差异(P>0.05);A组与B组、C组与D组比较无显著性差异(P>0.05);A组与C组、A组与D组、B组与C组、B组与D组两两比较,有显著性差异(P<0.05)。
与对照组相比,24小时A、B、C、D组细胞相对增殖率分别为:78.89±6.03%,82.33±4.44%,95.36±4.17%,97.92±5.83%。A、B组与对照组之间有显著性差异(P<0.05);C、D组与对照组之间无显著性差异(P>0.05);A组与B组、C组与D组比较,无显著性差异(P>0.05);A组与C组、A组与D组、B组与C组、B组与D组两两比较,有显著性差异(P<0.05)。
4不同烤瓷合金表面人牙龈成纤维细胞的MTT比色结果显示:与对照组相比,镍铬烤瓷合金组(A),钴铬烤瓷合金组(B),镍铬烤瓷合金镀金组(C),金铂烤瓷合金组(D)细胞相对增殖率分别为:69.39±3.71%,75.00±5.03%,86.72±3.55%,89.27±3.88%。A、B、C、D组与对照组之间均有显著性差异(P<0.05);A组与B组、C组与D组比较,无显著性差异(P>0.05);A组与C组、A组与D组、B组与C组、B组与D组两两比较,有显著性差异(P<0.05)。
5扫描电镜观察人牙龈成纤维细胞黏附行为:与对照组相比,镍铬烤瓷合金(A),钴铬烤瓷合金(B),镍铬烤瓷合金镀金(C),金铂烤瓷合金(D)表面附着细胞数目少,显示细胞极性,沿材料表面沟纹方向排列。A、B、C组细胞形态变化明显。D组细胞形态变化不明显。
结论:
1体外成功培养了人牙龈成纤维细胞。
2镍铬烤瓷合金和钴铬烤瓷合金提浸液对人牙龈成纤维细胞的增殖显示先促进后抑制作用;并且使细胞IL-6分泌水平升高。镍铬烤瓷合金镀金与金铂烤瓷合金提浸液不影响细胞细胞的增殖活性和IL-6的分泌水平。
3镍铬烤瓷合金、钴铬烤瓷合金、镍铬烤瓷合金镀金和金铂烤瓷合金均抑制人牙龈成纤维细胞的附着生长,影响细胞排列与形态。
4金铂烤瓷合金与镍铬烤瓷合金镀金对人牙龈成纤维细胞生物学行为的影响较小,镍铬与钴铬烤瓷合金对细胞的生物学行为影响较大。
Objective: This study is to investigate the effect of Ni-Cr, Co-Cr and Au-Pt ceramic alloys and Ni-Cr ceramic alloys with gold plating on the secretion of interleukin-6 (IL-6) by human gingival firoblasts (HGFs), and on the proliferation and attachment of HGF.
Methods:
1 Specimens of healthy gingival tissue were obtained from clinical operations, and primary culture of HGFs was performed using the tissue culture method. In order to identify the origin of the cultured cells, we performed immunocytochemical staining including vimentin and cytokeratin staining.
2 With untreated DMEM medium as negative control, we cultured HGFs in the eluates of various types of ceramic alloys separately. The amount of IL-6 secreted in the medium supermatant was measured by enzyme-linked immunosorbent assay (ELISA). In addition, we examined the influence of the eluates of various ceramic alloys on the growth and proliferation of HGFs.
3 HGFs were seeded onto the surface of various types of ceramic alloys as well as onto the cover slips, which served as negative control. Then we examined the adhesive growth and proliferating activity of human gingival firoblasts by Methylthiazoletrazolium (MTT) assay, and observe the cell material attachment with scanning electron microscope (SEM).
Results:
1 Immunocytochemistry result revealed that vimentin staining was positive and cytokeratin staining was negative. Therefore, the cultured cells were proved to be in accordance with the properties of fibroblasts from the mesoderm.
2 The result of ELISA measuring the secretion of IL-6 by HGFs, which is cultivated in eluates of various types of ceramic alloys, is listed as follows. At 6 hours, the average concentration level of IL-6 secreted in the medium supermatant for control group, Ni-Cr ceramic alloys (A), Co-Cr ceramic alloys (B), Ni-Cr ceramic alloys with gold plating (C) and Au-Pt ceramic alloys (D) is 133.75±12.50pg/ml,180.00±10.36pg/ml,168.50±10.08pg/ml,145.00±8.90pg/ml,and 138.75±10.31pg/ml respectively. Group A or group B is significantly different from the control (P<0.05). However, group C or group D is not significantly different from the control (P>0.05). Additionally, Group A is significantly different from group C or group D (P<0.05). Similarly, Group B is significantly different from group C or group D (P<0.05). Both of the differences between group A and B, and group C and D are not significant (P>0.05).
At 24 hours, the average concentration level of IL-6 secreted in the medium supermatant for control group and group A, group B, group C, group D is 189.50±10.12 pg/ml,228.75±8.54 pg/ml,223.75±7.50 pg/ml,200.50±9.57 pg/ml,and 196.50±10.79 pg/ml respectively. Group A or group B is significantly different from the control (P<0.05). However, group C or group D is not significantly different from the control (P>0.05). Additionally, Group A is significantly different from group C or group D (P<0.05). Similarly, Group B is significantly different from group C or group D (P<0.05). Both of the differences between group A and B, and group C and D are not significant (P>0.05).
3 The result of MTT assay measuring the proliferating activity of HGFs, which is cultivated in eluates of various types of ceramic alloys, is listed as follows.
At 6 hours, the average proliferating activity of HGFs for control group, Ni-Cr ceramic alloys (A), Co-Cr ceramic alloys (B), Ni-Cr ceramic alloys with gold plating (C) and Au-Pt ceramic alloys (D) is 113.07±4.16%,110.50±4.55%, 102.57±4.36%,and 99.00±3.17% respectively. Group A or group B is significantly different from the control (P<0.05). However, group C or group D is not significantly different from the control (P>0.05). Additionally, Group A is significantly different from group C or group D (P<0.05). Similarly, Group B is significantly different from group C or group D (P<0.05). Both of the differences between group A and B, and group C and D are not significant (P>0.05).
At 24 hours, the average proliferating activity of HGFs for control group and group A, group B, group C, group D is 78.89±6.03%, 82.33±4.44%, 95.36±4.17%, and 97.92±5.83% respectively. Group A or group B is significantly different from the control (P<0.05). However, group C or group D is not significantly different from the control (P>0.05). Additionally, Group A is significantly different from group C or group D (P<0.05). Similarly, Group B is significantly different from group C or group D (P<0.05). But no significant difference was observed between group A and group B, group C and group D (P>0.05).
4 The result of MTT assay measuring the proliferating activity of HGFs, which is cultivated onto the surface of various types of ceramic alloys, showed that the average proliferating activity of HGFs for control group, Ni-Cr ceramic alloys (A), Co-Cr ceramic alloys (B), Ni-Cr porcelain alloys with gold plating (C) and Au-Pt porcelain alloys (D) is 69.39±3.71%, 75.00±5.03%, 86.72±3.55%, and 89.27±3.88% respectively. Each of the group A, group B, group C, and group D is significantly different from the control (P<0.05). Additionally, Group A is significantly different from group C or group D (P<0.05). Similarly, Group B is significantly different from group C or group D (P<0.05). But no significant difference was observed between group A and group B, group C and group D (P>0.05).
5 The SEM result indicates that the number of HGFs attached to the surface of Ni-Cr ceramic alloys (A), Co-Cr ceramic alloys (B), Ni-Cr ceramic alloys with gold plating (C) and Au-Pt ceramic alloys (D) were less than that of HGFs attached to the surface of the control group. The alignment of the cells along the rills of the materials showed polarity. The morphological change of cells between group A, B, C and the control is significant. The morphological change of cells between group D and the control is not significant.
Conclusion:
1 The human gingival fibroblasts (HGFs) were cultured successfully in vitro.
2 The eluates of Ni-Cr and Co-Cr ceramic alloys can first promote the proliferation of HGFs, and then suppress it. Besides, they can stimulate the secretion of IL-6. But the eluates of Ni-Cr ceramic alloys with gold plating and Au-Pt porcelain alloys did not affect the proliferation of HGFs and the secretion of IL-6.
3 Ni-Cr, Co-Cr, Ni-Cr and Au-Pt ceramic alloys and Ni-Cr ceramic alloys with gold plating can suppress the growth of adhesive HGFs, and influence the cells alignment and morphology.
4 Au-Pt ceramic alloy and Ni-Cr ceramic alloy with gold plating have no obvious influnce on the biological behavior of HGFs, while Ni-Cr and Co-Cr ceramic alloys have significant influnce on that.
方法:
1于临床取得正常牙龈组织,采用组织块贴壁法进行人牙龈成纤维细胞的原代培养,运用细胞免疫组织化学染色方法通过波形丝蛋白、角蛋白染色鉴定细胞来源。
2用不同烤瓷合金提浸液分别对人牙龈成纤维细胞进行培养,以未处理的DMEM培养液作为阴性对照。通过酶联免疫吸附试验(Enzyme-linked immunosorbent assay,ELISA)抗体加心法测定细胞培养上清液中IL-6的分泌量;通过四甲基偶氮唑盐微量酶反应比色法(Methylthiazoletrazolium,MTT)检测烤瓷合金提浸液对人牙龈成纤维细胞生长与增殖的影响。
3以盖玻片接种细胞作为阴性对照,在不同烤瓷合金表面分别接种人牙龈成纤维细胞。采用MTT法检测人牙龈成纤维细胞在不同烤瓷合金表面的附着生长与增殖活性;扫描电镜观察细胞形态与排列附着情况。
结果:
1细胞免疫组织化学染色,波形丝蛋白染色阳性、角蛋白染色阴性,证实细胞符合中胚层来源的成纤维细胞特点。
2不同烤瓷合金提浸液培养人牙龈成纤维细胞,ELISA结果显示:
6小时细胞培养上清液中IL-6的浓度均值分别为:对照组:133.75±12.50pg/ml,镍铬烤瓷合金组(A):180.00±10.36 pg/ml,钴铬烤瓷合金组(B):168.50±10.08 pg/ml,镍铬烤瓷合金镀金组(C):145.00±8.90 pg/ml,金铂烤瓷合金组(D):138.75±10.31 pg/ml。A、B组与对照组之间有显著性差异(P<0.05);C、D组与对照组之间无显著性差异(P>0.05);A组与B组、C组与D组比较,无显著性差异(P>0.05);A组与C组、A组与D组、B组与C组、B组与D组两两比较,有显著性差异(P<0.05)。
24小时对照组与A、B、C、D组细胞上清液中IL-6的浓度均值分别为:189.50±10.12pg/ml,228.75±8.54pg/ml,223.75±7.50pg/ml , 200.50±9.57pg/ml , 196.50±10.79pg/ml。A、B组与对照组之间有显著性差异(P<0.05);C、D组与对照组之间无显著性差异(P>0.05);A组与B组、C组与D组比较,无显著性差异(P>0.05);A组与C组、A组与D组、B组与C组、B组与D组两两比较,有显著性差异(P<0.05)。
3不同烤瓷合金提浸液培养人牙龈成纤维细胞,MTT比色实验结果显示:
与对照组相比,6小时镍铬烤瓷合金组(A),钴铬烤瓷合金组(B),镍铬烤瓷合金镀金组(C),金铂烤瓷合金组(D)细胞相对增殖率均值分别为:113.07±4.16%,110.50±4.55%,102.57±4.36%,99.00±3.17%。A、B组与对照组之间有显著性差异(P<0.05);C、D组与对照组之间无显著性差异(P>0.05);A组与B组、C组与D组比较无显著性差异(P>0.05);A组与C组、A组与D组、B组与C组、B组与D组两两比较,有显著性差异(P<0.05)。
与对照组相比,24小时A、B、C、D组细胞相对增殖率分别为:78.89±6.03%,82.33±4.44%,95.36±4.17%,97.92±5.83%。A、B组与对照组之间有显著性差异(P<0.05);C、D组与对照组之间无显著性差异(P>0.05);A组与B组、C组与D组比较,无显著性差异(P>0.05);A组与C组、A组与D组、B组与C组、B组与D组两两比较,有显著性差异(P<0.05)。
4不同烤瓷合金表面人牙龈成纤维细胞的MTT比色结果显示:与对照组相比,镍铬烤瓷合金组(A),钴铬烤瓷合金组(B),镍铬烤瓷合金镀金组(C),金铂烤瓷合金组(D)细胞相对增殖率分别为:69.39±3.71%,75.00±5.03%,86.72±3.55%,89.27±3.88%。A、B、C、D组与对照组之间均有显著性差异(P<0.05);A组与B组、C组与D组比较,无显著性差异(P>0.05);A组与C组、A组与D组、B组与C组、B组与D组两两比较,有显著性差异(P<0.05)。
5扫描电镜观察人牙龈成纤维细胞黏附行为:与对照组相比,镍铬烤瓷合金(A),钴铬烤瓷合金(B),镍铬烤瓷合金镀金(C),金铂烤瓷合金(D)表面附着细胞数目少,显示细胞极性,沿材料表面沟纹方向排列。A、B、C组细胞形态变化明显。D组细胞形态变化不明显。
结论:
1体外成功培养了人牙龈成纤维细胞。
2镍铬烤瓷合金和钴铬烤瓷合金提浸液对人牙龈成纤维细胞的增殖显示先促进后抑制作用;并且使细胞IL-6分泌水平升高。镍铬烤瓷合金镀金与金铂烤瓷合金提浸液不影响细胞细胞的增殖活性和IL-6的分泌水平。
3镍铬烤瓷合金、钴铬烤瓷合金、镍铬烤瓷合金镀金和金铂烤瓷合金均抑制人牙龈成纤维细胞的附着生长,影响细胞排列与形态。
4金铂烤瓷合金与镍铬烤瓷合金镀金对人牙龈成纤维细胞生物学行为的影响较小,镍铬与钴铬烤瓷合金对细胞的生物学行为影响较大。
Objective: This study is to investigate the effect of Ni-Cr, Co-Cr and Au-Pt ceramic alloys and Ni-Cr ceramic alloys with gold plating on the secretion of interleukin-6 (IL-6) by human gingival firoblasts (HGFs), and on the proliferation and attachment of HGF.
Methods:
1 Specimens of healthy gingival tissue were obtained from clinical operations, and primary culture of HGFs was performed using the tissue culture method. In order to identify the origin of the cultured cells, we performed immunocytochemical staining including vimentin and cytokeratin staining.
2 With untreated DMEM medium as negative control, we cultured HGFs in the eluates of various types of ceramic alloys separately. The amount of IL-6 secreted in the medium supermatant was measured by enzyme-linked immunosorbent assay (ELISA). In addition, we examined the influence of the eluates of various ceramic alloys on the growth and proliferation of HGFs.
3 HGFs were seeded onto the surface of various types of ceramic alloys as well as onto the cover slips, which served as negative control. Then we examined the adhesive growth and proliferating activity of human gingival firoblasts by Methylthiazoletrazolium (MTT) assay, and observe the cell material attachment with scanning electron microscope (SEM).
Results:
1 Immunocytochemistry result revealed that vimentin staining was positive and cytokeratin staining was negative. Therefore, the cultured cells were proved to be in accordance with the properties of fibroblasts from the mesoderm.
2 The result of ELISA measuring the secretion of IL-6 by HGFs, which is cultivated in eluates of various types of ceramic alloys, is listed as follows. At 6 hours, the average concentration level of IL-6 secreted in the medium supermatant for control group, Ni-Cr ceramic alloys (A), Co-Cr ceramic alloys (B), Ni-Cr ceramic alloys with gold plating (C) and Au-Pt ceramic alloys (D) is 133.75±12.50pg/ml,180.00±10.36pg/ml,168.50±10.08pg/ml,145.00±8.90pg/ml,and 138.75±10.31pg/ml respectively. Group A or group B is significantly different from the control (P<0.05). However, group C or group D is not significantly different from the control (P>0.05). Additionally, Group A is significantly different from group C or group D (P<0.05). Similarly, Group B is significantly different from group C or group D (P<0.05). Both of the differences between group A and B, and group C and D are not significant (P>0.05).
At 24 hours, the average concentration level of IL-6 secreted in the medium supermatant for control group and group A, group B, group C, group D is 189.50±10.12 pg/ml,228.75±8.54 pg/ml,223.75±7.50 pg/ml,200.50±9.57 pg/ml,and 196.50±10.79 pg/ml respectively. Group A or group B is significantly different from the control (P<0.05). However, group C or group D is not significantly different from the control (P>0.05). Additionally, Group A is significantly different from group C or group D (P<0.05). Similarly, Group B is significantly different from group C or group D (P<0.05). Both of the differences between group A and B, and group C and D are not significant (P>0.05).
3 The result of MTT assay measuring the proliferating activity of HGFs, which is cultivated in eluates of various types of ceramic alloys, is listed as follows.
At 6 hours, the average proliferating activity of HGFs for control group, Ni-Cr ceramic alloys (A), Co-Cr ceramic alloys (B), Ni-Cr ceramic alloys with gold plating (C) and Au-Pt ceramic alloys (D) is 113.07±4.16%,110.50±4.55%, 102.57±4.36%,and 99.00±3.17% respectively. Group A or group B is significantly different from the control (P<0.05). However, group C or group D is not significantly different from the control (P>0.05). Additionally, Group A is significantly different from group C or group D (P<0.05). Similarly, Group B is significantly different from group C or group D (P<0.05). Both of the differences between group A and B, and group C and D are not significant (P>0.05).
At 24 hours, the average proliferating activity of HGFs for control group and group A, group B, group C, group D is 78.89±6.03%, 82.33±4.44%, 95.36±4.17%, and 97.92±5.83% respectively. Group A or group B is significantly different from the control (P<0.05). However, group C or group D is not significantly different from the control (P>0.05). Additionally, Group A is significantly different from group C or group D (P<0.05). Similarly, Group B is significantly different from group C or group D (P<0.05). But no significant difference was observed between group A and group B, group C and group D (P>0.05).
4 The result of MTT assay measuring the proliferating activity of HGFs, which is cultivated onto the surface of various types of ceramic alloys, showed that the average proliferating activity of HGFs for control group, Ni-Cr ceramic alloys (A), Co-Cr ceramic alloys (B), Ni-Cr porcelain alloys with gold plating (C) and Au-Pt porcelain alloys (D) is 69.39±3.71%, 75.00±5.03%, 86.72±3.55%, and 89.27±3.88% respectively. Each of the group A, group B, group C, and group D is significantly different from the control (P<0.05). Additionally, Group A is significantly different from group C or group D (P<0.05). Similarly, Group B is significantly different from group C or group D (P<0.05). But no significant difference was observed between group A and group B, group C and group D (P>0.05).
5 The SEM result indicates that the number of HGFs attached to the surface of Ni-Cr ceramic alloys (A), Co-Cr ceramic alloys (B), Ni-Cr ceramic alloys with gold plating (C) and Au-Pt ceramic alloys (D) were less than that of HGFs attached to the surface of the control group. The alignment of the cells along the rills of the materials showed polarity. The morphological change of cells between group A, B, C and the control is significant. The morphological change of cells between group D and the control is not significant.
Conclusion:
1 The human gingival fibroblasts (HGFs) were cultured successfully in vitro.
2 The eluates of Ni-Cr and Co-Cr ceramic alloys can first promote the proliferation of HGFs, and then suppress it. Besides, they can stimulate the secretion of IL-6. But the eluates of Ni-Cr ceramic alloys with gold plating and Au-Pt porcelain alloys did not affect the proliferation of HGFs and the secretion of IL-6.
3 Ni-Cr, Co-Cr, Ni-Cr and Au-Pt ceramic alloys and Ni-Cr ceramic alloys with gold plating can suppress the growth of adhesive HGFs, and influence the cells alignment and morphology.
4 Au-Pt ceramic alloy and Ni-Cr ceramic alloy with gold plating have no obvious influnce on the biological behavior of HGFs, while Ni-Cr and Co-Cr ceramic alloys have significant influnce on that.
引文
1 马轩祥主编.口腔修复学.人民卫生出版社,2003
2 冯海兰主编.口腔修复学.北京大学医学出版社,2005
3 陈治清主编.口腔材料学(第 3 版).人民卫生出版社,2006
4 Wataha JC, Malcolm C, Hanks CT. Correlation between cytotoxicity and elements released by dental casting alloys. [J] Int J. Prosthet Dent, 1995, 8(1): 9~14
5 Hoang AM, Chen D, Oates TW, et al. Development and characterization of a transformed human periodontal ligament cell line. J Periodontol, 1997, 68(11): 1054~1062
6 Ohazama A, Isatsu K, Hatayama J, et al. Periodontal tissue regeneration using fibrin tissue adhesion material in vitro and in vivo. Periodont Clin Investig, 1996, 18(1): 26~38
7 Feng F, Liu CM, Hsu WC, et al. Long-term effects of gingival fibroblast-coated hydroxylapatite graft on periodontal osseous defects. J Formos Med Assoc, 1995, 94(8): 494~498
8 司徒镇强, 吴军正. 细胞培养. 世界图书出版西安公司, 2004
9 郝和平,卜长生, 刘秦玉等. 医疗器械生物学评价标准实施指南. 中国标准出版社,2000
10 Candiano G, Bruschi M, Musante L, et al. Blue silver: a very sensitive colloidal Coomassie G-250 staining for proteome analysis. J Electrophoresis, 2004, 25(9): 1327~1333
11 王磊,彭式韫,刘青.白细胞介素 6 与牙周炎的关系[J]. 牙体牙髓牙周病学杂志,2003, 13 (1): 53~56
12 邓旭亮, 胡晓阳, 欧阳翔英, 等. 五种全冠合金铸造后的腐蚀性研究. 现代口腔医学杂志, 2002, 16 (1): 32~ 33
13 Wataha JC, Lewis JB, Volkmann KR, et al. Sublethal concentrations of Au (III), Pd (II), and Ni (II) differentially alter inflammatory cytokine secretion from activated monocytes. J Biomed Mater Res B Appl Biomater, 2004, 69(1): 11-17
14 Wataha JC, Ratanasathien S,Hanks CT, et al. In vitro IL-1 beta and TNF-alpha release from THP-1 monocytes in response to metal ions. J Dent Mater, 1996, 12(6): 322~327
15 Schmalz G, Schweikl H, Hiller KA. Release of prostaglandin E2, IL-6 and IL-8 from human oral epithelial culture models after exposure to compounds of dental materials. Eur J Oral Sci, 2000, 108(5): 442~448
16 Schmalz G, Schuster U, Schweikl H. Influence of metals on IL-6 release in vitro. J Biomaterials, 1998, 19(18):1689~1694
17 Wataha JC, Malcolm C. Effect of alloy surface composition on release of elements from dental casting alloys. J Oral Rehabil, 1996, 23(9): 583~589
18 石连水,朱洪水,朱嘉. 非贵金属烤瓷冠镀金的制作工艺. 口腔材料器械杂志. 2004, 13(2): 106~107
19 董正杰, 徐侃. 烤瓷冠修复后龈缘黑线产生的原因及防 治. 上海口腔医学, 2003, 12(6): 460~462
20 史言利, 牟月照, 赵宁, 等. 非贵金属烤瓷基底冠表面镀金修复前牙的近期疗效观察. 上海口腔医学, 2005, 14(1):11~13
21 范洪亮, 张纪刚, 杨志勇. 镀金技术在镍铬合金烤瓷修复中的临床应用. 临床口腔医学杂志, 2006, 22(1): 33~34
22 王俐, 焦艳军. 镍铬合金、含钛镍铬合金及其表面处理后材料的耐腐蚀性研究:[硕士学位论文]. 山西医科大学, 2006
23 陈志红, 刘丽. 镍铬合金、镍铬合金表面镀金和金合金在人工唾液中腐蚀行为的电化学研究:[硕士学位论文]. 浙江大学医学院, 2005
24 黄子勋著,实用电镀技术.化学工业出版社, 2003
25 Messer RL, Lucas LC. Cytotoxicity of nickel-chromium alloys: bulk alloys compared to multiple ion salt solutions. Dent Mater, 2000, 16(3): 207~212
26 Messer RL, Lucas LC. Evaluations of metabolic activities as biocompatibility tools: a study of individual ions' effects on fibroblasts. Dent Mater, 1999, 15(1): 1~6
27 Wataha JC, Lockwood PE, Schedle A, et al. Ag, Cu, Hg and Ni ions alter the metabolism of human monocytes during extended low-dose exposures. J Oral Rehabi1, 2002, 29(2): 133~139
28 孙平, 盛祖利, 张正仪等. 四种临床用烤瓷合金材料细胞毒性研究. 口腔医学, 2004, 24(6): 344~346
29 Sun ZL, Wataha JC, Hanks CT. Effects of metal ions on osteoblast-like cell metabolism and differentiation. J Biomed Mater Res, 1997, 34(1): 29~37
30 Mustafa K, Silva Lopez B, Hultenby K, et al. Attachment andproliferation of human oral fibroblasts to titanium surfaces blasted with TiO2 particles. A scanning electron microscopic and histomorphometric analysis. Clin Oral Implants Res, 1998: 9(3)
31 刘建军, 王忠义. 纯钛金属表面粗糙度对人牙龈成纤维细胞影响的体外实验研究:[硕士学位论文]. 第四军医大学, 2001
32 Chou L, Firth ID, Uitto VJ, et al. Effects of titanium substratum and grooved surface topography on metalloproteinase-2 expression in human fibroblasts. J Biomed Mater Res. 1998, 39(3): 437~445
1 International Organization for Standardization. ISO 10993: Biological Evaluation of Medical Devices. London: ISO, 1992
2 Cheung HS, Haak MH. Growth of osteoblasts on porous calcium phosphate ceramic: an in vitro model for biocompatibility study. Biomaterials, 1989, 10(1): 63~67
3 Wataha JC, Hanks CT, Sun Z. Effect of cell line on in vitro metal ion cytotoxicity. Dent Mater, 1994, 10(3): 156~61
4 Gretzer C, Eriksson AS, Allden B et al. Monocyte activation on titanium-sputtered polystyrene surfaces in vitro: the effect of culture conditions on interleukin-1 release. Biomaterials, 1996, 17(9): 851~858
5 Tobin BW, Leeper-Woodford SK, Hashemi BB, et al. Altered TNF-alpha, glucose, insulin, and amino acids in islets of Langerhans cultured in a microgravity model system. Am J Physiol Endocrinol Metab, 2001, 280 (1): 92~102
6 Oshima H, Nakamura M. A study on reference standard forcytotoxicity assay of biomaterials. Biomed Mater Eng, 1994, 4 (4): 327~332
7 Wataha JC, Craig RG, Hanks CT. Precision of and new methods for testing in vitro alloy cytotoxicity. Dent Mater, 1992, 8(1): 65~67
9 上海第二医科大学生物医学材料教研室编. 非直接接触血液的医用生物材料性能测试, 1985
10 Barka T, Andersson P. Histochemistry, Theory, Practice and Bibliography. Harper and Row Press, New York, 1963: 313~317
11 司徒镇强,吴军正.细胞培养. 世界图书出版公司2004, 72~73
12 杨抚华. 医学细胞生物学概论. 四川科学技术出版社, 1991
13 Messer RL, Lucas LC. Cytotoxicity of nickel-chromium alloys: bulk alloys compared to multiple ion salt solutions. Dent Mater, 2000, 16(3): 207~212
14 Wataha JC, Lewis JB, Volkmann KR, et al. Sublethal concentrations of Au (III), Pd (II), and Ni (II) differentially alter inflammatory cytokine secretion from activated monocytes. J Biomed Mater Res B Appl Biomater, 2004, 69(1): 11~17
15 Wataha JC, Lockwood PE, Schedle A, et al. Ag, Cu, Hg and Ni ions alter the metabolism of human monocytes during extended low-dose exposures. J Oral Rehabil, 2002, 29(2): 133~139
16 Wataha JC, Hanks CT, Craig RG. In vitro effects of metal ions on cellular metabolism and the correlation between these effects and the uptake of the ions. J Biomed Mater Res, 1994, 28(4): 427~433
17 Wataha JC, Hanks CT, Sun Z. In vitro reaction of macrophages to metal ions from dental biomaterials. Dent Mater, 1995, 11(4): 239~245
18 Sun ZL,Wataha JC, Hanks CT. Effects of metal ions on osteoblast-like cell metabolism and differentiation. J Biomed Mater Res, 1997, 34(1): 29~37
19 Wataha JC, Lockwood PE, Noda M, et al. Effect of toothbrushing on the toxicity of casting alloys. J Prosthet Dent, 2002, 87(1): 94~98
20 Bumgardner JD, Lucas LC. Ellular response to metallic ions released from nickel-chromium dental alloys. J Dent Res, 1995, 74(8): 1521~1527
21 Al-Hiyasat AS, Darmani H. The effects of recasting on the cytotoxicity of base metal alloys. J Prosthet Dent, 2005, 93(2): 158~163
22 Schedle A, Samorapoompichit P, Rausch-fan XH, etal. Response of HL-160 Fibroblasts, Human Gingival Fibroblasts, and Human Tissue Mast Cells toVarious Metal Cations. J Dent Res, 1995, 74(8): 1513~1520
23 邓旭亮,胡晓阳,欧阳翔英,等. 五种全冠合金铸造后的腐蚀性研究. 现代口腔医学杂志, 2002, 16(1): 32~33
24 Facckni F, Franceschetti P, Cerpelloni M, et al. In vivostudy on metal release from fixed orthodontic appliances and DNA damage in oral mucosa cells. Am J Orthod Dentofacial Orthop, 2003, 124(6): 687~694
25 Cortizo MC, De Mele MF, Cortizo AM. Metallic dental material biocompatibility in osteoblastlike cells: correlation with metal ion release. Biol Trace Elem Res, 2004, 100(2): 151~168
26 Kamagata Y, Miyasaka N, Inoue H, et al. Cytokine production in inflamed human gingival tissues interleukin-6. Nippon Shishubyo Gakkai Kaishi, 1989, 31(4): 1081~1087
27 Mehmet D, JUlide O, Ali Ugur U, et al. Influence of dental alloys and an all-ceramic material on cell viability and interleukin-1beta release in a three-dimensional cell culture model. Turk J Med Sci, 2005, 35(2): 203-208
28 Spyrou P, Papaioannou S, Hampson G, et al. Cytokine release by osteoblast-like cells cultured on implant discs of varying alloy compositions. Clin Oral Implants Res, 2002, 13(6): 623~630
29 Wataha JC, Ratanasathien S,Hanks CT, et al. In vitro IL-lβand TNF-α release from THP-1 monocytes in response to metal ions. J Dent Mater, 1996, 12(6): 322~327
30 Candiano G, Bruschi M, Musante L, et al. Blue silver: A very sensitive colloidal Coomassie G-250 staining for proteome analysis. J Electrophoresis, 2004, 25(9): 1327~ 1333
31 Schmalz G, Schuster U, Schweik H. Influence of metals onIL-6 release in vitro J. Biomaterials, 1998, 19(18): 1689 ~1694
32 Schmalz G, Schweikl H, Hiller KA. Release of prostanglandin E2 , IL-6 and IL-8 from human oral epithelial culture models after exposure to compounds of dental materials. Eur J Oral Sci, 2000, 108(5): 442~448
33 李岩涛,贺兰,李洁,等. 牙周炎龈组织肿瘤坏死因子 α的含量及其与牙周炎的关系[J]. 口腔医学,2002,22(1):12~14
34 Schmalz G, Arenholt-Bindslev D, Hiller KA, et al. Epithelium-fibroblast co-culture for assessing mucosal irritancy of metals used in dentistry. Eur J Oral Sci, 1997, 105(1): 86~91
35 Haynes DR, Boyle SJ, Rogers SD, et al. Variation in cytokines induced by particles from different prosthetic materials. Clin Orthop Relat Res, 1998, (352): 223~230
2 冯海兰主编.口腔修复学.北京大学医学出版社,2005
3 陈治清主编.口腔材料学(第 3 版).人民卫生出版社,2006
4 Wataha JC, Malcolm C, Hanks CT. Correlation between cytotoxicity and elements released by dental casting alloys. [J] Int J. Prosthet Dent, 1995, 8(1): 9~14
5 Hoang AM, Chen D, Oates TW, et al. Development and characterization of a transformed human periodontal ligament cell line. J Periodontol, 1997, 68(11): 1054~1062
6 Ohazama A, Isatsu K, Hatayama J, et al. Periodontal tissue regeneration using fibrin tissue adhesion material in vitro and in vivo. Periodont Clin Investig, 1996, 18(1): 26~38
7 Feng F, Liu CM, Hsu WC, et al. Long-term effects of gingival fibroblast-coated hydroxylapatite graft on periodontal osseous defects. J Formos Med Assoc, 1995, 94(8): 494~498
8 司徒镇强, 吴军正. 细胞培养. 世界图书出版西安公司, 2004
9 郝和平,卜长生, 刘秦玉等. 医疗器械生物学评价标准实施指南. 中国标准出版社,2000
10 Candiano G, Bruschi M, Musante L, et al. Blue silver: a very sensitive colloidal Coomassie G-250 staining for proteome analysis. J Electrophoresis, 2004, 25(9): 1327~1333
11 王磊,彭式韫,刘青.白细胞介素 6 与牙周炎的关系[J]. 牙体牙髓牙周病学杂志,2003, 13 (1): 53~56
12 邓旭亮, 胡晓阳, 欧阳翔英, 等. 五种全冠合金铸造后的腐蚀性研究. 现代口腔医学杂志, 2002, 16 (1): 32~ 33
13 Wataha JC, Lewis JB, Volkmann KR, et al. Sublethal concentrations of Au (III), Pd (II), and Ni (II) differentially alter inflammatory cytokine secretion from activated monocytes. J Biomed Mater Res B Appl Biomater, 2004, 69(1): 11-17
14 Wataha JC, Ratanasathien S,Hanks CT, et al. In vitro IL-1 beta and TNF-alpha release from THP-1 monocytes in response to metal ions. J Dent Mater, 1996, 12(6): 322~327
15 Schmalz G, Schweikl H, Hiller KA. Release of prostaglandin E2, IL-6 and IL-8 from human oral epithelial culture models after exposure to compounds of dental materials. Eur J Oral Sci, 2000, 108(5): 442~448
16 Schmalz G, Schuster U, Schweikl H. Influence of metals on IL-6 release in vitro. J Biomaterials, 1998, 19(18):1689~1694
17 Wataha JC, Malcolm C. Effect of alloy surface composition on release of elements from dental casting alloys. J Oral Rehabil, 1996, 23(9): 583~589
18 石连水,朱洪水,朱嘉. 非贵金属烤瓷冠镀金的制作工艺. 口腔材料器械杂志. 2004, 13(2): 106~107
19 董正杰, 徐侃. 烤瓷冠修复后龈缘黑线产生的原因及防 治. 上海口腔医学, 2003, 12(6): 460~462
20 史言利, 牟月照, 赵宁, 等. 非贵金属烤瓷基底冠表面镀金修复前牙的近期疗效观察. 上海口腔医学, 2005, 14(1):11~13
21 范洪亮, 张纪刚, 杨志勇. 镀金技术在镍铬合金烤瓷修复中的临床应用. 临床口腔医学杂志, 2006, 22(1): 33~34
22 王俐, 焦艳军. 镍铬合金、含钛镍铬合金及其表面处理后材料的耐腐蚀性研究:[硕士学位论文]. 山西医科大学, 2006
23 陈志红, 刘丽. 镍铬合金、镍铬合金表面镀金和金合金在人工唾液中腐蚀行为的电化学研究:[硕士学位论文]. 浙江大学医学院, 2005
24 黄子勋著,实用电镀技术.化学工业出版社, 2003
25 Messer RL, Lucas LC. Cytotoxicity of nickel-chromium alloys: bulk alloys compared to multiple ion salt solutions. Dent Mater, 2000, 16(3): 207~212
26 Messer RL, Lucas LC. Evaluations of metabolic activities as biocompatibility tools: a study of individual ions' effects on fibroblasts. Dent Mater, 1999, 15(1): 1~6
27 Wataha JC, Lockwood PE, Schedle A, et al. Ag, Cu, Hg and Ni ions alter the metabolism of human monocytes during extended low-dose exposures. J Oral Rehabi1, 2002, 29(2): 133~139
28 孙平, 盛祖利, 张正仪等. 四种临床用烤瓷合金材料细胞毒性研究. 口腔医学, 2004, 24(6): 344~346
29 Sun ZL, Wataha JC, Hanks CT. Effects of metal ions on osteoblast-like cell metabolism and differentiation. J Biomed Mater Res, 1997, 34(1): 29~37
30 Mustafa K, Silva Lopez B, Hultenby K, et al. Attachment andproliferation of human oral fibroblasts to titanium surfaces blasted with TiO2 particles. A scanning electron microscopic and histomorphometric analysis. Clin Oral Implants Res, 1998: 9(3)
31 刘建军, 王忠义. 纯钛金属表面粗糙度对人牙龈成纤维细胞影响的体外实验研究:[硕士学位论文]. 第四军医大学, 2001
32 Chou L, Firth ID, Uitto VJ, et al. Effects of titanium substratum and grooved surface topography on metalloproteinase-2 expression in human fibroblasts. J Biomed Mater Res. 1998, 39(3): 437~445
1 International Organization for Standardization. ISO 10993: Biological Evaluation of Medical Devices. London: ISO, 1992
2 Cheung HS, Haak MH. Growth of osteoblasts on porous calcium phosphate ceramic: an in vitro model for biocompatibility study. Biomaterials, 1989, 10(1): 63~67
3 Wataha JC, Hanks CT, Sun Z. Effect of cell line on in vitro metal ion cytotoxicity. Dent Mater, 1994, 10(3): 156~61
4 Gretzer C, Eriksson AS, Allden B et al. Monocyte activation on titanium-sputtered polystyrene surfaces in vitro: the effect of culture conditions on interleukin-1 release. Biomaterials, 1996, 17(9): 851~858
5 Tobin BW, Leeper-Woodford SK, Hashemi BB, et al. Altered TNF-alpha, glucose, insulin, and amino acids in islets of Langerhans cultured in a microgravity model system. Am J Physiol Endocrinol Metab, 2001, 280 (1): 92~102
6 Oshima H, Nakamura M. A study on reference standard forcytotoxicity assay of biomaterials. Biomed Mater Eng, 1994, 4 (4): 327~332
7 Wataha JC, Craig RG, Hanks CT. Precision of and new methods for testing in vitro alloy cytotoxicity. Dent Mater, 1992, 8(1): 65~67
9 上海第二医科大学生物医学材料教研室编. 非直接接触血液的医用生物材料性能测试, 1985
10 Barka T, Andersson P. Histochemistry, Theory, Practice and Bibliography. Harper and Row Press, New York, 1963: 313~317
11 司徒镇强,吴军正.细胞培养. 世界图书出版公司2004, 72~73
12 杨抚华. 医学细胞生物学概论. 四川科学技术出版社, 1991
13 Messer RL, Lucas LC. Cytotoxicity of nickel-chromium alloys: bulk alloys compared to multiple ion salt solutions. Dent Mater, 2000, 16(3): 207~212
14 Wataha JC, Lewis JB, Volkmann KR, et al. Sublethal concentrations of Au (III), Pd (II), and Ni (II) differentially alter inflammatory cytokine secretion from activated monocytes. J Biomed Mater Res B Appl Biomater, 2004, 69(1): 11~17
15 Wataha JC, Lockwood PE, Schedle A, et al. Ag, Cu, Hg and Ni ions alter the metabolism of human monocytes during extended low-dose exposures. J Oral Rehabil, 2002, 29(2): 133~139
16 Wataha JC, Hanks CT, Craig RG. In vitro effects of metal ions on cellular metabolism and the correlation between these effects and the uptake of the ions. J Biomed Mater Res, 1994, 28(4): 427~433
17 Wataha JC, Hanks CT, Sun Z. In vitro reaction of macrophages to metal ions from dental biomaterials. Dent Mater, 1995, 11(4): 239~245
18 Sun ZL,Wataha JC, Hanks CT. Effects of metal ions on osteoblast-like cell metabolism and differentiation. J Biomed Mater Res, 1997, 34(1): 29~37
19 Wataha JC, Lockwood PE, Noda M, et al. Effect of toothbrushing on the toxicity of casting alloys. J Prosthet Dent, 2002, 87(1): 94~98
20 Bumgardner JD, Lucas LC. Ellular response to metallic ions released from nickel-chromium dental alloys. J Dent Res, 1995, 74(8): 1521~1527
21 Al-Hiyasat AS, Darmani H. The effects of recasting on the cytotoxicity of base metal alloys. J Prosthet Dent, 2005, 93(2): 158~163
22 Schedle A, Samorapoompichit P, Rausch-fan XH, etal. Response of HL-160 Fibroblasts, Human Gingival Fibroblasts, and Human Tissue Mast Cells toVarious Metal Cations. J Dent Res, 1995, 74(8): 1513~1520
23 邓旭亮,胡晓阳,欧阳翔英,等. 五种全冠合金铸造后的腐蚀性研究. 现代口腔医学杂志, 2002, 16(1): 32~33
24 Facckni F, Franceschetti P, Cerpelloni M, et al. In vivostudy on metal release from fixed orthodontic appliances and DNA damage in oral mucosa cells. Am J Orthod Dentofacial Orthop, 2003, 124(6): 687~694
25 Cortizo MC, De Mele MF, Cortizo AM. Metallic dental material biocompatibility in osteoblastlike cells: correlation with metal ion release. Biol Trace Elem Res, 2004, 100(2): 151~168
26 Kamagata Y, Miyasaka N, Inoue H, et al. Cytokine production in inflamed human gingival tissues interleukin-6. Nippon Shishubyo Gakkai Kaishi, 1989, 31(4): 1081~1087
27 Mehmet D, JUlide O, Ali Ugur U, et al. Influence of dental alloys and an all-ceramic material on cell viability and interleukin-1beta release in a three-dimensional cell culture model. Turk J Med Sci, 2005, 35(2): 203-208
28 Spyrou P, Papaioannou S, Hampson G, et al. Cytokine release by osteoblast-like cells cultured on implant discs of varying alloy compositions. Clin Oral Implants Res, 2002, 13(6): 623~630
29 Wataha JC, Ratanasathien S,Hanks CT, et al. In vitro IL-lβand TNF-α release from THP-1 monocytes in response to metal ions. J Dent Mater, 1996, 12(6): 322~327
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