| 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.
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