BACKGROUND:The micro/nanostructured gradient biomimetic surface of implant materials can simulate the structure of the extracellular environment in human bone tissue,thereby achieving perfect bone integration function.However,further research is needed on the mechanisms by which the surface microstructure of bone implant materials regulates cell function and promotes osteogenesis. OBJECTIVE:To analyze the effect of titanium sheet microstructure surface on osteogenic differentiation of MC3T3-E1 osteoblasts. METHODS:(1)At a constant voltage of 5 V or 20 V,nanotube arrays of different diameters were prepared on the surface of titanium sheets by acid etching and anodic oxidation techniques,and were recorded as group R5 and group R20,respectively.The surface morphology,roughness,and hydrophilicity of pure titanium sheet(without acid etching or anodizing treatment)were measured in group R5 and group R20.(2)MC3T3-E1 osteoblasts of logarithmic growth stage were inoculated on the surface of pure titanium sheets,R5 group and R20 group respectively.After 24 hours of osteogenic induction culture,the expression of mechanical sensitive channel protein 1 was analyzed by RT-PCR and immunofluorescence staining.Osteoblast inducible base with or without the mechanosensitive channel protein 1 activator Yada1 was added,and alkaline phosphatase staining was performed after 7 days of culture.Alizarin red staining was performed after 14 days of culture. RESULTS AND CONCLUSION:(1)The surface of pure titanium sheets was smooth under scanning electron microscope.Relatively uniform and orderly nanotube arrays with average diameters of about 30 nm and 100 nm were observed on the surface of titanium sheets of groups R5 and R20,respectively.The results of scanning electron microscope were further verified by atomic force microscopy.The surface roughness of titanium sheet of group R5 was higher than that of pure titanium(P<0.05),and the water contact angle was lower than that of pure titanium(P<0.05).The surface roughness of titanium sheet in group R20 was higher than that in group R5(P<0.05),and the water contact angle was lower than that in group R5(P<0.05).(2)RT-PCR and immunofluorescence staining showed that the expression of mechanosensitive channel protein 1 in group R5 was higher than that in pure titanium group(P<0.05),and the expression of mechanosensitive channel protein 1 in group R20 was higher than that in group R5(P<0.05).Under the osteogenic induction,compared with the condition without Yada1,there were no significant changes in the activity of alkaline phosphatase and the deposition of calcified nodules in pure titanium group after Yada1 addition,while the activity of alkaline phosphatase and the deposition of calcified nodules in groups R5 and R20 after Yada1 addition were significantly increased(P<0.05).With or without Yada1,the alkaline phosphatase activity and calcified nodule deposition in group R5 were higher than those in pure titanium group(P<0.05),and the alkaline phosphatase activity and calcified nodule deposition in group R20 were higher than those in group R5(P<0.05).(3)The results show that the surface microstructure of titanium sheet can promote the osteogenic differentiation of osteoblast MC3T3-E1 by activating mechanosensitive channel protein 1.

Acute pancreatitis （AP） is one of the most clinically common acute digestive disorders characterized by quick onset，rapid progression，severe condition，and high mortality. If the disease is not timely intervened in the early stage，it can develop into severe AP in the later stage，which damages the long-term quality of life and brings serious economic burden to patients and their families. However， the pathogenesis of this disease is complex and has not been fully explained. The generation and development of AP is closely related to many signaling pathways. Among them，Toll-like receptor 4（TLR4），as a transmembrane signal transduction receptor，can mediate immune response and inflammatory response，and play a key role in the occurrence and development of AP. Traditional Chinese medicine（TCM）can regulate the TLR4 signaling pathway with multiple targets，multiple effects，and multiple administration methods to inhibit inflammatory response，and effectively intervene in the progression of AP， which has gradually become a new craze for preventing and treating AP. Many studies have shown that TCM has obvious advantages in the prevention and treatment of AP. It can effectively treat AP by regulating TLR4 signaling pathway，strengthening immune resistance and defense，and inhibiting inflammatory response. Despite of the research progress，there is still a lack of comprehensive review on TCM regulation of TLR4 signaling pathway in the treatment of AP. Therefore，the literature on TCM regulation of TLR4 signaling pathway published in recent years was systematically reviewed and elaborated，aiming to provide new ideas for the treatment of AP and further drug development.

Objective: To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts (HGFs) and to provide experimental evidence for surface modification of implant abutments. Methods: The samples were divided into an NC group (negative control, no other treatment on a smooth surface), an NM-1 group (nanomesh-1, electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage), and an NM-2 group (nanomesh-2, electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage). The surface morphologies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy (SEM). The surface hydrophilicities of the samples were measured with a contact angle measuring instrument. The proliferation of HGFs on the different samples were evaluated with CCK-8, and the expression of adhesion-related genes, including collagen Ⅰ (COL1A1), collagen Ⅲ (COL3A1), fibronectin 1 (FN1), focal adhesion kinase (FAK), vinculin (VCL), integrin α2 (ITGA2), and integrin β1 (ITGB1), on the different samples was measured with qRT-PCR. The expression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy (CLSM) after immunofluorescent staining. Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining. Results: SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups, with grid diameters of approximately 30 nm for the NM-1 group and approximately 150 nm for the NM-2 group. Compared with that of the NC group, the water contact angles of the NM-1 group and NM-2 groups were significantly lower (P<0.000 1). Cell proliferation in the NM-1 group was significantly greater than that in the NC group (P<0.01). Moreover, there was no significant difference in the water contact angles or cell proliferation between the NM-1 group and the NM-2 group. SEM revealed that HGFs were adhered well to the surfaces of all samples, while the HGFs in the NM-1 and NM-2 groups showed more extended areas, longer morphologies, and more developed pseudopodia than did those in the NC group after 24 h. qRT-PCR revealed that the expression levels of the adhesion-related genes COL1A1, COL3A1, FN1, FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups (P<0.01). The expression of vinculin protein in the NM-1 group was the highest, and the number of focal adhesions was greatest in the NM-1 group (P<0.01). The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers (P<0.000 1). Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion, proliferation, collagen fiber secretion and syntheses of HGFs, and electrochemical dealloying of Ti6Al4V with a grid diameter of approximately 30 nm obviously promoted HGF formation.

Objective To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts(HGFs)and to provide experimental evidence for surface modification of implant abutments.Methods The samples were divided into an NC group(negative control,no other treatment on a smooth surface),an NM-1 group(nanomesh-1,electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage),and an NM-2 group(nanomesh-2,electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage).The surface morpholo-gies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy(SEM).The surface hydrophilicities of the samples were measured with a contact angle measuring instrument.The prolif-eration of HGFs on the different samples were evaluated with CCK-8,and the expression of adhesion-related genes,in-cluding collagen Ⅰ(COL1A1),collagen Ⅲ(COL3A1),fibronectin 1(FN1),focal adhesion kinase(FAK),vinculin(VCL),integrin α2(ITGA2),and integrin β1(ITGB1),on the different samples was measured with qRT-PCR.The ex-pression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy(CLSM)after immuno-fluorescent staining.Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining.Results SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups,with grid diameters of approximately 30 nm for the NM-1 group and approxi-mately 150 nm for the NM-2 group.Compared with that of the NC group,the water contact angles of the NM-1 group and NM-2 groups were significantly lower(P＜0.000 1).Cell proliferation in the NM-1 group was significantly greater than that in the NC group(P＜0.01).Moreover,there was no significant difference in the water contact angles or cell prolifer-ation between the NM-1 group and the NM-2 group.SEM revealed that HGFs were adhered well to the surfaces of all samples,while the HGFs in the NM-1 and NM-2 groups showed more extended areas,longer morphologies,and more de-veloped pseudopodia than did those in the NC group after 24 h.qRT-PCR revealed that the expression levels of the ad-hesion-related genes COL1A1,COL3A1,FN1,FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups(P＜0.01).The expression of vinculin protein in the NM-1 group was the highest,and the num-ber of focal adhesions was greatest in the NM-1 group(P＜0.01).The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers(P＜0.000 1).Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion,proliferation,colla-gen fiber secretion and syntheses of HGFs,and electrochemical dealloying of Ti6Al4V with a grid diameter of approxi-mately 30 nm obviously promoted HGF formation.

Objective To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts(HGFs)and to provide experimental evidence for surface modification of implant abutments.Methods The samples were divided into an NC group(negative control,no other treatment on a smooth surface),an NM-1 group(nanomesh-1,electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage),and an NM-2 group(nanomesh-2,electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage).The surface morpholo-gies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy(SEM).The surface hydrophilicities of the samples were measured with a contact angle measuring instrument.The prolif-eration of HGFs on the different samples were evaluated with CCK-8,and the expression of adhesion-related genes,in-cluding collagen Ⅰ(COL1A1),collagen Ⅲ(COL3A1),fibronectin 1(FN1),focal adhesion kinase(FAK),vinculin(VCL),integrin α2(ITGA2),and integrin β1(ITGB1),on the different samples was measured with qRT-PCR.The ex-pression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy(CLSM)after immuno-fluorescent staining.Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining.Results SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups,with grid diameters of approximately 30 nm for the NM-1 group and approxi-mately 150 nm for the NM-2 group.Compared with that of the NC group,the water contact angles of the NM-1 group and NM-2 groups were significantly lower(P＜0.000 1).Cell proliferation in the NM-1 group was significantly greater than that in the NC group(P＜0.01).Moreover,there was no significant difference in the water contact angles or cell prolifer-ation between the NM-1 group and the NM-2 group.SEM revealed that HGFs were adhered well to the surfaces of all samples,while the HGFs in the NM-1 and NM-2 groups showed more extended areas,longer morphologies,and more de-veloped pseudopodia than did those in the NC group after 24 h.qRT-PCR revealed that the expression levels of the ad-hesion-related genes COL1A1,COL3A1,FN1,FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups(P＜0.01).The expression of vinculin protein in the NM-1 group was the highest,and the num-ber of focal adhesions was greatest in the NM-1 group(P＜0.01).The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers(P＜0.000 1).Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion,proliferation,colla-gen fiber secretion and syntheses of HGFs,and electrochemical dealloying of Ti6Al4V with a grid diameter of approxi-mately 30 nm obviously promoted HGF formation.

Objective To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts(HGFs)and to provide experimental evidence for surface modification of implant abutments.Methods The samples were divided into an NC group(negative control,no other treatment on a smooth surface),an NM-1 group(nanomesh-1,electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage),and an NM-2 group(nanomesh-2,electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage).The surface morpholo-gies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy(SEM).The surface hydrophilicities of the samples were measured with a contact angle measuring instrument.The prolif-eration of HGFs on the different samples were evaluated with CCK-8,and the expression of adhesion-related genes,in-cluding collagen Ⅰ(COL1A1),collagen Ⅲ(COL3A1),fibronectin 1(FN1),focal adhesion kinase(FAK),vinculin(VCL),integrin α2(ITGA2),and integrin β1(ITGB1),on the different samples was measured with qRT-PCR.The ex-pression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy(CLSM)after immuno-fluorescent staining.Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining.Results SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups,with grid diameters of approximately 30 nm for the NM-1 group and approxi-mately 150 nm for the NM-2 group.Compared with that of the NC group,the water contact angles of the NM-1 group and NM-2 groups were significantly lower(P＜0.000 1).Cell proliferation in the NM-1 group was significantly greater than that in the NC group(P＜0.01).Moreover,there was no significant difference in the water contact angles or cell prolifer-ation between the NM-1 group and the NM-2 group.SEM revealed that HGFs were adhered well to the surfaces of all samples,while the HGFs in the NM-1 and NM-2 groups showed more extended areas,longer morphologies,and more de-veloped pseudopodia than did those in the NC group after 24 h.qRT-PCR revealed that the expression levels of the ad-hesion-related genes COL1A1,COL3A1,FN1,FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups(P＜0.01).The expression of vinculin protein in the NM-1 group was the highest,and the num-ber of focal adhesions was greatest in the NM-1 group(P＜0.01).The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers(P＜0.000 1).Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion,proliferation,colla-gen fiber secretion and syntheses of HGFs,and electrochemical dealloying of Ti6Al4V with a grid diameter of approxi-mately 30 nm obviously promoted HGF formation.

Objective To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts(HGFs)and to provide experimental evidence for surface modification of implant abutments.Methods The samples were divided into an NC group(negative control,no other treatment on a smooth surface),an NM-1 group(nanomesh-1,electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage),and an NM-2 group(nanomesh-2,electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage).The surface morpholo-gies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy(SEM).The surface hydrophilicities of the samples were measured with a contact angle measuring instrument.The prolif-eration of HGFs on the different samples were evaluated with CCK-8,and the expression of adhesion-related genes,in-cluding collagen Ⅰ(COL1A1),collagen Ⅲ(COL3A1),fibronectin 1(FN1),focal adhesion kinase(FAK),vinculin(VCL),integrin α2(ITGA2),and integrin β1(ITGB1),on the different samples was measured with qRT-PCR.The ex-pression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy(CLSM)after immuno-fluorescent staining.Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining.Results SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups,with grid diameters of approximately 30 nm for the NM-1 group and approxi-mately 150 nm for the NM-2 group.Compared with that of the NC group,the water contact angles of the NM-1 group and NM-2 groups were significantly lower(P＜0.000 1).Cell proliferation in the NM-1 group was significantly greater than that in the NC group(P＜0.01).Moreover,there was no significant difference in the water contact angles or cell prolifer-ation between the NM-1 group and the NM-2 group.SEM revealed that HGFs were adhered well to the surfaces of all samples,while the HGFs in the NM-1 and NM-2 groups showed more extended areas,longer morphologies,and more de-veloped pseudopodia than did those in the NC group after 24 h.qRT-PCR revealed that the expression levels of the ad-hesion-related genes COL1A1,COL3A1,FN1,FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups(P＜0.01).The expression of vinculin protein in the NM-1 group was the highest,and the num-ber of focal adhesions was greatest in the NM-1 group(P＜0.01).The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers(P＜0.000 1).Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion,proliferation,colla-gen fiber secretion and syntheses of HGFs,and electrochemical dealloying of Ti6Al4V with a grid diameter of approxi-mately 30 nm obviously promoted HGF formation.

Objective To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts(HGFs)and to provide experimental evidence for surface modification of implant abutments.Methods The samples were divided into an NC group(negative control,no other treatment on a smooth surface),an NM-1 group(nanomesh-1,electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage),and an NM-2 group(nanomesh-2,electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage).The surface morpholo-gies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy(SEM).The surface hydrophilicities of the samples were measured with a contact angle measuring instrument.The prolif-eration of HGFs on the different samples were evaluated with CCK-8,and the expression of adhesion-related genes,in-cluding collagen Ⅰ(COL1A1),collagen Ⅲ(COL3A1),fibronectin 1(FN1),focal adhesion kinase(FAK),vinculin(VCL),integrin α2(ITGA2),and integrin β1(ITGB1),on the different samples was measured with qRT-PCR.The ex-pression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy(CLSM)after immuno-fluorescent staining.Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining.Results SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups,with grid diameters of approximately 30 nm for the NM-1 group and approxi-mately 150 nm for the NM-2 group.Compared with that of the NC group,the water contact angles of the NM-1 group and NM-2 groups were significantly lower(P＜0.000 1).Cell proliferation in the NM-1 group was significantly greater than that in the NC group(P＜0.01).Moreover,there was no significant difference in the water contact angles or cell prolifer-ation between the NM-1 group and the NM-2 group.SEM revealed that HGFs were adhered well to the surfaces of all samples,while the HGFs in the NM-1 and NM-2 groups showed more extended areas,longer morphologies,and more de-veloped pseudopodia than did those in the NC group after 24 h.qRT-PCR revealed that the expression levels of the ad-hesion-related genes COL1A1,COL3A1,FN1,FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups(P＜0.01).The expression of vinculin protein in the NM-1 group was the highest,and the num-ber of focal adhesions was greatest in the NM-1 group(P＜0.01).The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers(P＜0.000 1).Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion,proliferation,colla-gen fiber secretion and syntheses of HGFs,and electrochemical dealloying of Ti6Al4V with a grid diameter of approxi-mately 30 nm obviously promoted HGF formation.

Objective To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts(HGFs)and to provide experimental evidence for surface modification of implant abutments.Methods The samples were divided into an NC group(negative control,no other treatment on a smooth surface),an NM-1 group(nanomesh-1,electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage),and an NM-2 group(nanomesh-2,electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage).The surface morpholo-gies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy(SEM).The surface hydrophilicities of the samples were measured with a contact angle measuring instrument.The prolif-eration of HGFs on the different samples were evaluated with CCK-8,and the expression of adhesion-related genes,in-cluding collagen Ⅰ(COL1A1),collagen Ⅲ(COL3A1),fibronectin 1(FN1),focal adhesion kinase(FAK),vinculin(VCL),integrin α2(ITGA2),and integrin β1(ITGB1),on the different samples was measured with qRT-PCR.The ex-pression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy(CLSM)after immuno-fluorescent staining.Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining.Results SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups,with grid diameters of approximately 30 nm for the NM-1 group and approxi-mately 150 nm for the NM-2 group.Compared with that of the NC group,the water contact angles of the NM-1 group and NM-2 groups were significantly lower(P＜0.000 1).Cell proliferation in the NM-1 group was significantly greater than that in the NC group(P＜0.01).Moreover,there was no significant difference in the water contact angles or cell prolifer-ation between the NM-1 group and the NM-2 group.SEM revealed that HGFs were adhered well to the surfaces of all samples,while the HGFs in the NM-1 and NM-2 groups showed more extended areas,longer morphologies,and more de-veloped pseudopodia than did those in the NC group after 24 h.qRT-PCR revealed that the expression levels of the ad-hesion-related genes COL1A1,COL3A1,FN1,FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups(P＜0.01).The expression of vinculin protein in the NM-1 group was the highest,and the num-ber of focal adhesions was greatest in the NM-1 group(P＜0.01).The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers(P＜0.000 1).Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion,proliferation,colla-gen fiber secretion and syntheses of HGFs,and electrochemical dealloying of Ti6Al4V with a grid diameter of approxi-mately 30 nm obviously promoted HGF formation.

Objective To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts(HGFs)and to provide experimental evidence for surface modification of implant abutments.Methods The samples were divided into an NC group(negative control,no other treatment on a smooth surface),an NM-1 group(nanomesh-1,electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage),and an NM-2 group(nanomesh-2,electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage).The surface morpholo-gies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy(SEM).The surface hydrophilicities of the samples were measured with a contact angle measuring instrument.The prolif-eration of HGFs on the different samples were evaluated with CCK-8,and the expression of adhesion-related genes,in-cluding collagen Ⅰ(COL1A1),collagen Ⅲ(COL3A1),fibronectin 1(FN1),focal adhesion kinase(FAK),vinculin(VCL),integrin α2(ITGA2),and integrin β1(ITGB1),on the different samples was measured with qRT-PCR.The ex-pression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy(CLSM)after immuno-fluorescent staining.Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining.Results SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups,with grid diameters of approximately 30 nm for the NM-1 group and approxi-mately 150 nm for the NM-2 group.Compared with that of the NC group,the water contact angles of the NM-1 group and NM-2 groups were significantly lower(P＜0.000 1).Cell proliferation in the NM-1 group was significantly greater than that in the NC group(P＜0.01).Moreover,there was no significant difference in the water contact angles or cell prolifer-ation between the NM-1 group and the NM-2 group.SEM revealed that HGFs were adhered well to the surfaces of all samples,while the HGFs in the NM-1 and NM-2 groups showed more extended areas,longer morphologies,and more de-veloped pseudopodia than did those in the NC group after 24 h.qRT-PCR revealed that the expression levels of the ad-hesion-related genes COL1A1,COL3A1,FN1,FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups(P＜0.01).The expression of vinculin protein in the NM-1 group was the highest,and the num-ber of focal adhesions was greatest in the NM-1 group(P＜0.01).The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers(P＜0.000 1).Conclusion The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion,proliferation,colla-gen fiber secretion and syntheses of HGFs,and electrochemical dealloying of Ti6Al4V with a grid diameter of approxi-mately 30 nm obviously promoted HGF formation.