Objective:To investigate the effect of microgroove surfaces of titanium on the adhesion and cell cycle progression of human gingival fibroblasts(HGFs).Methods:Microgroove titanium surfaces were fabricated on silicon plate by photolithography with parallel grooves:1 5,30 and 60 μm in width and 5 μm and 1 0 μm in depth,the groups were denoted as T1 5 /5,T1 5 /1 0,T30 /5,T30 /1 0, T60 /5 and T60 /1 0,respectively.Smooth titanium surfaces (T0)were used as the controls.Surface topography were observed by ES-EM.HGFs were cultured on the microgroove surfaces.Morphology of “contact guidance”was observed by immunofluorescence tech-nique.Cell cycle distribution was analyzed by Flow cytometry.Results:HGFs on the microgroove surfaces had “contact guidance”par-allel to the microgrooves,whereas the cells on T0 were oriented randomly.T60 /1 0 group had the highest percentage of S phase cells, followed by T30 group and T1 5 group,but still higher than that in the control group.In groups with higher groove width (T60 group and T30 group),the increase of groove depth benefited the increase of S phase percentage,while in T1 5 group,the increase of groove width decreased the S phase percentage.Conclusion:Surfaces of microgrooves with different dimensions achieved “contact guidance”for the cultured HGFs.The surfaces with increasing groove width and depth benefit the cell cycle progression.

Objective:To investigate the effects of microgroove surface morphology on the adhesion and proliferation of the human gin-gival fibroblasts(HGFs).Methods:Microgroove surfaces of titanium were fabricated by photolithography with parallel grooves of 15,30 or 60 μm in width and 5 μm or 10 μm in depth.The groups of the samples were denoted as T15 /5,T15 /10,T30 /5,T30 /10,T60 /5 and T60 /10.Smooth titanium surface(T0)was used as the control.The surface topography was observed by enviroment SEM(ES-EM).HGFs were cultured on the topographically modified surfaces.Morphology was observed by SEM.Cell proliferation was examined by CCK-8 kit.Results:HGFs on the microgroove surfaces had “contact guidance”parallel to the microgrooves,whereas the cells on T0 were oriented randomly.Cell proliferation was promoted and kept for longer period on T60 /10 surface.Conclusion:Surfaces of mi-crogrooves with increasing groove width and depth may achieve “contact guidance”for HGFs and promote cell proliferation.

Objective To compared the difference incontact guidance activity among microgroove surfaces with different sizes of human gingival fibroblast(HGF),with the hope of providing basis for size selection of microgroove for transmucosal part of dental implant.Methods Basing on the size of HGF,microgroove titanium surfaces were fabricated by photolithography with parallel grooves:15,30 or 60 μm in width and 5 or 10 μm in depth.The groups that used different microgroove surfaces were denoted as T15/5,T15/10,T30/5,T30/10,T60/5,and T60/10.Group T0(the control meanwhile was a sputter of titanium on a simple planar silicon substrate).The morphology that HGF arranged along the groove was analyzed by immunofluorescence staining.Difference in contact guidance activity was quantitatively compared basing on the consistency of nucleus arrangement and deformation ratio.Results Microgroove groups had significantly higher consistency of nucleus arrangement and deformation ratio compared to the control group,with T60/10 had the highest consistency of 0.937±0.024,and T15/5 had the lowest consistency of 0.660± 0.016 and T60/10 had the highest deformation ratio of 3.555±0.205,and T15/5 had the lowest deformation ratio of 1.819±0.011.Conclusions Microgroove surfaces of all the different sizes show contact guidance activity on HGF,and the contact guidance activity increases with the increase of width and depth.

Objective:To study the effects of titania nanotubes with three different diameters on human gingival fibroblast (HGF).Methods:Three groups of specimens were prepared. Titania nanotubes with diameters of 30, 100, and 200 nm were synthesized on titanium surfaces through electrochemical anodization at 10, 30, and 60 V, respectively. Specimens were assigned into the three groups according to the diameter of the titania nanotubes. Pure smooth titanium without any treatment was set as the control group. HGF were seeded on the surface of the samples. The cell morphology on the specimens was observed with immunofluorescence staining after 2 h, the cell adhesion after 2 d and cell proliferation after 1, 3, and 7 d were detected using methyl thiazolyl tetrazolium assay, and the secretion of type Ⅰ collagen after 7 d was determined using enzyme-linked immunosorbent assay (each group has three samples for each experiment).Results:HGF on the control group exhibited an oval shape without noticeable extensions. HGF on titania nanotubes with a diameter of 30 nm and titania nanotubes with a diameter of 100 nm elongated further and were arranged orderly. HGF on titania nanotubes with a diameter of 200 nm were sparsely distributed without noticeable extensions. Titania nanotubes with a diameter of 30 nm and titania nanotubes with a diameter of 100 nm could enhance the cell attachment (0.603±0.021 and 0.773±0.045), and secretion of type Ⅰ collagen [(36.5±9.5) and (47.7±4.5) μg/ml, respectively] compared with the control group whose cell attactment was 0.427±0.057, and secretion of type Ⅰ collagen was (22.2±5.9) μg/ml ( P<0.05). Furthermore, titania nanotubes with a diameter of 100 nm showed more cell attchment than titania nanotubes with a diameter of 30 nm did ( P<0.05). Ttania nanotubes with a diameter of 200 nm clearly impaired the cell adhesion (0.250±0.046) and secretion of type Ⅰ collagen [(10.1±3.7) μg/ml] compared with the control group ( P<0.05). At each time point, titania nanotubes with a diameter of 100 nm showed the highest cell proliferation, and the amount of cell proliferation was significantly higher than that on the titania nanotubes with a diameter of 200 nm and the control group at each time point ( P<0.05), and was also significantly higher than that on the titania nanotubes with a diameter of 30 nm at day three ( P<0.05). At each time point, titania nanotubes with a diameter of 200 nm showed the lowest cell proliferation, which was significantly lower than that on the control group at each time point ( P<0.05), except that there was no significant difference in the amount of cell proliferation between titania nanotubes with a diameter of 200 nm and the control group at day one ( P>0.05). Conclusions:Titania nanotubes with a diameter of 100 nm can improve the HGF attachment, proliferation, and secretion of type Ⅰ collagen.

Objective: To study the effect of microgroove surface modification of titanium and zirconia on the biological behavior of gingival fibroblasts in order to find suitable surface materials for the transmucosal part of the dental implant. Methods: Twenty specimens were divided into four groups: smooth titanium (Ti-S), smooth zirconia (ZC-S), microgroove titanium (Ti-MG) and microgroove zirconia (ZC-MG) (five specimens in each group). Microgroove modification of titanium and zirconia surfaces was carried out by using fine machining chip system in the last two groups. The width of groove ridge was 60 μm, the width of groove was 60 μm, the depth of groove was 10 μm. The surface morphologies (the groove width and depth) were observed by scanning electron microscope (SEM), the surface roughness, static contact angle and elemental of specimens in each group were detected by SEM, atomic force microscope (AFM), optical contact angle measuring device and energy-dispersion X-ray analysis (EDX). Morphology of human gingival fibroblast (HGF) that arranged along the groove was analyzed using laser scanning confocal microscope by immunofluorescence staining. Differences in cell proliferation were analyzed and compared using cell counting kit. Expression level of intergrin α₅, β₁ and collagen Ⅰ mRNA were compared among different groups by quantitative real-time PCR for 6 h and 3 d. Results: The surface roughness of smooth titanium group and smooth zirconia group was (63.23± 2.55) and (26.78±3.11) nm, respectively. Microgroove zirconia group showed the best hydrophilicity: the static contact angle was 51.2°±2.0°. HGF was arranged along the groove surface, and cell proliferation results showed that proliferation on microgroove zirconia was more significant than that on other groups from 6 h to 7 d (P<0.05). Intergrin α₅ mRNA has the highest expression in microgroove zirconia (P<0.05) in the early adhesion (6 h), and there was no significant difference in the surface expression of intergrin β₁ and collagen Ⅰ mRNA in the early adhesion (6 h) of each group. However, in the late adhesion (3 d), intergrin α₅, β₁ and collagenⅠ mRNA expression in microgroove surface groups were higher than those of the smooth groups (P<0.05). Conclusions Microgroove zirconia surface has small roughness and good hydrophilicity, which can guide HGF to line up in the groove, and this is beneficial to the HGF proliferation and the expression of structural proteins and functional proteins.

Objective:To compare the effects of gelatin(Gel)and polydopamine(PDA)modification of polycaprolactone(PCL)on the biological behaviour and osteogenic function of osteoblasts.Methods:PCL electrospun membranes were prepared by electrostatic spinning technique,PCL surface was modified by Gel and PDA respectively as G/PCL and D/PCL with chemical self-assembly tech-nique,and the physicochemical properties of the electrospun membranes were characterized by scanning electron microscopy(SEM),Fourier infrared spectroscopy(FTIR),X-ray photoelectron spectroscope(XPS)and contact angle measurement.The MC3T3-E1 cell adhesion morphology was observed by SEM,immunofluorescence staining followed by confocal microscopy(CLSM),cell proliferation at 1,3 and 5 d was tested by CCK-8 assay,alkaline phosphatase(ALP)staining,alizarin red staining and qRT-PCR were used to detect osteogenic gene expression of the cells.Results:A coating of PDA particles was observed on the surface of D/PCL film.FTIR and XPS showed that the characteristic peaks of Gel and PDA,and there was no obvious droplets on the surface of G/PCL and D/PCL ob-served by contact angle test.Cell density of G/PCL group was higher,the adhesion morphology was good and pseudopods were obvi-ous.CCK-8 assay showed the highest proliferation of the cells on G/PCL(P＜0.05).ALP and alizarin red staining of the cells were stronger in D/PCL group than in the other 2 groups.qRT-PCR results showed that the mRNA expression of ALP,COL-1,RUNX2 and OCN was higher in the D/PCL group than in the other 2 groups.Conclusion:Both Gel and PDA modification can enhance the cell adhesion,proliferation and osteogenic properties of PCL scaffolds,Gel modification may induce a more pronounced proliferative effect and PDA modification more pronounced osteogenic effect.

Objective The aim of this study was to evaluate the effects of collagen modification on the osteogenic performance of different surface-modified titanium,including alkaline etching,alkaline etching followed by silaniza-tion,and alkaline etching followed by dopamine modifi-cation.The proliferation,adhesion,and osteogenic differ-entiation abilities of MC3T3-E1 cells on the surfaces with collagen modification were analyzed and compared.Methods Collagen was immobilized on the surfaces of pure titanium(Ti-C),alkaline-etched titanium(Ti-Na-C),alkaline-etched and silanized titanium(Ti-A-C),and alkaline-etched and dopamine-modified titanium(Ti-D-C),with pure titanium(Ti)as the control group.The surface morphology was observed by scanning electron microscopy(SEM),and the surface elemental composition was analyzed by X-ray photoelectron spectroscopy(XPS).Contact angle measurements were conducted to evaluate the hydrophilicity of the surfaces.MC3T3-E1 cells were cultured on the surfaces,and their proliferation,adhesion,and osteogenic differentiation abilities were assessed using CCK-8 assay,laser scanning confocal microscope,alkaline phosphatase(ALP)staining,Alizarin red staining and quantitative analysis,as well as real-time quan-titative polymerase chain reaction(RT-qPCR)to evaluate the mRNA expression levels of osteogenic-related genes,includ-ing ALP,typeⅠcollagen(COL-1),osteocalcin(OCN),osteopontin(OPN).Results SEM and XPS results confirmed the successful immobilization of collagen on the titanium surfaces,with the Ti-Na-C group exhibiting a higher amount of col-lagen modification.Contact angle measurements showed improved hydrophilicity of the surfaces after collagen modifica-tion.CCK-8 results indicated good compatibility of the materials with MC3T3-E1,with enhanced cell proliferation on the collagen-modified surfaces.Cell fluorescence staining revealed better cell spreading on the collagen-modified surfaces,and ALP and Alizarin red staining results suggested that the Ti-Na-C group exhibited the best osteogenic performance,with significantly higher absorbance values in the Alizarin red quantification analysis.RT-qPCR analysis showed that the Ti-Na-C group had the highest expression of the osteogenic-related gene OPN.Conclusion Among the different colla-gen modification approaches employed in this study,collagen modification on alkaline-etched titanium surfaces showed the most conducive effects on MC3T3-E1 cell adhesion,spreading,proliferation,and osteogenic differentiation.This ap-proach can be considered as the optimal collagen modification strategy for enhancing osteogenesis on titanium surfaces.

To study the effect of microgroove surface modification of titanium and zirconia on the biological behavior of gingival fibroblasts in order to find suitable surface materials for the transmucosal part of the dental implant. Methods Twenty specimens were divided into four groups: smooth titanium (Ti?S), smooth zirconia (ZC?S), microgroove titanium (Ti?MG) and microgroove zirconia (ZC?MG) (five specimens in each group). Microgroove modification of titanium and zirconia surfaces was carried out by using fine machining chip system in the last two groups. The width of groove ridge was 60 μm, the width of groove was 60 μm, the depth of groove was 10 μm. The surface morphologies (the groove width and depth) were observed by scanning electron microscope (SEM), the surface roughness, static contact angle and elemental of specimens in each group were detected by SEM, atomic force microscope (AFM), optical contact angle measuring device and energy?dispersion X?ray analysis (EDX). Morphology of human gingival fibroblast (HGF) that arranged along the groove was analyzed using laser scanning confocal microscope by immunofluorescence staining. Differences in cell proliferation were analyzed and compared using cell counting kit. Expression level of intergrin α5, β1 and collagenⅠ mRNA were compared among different groups by quantitative real?time PCR for 6 h and 3 d. Results The surface roughness of smooth titanium group and smooth zirconia group was (63.23± 2.55) and (26.78±3.11) nm, respectively. Microgroove zirconia group showed the best hydrophilicity: the static contact angle was 51.2°±2.0°. HGF was arranged along the groove surface, and cell proliferation results showed that proliferation on microgroove zirconia was more significant than that on other groups from 6 h to 7 d (P<0.05). Intergrin α5 mRNA has the highest expression in microgroove zirconia (P<0.05) in the early adhesion (6 h), and there was no significant difference in the surface expression of intergrin β1 and collagenⅠmRNA in the early adhesion (6 h) of each group. However, in the late adhesion (3 d), intergrin α5, β1 and collagenⅠmRNA expression in microgroove surface groups were higher than those of the smooth groups (P<0.05). Conclusions Microgroove zirconia surface has small roughness and good hydrophilicity, which can guide HGF to line up in the groove, and this is beneficial to the HGF proliferation and the expression of structural proteins and functional proteins.

OBJECTIVES: This study aimed to investigate the effect of new biomimetic micro/nano surfaces on the osteoclastic differentiation of RAW264.7 macrophages by simulating natural osteons for the design of concentric circular structures and modifying graphene oxide (GO). METHODS: The groups were divided into smooth titanium surface group (SS), concentric microgrooved titanium surface group (CMS), and microgroove modified with GO group (GO-CMS). The physicochemical properties of the material surfaces were studied using scanning electron microscopy (SEM), contact-angle measurement, atomic force microscopy, X-ray photoelectron spectroscopy analysis, and Raman spectroscopy. The effect of the modified material surface on the cell biological behavior of RAW264.7 was investigated by cell-activity assay, SEM, and laser confocal microscopy. The effect on the osteoclastic differentiation of macrophages was investiga-ted by tartrate-resistant acid phosphatase (TRAP) immunofluorescence staining and quantitative real-time polymerase chain reaction (qRT-PCR) experiments. RESULTS: Macrophages were arranged in concentric circles along the microgrooves, and after modification with GO, the oxygen-containing groups on the surface of the material increased and hydrophilicity increased. Osteoclasts in the GO-CMS group were small in size and number and had the lowest TRAP expression. Although it promoted the proliferation of macrophages in the GO-CMS group, the expression of osteoclastic differentiation-related genes was lower than that in the SS group, and the difference was statistically significant (P<0.05). CONCLUSIONS Concentric circular microgrooves restricted the fusion of osteoclasts and the formation of sealing zones. Osteomimetic concentric microgrooves modified with GO inhibited the osteoclastic differentiation of RAW 264.7 macrophages.
Graphite/pharmacology* ; Titanium/pharmacology* ; Haversian System ; Macrophages ; Cell Differentiation ; Oxides/pharmacology* ; Surface Properties