1.The effects of microgroove surface form of titanium on the adhesion and cell cycle progression of human gingival fibroblasts
Yingzhen LAI ; Shan LIN ; Jiang CHEN
Journal of Practical Stomatology 2016;32(1):89-95
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.
2.Effect of microgroove surface morphology on the adhesion and proliferation of human gingival fibroblasts
Yingzhen LAI ; Jiang CHEN ; Shan LIN
Journal of Practical Stomatology 2015;(4):515-519
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.
3. Effects of different implant materials and surface microgrooves on the biological behavior of gingival fibroblasts
Yingzhen LAI ; Xueguan LU ; Yihuang CAI
Chinese Journal of Stomatology 2019;54(10):676-682
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 (
4.Comparative study on contact guidance activity of human gingival fibroblasts on microgroove surfaces.
Chinese Journal of Stomatology 2015;50(1):33-37
OBJECTIVETo compared the difference in contact 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.
METHODSBasing 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.
RESULTSMicrogroove 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.
CONCLUSIONSMicrogroove 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.
Cell Adhesion ; physiology ; Dental Implants ; Fibroblasts ; cytology ; physiology ; Gingiva ; cytology ; Humans ; Surface Properties ; Titanium
5.Effect of graphene-oxide-modified osteon-like concentric microgrooved surface on the osteoclastic differentiation of macrophages.
Hong WANG ; Qinglin WU ; Yingzhen LAI ; Yihuang CAI
West China Journal of Stomatology 2023;41(2):165-174
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*
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Titanium/pharmacology*
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Haversian System
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Macrophages
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Cell Differentiation
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Oxides/pharmacology*
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Surface Properties