Adhesion Of Human Gingival Fibroblasts Research Articles

Impact of Composition and Autoclave Sterilization on the Mechanical and Biological Properties of ECM-Mimicking Cryogels

Cryogels represent a valid strategy as scaffolds for tissue engineering. In order to adequately support adhesion and proliferation of anchorage-dependent cells, different polymers need to be combined within the same scaffold trying to mimic the complex features of a natural extracellular matrix (ECM). For this reason, in this work, gelatin (Gel) and chondroitin sulfate (CS), both functionalized with methacrylic groups to produce CSMA and GelMA derivatives, were selected to prepare cryogel networks. Both homopolymer and heteropolymer cryogels were produced, via radical crosslinking reactions carried out at −12 °C for 2 h. All the scaffolds were characterized for their mechanical, swelling and morphological properties, before and after autoclave sterilization. Moreover, they were evaluated for their biocompatibility and ability to support the adhesion of human gingival fibroblasts and tenocytes. GelMA-based homopolymer networks better withstood the autoclave sterilization process, compared to CSMA cryogels. Indeed, GelMA cryogels showed a decrease in stiffness of approximately 30%, whereas CSMA cryogels of approximately 80%. When GelMA and CSMA were blended in the same network, an intermediate outcome was observed. However, the hybrid scaffolds showed a general worsening of the biological performance. Indeed, despite their ability to withstand autoclave sterilization with limited modification of the mechanical and morphological properties, the hybrid cryogels exhibited poor cell adhesion and high LDH leakage. Therefore, not only do network components need to be properly selected, but also their combination and ability to withstand effective sterilization process should be carefully evaluated for the development of efficient scaffolds designed for tissue engineering purposes.

Evaluation of Adhesion and Viability of Human Gingival Fibroblasts on Strontium-Coated Titanium Surfaces: an in vitro Study.

Applying multifunctional coatings employing strontium (Sr) ions on titanium (Ti) surfaces is a useful and biocompatible method to improve osseointegration and prevent tissue infections through antimicrobial activity. Nonetheless, the effectiveness of Sr coating on the adhesion and viability of human gingival fibroblasts (HGFs) to Ti surfaces remains unclear. The study aimed to evaluate the effect of Sr coating on the adhesion and viability of HGFs to Ti surfaces. The Ti wafers were divided into two groups based on Sr coating: uncoated Ti (control) and Sr-coated Ti. The Magnetron sputtering technique was used for Sr coating on Ti surfaces. The HGFs were seeded onto the surfaces and cultured for 48 and 96 hours before the cell adhesion and viability of the attached HGFs were assessed. The adhesion of HGFs was analyzed using the attached cell numbers at 48 h and 96 h, and the morphology at 24 h and 72 h. The cytotoxic effect on HGFs was assessed after 24 and 72 hours of incubation using cell viability assay. Student's t-test was used for statistical analysis. The number of cells attached to Sr-coated surfaces was significantly greater than those attached to uncoated Ti surfaces after 48 hours (P<0.0001) and 96 hours (P=0.0002). Sr-coated and uncoated Ti surfaces were not cytotoxic to HGFs, with the cell viability ranging from 92% to 105% of the untreated control HGFs. There were no significant differences in cell viability between Sr-coated and uncoated Ti surfaces at 24 hours (P=0.3675) and 72 hours (P=0.0982). Sr-coated Ti surfaces induce adhesion of HGFs compared to uncoated Ti surfaces. Further, Sr-coated and uncoated Ti surfaces show no cytotoxic effect on the attached HGFs.

Effects and underlying mechanism of micro-nano-structured zirconia surfaces on biological behaviors of human gingival fibroblasts under inflammatory conditions

Zirconia is one of the most commonly used materials for abutments of dental implants, especially in the anterior region. Soft tissue integration to the zirconia abutment surface remains a challenge. Peri-implant soft tissue integration serves as a physiological barrier, attenuating pathogen penetration and preventing peri‑implant disease. The surface microstructure of zirconia has significant effects on the biological behaviors of human gingival fibroblasts (HGFs), but the effects under inflammatory conditions are still unclear. In this study, we established two micro-nano structures on zirconia surfaces using a femtosecond laser, including microgrooves with widths of 30 µm (G3) and 60 µm (G6) and depths of 5 µm, and nanoparticles inside the microgrooves. Polished surfaces were used as controls. HGFs were seeded onto the three groups of zirconia specimens and stimulated with lipopolysaccharide. The HGFs on micro-nano-structured zirconia surfaces exhibited lower inflammatory responses and higher cell adhesion, proliferation, and migration under inflammatory conditions compared with the polished surfaces. Additionally, the G3 group exhibited lower inflammatory responses and higher cell adhesion and migration than the G6 group. The micro-nano-structured zirconia surface exhibited decreased neutrophil infiltration and increased M2-type macrophage polarization in vivo. To explore the molecular mechanism, RNA sequencing and gene silencing were utilized, which revealed two critical target genes regulated by the G3 group. Overall, we proposed an innovative micro-nano-structured zirconia surface that reduced the in vitro and in vivo inflammatory responses and promoted HGF adhesion, migration, and proliferation under inflammatory conditions, in which TRAFD1 and NLRC5 were the underlying key genes. Statement of significanceZirconia is one of the most commonly used materials for abutments, especially in the anterior region. The surface microstructure of zirconia has significant effects on the biological behaviors of human gingival fibroblasts (HGFs), but few studies have investigated these effects under inflammatory conditions, and the mechanism remains unclear. In this study, we developed an innovative micro-nano-structured zirconia surface using a femtosecond laser, which reduces the in vitro and in vivo pro-inflammatory responses and promotes HGFs adhesion, migration, and proliferation under inflammatory conditions compared with the polished zirconia surface. The potential underlying mechanism was also investigated. This work has provided some theoretical basis for the micro-nano-structured zirconia surface in potentially reducing the inflammation and enhancing peri‑implant soft-tissue integration under inflammatory conditions.

Cellular experimental study and mechanism of surface modification of 3D printed titanium materials to modulate the sealing and antimicrobial properties of oral implants

Investigating the role of surface-modified 3D printed titanium implants in modulating oral soft tissue closure and antimicrobial properties. Eighteen 6-month-old healthy adult New Zealand White rabbits were selected as experimental animals, the left and right mandibular teeth of each experimental animal were randomly divided into two groups, and the bilateral first premolar teeth were extracted after general anesthesia, and one implant was implanted into each of the magnesium/zinc ionized surface-treated group (experimental group) and the surface-untreated group (control group), with a total of 36 implants, i.e., 18 implants for the experimental group and 18 implants for the control group. Bilateral mandibular samples from both groups were removed after 2 and 4 weeks of implantation to investigate the attachment and spreading of human gingival fibroblasts (HGFs) on the surface of the samples, their proliferative and migratory behaviors as well as the expression levels of related gene proteins (CoL-1 and TGF-β), and HE staining of the soft tissues of the samples and inflammatory cell counts. After 2 and 4 weeks of implantation, the experimental group’s HGFs adhesion and spreading were significantly better than that of the control group, the migration and proliferation of HGFs were significantly greater than that of the control group ( p &lt; .05), and the expression of CoL-1 and TGF-β genes was significantly higher than that of the control group ( p &lt; .05), and the inflammatory cell count was significantly smaller than that of the control group ( p &lt; .05). Surface modification by magnesium and zinc ions enhances the sealing performance and antimicrobial performance of 3D printed titanium implants in oral soft tissues, with significant effects.

Piranha-etched titanium nanostructure reduces biofilm formation in vitro

ObjectivesNano-modified surfaces for dental implants may improve gingival fibroblast adhesion and antibacterial characteristics through cell-surface interactions. The present study investigated how a nanocavity titanium surface impacts the viability and adhesion of human gingival fibroblasts (HGF-1) and compared its response to Porphyromonas gingivalis with those of marketed implant surfaces.Material and methodsCommercial titanium and zirconia disks, namely, sandblasted and acid-etched titanium (SLA), sandblasted and acid-etched zirconia (ZLA), polished titanium (PT) and polished zirconia (ZrP), and nanostructured disks (NTDs) were tested. Polished titanium disks were etched with a 1:1 combination of 98% H2SO4 and 30% H2O2 (piranha etching) for 5 h at room temperature to produce the NTDs. Atomic force microscopy was used to measure the surface topography, roughness, adhesion force, and work of adhesion. MTT assays and immunofluorescence staining were used to examine cell viability and adhesion after incubation of HGF-1 cells on the disk surfaces. After incubation with P. gingivalis, conventional culture, live/dead staining, and SEM were used to determine the antibacterial properties of NTD, SLA, ZLA, PT, and ZrP.ResultsEtching created nanocavities with 10–20-nm edge-to-edge diameters. Chemical etching increased the average surface roughness and decreased the surface adherence, while polishing and flattening of ZrP increased adhesion. However, only the NTDs inhibited biofilm formation and bacterial adherence. The NTDs showed antibacterial effects and P. gingivalis vitality reductions. The HGF-1 cells demonstrated greater viability on the NTDs compared to the controls.ConclusionNanocavities with 10–20-nm edge-to-edge diameters on titanium disks hindered P. gingivalis adhesion and supported the adhesion of gingival fibroblasts when compared to the surfaces of currently marketed titanium or zirconia dental implants.Clinical relevanceThis study prepared an effective antibacterial nanoporous surface, assessed its effects against oral pathogens, and demonstrated that surface characteristics on a nanoscale level influenced oral pathogens and gingival fibroblasts.Clinical trial registration: not applicable

Promoted Abutment-Soft Tissue Integration Around Self-Glazed Zirconia Surfaces with Nanotopography Fabricated by Additive 3D Gel Deposition.

Improving the biological sealing around dental abutments could promote the long-term success of implants. Although titanium abutments have a wide range of clinical applications, they incur esthetic risks due to their color, especially in the esthetic zone. Currently, zirconia has been applied as an esthetic alternative material for implant abutments; however, zirconia is purported to be an inert biomaterial. How to improve the biological activities of zirconia has thus become a popular research topic. In this study, we presented a novel self-glazed zirconia (SZ) surface with nanotopography fabricated by additive 3D gel deposition and investigated its soft tissue integration capability compared to that of clinically used titanium and polished conventional zirconia surfaces. Three groups of disc samples were prepared for in vitro study and the three groups of abutment samples were prepared for in vivo study. The surface topography, roughness, wettability and chemical composition of the samples were examined. Moreover, we analyzed the effect of the three groups of samples on protein adsorption and on the biological behavior of human gingival keratinocytes (HGKs) and human gingival fibroblasts (HGFs). Furthermore, we conducted an in vivo study in which the bilateral mandibular anterior teeth of rabbits were extracted and replaced with implants and corresponding abutments. The surface of SZ showed a unique nanotopography with nm range roughness and a greater ability to absorb protein. The promoted expression of adhesion molecules in both HGKs and HGFs was observed on the SZ surface compared to the surfaces of Ti and PCZ, while the cell viability and proliferation of HGKs and the number of HGFs adhesion were not significant among all groups. In vivo results showed that the SZ abutment formed strong biological sealing at the abutment-soft tissue interface and exhibited markedly more hemidesmosomes when observed with a transmission electron microscope. These results demonstrated that the novel SZ surface with nanotopography promoted soft tissue integration, suggesting its promising application as a zirconia surface for the dental abutment.

Evaluation of cell morphology and adhesion capacity of human gingival fibroblasts on titanium discs with different roughened surfaces: an in vitro scanning electron microscope analysis and cell culture study.

Implantoplasty is an option in peri-implantitis treatment. What is known about the effects of implantoplasty on peri-implant soft tissue adhesion and cell behaviours is limited. This study aimed to evaluate the morphological features and adhesion capacity of human gingival fibroblast (HGF) cells onto sand-blasted, large-grit, acid-etched (SLA®) titanium (Ti) discs surfaces roughened with different implantoplasty protocols. The study included a total of 48 Ti discs divided into four groups (n = 12 per group): Group I: machined, smooth surface discs; Group II: SLA® surface discs; Group III: SLA® surface discs roughened with diamond bur sequence (40 and 15-μm grit); Group IV: SLA® surface discs roughened with diamond bur sequence (125 and 40-μm grit). Following polishing procedure, the surface roughness value of discs was assessed by a profilometer and scanning electron microscope. HGFs were cultured on Ti discs and cell adhesion was examined after the 24th, 48th, and 72nd hours. Statistical significance was set at the p ≤ 0.05 level. Scanning electron microscope analyses of the discs revealed that fibroblasts exhibited well-dispersion and a firm attachment in all groups. The cells in group I and II had thin and long radial extensions from the areas where the nucleus was located to the periphery; however, attached cells in group III and IV showed more spindle-shaped morphology. The surface roughness parameters of the test groups were lower than those of the SLA®. The SLA® group showed the highest HGF adhesion (group II) (p ≤ 0.05). HGF adhesion in group IV was greater compared to group III, but less than group I. This study showed that the characteristics of the burs applied in the implantoplasty protocol are determinant for the surface roughness and fibroblast adhesion occurs on surfaces with decreased roughness following implantoplasty. Consequently, it should be kept in mind that the surface properties of the implant may affect the adherent cell morphology and adhesion.

Focal adhesion formation of primary human gingival fibroblast on hydrothermally and in-sol-made TiO2 -coated titanium.

Optimal cell adhesion of the gingival fibroblasts to dental implants is important for maintaining good implant integration. The aim of this study was to discover, if the nanoporous TiO2 -coating on titanium alloy substrates is able to increase the cell adhesion of the human gingival fibroblasts (HGF). The study consisted of three differently produced titanium groups: hydrothermally produced TiO2 -coating (HT), novel TiO2 -coating made in sol (SOL), and noncoated control group. Primary HGF cells were initiated from gingival biopsies from patients having a third molar extraction. HGF were cultivated on titanium discs for 2 and 24 h to determine the initial attachment with confocal microscope. The cell spreading and adhesion protein signals were measured. In addition, expression of adhesion proteins vinculin, paxillin, and focal adhesion kinase (FAK) were measured after 3 days of cultivation by using Western Blotting. Higher protein levels of paxillin, vinculin, and FAK were induced on both coated discs compared to noncoated discs. The difference was statistically significant (p < 0.05) concerning expression of paxillin. The cell spreading was significantly larger on SOL discs after 2 and 24 h when comparing to noncoated controls. The confocal microscope analyses revealed significantly higher adhesion protein signals on both HT- and SOL-coated titanium compared to control group. This study showed, that both methods to produce TiO2 -coatings are able to increase HGF adhesion protein expression and cell spreading on titanium surface. Accordingly, the coatings can potentially improve the gingival attachment to titanium implant surfaces.

Effects of TiS2 on Inhibiting Candida albicans Biofilm Formation and Its Compatibility with Human Gingival Fibroblasts in Titanium Implants.

Titanium is widely used in medical devices, such as dental and orthopedic implants, due to its excellent mechanical properties, low toxicity, and biocompatibility. However, the titanium surface has the risk of microbial biofilm formation, which results in infections from species such as Candida albicans (C. albicans). This kind of biofilm prevents antifungal therapy and complicates the treatment of infectious diseases associated with implanted devices. It is critical to developing a feasible surface to decrease microbial growth while not interfering with the growth of the host cells. This study reports the influence of titanium surface modification to titanium disulfide (TiS2) on inhibiting C. albicans biofilm formation while allowing the attachment of human gingival fibroblasts (HGFs) on their surface. The surface of titanium parts is directly converted to structured titanium and TiS2 using direct laser processing and crystal growth methods. C. albicans adhesion and colonization are then investigated on these surfaces by the colony counting assay and reactive oxygen species (ROS) assay, using 2',7'-dichlorofluorescin diacetate (DCFH-DA) and microscopy images. Also, the viability and adhesion of HGFs on these surfaces are investigated to show their adhesion and biocompatibility. Titanium samples with the TiS2 surface show both C. albicans biofilm inhibition and HGF attachment. This study provides insight into designing and manufacturing titanium biomedical implants.

Enhancing cell adhesive and antibacterial activities of glass-fibre-reinforced polyetherketoneketone through Mg and Ag PIII.

Glass-fibre-reinforced polyetherketoneketone (PEKK-GF) shows great potential for application as a dental implant restoration material; however, its surface bioinertness and poor antibacterial properties limit its integration with peri-implant soft tissue, which is critical in the long-term success of implant restoration. Herein, functional magnesium (Mg) and silver (Ag) ions were introduced into PEKK-GF by plasma immersion ion implantation (PIII). Surface characterization confirmed that the surface morphology of PEKK-GF was not visibly affected by PIII treatment. Further tests revealed that PIII changed the wettability and electrochemical environment of the PEKK-GF surface and enabled the release of Mg2+ and Ag+ modulated by Giavanni effect. In vitro experiments showed that Mg/Ag PIII-treated PEKK-GF promoted the proliferation and adhesion of human gingival fibroblasts and upregulated the expression of adhesion-related genes and proteins. In addition, the treated samples inhibited the metabolic viability and adhesion of Streptococcus mutans and Porphyromonas gingivalis on their surfaces, distorting bacterial morphology. Mg/Ag PIII surface treatment improved the soft tissue integration and antibacterial activities of PEKK-GF, which will further support and broaden its adoption in dentistry.

Vitamin C stimulates RNA expression of human gingival fibroblasts proliferation and adhesion in cigarette smokers: An in vitro study

BackgroundSmoking and the severity of periodontal disease have long been associated. In Saudi Arabia, tobacco smoking is rising, contributing to the increased demand for products that counter its detrimental effects. The antioxidant properties of vitamin C (vit C) make it a powerful countermeasure to tobacco toxicity. Observation of these effects on human gingival fibroblasts (hGFs) would suggest use of vitamin C in future dental applications. AimTo examine the proliferation, adhesion, and expression of extracellular RNA in human gingival fibroblasts extracted from cigarette smokers when compared to never-smokers, in association with vitamin C. Materials and MethodsHuman gingival fibroblasts were extracted from Periodontal free sites of healthy adult male participants. Group 1; consisted of Heavy cigarette smokers (n = 1) while group 2 was never-smokers (n = 1). Collected cells were cultured and subcultured in supplemented growth medium. Vitamin C was then induced in the medium at the experimental sixth passage. RNA expression analysis using quantitative reverse transcriptase-polymerase chain reaction was performed to analyze the adhesion, proliferation, and extracellular matrix expression. ResultsExpression of the adhesion gene (CD44) in the smoker group was significantly downregulated than never-smoker group (p-value = 0.024). After the induction of vitamin C, the smoker samples showed a significant improvement in their gene expression levels. The extracellular genes involved in this study (COL1A1, LAMA3, and TGFB3) were significantly affected by the smoking status. In addition, the proliferation of MK167 and CCNB1 genes in smokers and never-smokers was increased. ConclusionCigarette smoking affects the overall properties of human gingival fibroblasts’ adhesion, proliferation, and extra-cellular matrix formation. Furthermore, the addition of vitamin C affects these cellular properties in a positive manner.

Comparison between Single and Multi-LED Emitters for Photodynamic Therapy: An In Vitro Study on Enterococcus faecalis and Human Gingival Fibroblasts

Aim of the study: The aim was to evaluate the effects of two LED devices, TL-01 and TL-03 in photodynamic therapy (PDT), on Enterococcus faecalis and on human gingival fibroblasts (HGFs). TL-01, characterized by a single emitter, irradiates one periodontal site at a time, whereas the multi-led device (TL-03) irradiates all vestibular sites of a single arch at a time. Methods: E. faecalis bacterial suspensions and HGFs were incubated for 45 min with Aladent gel (ALAD) containing 5-aminolevulinic acid and then exposed to LED devices (ALAD-PDT), having different distance and timing of irradiation (TL-01 N (0.5 mm, for 7 min), TL-03 N (0.5 mm, 15 min) and TL-03 F (30.0 mm, 15 min)). For bacterial suspension, the colony forming units and the live/dead staining were evaluated after 24 h, while the protoporphyrin IX (PpIX) content was monitored in all phases of the experimentation. For HGFs, the cell viability, proliferation, cell morphology, and adhesion were evaluated at 24 h. Results: Both TL-01 and TL-03 showed a significant reduction of bacterial load. The photoinactivation was inversely proportional to the PpIX accumulation. TL-01 and TL-03 promoted proliferation and adhesion of HGFs. Conclusions: Both tested devices for ALAD-PDT were equally effective in significantly reducing Enterococcus faecalis growth and in promoting HGFs proliferation and adhesion, in vitro.

Effect of a Nanostructured Titanium Surface on Gingival Cell Adhesion, Viability and Properties against P. gingivalis.

Objectives: The transgingival part of titanium implants is either machined or polished. Cell-surface interactions as a result of nano-modified surfaces could help gingival fibroblast adhesion and support antibacterial properties by means of the physico-mechanical aspects of the surfaces. The aim of the present study was to determine how a nanocavity titanium surface affects the viability and adhesion of human gingival fibroblasts (HGF-1). Additionally, its properties against Porphyromonas gingivalis were tested. Material and Methods: Two different specimens were evaluated: commercially available machined titanium discs (MD) and nanostructured discs (ND). To obtain ND, machined titanium discs with a diameter of 15 mm were etched with a 1:1 mixture of 98% H2SO4 and 30% H2O2 (piranha etching) for 5 h at room temperature. Surface topography characterization was performed via scanning electron microscopy (SEM) and atomic force microscopy (AFM). Samples were exposed to HGF-1 to assess the effect on cell viability and adhesion, which were compared between the two groups by means of MTT assay, immunofluorescence and flow cytometry. After incubation with P. gingivalis, antibacterial properties of MD and ND were determined by conventional culturing, live/dead staining and SEM. Results: The present study successfully created a nanostructured surface on commercially available machined titanium discs. The etching process created cavities with a 10–20 nm edge-to-edge diameter. MD and ND show similar adhesion forces equal to about 10–30 nN. The achieved nanostructuration reduced the cell alignment along machining structures and did not negatively affect the proliferation of gingival fibroblasts when compared to MD. No differences in the expression levels of both actin and vinculin proteins, after incubation on MD or ND, were observed. However, the novel ND surface failed to show antibacterial effects against P. gingivalis. Conclusion: Antibacterial effects against P. gingivalis cannot be achieved with nanocavities within a range of 10–20 nm and based on the piranha etching procedure. The proliferation of HGF-1 and the expression levels and localization of the structural proteins actin and vinculin were not influenced by the surface nanostructuration. Further studies on the strength of the gingival cell adhesion should be performed in the future. Clinical relevance: Since osseointegration is well investigated, mucointegration is an important part of future research and developments. Little is known about how nanostructures on the machined transgingival part of an implant could possibly influence the surrounding tissue. Targeting titanium surfaces with improved antimicrobial properties requires extensive preclinical basic research to gain clinical relevance.

Er:YAG laser irradiation enhances bacterial and lipopolysaccharide clearance and human gingival fibroblast adhesion on titanium discs

To investigate the effect of Er:YAG laser treatment on lipopolysaccharide (LPS) clearance and fibroblast adhesion on titanium disks. Grade IV titanium discs (n = 216) were used and allocated to 6 groups. Group 1 was the negative control without Porphyromonas gingivalis inoculation. Discs in Groups 2–6 were incubated with P. gingivalis to form a biofilm. Group 3 received 0.12% chlorhexidine irrigation and Group 4 received titanium curettage to remove the biofilm. Group 5 was treated with Er:YAG laser irradiation and Group 6 was treated with titanium curettage plus Er:YAG laser irradiation. The contact angle and surface roughness were measured after the various treatments. The surface microstructure and residual bacteria were examined using scanning electron microscopy and confocal laser scanning microscopy, respectively. Residual LPS was examined using a limulus amoebocyte lysate assay and human gingival fibroblast adhesion was quantified using fluorescent microscopy. Curettage plus Er:YAG laser irradiation was the most effective method for removing bacteria and LPS. No significant difference in the amount of fibroblast adhesion was found between the control and Group 6. Combined use of Er:YAG laser irradiation and curettage optimizes LPS clearance and fibroblast adhesion on titanium discs.

Customized protein modification improves human gingival fibroblasts adhesion on SiO2

SiO2-based materials have great potential for dental implant abutment in the future, but the SiO2 itself didn't support cell adhesion well, which could hinder the gingival tissue from forming a favorable seal around the abutment. In this study, SiO2 showed inferior human gingival fibroblasts (HGF) adhesion than ZrO2, the classic abutment material, which was caused by the serum protein affinity difference. By mass spectrometry, fibronectin (FN) and vitronectin (FN) in serum were found to mediate cell adhesion on materials. Single- molecule tracking and molecule dynamics simulation revealed that the affinity between FN and SiO2 was weaker than that of ZrO2, and the cell adhesion on FN-coated SiO2 was not improved as well as that on ZrO2, while the affinity of VN and SiO2 was comparable with that of VN and ZrO2 and the VN-coated SiO2 facilitated the HGF adhesion as effectively as VN-coated ZrO2. Here, the customized protein modification improved the cell adhesion on SiO2 and promoted the development of the SiO2-based dental materials.

Titanium Substratum Roughness as a Determinant of Human Gingival Fibroblast Fibronectin and α-Smooth Muscle Actin Expression.

The most appropriate surface treatment to enhance gingival connective tissue formation on the abutment of dental implants remains undefined, with healing associated with a scar-like response. We have previously shown that topographies with an arithmetic average of the absolute profile height deviations (Ra) = 4.0 induces an anti-fibrotic phenotype in human gingival fibroblasts (HGFs) by causing nascent adhesion formation. With bacterial colonization considerations, we hypothesized that a lower Ra could be identified that would alter adhesion stability and promote a matrix remodeling phenotype. Focal adhesions (FAs) area decreased with increasing roughness, although no differences in cell attachment or proliferation were observed. Alpha smooth muscle actin (α-SMA) protein levels were significantly reduced on Ra = 3.0 and 4.0 vs. 0.1 (p < 0.05), with incorporation of α-SMA into stress fibers most prominent on Ra = 0.1. Fibronectin protein levels were reduced on 3.0 and 4.0 vs. 0.1 (p < 0.05), and Ra = 1.5 and deeper significantly altered fibronectin deposition. Addition of exogenous TGF-β3 increased HGF adhesion size on 0.1 surfaces, but not on any other topography. We conclude that Ra = 1.5 is sufficient to reduce adhesion size and inhibit α-SMA incorporation into stress fibers in HGFs, but 3.0 is required in the presence of exogenous TGF-β3. Our findings have implications for inhibiting fibrotic tissue formation surrounding percutaneous devices such as dental implants.

Mg-Fe layered double hydroxides modified titanium enhanced the adhesion of human gingival fibroblasts through regulation of local pH level

The durability of dental implants is closely related to osseointegration and surrounding soft tissue sealing. Appropriate local pH favors fibroblasts adhesion and contributes to soft tissue sealing. Layered double hydroxides (LDHs) are characterized by adjustable alkalinity, offering a possibility to investigate the influence of pH on cellular behaviors. Herein, we fabricated MgFe LDHs modified titanium. During calcination, the local pH value of LDHs increase, without altering other physics and chemical properties via OH− exchange mechanism. In vitro studies showed that LDHs films calcined at 250 °C for 2 h provide a local pH of 10.17, which promote early adhesion, proliferation, and type I collagen expression of human gingival fibroblasts (hGFs) through the formation of focal adhesion complex and activation of focal adhesion kinase related signaling pathways. In conclusion, endowing the titanium surface with appropriate alkalinity by MgFe LDHs films enhances the adhesion of hGFs, providing a new strategy of designing multifunctional biomaterials for soft tissue sealing around dental implants.

The Early Adhesion Effects of Human Gingival Fibroblasts on Bovine Serum Albumin Loaded Hydrogenated Titanium Nanotube Surface

The soft tissue sealing at the transmucal portion of implants is vital for the long-term stability of implants. Hydrogenated titanium nanotubes (H2-TNTs) as implant surface treatments were proved to promote the adhesion of human gingival fibroblasts (HGFs) and have broad usage as drug delivery systems. Bovine serum albumin (BSA) as the most abundant albumin in body fluid was crucial for cell adhesion and was demonstrated as a normal loading protein. As the first protein arriving on the surface of the implant, albumin plays an important role in initial adhesion of soft tissue cells, it is also a common carrier, transferring and loading different endogenous and exogenous substances, ions, drugs, and other small molecules. The aim of the present work was to investigate whether BSA-loaded H2-TNTs could promote the early adhesion of HGFs; H2-TNTs were obtained by hydrogenated anodized titanium dioxide nanotubes (TNTs) in thermal treatment, and BSA was loaded in the nanotubes by vacuum drying; our results showed that the superhydrophilicity of H2-TNTs is conducive to the loading of BSA. In both hydrogenated titanium nanotubes and non-hydrogenated titanium nanotubes, a high rate of release was observed over the first hour, followed by a period of slow and sustained release; however, BSA-loading inhibits the early adhesion of human gingival fibroblasts, and H2-TNTs has the best promoting effect on cell adhesion. With the release of BSA after 4 h, the inhibitory effect of BSA on cell adhesion was weakened.

Improved response of human gingival fibroblasts to titanium coated with micro-/nano-structured tantalum

ObjectivesThis study aims to evaluate the ability of tantalum-coated titanium to improve human gingival fibroblasts’ adhesion, viability, proliferation, migration performance, and the potential molecular mechanisms.Materials and methodsTitanium plates were divided into two groups: (1) no coating (Ti, control), (2) Tantalum-coated titanium (Ta-coated Ti). All samples were characterized by scanning electronic microscopy, surface roughness, and hydrophilicity. Fibroblasts’ performance were analyzed by attached cell number at 1 h, 4 h, and 24 h, morphology at 1 h and 4 h, viability at 1 day, 3 days, 5 days, and 7 days, recovery after wounding at 6 h, 12 h, and 24 h. RT-PCR, western blot were applied to detect attachment-related genes’ expression and protein synthesis at 4 h and 24 h. Student’s t test was used for statistical analysis.ResultsTantalum-coated titanium demonstrates a layer of homogeneously distributed nano-grains with mean diameter of 25.98 (± 14.75) nm. It was found that after tantalum deposition, human gingival fibroblasts (HGFs) adhesion, viability, proliferation, and migration were promoted in comparison to the control group. An upregulated level of Integrin β1 and FAK signaling was also detected, which might be the underlying mechanism.ConclusionIn the present study, adhesion, viability, proliferation, migration of human gingival fibroblasts are promoted on tantalum-coated titanium, upregulated integrin β1 and FAK might contribute to its superior performance, indicating tantalum coating can be applied in transmucosal part of dental implant.Clinical significanceTantalum deposition on titanium surfaces can promote human gingival fibroblast adhesion, accordingly forming a well-organized soft tissue sealing and may contribute to a successful osseointegration.

Co-implantation of magnesium and zinc ions into titanium regulates the behaviors of human gingival fibroblasts

Soft tissue sealing around implants acts as a barrier between the alveolar bone and oral environment, protecting implants from the invasion of bacteria or external stimuli. In this work, magnesium (Mg) and zinc (Zn) are introduced into titanium by plasma immersed ion implantation technology, and their effects on the behaviors of human gingival fibroblasts (HGFs) as well as the underlying mechanisms are investigated. Surface characterization confirms Mg and Zn exist on the surface in metallic and oxidized states. Contact angle test suggests that surface wettability of titanium changes after ion implantation and thus influences protein adsorption of surfaces. In vitro studies disclose that HGFs on Mg ion-implanted samples exhibit better adhesion and migration while cells on Zn ion-implanted samples have higher proliferation rate and amounts. The results of immunofluorescence staining and real-time reverse-transcriptase polymerase chain reaction (RT-PCR) suggest that Mg mainly regulates the motility and adhesion of HGFs through activating the MAPK signal pathway whereas Zn influences HGFs proliferation by triggering the TGF-β signal pathway. The synergistic effect of Mg and Zn ions ensure that HGFs cultured on co-implanted samples possessed both high proliferation rate and motility, which are critical to soft tissue sealing of implants.