• The Journal of Korean Academy of Prosthodontics
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• v.55 no.4
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• pp.361-371
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• 2017

Purpose: We aimed to investigate the gene expression of human gingival fibroblasts on microgroove surface using DNA microarray. Materials and methods: Microgrooves were applied on grade II titanium discs to have 0/$0{\mu}m$ (NE0, control group), 60/$10{\mu}m$ (E60/10, experimental group) of respective width/depth by photolithography. The entire surface of the microgrooved Ti substrata was further acid etched and used as the two experimental groups in this study. Human gingival fibroblasts were cultured in the experimental group and the control group, and total RNA was extracted. The oligonucleotide microarray was performed to confirm the changes of various gene expression levels between experimental group and control group. Changes of gene expression level were determined at the pathway level by mapping the expression results of DNA chips, using the KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis. Results: Gene expression levels on E60/10 and NE0 were analyzed, there were 123 genes showing significant differences in expression more than 1.5 times on E60/10 microgrooved surface compared to NE0 surface, and 19 genes showing significant differences in expression more than 2 times. The KEGG pathway analysis confirmed the changes in gene expression levels under experimental conditions. Cell signaling, proliferation, and activity among the various gene expression results were identified. Conclusion: Microgrooved surfaces induce gene expression changes and related cell signaling. According to the results of this study, microgrooves can be used as the surface of various biomaterials which need to improve cell activity through gene expression changes and activation of cell signaling.

• Journal of Periodontal and Implant Science
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• v.28 no.4
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• pp.703-721
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• 1998

In order to observe the effects of Nicotine and NNK on cultured human gingival fibroblast, several factors were examined including mutagenicity, the number of cells attached culture plate surface through MTT test, the abundance of collagen & collagenase in mRNA level and collagenolytic activity in extracellular matrix. The results were as follows; 1. Regardless of the co-existence of S9, Nicotine did not show the mutagenicity by itself and NNK by itself showd the same result; However, dose related mutagenicity was shown in NNK with S9. 2. The number of fibroblasts attached cultured plate surface was measured by MTT procedure. The number of cells in Non-smokers increased at all time periods as compared to those of smoker. 3. Non-smoker's fibroblast treated by NNK or Nicotine was dosedependently dosedependently decreased in the number of cells when compared to untreated control. In higher dose, Nicotine showed the cellular toxicity, but NNK did not. 4. No change in the abundance of mRNA for pro${\alpha}1$ and pro${\alpha}2$ was shown in Nicotine treated group but in gingival fibroblasts following treatment with NNK, the abundance of mRNA for pro${\alpha}1$, but not pro${\alpha}2$ collagen was decreased. 5. The abundance of mRNA for collagenase was decreased when NNK was treated but no change occurred in Nicotine treated group. 6. The effect of NNK and Nicotine in collagenolytic activity showed that, collagenase activity exclusively react to type I collagen, was increased in both group, but gelatinase exclusively react to type IV collagen was not influenced at all. Collagenase activity of smoker's fibroblast was also increased as much as Nicotine and NNK group. The findings suggest that both of Nicotine and NNK lead gingival fibroblast to decrease in the abundance of collagen. And it seems to be that Nicotine and NNK have independent pathway toward the gingival fibroblast.

• The Journal of Advanced Prosthodontics
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• v.7 no.2
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• pp.138-145
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• 2015

PURPOSE. The objective of this study was to conduct an in vitro comparative evaluation of polished and laser-dimpled titanium (Ti) surfaces to determine whether either surface has an advantage in promoting the attachment of epithelial-like cells and fibroblast to Ti. MATERIALS AND METHODS. Forty-eight coin-shaped samples of commercially pure, grade 4 Ti plates were used in this study. These discs were cleaned to a surface roughness (Ra: roughness centerline average) of 180 nm by polishing and were divided into three groups: SM (n=16) had no dimples and served as the control, SM15 (n=16) had $5-{\mu}m$ dimples at $10-{\mu}m$ intervals, and SM30 (n=16) had $5-{\mu}m$ dimples at $25-{\mu}m$ intervals in a $2{\times}4mm^2$ area at the center of the disc. Human gingival squamous cell carcinoma cells (YD-38) and human lung fibroblasts (MRC-5) were cultured and used in cell proliferation assays, adhesion assays, immunofluorescent staining of adhesion proteins, and morphological analysis by SEM. The data were analyzed statistically to determine the significance of differences. RESULTS. The adhesion strength of epithelial cells was higher on Ti surfaces with $5-{\mu}m$ laser dimples than on polished Ti surfaces, while the adhesion of fibroblasts was not significantly changed by laser treatment of implant surfaces. However, epithelial cells and fibroblasts around the laser dimples appeared larger and showed increased expression of adhesion proteins. CONCLUSION. These findings demonstrate that laser dimpling may contribute to improving the peri-implant soft tissue barrier. This study provided helpful information for developing the transmucosal surface of the abutment.

• Journal of Periodontal and Implant Science
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• v.26 no.1
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• pp.89-102
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• 1996

The goal of periodontal therapy is to regenerate the loss of periodontal attachment appratus. Current theories suggest the cells of the periodontium have the capacity, when appropriately triggered, to actively participate in restoring connective tissues, including mineralized tissues. This study was performed to define the hard tissue regeneration effect of periodontal ligament(PDL) cells in vitro and the effect of rate of the composition in gingival fibroblasts(GF) on the hard tissue regeneration capacity of PDL cells. For this study, Cell growth rate, alkaline phosphatase(Al.Pase) levels and the ability to produce mineralized nodules in co-culture of PDL cells and GF were examined. The results were as follows : 1. At 7 and 15 days, Cell growth of co-culture of PDL and GF(50 : 50) was greater than that of PDL cells or GF alone(P>0.05). 2. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(p<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P>0.05) 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group 1(PDL 100%), 2(PDL 70% : GF 30%), and 3(PDL 50% : GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30% GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05) From the above results, it is assumed that the co-culture of PDL cells and GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density.

• Journal of Periodontal and Implant Science
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• v.27 no.1
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• pp.129-150
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• 1997

The purpose of this study was to evaluate the effects of tetracycline(TC}, flurbiprofen, and PDGF-BB loaded biodegradable membranes on the cell-attachment, the activity of loaded PDGF-BB, in vivo release kinetics, and guided bone regenerative potentials. To evaluate the cell attachment to membranes, the number of gingival fibroblasts attached to each membrane(10% TC, 10% flurbiprofen, $200ng/cm^2$ PDGF-BB loaded membranes, drug-unloaded membrane) was counted by coulter counter and the morphologic pattern of attached cells was examined under SEM. To determine whether the activity of loaded PDGF-BB is sustained, the cellular growth and survival rate of gingival fibroblasts was used for both standard PDGF-BB and loaded PDGF-BB. For evaluation of in vivo release kinetics, drug-loaded membranes were implanted on the dorsal skin of the rats. On 1, 3, 7, 10, 14, 21, and 28 days after implantation, the amount of remaining drugs were measured by HPLC assay for TC and flurbiprofen, and by ${\gamma}-scintillation$ counter for $PDGF-BB^{1125}$. For evaluation of guided regenerative potential, the amount of new bone in the calvarial defect(5mm in diameter) of the rat was measured by histomorphometry 1 and 2 weeks after implantation of membranes. The number of cells attached to the PDGF-BB loaded membrane was largest as compared with the other mernbranes.(p< 0.05) The activity of loaded PDGF-BB was not significantly different from the activity of standard PDGF-BB.(p<0.05) After initial burst release of drug during the first 24 hours, drugs were gradually released for 4 weeks. Especially the release rate of PDGF-BB was nearly constant during 4 weeks. PDGF-BB loaded membranes(200, $400ng/cm^2$) were effective in guided bone regeneration as compared with drug-unloaded membrane. These results implicate that drug-loaded biodegradable membranes might be a useful for guided bone regeneration.

• Journal of Periodontal and Implant Science
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• v.45 no.3
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• pp.120-126
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• 2015

Purpose: With the significance of stable adhesion of alveolar bone and peri-implant soft tissue on the surface of titanium for successful dental implantation procedure, the purpose of this study was to apply microgrooves on the titanium surface and investigate their effects on peri-implant cells and tissues. Methods: Three types of commercially pure titanium discs were prepared; machined-surface discs (A), sandblasted, large-grit, acid-etched (SLA)-treated discs (B), SLA and microgroove-formed discs (C). After surface topography of the discs was examined by confocal laser scanning electron microscopy, water contact angle and surface energy were measured. Human gingival fibroblasts (hGFs) and murine osteoblastic cells (MC3T3-E1) were seeded onto the titanium discs for immunofluorescence assay of adhesion proteins. Commercially pure titanium implants with microgrooves on the coronal microthreads design were inserted into the edentulous mandible of beagle dogs. After 2 weeks and 6 weeks of implant insertion, the animal subjects were euthanized to confirm peri-implant tissue healing pattern in histologic specimens. Results: Group C presented the lowest water contact angle ($62.89{\pm}5.66{\theta}$), highest surface energy ($45{\pm}1.2mN/m$), and highest surface roughness ($Ra=22.351{\pm}2.766{\mu}m$). The expression of adhesion molecules of hGFs and MC3T30E1 cells was prominent in group C. Titanium implants with microgrooves on the coronal portion showed firm adhesion to peri-implant soft tissue. Conclusions: Microgrooves on the titanium surface promoted the adhesion of gingival fibroblasts and osteoblastic cells, as well as favorable peri-implant soft tissue sealing.

• Restorative Dentistry and Endodontics
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• v.29 no.4
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• pp.370-377
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• 2004

The purpose of this study was to regenerate human dental pulp tissues similar to native pulp tissues. Using the mixture of type I collagen solution, primary cells collected from the different tissues (pulp, gingiva, and skin) and NIH 3T3 ($1{\;}{\times}{\;}10^5{\;}cells/ml/well$) were cultured at 12-well plate at $37^{\circ}C$ for 14 days. Standardized photographs were taken with digital camera during 14 days and the diameter of the contracted collagen gel matrix was measured and statistically analyzed with student t-test. As one of the pulp tissue engineering, normal human dental pulp tissue and collagen gel matrix cultured with dental pulp cells for 14 days were fixed and stained with Hematoxyline & Eosin. According to this study, the results were as follows: 1. The contraction of collagen gel matrix cultured with pulp cells for 14 days was significantly higher than other fibroblasts (gingiva, skin) (p < 0.05), 2. The diameter of collagen gel matrix cultured with pulp cells was reduced to 70.4% after 7 days, and 57.1% after 14 days. 3. The collagen gel without any cells did not contract, whereas the collagen gel cultured with gingiva and skin showed mild contraction after 14 days (88.1% and 87.6% respectively). 4. The contraction of the collagen gel cultured with NIH 3T3 cells after 14 days was higher than those cultured with gingival and skin fibroblasts, but it was not statistically significant (72.1%, p > 0.05). 5. The collagen gel matrix cultured with pulp cells for 14 days showed similar shape with native pulp tissue without blood vessels. This approach may provide a means of engineering a variety of other oral tissue as well and these cell behaviors may provide information needed to establish pulp tissue engineering protocols.

• Journal of Korean society of Dental Hygiene
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• v.19 no.2
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• pp.173-181
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• 2019

Objectives: The resin infiltration technique is a promising alternative therapy for arresting the early dental caries. However, there are very few reports on the safety and biocompatibility of this technique. We evaluated various properties of resin infiltrant (RI) based on a triethylene glycol dimethacrylate (TEGDMA).The water sorption (Wsp) and water solubility (Wsl) was assessed. Additionally, the cytotoxicity of RI against both animal and human fibroblast cell lines was investigated. Methods: The RI of the $Icon^{(R)}$, the first product developed for resin infiltration, is mainly composed of TEGDMA in the resin matrix. The Wsp and Wsl for the RI were measured in accordance with ISO 4049 specifications. Fourier-transform infrared spectroscopy (FTIR) was used for analyzing the polymerization before and after curing of RI. The cytotoxicity of RI against the mouse fibroblasts (L929) and human gingival fibroblasts (hTERT-hNOF) was evaluated using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and the data were analyzed using one-way analysis of variance. Results: Wsp and Wsl of the RI specimens were $53.37{\mu}g/mm^3$ and $10.6{\mu}g/mm^3$, respectively. FTIR analysis revealed a slightly higher degree of curing with longer irradiation time. The degree of conversion for RI was high (80.9%) after 40 seconds of light curing. There was a significant decrease in the viability of L929 and hTERT-hNOF cells at RI extraction solution concentrations above 50%, respectively, compared to that in the negative control (p< 0.05). Conclusions: Even though the RI exhibited positive effect on the early prevention of dental caries, the clinicians should also consider the toxicity of RI on periodontal tissues.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.4
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• pp.359-366
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• 2007

Purpose: In spite of various advantages of hyaluronic acid in wound healing, there are few research about wound healing process and period in oral and maxillofacial surgery. So, We evaluated the effects of local application of hyaluronic acid gel on wound healing of the oral mucosa using this animal model. Materials and methods: Young adult New Zealand White rabbits, weighting between 2.5 and 3.0kg, were used. Almost uniform round ulcers could be created on the gingival of the rabbits by chemical injury with acetic acid. In experimental group, hyaluronic acid gel was applied daily until the wounds healed and in control group, ulcer lesion was not any treatment. The lengths of ulcers were measured the longest and shortest axes of the ulcers and calculated the areas of ulcer. For histological examination, specimens were made, and observed under a light microscope. Results: The results showed that the number of fibroblasts, new blood vessels and the epithelial thickness from experimental group was higher than from control group. Hyaluronic acid promoted proliferation of the fibroblast, keratinocytes isolated from gingival tissue of rabbits in vitro. Topical application of hyaluronic acid accelerated healing of ulcers created in rabbits. Conclusion: The hyaluronic acid may be effective for wound healing of oral mucosal lesions.

• Journal of Periodontal and Implant Science
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• v.27 no.1
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• pp.151-163
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• 1997

The purpose of this study was to evaluate on the biodegradability, biocompatibility and tissue regenerative capacity of synthetic biodegradable $mernbranes-Resolut^{(R)}$, $Guidor^{(R)}$ and $Biomesh^{(R)}$. To evaluate the cell attachment on the membranes, in vitro, the number of gingival fibroblasts attached to each membrane was counted by hemocytometer. Cytotoxicity test for the membranes was performed by MTT test with gingival fibroblast For evaluation of guided- bone regenerative potential, the amount of new bone formation in the rat calvarial defects(5mm in diameter) beneath the membranes was observed for two weeks and examined of the specimens by Massons trichrome staining. Biodegradability was observed for 2, 4, 8 and 12 weeks after implantation of each materials under the skin of rats and examined the specimens with H & E staining. The number of cell attachment were the greatest in $Biomesh^{(R)}$ and followed by $Resolut^{(R)}$. Cell viability of three membranes was almost similar levels. Biodegradability of $Resolut^{(R)}$ was the highest among three membrane and the potential of guided bone regeneration was the greatest in the $Biomesh^{(R)}$ and $Resolut^{(R)}$ was followed. These results suggested that commercially available biodegradable membranes were non-toxic and highly potential to guided bone regeneration.