• Journal of Periodontal and Implant Science
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• 제31권1호
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• pp.163-180
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• 2001

The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of periodontal ligament cells. Primary human periodontal ligament cells were cultured in dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells of 4th to 7th passage were inoculated in the multiwell plates coated with chitosan in concentration of 0.22, 0.2, and $2mg/m{\ell}$. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized modules was evaluated after 21 days of culture. The results were as follows : 1. The morphology of periodontal ligament cells on the chitosan coating was round or spheric. Round cells were aggregated after 6 hours of culture. Aggregated cells on the chitosan coated surface showed nodule-like appearance after 24 hours of culture and not achieved confluency at 7 days. 2. During early period of culture, the attachment of periodontal ligament cells were inhibited by chitosan coating. Inhibition of cell attachment tended to increase with the concentration of chitosan. 3. At the chitosan concentration of 0.02 and $0.2mg/m{\ell}$, periodontal ligament cells were more rapidly proliferated at 7 days, compared to the control group. At the concentration of $2mg/m{\ell}$, the proliferation of periodontal ligament cells was inhibitied(p<0.01). 4. Alkaline phosphatase activity of periodontal ligament cells was increased in chitosan coated group, especially at the concentration of $0.02mg/m{\ell}$after 4 days of culture.5. Periodontal ligament cells produced mineralized nodules on chitosan coated wells without the addition of mineralized nodule forming materials (ascorbic acid, ${\beta}-glycerophosphat$, dexamethasone). With the addition of mineralized nodule forming materials, periodontal ligament cells produced more mineralized nodules at the concentration of $0.02mg/m{\ell}$, compared to the control. In summary, the attachment, proliferation, cell activity, and alkaline phosphatase activity of periodontal ligament cells depended on the concentration of coated chitosan. Chitosan stimulated mineralized nodule formation by periodontal ligament cells. At the appropriate concentration($0.02mg/m{\ell}$), chitosan could increase alkaline phosphatase activity and stimulate the formation of mineralized nodule by periodontal ligament cells. These results suggest that chitosan can be used as an adjunct for bone graft material, and the matrix of tissue engineering for periodontal regeneration, especially bone regeneration.

• The Journal of Advanced Prosthodontics
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• 제6권6호
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• pp.512-520
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• 2014

PURPOSE. The purpose of this study was to assess the surface characteristics and the biocompatibility of zirconium (Zr) coating on Ti-6Al-4V alloy surface by radio frequency (RF) magnetron sputtering method. MATERIALS AND METHODS. The zirconium films were developed on Ti-6Al-4V discs using RF magnetron sputtering method. Surface profile, surface composition, surface roughness and surface energy were evaluated. Electrochemical test was performed to evaluate the corrosion behavior. Cell proliferation, alkaline phosphatase (ALP) activity and gene expression of mineralized matrix markers were measured. RESULTS. SEM and EDS analysis showed that zirconium deposition was performed successfully on Ti-6Al-4V alloy substrate. Ti-6Al-4V group and Zr-coating group showed no significant difference in surface roughness (P>.05). Surface energy was significantly higher in Zr-coating group than in Ti-6Al-4V group (P<.05). No difference in cell morphology was observed between Ti-6Al-4V group and Zr-coating group. Cell proliferation was higher in Zr-coating group than Ti-6Al-4V group at 1, 3 and 5 days (P<.05). Zr-coating group showed higher ALP activity level than Ti-6Al-4V group (P<.05). The mRNA expressions of bone sialoprotein (BSP) and osteocalcin (OCN) on Zr-coating group increased approximately 1.2-fold and 2.1-fold respectively, compared to that of Ti-6Al-4V group. CONCLUSION. These results suggest that zirconium coating on Ti-6Al-4V alloy could enhance the early osteoblast responses. This property could make non-toxic metal coatings on Ti-6Al-4V alloy suitable for orthopedic and dental implants.

• The Journal of Advanced Prosthodontics
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• 제6권3호
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• pp.157-164
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• 2014

PURPOSE. This study focused on in vitro cell differentiation and surface characteristics in a magnesium coated titanium surface implanted on using a plasma ion source. MATERIALS AND METHODS. 40 commercially made pure titanium discs were prepared to produce Ti oxide machined surface (M) and Mg-incorporated Ti oxide machined surface (MM). Surface properties were analyzed using a scanning electron microscopy (SEM). On each surface, alkaline phosphatase (ALP) activity, alizarin red S staining for mineralization of MC3T3-E1 cells, and quantitative analysis of osteoblastic gene expression, were evaluated. Actin ring formation assay and gene expression analysis of TRAP and GAPDH performing RT-PCR were performed to characterize osteoclast differentiation on mouse bone marrow-derived macrophages (BMMs). RESULTS. MM showed similar surface morphology and surface roughness with M, but was slightly smoother after ion implantation at the micron scale. M was more hydrophobic than MM. No significant difference between surfaces on ALP activity at 7 and 14 days were observed. Real-time PCR analyses showed similar levels of mRNA expression of the osteoblast phenotype genes; osteopontin (OPN), osteocalcin (OCN), bone sialoprotein (BSP), and collagen 1 (Col 1) in cell grown on MM at 7, 14 and 21 days. Alizarin red S staining at 21 days showed no significant difference. BMMs differentiation increased in M and MM. Actin ring formation assay and gene expression analysis of TRAP showed osteoclast differentiation to be more active on MM. CONCLUSION. Both M and MM have a good effect on osteoblastic cell differentiation, but MM may speed the bone remodeling process by activating on osteoclast differentiation.

• The Journal of Advanced Prosthodontics
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• 제3권3호
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• pp.152-160
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• 2011

PURPOSE. This study was to investigate the effects of recombinant human platelet-derived growth factor (rhPDGF-BB) and heparin to titanium surfaces for enhancement of osteoblastic functions and inhibition of inflammation activity. MATERIALS AND METHODS. The anodized titanium discs, not coated with any material, were used as a control group. In heparinized-Ti group, dopamine was anchored to the surface of Ti substrates, and coated with heparin. In PDGF-Ti group, rhPDGF-BB was immobilized onto heparinized Ti surface. The surface morphologies were investigated by the scanning electron microscope in each group. The release kinetics of rhPDGF-BB were analyzed, and cytotoxicity tests for each group were conducted. The biocompatibilities were characterized by measuring cell proliferation, alkaline phosphatase activity, and calcium deposition using MG-63 cells. Statistical comparisons were carried out by one-way ANOVA tests. Differences were considered statistically significant at $^*$P<.05 and $^{**}$P<.001. RESULTS. The combination of rhPDGF-BB and heparin stimulated alkaline phosphatase activity and OCN mRNA expression in osteoblastic cells ($^*$P<.05 and $^{**}$P<.001). MG-63 cells grown on PDGF-Ti had significantly higher amounts of calcium deposition than those grown on anodized Ti ($^{**}$ P<.001). Heparinized Ti was more anti-inflammatory compared to anodized Ti, when exposed to lipopolysaccharide using the transcript levels of TNF-${\alpha}$ and IL-6 of proinflammatory cytokine ($^*$P<.05 and $^{**}$P<.001). CONCLUSION. The result of this study demonstrated that the incorporation of rhPDGF-BB and heparin onto Ti surface enhanced osteoblastic functions and inhibited inflammation.

• 공업화학
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• 제32권4호
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• pp.461-466
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• 2021

본 논문에서는 이동성이 좋고 경제적이며, 간편하게 일회용 진단칩으로 제작 가능한 스크린 프린팅 한 탄소칩 전극[screen printed carbon electrode (SPCE)] 기반의 전압전류법 나노물질 융합형 바이오센서를 제작하여 폐암 조기진단에 활용 가능한 단백질 표지 인자 중에 하나인 heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) 단백질의 농도를 정량 분석하고자 하였다. 먼저 SPCE 표면에 금 나노입자를 전기적으로 증착한 후 크로스링커를 이용하여 hnRNP A1에 특이적으로 결합할 수 있는 바이오리셉터인 DNA 압타머를 고정하였다. Ethanolamine을 블로킹 시약으로 사용하여 압타머와 함께 센서 표면에 고정하여 그 표면을 처리함으로써 비특이적인 생물질의 흡착에 의한 방해 신호를 최소화하고자 하였다. DNA칩과 hnRNP A1 용액을 접촉하여 DNA와 hnRNP A1을 결합시킨 후 alkaline phosphatase (ALP) 효소로 접합한 hnRNP A1 항체(anti-hnRNP A1)을 센서칩 표면으로 주입하여 샌드위치 복합체를 형성하고, 이를 기질인 4-aminophenyl phosphate (APP)와 효소-기질 특이적 산화 반응에 의한 전류 변화를 순환 전압전류법과 시차 펄스전압전류법으로 측정하여 단백질의 농도를 정량적으로 분석하였다. 상기 산화 반응에 의한 피크 전류 변화는 순환전압전류법과 시차 펄스 전압전류법을 사용할 때 -0.05와 -0.17 V (vs. Ag/AgCl) 전위 값에서 각각 일어났다. 개발한 나노바이오센서를 실제 정상인 혈청 시료 분석에 적용 가능함을 보여줌으로써 혈청 한 방울로 폐암의 조기진단 가능성을 제시하고자 하였다.

• Ecology and Resilient Infrastructure
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• 제5권2호
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• pp.82-87
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• 2018

Proliferation of algae on the surface of concrete or mortar in aquatic habitat has a negative impact on maintenance of concrete-based structures. Growth of algae may decrease stability of structure by bio-deterioration. In this study, we developed a functional mortar for restraining bio-deterioration by using $Cu^{2+}$ ion. The mortar contains soluble glass beads made of $Cu^{2+}$ ion, which can dissolve into water slowly. Mortars prepared with different ratio of glass beads (0, 2, 5, 10, and 15%) were placed in a culture medium with algae and incubated over a month period. Water chemistry, chlorophyll-a, and extracellular enzyme activities were measured. The incubation was conducted in both freshwater and seawater conditions, to assess applicability to both aquatic conditions. Overall, mortar with Cu glass exhibited lower chlorophyll-a content, suggesting that the functional mortar reduced algal growth. DOC concentration increased because debris of dead algae increased. Cu glass also decreased phosphatase activity, which is involved in the regeneration of inorganic P from organic moieties. Since, P is often a limiting nutrient for algal production, algal growth may be inhibited. Activities of ${\beta}$-glucosidase and N-acetylglucosaminidase were not significantly affected because carbon and nitrogen mineralization may not be influenced by the Cu glass beads. Our study suggests that functional mortar with Cu glass beads may reduce the growth of algae on the surface, while it has little environmental impact.

• 대한치과교정학회지
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• 제26권2호
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• pp.163-174
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• 1996

본 실험은 서로 다른 기전으로 골흡수를 억제한다고 추정되는 약제인 bisphosphonate와 indomethacin을 백서에 투여한 후 교정력을 이용한 치아이동시 약제가 골개조에 미치는 영향을 밝히고자 시행되었다. 동일한 조건에서 사육된 체중 260-300g의 웅성백서(Sprague-Dawley계)를 대조군, bisphosphonate 투여군 및 indomethacin 투여군으로 분류하고 각 군은 다시 장치를 장착한 실험측과 장치를 장착하지 않은 대조측으로 분류하였다. Bisphosphonate(6.3mg/kg,$2.52x10^{-2}mol/L$)와 indomethacin(9mg/kg, $2.52x10^{-2}mol/L$)은 교정장치 장착 6시간 전, 1시간 전 및 24시간 후에 복강내 주사하였으며, 교정력이 가해진 시점으로부터 72시간이 경과한 후 파골세포수를 측정하고 조직학적인 성상을 관찰하였다. 또한 혈액을 채취한 후 혈청 acid phosphatase 및 lactate dehydrogenase 양을 측정하여 다음과 같은 성적을 얻었다. 실험측의 파골세포수는 장치장착 1시간 전에 투여한 bisphosphonate군과 indomethacin군에서 가장 적게 나타났으며, 다른 시간의 약물 투여군은 대조군과 차이가 없었다. 대조측의 파골세포수는 실험측보다 현저히 적게 나타났으며, 대조군 및 약물투여군간에 차이가 없었다. 대조군과 6시간 전 및 24시간 후에 투여한 indomethacin군은 심한 골흡수 양상을 보인 반면, 1시간전 indomethacin군 및 모든 bisphosphonate군에서는 골흡수의 정도가 적었다. 파골세포의 주름변연과 투명대는 대조군 및 indomethacin군에서 가장 뚜렷하게 나타났으며, bisphosphonate 투여군에서는 일부 파골세포들이 무딘 세포돌기만을 내거나 골표면에 부착하지 않고 있는 경우가 많았다. Bisphosphonate와 indomethacin 투여군 모두에서 acid phosphatase 및 lactate dehydrogenase 값이 대조군보다 낮았으며, acid phosphatase 값은 bisphosphonate군이 indomethacin군보다 낮았으나 lactate dehydrogenase 값은 차이가 없었다. 이상의 결과로 bisphosphonate는 파골세포의 수 및 대사활동을 감소시키며 indomethacin은 파골세포의 수를 감소시킴을 알 수 있었다. 또한 두 약물을 비교하면 bisphosphonate는 indomethacin에 비해 골흡수 억제효과가 더 크며 투여시간에 따른 제약도 더 적은 것으로 사료된다.

• The Journal of Advanced Prosthodontics
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• 제6권2호
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• pp.96-102
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• 2014

PURPOSE. This study was performed to characterize the effects of zirconia coated with calcium phosphate and hydroxyapatite compared to smooth zirconia after bone marrow-derived osteoblast culture. MATERIALS AND METHODS. Bone marrow-derived osteoblasts were cultured on (1) smooth zirconia, (2) zirconia coated with calcium phosphate (CaP), and (3) zirconia coated with hydroxyapatite (HA). The tetrazolium-based colorimetric assay (MTT test) was used for cell proliferation evaluation. Scanning electron microscopy (SEM) and alkaline phosphatase (ALP) activity was measured to evaluate the cellular morphology and differentiation rate. X-ray photoelectron spectroscopy (XPS) was employed for the analysis of surface chemistry. The genetic expression of the osteoblasts and dissolution behavior of the coatings were observed. Assessment of the significance level of the differences between the groups was done with analysis of variance (ANOVA). RESULTS. From the MTT assay, no significant difference between smooth and surface coated zirconia was found (P>.05). From the SEM image, cells on all three groups of discs were sporadically triangular or spread out in shape with formation of filopodia. From the ALP activity assay, the optical density of osteoblasts on smooth zirconia discs was higher than that on surface treated zirconia discs (P>.05). Most of the genes related to cell adhesion showed similar expression level between smooth and surface treated zirconia. The dissolution rate was higher with CaP than HA coating. CONCLUSION. The attachment and growth behavior of bone-marrow-derived osteoblasts cultured on smooth surface coated zirconia showed comparable results. However, the HA coating showed more time-dependent stability compared to the CaP coating.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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• pp.275-275
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• 2013

Osteoblast is one of cells related with osseointegration and many research have conducted the adhesion of osteoblast onto the surface of implant. In the osseointegration, biocompatibility of the implant and cell adhesion to the surface are important factors. The researches related to cell adhesion have a direction from micro-scaled surface roughness to nano-scaled surface roughness with advancing nanotechnology. A cell reacts and sense to stimuli from extracellular matrix (ECM) and topography of the ECM [1]. Thus, for better osseointegration, we should provide an environment similar to ECM. In this study, we synthesize TiO2 nanowires using hydrothermal reaction because TiO2 provides inertness to titanium on its surface and enables it used as an implant material for the orthopedic treatment such as fixation of the bone fracture [2]. Ti substrate is immersed into NaOH aqueous solution. The solution are heated at $140{\sim}200^{\circ}C$ for various time (10~720 minutes). After heat treatment, we take out the sample and immerse it into HCl aqueous solution for 1 hour. The acid treated sample is heated again at $500^{\circ}C$ for 3 hours [3]. Then, we culture osteoblast on the TiO2 nanowires. For investigating cell adhesion onto nanostructured surface, we conduct several tests such as MTT assay, ALP (Alkaline phosphatase) activity assay, measuring calcium expression, and so on. These preliminary results of the cell culture on the nanowires are foundation for investigating cell-material interaction especially with nanostructure interaction.

• Journal of Industrial and Engineering Chemistry
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• 제67권
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• pp.244-254
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• 2018

Surface modified poly ${\text\tiny{L}}$-lactic acid (PLLA) samples with hydroxyapatite (HA), heparin and bone morphogenetic protein-2 (BMP-2) mediated by polydopamine (pDA) coating (PLLA/pDA/HA/Hep/BMP-2) were prepared, and their effects on the enhancements of bone formation and osseointegration were evaluated in vitro and in vivo as compared to PLLA, PLLA/pDA/HA, and PLLA/pDA/Hep/BMP-2. The changes in surface chemical compositions, morphologies and wettabilities were observed by X-ray photoelectron spectroscopy (XPS), field-emission scanning electron microscopy (FE-SEM), atomic force microscopy (AFM) and water contact angle measurements. Pre-coating of HA particles with pDA provided uniform and homogeneous anchoring of particles to PLLA surface. In addition, the strong ionic interaction between heparin and pDA led PLLA surface readily heparinized for loading of BMP-2. In vitro experiments revealed that the levels of alkaline phosphatase (ALP) activity, calcium deposition, and osteocalcin (OCN) gene expression were higher in MG-63 human osteosarcoma cell lines grown on PLLA/pDA/HA/Hep/BMP-2 than on control PLLA, PLLA/pDA/HA, and PLLA/pDA/Hep/BMP-2. In vivo studies using micro-computed tomography (micro-CT) also showed that PLLA/pDA/HA/Hep/BMP-2 screw exhibited greatest value of bone volume (BV) and bone volume/tissue volume (BV/TV) among samples. Histological evaluations with H&E and Von Kossa staining demonstrated that a combination of HA and BMP-2 contributed to the strong osseointegration.