• The Korean Journal of Zoology
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• v.32 no.4
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• pp.412-419
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• 1989

Monoclonal antibodies (MAbs) reacting with the plasmalemma of Amoeba proteus were produced. Specificity of the 3 MAbs was determined by transfer blotting of the SDS polvacryfamide gel. AMS antibody reacted with the mucopolysaccharide bands of the spacer gel, 220 KD and 50 KD proteins of the resolving gel. The maior glycoprotein bands (175 KD, 165 KD) and 50 KD protein of the plasmalemma were recognized by AUG antibody. A third, AMP antibody reacted with the 50 KD protein only. In immunofluorescence microscopy of the enzyme treated cells, the antigens of these MAbs were sensitive to proteases, but not sensitive to neuraminidase. In the assay of cell to substratum attachment after binding with the antibody, AMG and AMP antibodies exerted no effect, but AMS hindered the attachment and cell spreading. Thus the effective components of the plasmalemma in cell to substratum attachment appear to be the mucopolysaccharides and 220 KD protein. The membranes of latex particle infested phagosomes did not show any distinction from the plasmalemma in fluorescence microscopy. Phagosome membranes of amoebae appear to be derived from the plasma membrane without selection in terms of the antigen composition. Amoeba Proteus의 세포막과 반응하는 단세포군 항체를 생산하였다. SDS polyacrylamide gel을 transfer blotting하여 이들 항체의 반응 특이성을 조사해 본 결과 AMS 단항체는 PAS로 염색되는 spacer gel의 mucopolysaccharide 린드, resolving gel의 220 KD 및 50 KD 단백질과 반응하였으며, 세포막의 주요 당단백질인 175 KD 및 165 KD 빈드와 50 KD 단백질은 AMG 단항체에 의해서 인지되었다. 그리고 AMP단항체는 공통인 50 KD 단백질과 특이하게 반응하였다. 효소처리한 아메바의 면역형광칠미경적 조사에서 이들 항체에 대한 항원분자들은 모두 단백질분해효소에 민감하였으며 neuraminidase에 대해서는 변화가 없었다. 이들 항체를 결합시킨 아메바의 용기표면 부착 가능성을 분석한 결과 AMP 및 AMG 단항체는 아무런 영향을 미치지 못하였으며 AMS 단항체는 세포의 용기표면 부착 및 세포의 펴짐을 저해하였다. 따라서 아메바의 용기표면 부착은 mucopolysaccharide 및 220 KD 단백질에 의해서 매게되는 것으로 나타났다. 그리고 latex particle을 담고 있는 식포막은 면역형광형미경적 조사에서 세포막과 차이가 없었다. 따라서 겐포막은 항원 조성에 있어서 비 선택적으로 세포막에서 유도되는 것으로 나타났다.

• Proceedings of the Materials Research Society of Korea Conference
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• 2009.11a
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• pp.41.2-41.2
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• 2009

Electrospinning of Polyvinylalcohol (PVA), Gelatin (GE), and PVA/GE blend solutions in acetic acid were investigated to fabricate biodegradable for tissue engineering. The morphology of the electrospun nanofibers was investigated with a field emission scanning electron microscope. The fibers have average diameters in the range 50-150 nm. The miscibility of PVA/GE blend fibers was examined by differential scanning calorimetry.The PVA and GE were immiscible in the as-spun nanofibrous structure. X-ray diffraction (XRD) determined the crystallinity of the membrane and tensile strength for evaluation physical properties. An in vitro study of PVA/GE blend nanofibers was conducted. To assay the cytocompatibility and cell behavior on the PVA/GE blend nanofibrous scaffolds, cell attachment and spreading of fibroblasts seeded on the scaffolds were studied. Our results indicate that thePVA/GE blend nanofibrous matrix, particularly the one that contained 20% PVA and 80% GE could be a good candidate for tissue engineering scaffolds, because it has an excellent cell attachment and spreading for fibroblast cell.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.34 no.6
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• pp.635-639
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• 2008

Objectives : the effect of different sterilization methods on the surface morphology of PPDO-hybrid-PLGA nanofiber scaffold and attachments of PC12 cell were investigated. Methods : Poly (p-dioxone)-hybrid-Poly (lactide-glycolide) (PPDO-hybrid-PLGA) nanofiber scaffold, fabricated in a tube form with 1.5 mm internal diameter, 0.2 mm thickness and 5 mm length, was prepared using electrospinning method. To study the surface morphology using SEM, The study group and control group in respective were; Control:Non-sterilized scaffold, Group I:scaffold sterilized with 70% Alcohol, Group II: scaffold sterilized with Ethylene Oxide at $65^{\circ}C$, and Group III: scaffold sterilized with Ethylene Oxide at $37^{\circ}C$. To investigate viability of the PC12 cell on the scaffold, The study group and control group in respective were; Control: sterilized with 70% Alcohol, Group I: sterilized with Ethylene Oxide at $65^{\circ}C$, and Group II: sterilized with Ethylene Oxide at $37^{\circ}C$. Results : 1. The surface morphology was slightly changed in Group I, II and Group III, compared with control. 2. The attachment of PC12 cells in Group I, II was not higher than in control Discussion : The attachment of PC12 cell is not influenced by different sterilization methods.

• Restorative Dentistry and Endodontics
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• v.40 no.4
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• pp.290-298
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• 2015

Objectives: Sodium hypochlorite (NaOCl) is an excellent bactericidal agent, but it is detrimental to stem cell survival, whereas intracanal medicaments such as calcium hydroxide ($Ca[OH]_2$) promote the survival and proliferation of stem cells. This study evaluated the effect of sequential NaOCl and $Ca(OH)_2$ application on the attachment and differentiation of dental pulp stem cells (DPSCs). Materials and Methods: DPSCs were obtained from human third molars. All dentin specimens were treated with 5.25% NaOCl for 30 min. DPSCs were seeded on the dentin specimens and processed with additional 1 mg/mL $Ca(OH)_2$, 17% ethylenediaminetetraacetic acid (EDTA) treatment, file instrumentation, or a combination of these methods. After 7 day of culture, we examined DPSC morphology using scanning electron microscopy and determined the cell survival rate with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. We measured cell adhesion gene expression levels after 4 day of culture and odontogenic differentiation gene expression levels after 4 wk using quantitative real-time polymerase chain reaction. Results: DPSCs did not attach to the dentin in the NaOCl-treated group. The gene expression levels of fibronectin-1 and secreted phosphoprotein-1 gene in both the $Ca(OH)_2$- and the EDTA-treated groups were significantly higher than those in the other groups. All $Ca(OH)_2$-treated groups showed higher expression levels of dentin matrix protein-1 than that of the control. The dentin sialophosphoprotein level was significantly higher in the groups treated with both $Ca(OH)_2$ and EDTA. Conclusions: The application of $Ca(OH)_2$ and additional treatment such as EDTA or instrumentation promoted the attachment and differentiation of DPSCs after NaOCl treatment.

• Journal of Periodontal and Implant Science
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• v.33 no.2
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• pp.193-213
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• 2003

The purpose of this study is to evaluate the phenomenon of attachment and spreading of the cultured rat　calvarial cell inoculated on their surface of different kinds of biodegradable membrane which had been used on tissue regeneration on periodontal defects by using scanning electron microscope. In this experiment 30 Sprague-Dawley male rats (mean BW 150gm) were used to harvest abundant number of cell in the short period. The rats were sacrificed by decapitatioan to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Biodegradable barrier membrane were collected with collagen type, and were divided into 3 different kind of surface such as scattered, polarized and fine-net type as their surface texture. Microcover plate which usually used for cell culture was used as control for smooth surface. All the membrane were seeded with cultured calvarial cell on their surface. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. After the culture as designed time, all the membrane were washed with 0.1 M Phosphate Buffered saline and fuxed with 2.5% Glutaraldehyde. And all specimen were treated with $OsO_4$, and Tannic acid before drying the cell for coating the cell with gold. Scanning Electron Microscope was used to observation. The following results were obtained. I. During the whole period of experiment, the phenomenon of cell attachment and spreading were revealed similar pattern to compare with smooth surface culture plate and ordinary culture dish. 2. The shape of cell attachment and spreading on the surface of barrier membrane were observed no remarked difference pattern between smooth surface culture plate and ordinary culture dish. 3. The cytoplasmic process of cultured calvaria cell extent to the deep portion of barrier membrane like as their own proper shape. 4. There were no remarkable relationships between the degree of cultured cell spreading and surface structure of barrier membrane. 5. Slight starified layer of cultured calvaria cell were observed on the scattered type of resorbable membrane, Conclusively, this study thus suggest that cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of carrier for many cell which could be used as new tissue regeneration, and those tissue engeering technique may become an new method in the approach to the repair of bone defects.

• KSBB Journal
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• v.4 no.2
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• pp.94-98
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• 1989

In vitro attachment experiments of bacteria to surface of host plant cell were carried out using C14 labeled cells of A. rhizogenes strain A4 and carrot protoplasts isolated from suspension culture of cells. Protoplasts were cocultivated with A. rhizogenes at various times after their isolation. Attachment kinetics showed that adherence of bacteria to protoplasts attained a maximum level within 120mins of co-cultivation. Maximum attachment occured at pH 6.0 and 24-35$^{\circ}C$. Bacterial attachment was observed at botg carrot cells with and without primary cell wall. The inhibition of transformation on the carrot root discs by A. rhizogenes was observed when non-related strain and heat inactivated bacterial strain cells were pretreated.

• KSBB Journal
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• v.4 no.1
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• pp.18-20
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• 1989

Solid gelatin microcarrier with the size distribution between $100{\mu}{\textrm}{m}$ and $400{\mu}{\textrm}{m}$ was prepared for the attachment and growth experiment for anchorage-dependent animal cell lines, i.e., L 929 and BHK 21. The growth and the maximum cell densities on this gelatin based and polyacrylamide (PAA) microcarriers were compared with those on the commercial dextran based Cytodex 3 microcarrier. Both cell lines showed good comparable attachment and growth patterns on the above three microcarriers. The mouse fibroblast, L 929 showed about the same maximum cell density on PAA, gelatin and Cytodex 3 MC'S BUT BHK 21, the baby hamster kidney cell line, showed the best result on Cytodex 3, which was about $4\times10^6$ cells/ml with the microcarrier concentration of 10 g/1.

• Biomaterials and Biomechanics in Bioengineering
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• v.3 no.1
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• pp.59-69
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• 2016

A surface resisting protein adsorption and cell adhesion is highly desirable for many biomedical applications such as diagnostic devices, biosensors and blood-contacting devices. In this study, a surface conjugated with sulfobetaine molecules was fabricated via the click reaction for the anti-fouling purpose. An alkyne-containing substrate (Alkyne-PPX) was generated by chemical vapor deposition of 4-ethynyl-[2,2]paracyclophane. Azide-ended mono-sulfobetaine molecules were synthesized and then conjugated on Alkyne-PPX via the click reaction. The protein adsorption from 10% serum was reduced by 57%, while the attachment of L929 cells was reduced by 83% onto the sulfobetaine-PPX surface compared to the protein adsorption and cell adhesion on Alkyne-PPX. In conclusion, we demonstrate that conjugation of mono-sulfobetaine molecules via the click chemistry is an effective way for reduction of non-specific protein adsorption and cell attachment.

• Journal of Periodontal and Implant Science
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• v.25 no.3
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• pp.539-556
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• 1995

Periodontal therapeutic modalities should be re-establishing and regenerating the periodontal tissue previously lost to the disease. To achieve periodontal regeneration, periodontal ligament cells must selective migrate to the deneded root surface, attached and proliferated it. Local pH concentration is one of the most factors that periodontal regeneration. The aims of this study were to examine on biological effects of pH to the human periodontal ligament cells in vitro, especially on the cell morphology, attachment, activity, vitality and viability. Human periodontal ligament cells were cultured from extracted tooth for non-periodontal reason. Immediately after extraction, any soft tissue adhering to the cervical parts of the roots was carefully removed with a sterile curette. To produce different pH levels in the media, Eagle's MEM was adjusted from pH 6.6 to 8.2 in 0.2 intervals with 1 M NaOH and 1 N HCl. After cultivation, Then, Periodontal ligament cells were cultured at pH ranging from 6.6-8.2. attachment assay was done at 1, 2 day incubation and activity assay was done at 1, 2, 3 day incubation. The experiments were evaluated by scaning electron microscopic techniques (HITACHIX-650 Scaning Electron Microanalyzer, Tokyo, Japan), MTT assay, and the cultured periodontal ligament cells were fixed in neutral formalin for 24 hours and immunohistochemically processed by PCNA for proliferating ability. The surviving cells in the medium showed slightly increased volume and widening intercellular distances at low concentration of pH than control group (pH 7.4), and apparently shrinkage at high concentration of pH than control group (pH 7.4). The results of the statistical analysis from the experiment on attachment, vitality and viability were as follows. Attachment of periodontal ligament cells at 1st and 2nd day, similar attachment rate of low concentration pH compared with control value (pH 7.4). But above pH 8.0, attachment rate were statistically significant decrease from control value(P<0.05). Periodontal ligament cell's activities were maximum at pH 7.6 by MTT assay. Similar with control value at low concentration of pH. But, the activities were statistically significant decrease at high concentraration of pH(P<0.05). Cellular proliferating rate (PCNA index) were statistically significant decrease from control value at low and high concentration of pH(p<0.05). This results suggested that hjgh concentration pH, in other words, alkali pH was cytotoxic effects on human periodontal ligament cells in vitro.

• The Journal of Advanced Prosthodontics
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• v.7 no.4
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• pp.278-287
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• 2015

PURPOSE. The aim of this study was to compare the color stability, water sorption and cytotoxicity of thermoplastic acrylic resin for the non-metal clasp dentures to those of thermoplastic polyamide and conventional heat-polymerized denture base resins. MATERIALS AND METHODS. Three types of denture base resin, which are conventional heat-polymerized acrylic resin (Paladent 20), thermoplastic polyamide resin (Bio Tone), thermoplastic acrylic resin (Acrytone) were used as materials for this study. One hundred five specimens were fabricated. For the color stability test, specimens were immersed in the coffee and green tee for 1 and 8 weeks. Color change was measured by spectrometer. Water sorption was tested after 1 and 8 weeks immersion in the water. For the test of cytotoxicity, cell viability assay was measured and cell attachment was analyzed by FE-SEM. RESULTS. All types of denture base resin showed color changes after 1 and 8 weeks immersion. However, there was no significant difference between denture base resins. All specimens showed significant color changes in the coffee than green tee. In water sorption test, thermoplastic acrylic resin showed lower values than conventional heat-polymerized acrylic resin and thermoplastic polyamide resin. Three types of denture base showed low cytotoxicity in cell viability assay. Thermoplastic acrylic resin showed the similar cell attachment but more stable attachment than conventional heat-polymerized acrylic resin. CONCLUSION. Thermoplastic acrylic resin for the non-metal clasp denture showed acceptable color stability, water sorption and cytotoxicity. To verify the long stability in the mouth, additional in vitro studies are needed.