Biodegradable barrier membrane has been demonstrated to have guided bone regeneration capacity on the animal study. The purpose of this study is to evaluate the effects of cultured calvarial cell inoculated on the biodegradable barrier membrane for the regeneration of the artificial bone defect. In this experiment 35 Sprague-Dawley male rats(mean BW 150gm) were used. 30 rats were divided into 3 groups. In group I, defects were covered periosteum without membrane. In group II, defects were repaired using biodegradable barrier membrane. In group III, the defects were repaired using biodegradable barrier membrane seeded with cultured calvarial cell. Every surgical procedure were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). After anesthesia, 5 rats were sacrificed by decapitation 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. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. The membrane were inserted on the artificial bone defect after 3 days of culture. A single 3-mm diameter full-thickness artificial calvarial defect was made in each animal by using with bone trephine drill. After the every surgical intervention of animal, all of the animals were sacrificed at 1, 2, 3 weeks after surgery by using of perfusion technique. For obtaining histological section, tissues were fixed in 2.5% Glutaraldehyde (0.1M cacodylate buffer, pH 7.2) and Karnovsky's fixative solution, and decalcified with 0.1M disodium ethylene diaminetetraacetate for 3 weeks. Tissue embeding was performed in paraffin and cut parallel to the surface of calvaria. Section in 7${\mu}m$ thickness of tissue was done and stained with Hematoxylin-Eosin. All the specimens were observed under the light microscopy. The following results were obtained. 1 . During the whole period of experiment, fibrous connective tissue was revealed at 1week after surgery which meant rapid soft tissue recovery. The healing rate of defected area into new bone formation of the test group was observed more rapid tendency than other two groups. 2 . The sequence of healing rate of bone defected area was as follows ; test group, positive control, negative control group. 3 . During the experiment, an osteoclastic cell around preexisted bone was not found. New bone formation was originated from the periphery of the remaing bone wall, and gradually extended into central portion of the bone defect. 4 . The biodegradable barrier membrane was observed favorable biocompatibility during this experimental period without any other noticeable foreign body reaction. And mineralization in the newly formed osteoid tissue revealed relatively more rapid than other group since early stage of the healing process. Conclusively, the cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of regeneration of artificial bone defects of alveolar bone. This study thus demonstrates a tissue-engineering the approach to the repair of bone defects, which may have clinical applications in clinical fields of the dentistry including periodontics.
Journal of Korean Society of Environmental Engineers
/
v.27
no.1
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pp.101-108
/
2005
Because the stone cultural properties located outdoors, they have been altered and deteriorated in external appearance due to environmental factors such as acid rain, extreme change in temperature, and salts. Damage to stone cultural properties is accelerated particularly due to recent industrial development and environmental pollution. An experimental study was conducted to evaluate the effect of environmental contaminants on the weathering of granite. And as part of the developing of conservation method, $TiO_2$ catalyst was prepared and tested. When fresh granite was dipped into the salt and acid solutions, dissolution rate of eight minerals (Si, Mg, Ca, Na, K, Fe, Mn, Al) are abruptly increased at initial stage of reaction and then increased steadily until 100 cycles. After salt and acid solution experiments, the mineral compositions of the granite surface were lower then that of the fresh granite and density of the weathered granite was steadily decreased from $2.60\;g/cm^3$ to $2.56\;g/cm^3$, but Poissions ratio and absorption ratio were slightly increased. It was expected at stone cultural assets could be weathered by salts and acid rain. In the case of $TiO_2$ was coated to the granite, the dissolution rate of minerals and absorption ratio of $TiO_2$ coated granite were decreased. Therefore, the $TiO_2$ coating method tested in this study considered to be a viable method to assist in the conservation of stone cultural properties from environmental contaminants.
The turnover of collagen is controlled by the balance between collagen synthesis and degradation. The production of collagenase (matrix metalloproteinase-1) and its inhibitor, tissue inhibitor of matrix metallopmteinase-1 (TIMP-1) are one of the substances which regulate this balance. The periodontal ligament fibroblast plays an important role in collagen metabolism during orthodontic treatment and is believed to be an origin of the osteoblast in the alveolar bone. The collagenase secreted by the periodontal ligament fibroblast and the osteoblast initiates the bone resorption by removing the osteoid layer in the alveloar bone. The interleukin-$1{\beta}$ is secreted by the macrophage during orthodontic treatment. The present study was undertaken to assess the effect of mechanical stress and interleukin-$1{\beta}$ on the expression of collagenase and TIMP-1 in the periodontal ligament fibroblasts using reverse transcription polymerase chain reaction and immunohistochemical staining. The periodontal ligament fibroblasts were stitched by placing the $Petriperm dish^{\circledR}$ dish on the top of spheroidal convex watch glass ($5\%$ surface increase) and tented with interleukin-$1{\beta}$ (1.0 ng/ml), or treated with both of them. Treatment with mechanical stress and/or interleukin-$1{\beta}$ resulted in increased collagenase mRNA expression. The mechanical stress treated group (1.61, 1.62, 1.37 fold increase), the interleukin-$1{\beta}$, tented group (1.68, 1.60, 3.78 fold increase), the mechanical stress and interleukin-$1{\beta}$ treated group (1.89, 1.72, 5.48 fold increase) induced increases in collagenase mRNA compared with the control group after 2, 4, 8 hours respectively. But TIMP-1 mRNA expressions at experimental groups were decreased after 2, 4 hours and increased after 8 hours. The mechanical stress treated group (0.16, 0.49 fold decrease and 3.77 fold increase), the interleukin-$1{\beta}$ treated group (0.15,0.44 fold decrease and 4.46 fold increase), the mechanical stress and interleukin-$1{\beta}$ tented group (0.15, 0.69 fold decrease and 4.81 fold increase) induced changes in TIMP-1 mRNA compared with the control group after 2, 4, 8 hours, respectively. Immunohistochemical stain showed that increased collagenase and TIMP-1 staining of the mechanical stress tented group, the interleukin-$1{\beta}$ treated group, and the mechanical stress and interleukin-$1{\beta}$ treated group compared with that of the control group after 8 hours. These findings suggest that mechanical stress and interleukin-$1{\beta}$ regulate expression of collagenase and TIMP-1.
In infectious disease, invasion of host tissue by bacteria or their products frequently induces a wide variety of inflammatory and immunopathologic reaction. Evidence indicates that cytokines are involved in the initiation and progression of chronic inflammatory diseases, such as periodontitis. Interleukin-6, which is a multifunctional cytokine, has important roles in acute and chronic inflammation and may also be implicated in bone resorption. Periodontal diseases are characterized by chronic inflammation of the periodontium with alveolar bone resoption. A principal driving force behind this response appears to lie in the immune system's response to bacteria. Many of the cell components which have been shown to function as virulence factors in gram-negative bacteria are associated with the bacterial surface. Of these, lipopolysaccharide has been characterized as one that mediates a number of biological activities which can lead to the destruction of host tissue. Non-steroidal antiinflammatory drug is used for reduce inflammation, and most of NSAIDs inhibit prostaglandine $E_2$ production, but it is shown that $PGE_2$ production is stimulated by IL-1 in recent study. So, the influence of other cytokines except $PGE_2$ on periodontium can not be avoided. Therefore, new antiinflammatory drug is needed. Rhizoma coptidis is used in oriental medicine for anti-inflammation and antiseptics. In this present study, we examined the IL-6 release in periodontal ligament cells treated with the lipopolysaccharide, and also the effect of rhizoma coptidis on cellular activity and IL-6 production of periodontal ligament cells. To evaluate the effect of rhizoma coptidis on cellular activity, the cells were seeded at a cell density of $1{\times}10^4$ cells/well in 24-well culture plates. After one day incubation, 1-6, 10-9 and 10-12 g/ml of rhizoma coptidis and 5, $10{\mu}g/ml$ of LPS were added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay were carried out. To evaluate the effect of rhizoma coptidis on IL-6 production, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates. After one day incubation, 10-9 g/ml of rhizoma coptidis and 5, $10{\mu}g/ml$ of LPS were added to the each well and incubated for 3, 6, 12 and 24 hours. Then, amounts of IL-6 production is measured by IL-6 ELISA kit used. The results were as follows : 1. Rhizoma coptidisrbelow to ($10^{-6}g/ml$) significantly increaed cellular activity of periodontal ligament cells than control. 2. Rhizoma coptidist ($10^{-9}g/ml$) significantly increased cellular activity of LPS($5{\mu}g/ml$)-treated periodontal ligament cells than control. 3. LPS(5 and $10{\mu}g/ml$) significantly increased IL-6 production of periodontal ligament cells than control. 4. Rhizoma coptidis($10^{-9}g/ml$) decreased IL-6 production of LPS ($5{\mu}g/ml$)-treated periodontal.ligarnent cells than LPS only tested group. These findings suggest that stimulation of the IL-6 release of periodontal ligament cells by LPS may have a role in the progression of inflammation and alveolar bone resoption in periodontal disease, and that inhibition of the IL-6 release of cells and stimulation of cellular activity by rhizoma coptidis may help the periodontal regeneration.
Kim, Soo-Jin;Min, Byoung-Hoon;Lee, Haeng-Sook;Lee, Byoung-Wook;Joo, Kyoung-Hwan
Applied Microscopy
/
v.39
no.2
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pp.167-173
/
2009
Capillaria hepatica is a parasitic nematode which causes hepatic capillariasis in rodents and other mammals, including man. Rat species of the genus Rattus are main primary host and rates of genus Rattus of up to 100% have been reported. Infection to reservoir and other mammalian hosts occur incidentally due to ingestion of water or food contaminated with C. hepatica embryonated eggs. The worms mature exclusively inside the liver, but they die and disassemble soon after egg spawning in rats. Dead worms and their eggs cause immune response of focal necrosis and inflammation within the liver. C. hepatica adult with a thin and long body is similar to capillary. The members of Order Trichurida are characterized by having a stichosome and the bacillary bands in front of the body. As already mentioned, the adult C. hepatica residesin the liver, where it deposits groups of eggs, and finally die in the encapsulated tissue of the liver. They produce eggs that elicit a marked granulomatous reaction that eventually destroy the worms. And the adult worms were mixed with eggs. So the complete isolation of the worm and observation of intact ultrastructure is very difficult. In this study, integument structure of C. hepatica isolated from the liver of mouse at 7 weeks after inoculation of embryonated eggs were observed with scanning and transmission electron microscopy. As a results, body length of isolated C. hepatica was about 99 mm. Cuticle, bacillary band and bacillary pore were obtained in the integument of worm. Bacillary pore across cuticular surface of the worm were observed. According to the existence of cap material, external forms of bacillary pore can be divided into three types such as flat, ingression, and ingression with the cap material type. The complete isolation of the worm and observation of ultrastructure of integument will provide the fundamental data which is important in the nematode research including C. hepatica.
Namkung, Su Min;Choi, Jeong Su;Park, Ji Hyang;Yang, Man Gil;Lee, Min Woo;Kim, Suhng Wook
Korean Journal of Clinical Laboratory Science
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v.49
no.3
/
pp.220-226
/
2017
Dopamine (DA) and serotonin (5-Hydroxytryptamine, 5-HT) are neurotransmitters and hormones that exist in small amounts but have important role in the body. Serum and 24-hour urine are used as specimens, and are usually examined by HPLC-MS. In this study, we tried to detect DA and 5-HT by competitive ELISA using antigen-antibody (Ab) reaction. After immobilizing $5{\mu}g/mL$ BSA conjugate on a 96-well surface, hormone and primary Ab, which are respectively diluted to different concentrations, were treated. Then, HRP-conjugated secondary Ab and TMB were added to measure absorbance. The regression equation and $R^2$ value were calculated based on absorbance, and sensitivity of Ab to hormone as well as the correlation between hormone concentration and absorbance were determined. In DA ELISA, $R^2$, the correlation between the concentration of hormone and absorbance, was the highest by 0.91 when anti-dopamine Ab was diluted 6,000 times and 7,000 times. In 5-HT ELISA, $R^2$ was bigger than 0.90 in every concentration except 3,000 times and 6,000 times. Both DA and 5-HT were not effectively detected at low concentrations (less than $1.0{\times}10^{-7}M$); and because reference value of serum DA is lower than this, HPLC-MS was required to detect serum DA. However, competitive ELISA may be effective in detecting 24-hour urine DA, serum, and 24-hour 5-HT. Further studies are needed to detect hormones more accurately at lower concentrations.
We prepared a new a SOS(silicon-on-silicide) wafer pair which is consisted of Si(100)/1000$\AA$-NiSi Si (100) layers. SOS can be employed in MEMS(micro- electronic-mechanical system) application due to low resistance of the NiSi layer. A thermally evaporated $1000\AA$-thick Ni/Si wafer and a clean Si wafer were pre-mated in the class 100 clean room, then annealed at $300~900^{\circ}C$ for 15hrs to induce silicidation reaction. SOS wafer pairs were investigated by a IR camera to measure bonded area and probed by a SEM(scanning electron microscope) and TEM(transmission electron microscope) to observe cross-sectional view of Si/NiSi. IR camera observation showed that the annealed SOS wafer pairs have over 52% bonded area in all temperature region except silicidation phase transition temperature. By probing cross-sectional view with SEM of magnification of 30,000, we found that $1000\AA$-thick uniform NiSi layer was formed at the center area of bonded wafers without void defects. However we observed debonded area at the edge area of wafers. Through TEM observation, we found that $10-20\AA$ thick amourphous layer formed between Si surface and NiSix near the counter part of SOS. This layer may be an oxide layer and lead to degradation of bonding. At the edge area of wafers, that amorphous layer was formed even to thickness of $1500\AA$ during annealing. Therefore, to increase bonding area of Si NiSi ∥ Si wafer pairs, we may lessen the amorphous layers.
This study was performed to determine the optimum coagulant dosing amount for effective treatment of raw water. The removal rate of turbidity and the variations of water qualities according to various dosage of coagulants such as Alum, PAC and PACS were investigated. The optimum coagulant dosing amount to make the lowest turbidity of water were 35mg/ι t of Alum, 30mg/ι of PAC and 10mg/ι of PACS in case of 5 NTU of raw water turbidity, and 30mg/ι of Alum, 25mg/ι of PAC and 10mg/ι of PACS in case of 10 NTU of that, respectively. The removal rates of turbidity at 4 min. and 8 min. of settling time were 10 and 72% of Alum, 44 and 62% of PAC and 25 and 55% of PACS in case of 5 NTU, and 52 and 70% of Alum, 90 and 95% of PAC and 10 and 28% of PACS in case of 10 NTU, respectively. Judging from the settling capability of floc., the reaction time of floe. formation and removal efficiency of turbidity, PAC was evaluated as more effective coagulant than Alum and PACS. Also PAC was regarded as the most effective coagulant when the water supply was changed sharply and the fluctuation of the surface loading occured with wide and sharp in settling basin. pH and alkalinity of the water were decreased with increasing coagulants dosage. But pH and alkalinity were not decreased below 5.8 which is the standard for drinking water quality, and 10mg/ι which is the limit concentration of floc. breakage, respectively. Residual Al of the treated water was decreased with increasing coagulants dosage in case of 5 and 10NTU of raw water turbidity. $KMnO_4$ consumption of the water was decreased with increasing coagulants dosage. The reduction rate of $KMnO_4$ consumption at the optimum coagulants dosage were 39% of Alum. 18% of PAC and 11% of PACS in case of 5 NTU of raw water turbidity, and 42% of Alum, 27% of PAC and 36% of PACS in case of 10 NTU of that, respectively. Any relationship was not found between the removal rate of turbidity and KMnO$_4$ consumption. TOC of the water was a bit decreased with increasing coagulants dosage up to 30mg/ι but not changed above 30mg/ι of coagulants dosage. The degree of TOC reduction was increased in the order of Alum, PAC and PACS treatment. Zeta potential of the colloidal floe. at the optimum coagulants dosage was in the range of -20~-15mV in case of 5 NTU of raw water turbidity and 0~0.5mV in case of 10 NTU of that. respectively. Although the kinds and dosages of coagulants were different, zeta potential range were fixed under the conditions of the best coagulation efficiency.
This study was conducted to investigate changes in inorganic phosphorus fractions in phosphorus accumulated plastic film house soils under cropping condition. Pot experiment was conducted with surface soils taken from plastic film house fields cultivated for 3, 8 and 16 years. Phosphorus fertilizer was applied at the rates of 0 (P0), $100kg\;ha^{-1}$ (recommended application rate. P1) and $200kg\;ha^{-1}$ (P2) as fused phosphate. Crops were grown sequentially in the order of corn-Chinese cabbage-Chinese cabbage-corn for 3 years. The content of easily soluble-P fraction was decreased with cropping. There were significantly positive relationship not only between the contents of easily soluble-P and the amounts of soil available P ($r=0.839^{**}{\sim}0.952^{**}$ for Bray-1 P, $r=0.895^{**}{\sim}0.967^{**}$ for Lancaster P, and $r=0.491^{**}{\sim}0.821^{**}$ for Olsen P), but also between the amounts of P absorbed by plants and the amounts of easily soluble P decreased ($r=0.644^{**}{\sim}0.822^{**}$). The decrease of easily soluble-P during cropping period could be described by a first-order reaction. The number of cropping times needed to decrease the content of easily soluble-P to an index level of $0.2mg\;kg^{-1}$, which is commonly reported as the desired concentration for soil P, was predicted in the range of 26~33 cropping times by using the equations. Regardless of P fertilization, the proportion of Al-P to total P was little varied during cropping period, but the proportion of Fe-P to total P increased with cropping. Although the content of Ca-P was high before cropping, the proportion of Ca-P to total P was increased with cropping. The proportion of reductant soluble-P to total P was little varied for P fertilizer treatment but was increased for no P fertilizer treatment. The residual-P was decreased during cropping period due to the absorption by crops and the conversion to other inorganic P fractions.
Kim, Ji-Young;Lee, Hye Jeen;Kim, Me Jin;Kim, Young Ho;Jung, Ji A;Yang, Seung;Hwang, Il Tae;Lee, Hae Ran
Clinical and Experimental Pediatrics
/
v.51
no.4
/
pp.377-382
/
2008
Purpose : The aim of this study is to assess the usefulness of skin test by an inactivated, 1/50 diluted solution of attenuated varicella vaccine in evaluating the immune status to varicella. Methods : Total 41 subjects (22 males, 19 females, aged 1-32 years) were enrolled from July to August, 2005. Past medical history including varicella infection, varicella vaccination were investigated through questionnaires. The skin test solution was prepared from solution of attenuated varicella vaccine(Oka strain) which was inactivated by exposure to room temperature for 10 days and diluted at 1/50 with normal saline. Skin test was done by injecting 0.1 mL of the solution intradermally into the volar surface of the right forearm and sterile normal saline was used as a control on the left forearm. Positive reaction was defined when the transverse diameter of the induration was 5 mm or more. Serum varicella zoster virus specific IgG antibody test by ELISA (enzyme-linked immunosorbent assay) was done. Results : In adults, the sensitivity of the varicella zoster virus skin test compared to ELISA was 94.7% and the positive predictive value was 100%. In children, both the positive predictive value and specificity were 100% but the sensitivity and the negative predictive value were 50% and 30.7% respectively. Children showed smaller skin test reactivity compared to adults. Conclusion : The varicella zoster virus skin test using inactivated, 1/50 diluted solution of attenuated varicella vaccine was proved as one of the useful tools for evaluating the immunity and susceptibility of the varicella zoster virus.
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