• Korean Chemical Engineering Research
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• v.43 no.1
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• pp.103-109
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• 2005

Isolation and primary culture technique of rabbit oral keratinocytes, and the study for effect of environmental factors on the cell growth were carried out in T75-flask. $1.92{\pm}0.59{\times}10^6$ viable cells were isolated by trypsin enzymatic digestion method from $0.25cm^2$ biopsy of rabbit oral mucosa. Primary culture with 10 mL of K-SFM containing 50 mg/L BPE, $5.0{\mu}g/L$ EGF and 0.15 mM $Ca^{2+}$ showed confluence after 8 days and doubling time was 2.54 days. Effect of medium types, medium volume and supplement types on the cell growth was investigated after the cultured keratinocytes had been harvested from primary confluence. Serum addition showed adverse effect and the increase of serum concentration didn't have an effect on the cell growth. The increase of medium volume decreased the cell growth. The increase of calcium concentration increased the cell growth and 2.0 mM was optimum value. In conclusion, when rabbit oral keratinocytes was cultured in T75-flask, the most effective conditions was to use 10 mL of K-SFM containing 50 mg/L BPE, $5.0{\mu}g/L$ EGF and 2.0 mM $Ca^{2+}$, and doubling time was 1.32 days. This study can provide the useful informations to develop a process and design a bioreactor for the culture of keratinocytes in human body like skin and cornea, as well as mucosa.

• Journal of the korean academy of Pediatric Dentistry
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• v.25 no.2
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• pp.400-420
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• 1998

We already know that it is very difficult to obtain an "isolated field" for direct bonding during the surgical exposure of unerupted teeth. The aim of this in-vitro study is to simulate the clinical situation of forced eruption and to evaluate the tensile strengths of preligatured button with several types of contamination which can happen during the surgical exposure of unerupted teeth. Four orthodontic direct bonding systems were used. ($Ortho-One^{TM}$, $Rely-a-Bond^{(R)}$, $Ortho-Two^{TM}$, Phase $II^{(R)}$) Each material was divided into four groups(n=20) : Group 1. (Control, no contamination), Group 2. (Rinse etching agent with saline instead of water), Group 3. (Blood contamination of etched surface for 30 seconds), Group 4. (Blood contamination of primed surface for 30 seconds) 320 bovine anterior permanent teeth were divided into the above mentioned 16 groups. Enamel surface was flattened and ground under water coolant. Pre-ligatured buttons were prepared to the same form. (Cut 0.25 ligature wire 10 cm in length. Twist the ligature wire 30 times clockwise. Mark the wire 15mm and 35mm points from button. Make a loop sticking two points together and twist the loop 6 times counterclockwise.) The bonded specimens were stored at $37^{\circ}C$ saline solution for 3 days. Then the tensile strength of each sample was measured with Instron universal testing machine, crosshead speed of 0.5mm/min. The following results were obtained: 1. As compared to control groups (Group 1) of each material, Rely-a-Bond had a significantly lower mean tensile strengths than other material. (p<0.01) 2. In Group 2. of Ortho-One and Rely-a-Bond, the mean tensile strengths decreased about 7.7% and 11.1%, respectively with statistical significances. (p<0.05) 3. In Group 2. of Ortho-Two and Phase II, the mean tensile strengths did not decrease. 4. In Group 3. of Ortho-One, Rely-a-Bond, Ortho-Two, and Phase II, the mean tensile strengths decreased about 60.8%, 56.1%, 60.2%, and 46.0%, respectively with statistical significances. (p<0.01) 5. In Group 4. of Ortho-One and Rely-a-Bond, the mean tensile strengths did not decrease. 6. In Group 4. of Ortho-Two and Phase II, the mean tensile strengths were decreased about 20.95% and 22.28%, respectively with statistical significances. (p<0.01) There were formations of a hump shaped mass from bonding resin under blood contamination which disturbed direct bonding procedure. According to Reynolds, the proper bond strength for clinical manipulation should be at least 45N or about 4.5Kg.F. According to these results, it can be concluded that Ortho-One could be used during surgical exposure of unerupted teeth. In any case, blood contamination of the etched surface should be avoided, but the blood contamination of primed surface of Ortho-One may not decrease bond strength. Just 'blowing-out' is enough to remove blood from primed surface of Ortho-One. You can verify the clean surface of the primer of Ortho-One after blowing out the blood contamination.

• Journal of the korean academy of Pediatric Dentistry
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• v.34 no.1
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• pp.36-42
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• 2007

To compare the survival rate of periodontal ligament cells preserved in storage media with good availability at the time of an avulsion injury, periodontal ligament cells were incubated in ${\alpha}-MEM$ culture medium containing 10% FBS in condition of $37^{\circ}C$, 5% $CO_2$. These cells were then cultured in HBSS, ${\alpha}-MEM$, milk(S co., P. co.) and tap water at the temperature of 4, 25, $37^{\circ}C$ each in 60 min. The groups were measured by MTT assay. The results were as follows : 1. Among the storage media at $4^{\circ}C$, ${\alpha}-MEM$ and P-milk had the highest preserving ability of periodontal ligament cells, while that of HBSS S-milk and tap was low in order. 2. Among the storage media at $25^{\circ}C$, ${\alpha}-MEM$ had the highest preserving ability of periodontal ligament cells, while that of P-milk, HBSS, S-milk, tap water was low in order. 3. Among the storage media at $37^{\circ}C$, the preserving ability of periodontal ligament cells was very high in ${\alpha}-MEM$, P-milk, HBSS and S-milk, it's lowest in tap water. 4. The preserving ability of periodontal ligament cells in ${\alpha}-MEM$ was high at $4^{\circ}C$ and it's low in order of $25^{\circ}C$, $37^{\circ}C$, but in HBSS was high at $4^{\circ}C$ and it's low at $25^{\circ}C$, $37^{\circ}C$ 5. The preserving ability of periodontal ligament cells in S-milk and P-milk was high at $4^{\circ}C$, $25^{\circ}C$ and it s low at $37^{\circ}C$. In conclusion, HBSS is the storage medium of choice in an avulsion, but in this study it is preferable to choose milk at $4^{\circ}C$ for tooth since it is easy to get and affect cell viability.

• Tuberculosis and Respiratory Diseases
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• v.48 no.4
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• pp.487-499
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• 2000

Background : Acute lung injury is an hypoxic respiratory failure resulting from damage to the alveolar-capillary membrane, which can be developed by a variety of systemic inflammatory diseases. In this study the therapeutic effects of intra-tracheal pulmonary surfactant instillation was evaluated in the intratracheal endotoxin induced acute lung injury model of a rat. Methods : Twenty Sprague-Dawley rats were divided into 4 groups, and normal saline (2 ml/kg, for group 1) or LPS (5 mg/kg, for group 2, 3, and 4) was instilled into the trachea respectively. Either normal saline (2 ml/kg, for group 1 & 2, 30 min later) or bovine surfactant (15 mg/kg, 30 min later for group 3, 5 hr later for group 5) was instilled into the trachea. The therapeutic effect of intratracheal surfactant therapy was evaluated with one chamber body plethysmography (respiratory frequency, tidal volume and enhanced pause), ABGA, BAL fluid analysis (cell count with differential, protein concentration) and pathologic examination of the lung. Results : Intratracheal endotoxin instillation increased the respiration rate decreased the tidal volume and int creased the Penh in all group of rats. Intratracheal instillation of surfactant decreased Penh, increased arterial oxygen tension, decreased protein concentration of BAL fluid and decreased lung inflammation at both times of administration (30 minute and 5 hour after endotoxin instillation). Conclusion : Intratracheal instillation of surfactant can be a beneficial therapeutic modality as discovered in the acute lung injury model of rats induced by intratracheal LPS intillation. It deserves to be evaluated for treatment of human acute lung injury.

• Tuberculosis and Respiratory Diseases
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• v.63 no.6
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• pp.497-506
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• 2007

Background: The present investigation was performed in rats and isolated human neutrophils in order to confirm the presumptive role of the positive feedback loop of cytosolic phospholipase $A_2$ ($cPLA_2$) activation by plateletactivating factor (PAF). Methods: The possible formation of the positive feedback loop of the $cPLA_2$ activation and neutrophilic respiratory burst was investigated in vivo and in vitro by measurement of the parameters denoting acute lung injury. In addition, morphological examinations and electron microscopic cytochemistry were performed for the detection of free radicals in the lung. Results: Five hours after intratracheal instillation of PAF ($5{\mu}g/rat$), the lung leak index, lung myeloperoxidase (MPO) activity, the number of neutrophils and the concentration of cytokine-induced neutrophil chemoattractant (CINC) in bronchoalveolar lavage fluid were increased by PAF as compared with those of control rats. The NBT assay and cytochrome-c reduction assay revealed an increased neutrophilic respiratory burst in isolated human neutrophils following exposure to PAF. Lung and neutrophilic $cPLA_2$ activity were increased following PAF exposure and exposure to hydrogen peroxide increased $cPLA_2$ activity in the lung. Histologically, inflammatory findings of the lung were observed after PAF treatment. Remarkably, as determined by $CeCl_3$ cytochemical electron microscopy, increased production of hydrogen peroxide was identified in the lung after PAF treatment. Conclusion: PAF mediates acute oxidative lung injury by the activation of $cPLA_2$, which may provoke the generation of free radicals in neutrophils.

• Journal of Life Science
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• v.28 no.10
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• pp.1220-1232
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• 2018

The purpose of this animal study was to evaluate, by histological analysis, bone regeneration in rabbit maxillary sinuses with an anorganic bovine graft (Bio-Oss) and a ${\beta}-tricalcium$ phosphate (${\beta}-TCP$) grafting. Bilateral sinus augmentation procedures were performed in 12 adult male rabbits. Rectangular replaceable bony windows were made with a piezoelectric thin saw insert. In the Bio-Oss group, Bio-Oss was grafted and in the ${\beta}-TCP$ group, ${\beta}-TCP$ was grafted and covered by replaceable bony windows. The animals were sacrificed at 2, 4, and 8 weeks after the surgical procedure. The augmented sinuses were evaluated by histomorphometric analysis using hematoxylin-eosin, Masson trichrome, and tartrate-resistant acid phosphatase stains and also by immunohistochemical analysis of proliferating cell nuclear antigen (PCNA), type I collagen, and osteocalcin content. Histologically, new bone formation was found on the surface of Bio-Oss and ${\beta}-TCP$ particles from 2 weeks and continued to 8 weeks. Significant higher new bone formation was revealed in the ${\beta}-TCP$ group than in the Bio-Oss group at 8 weeks. The amount of graft materials was significantly decreased in the ${\beta}-TCP$ group and the number of osteoclasts was significantly increased in the ${\beta}-TCP$ group from 4 to 8 weeks. Immunoreactivity to PCNA was reduced at 8 weeks. The expression of type I collagen was significantly increased in the ${\beta}-TCP$ group at 2 weeks, but was significantly increased in the Bio-Oss group at 8 weeks. Immunoreactivity to osteocalcin was increased from 2 to 8 weeks. These histological results can help in the selection of graft materials for implants. Both Bio-Oss and ${\beta}-TCP$ are proven graft materials, however, these results indicate that ${\beta}-TCP$ showed better bone regeneration results in rabbit maxillary sinus augmentation.

• Restorative Dentistry and Endodontics
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• v.31 no.6
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• pp.452-459
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• 2006

Competition will usually develop between the opposing walls as the restorative resin shrinks during polymerization. Magnitude of this phenomenon may be depended upon cavity configuration and volume. The purpose of this sturdy was to evaluate the effect of cavity configuration and volume on microleakage of composite resin restoration that has margins on the enamel site only. The labial enamel of forty bovine teeth was ground using a model trimmer to expose a flat enamel surface. Four groups with cylindrical cavities were defined, according to volume and configuration factor(Depth x Diameter / C-factor) - Group I : 1.5 mm ${\times}$ 2.0 mm / 4.0, Group II : 1.5 mm ${\times}$ 6.0 mm / 2.0, Group III : 2.Omm ${\times}$ 1.72 mm / 5.62, Group IV : 2.0 mm ${\times}$ 5.23 mm / 2.54. After treating with fifth-generation one-bottle adhesive - BC Plus$^{TM}$ (Vericom, AnYang, Korea), cavities were bulk flted with microhybrid composite resin - Denfill$^{TM}$ (Vericom). Teeth were stored in distilled water for one day at room temperature and were finished and polished with Sof-Lex system. Specimens were thermocycled 500 times between 5$^{\circ}$C and 55$^{\circ}$C for 30 second at each temperature. Teeth were isolated with two layers of nail varnish except the restoration surface and 1 mm surrounding margins. Electrical conductivity (${\mu}$A) was recorded in distilled water by electrochemical method. Microleakage scores were compared and analyzed using two-way ANOVA at 95% level. The results were as follows: 1. Small cavity volume showed lower microleakage score than large one, however, there was no statistically significant difference. 2. There was no relationship between cavity configuration and microleakage. Factors of cavity configuration and volume did not affect on microleakage of resin restorations with enamel margins only.

• Restorative Dentistry and Endodontics
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• v.24 no.4
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• pp.570-577
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• 1999

Intuitively, higher bond strengths should result in less leakage. However, the relationship between bond strengths and microleakage value is complex and not clearly understood. The purpose of this study was to evaluate the relationship between tensile bond strengths and microleakage values in the same restorations to understand the behavior of resin bonding to tooth structure. One-hundred and twenty enamel or dentin specimens from freshly extracted bovine mandibular incisors were used. The specimen was treated with 32% phosphoric acid for 15 seconds and rinsed for 20 seconds. the teeth were divided into four groups by means of wet bonding technique or dry bonding. One-Step$^{TM}$ adhesive were applied to the specimen. The specimens were immersed in 2% methylene blue solution for 7 days, and tensile bond strength and microleakage were measured. The results were as follows: 1. Significant negative correlation was found between bond strengths and micro leakage values. Hence, higher bond strengths seem to be associated with lower microleakage, and vice versa (r=-0 50, p<0.05). 2. The Enamel/Wet group showed significantly higher bond strength than Enamel/Dry one, and Dentin/Wet group showed higher strength than Dentin/Dry one (p<0.05). 3. Microleakage was significantly less ill wet bonding than in dry one at dentin (p<0.05), however, there was no significant difference between wet and dry bonding at enamel (p>0.05).

• Journal of Plant Biotechnology
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• v.33 no.3
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• pp.195-207
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• 2006

Stem cells are the primordial, initial cells which usually divide asymmetrically giving rise to on the one hand self-renewals and on the other hand progenitor cells with potential for differentiation. Zygote (fertilized egg), with totipotency, deserves the top-ranking stem cell - he totipotent stem cell (TSC). Both the ICM (inner cell mass) taken from the 6 days-old human blastocyst and ESC (embryonic stem cell) derived from the in vitro cultured ICM have slightly less potency for differentiation than the zygote, and are termed pluripotent stem cells. Stem cells in the tissues and organs of fetus, infant, and adult have highly reduced potency and committed to produce only progenitor cells for particular tissues. These tissue-specific stem cells are called multipotent stem cells. These tissue-specific/committed multipotent stem cells, when placed in altered environment other than their original niche, can yield cells characteristic of the altered environment. These findings are certainly of potential interest from the clinical, therapeutic perspective. The controversial terminology 'somatic stem cell plasticity' coined by the stem cell community seems to have been proved true. Followings are some of the recent knowledges related to the stem cell. Just as the tissues of our body have their own multipotent stem cells, cancerous tumor has undifferentiated cells known as cancer stem cell (CSC). Each time CSC cleaves, it makes two daughter cells with different fate. One is endowed with immortality, the remarkable ability to divide indefinitely, while the other progeny cell divides occasionally but lives forever. In the cancer tumor, CSC is minority being as few as 3-5% of the tumor mass but it is the culprit behind the tumor-malignancy, metastasis, and recurrence of cancer. CSC is like a master print. As long as the original exists, copies can be made and the disease can persist. If the CSC is destroyed, cancer tumor can't grow. In the decades-long cancer therapy, efforts were focused on the reducing of the bulk of cancerous growth. How cancer therapy is changing to destroy the origin of tumor, the CSC. The next generation of treatments should be to recognize and target the root cause of cancerous growth, the CSC, rather than the reducing of the bulk of tumor, Now the strategy is to find a way to identify and isolate the stem cells. The surfaces of normal as well as the cancer stem cells are studded with proteins. In leukaemia stem cell, for example, protein CD 34 is identified. In the new treatment of cancer disease it is needed to look for protein unique to the CSC. Blocking the stem cell's source of nutrients might be another effective strategy. The mystery of sternness of stem cells has begun to be deciphered. ESC can replicate indefinitely and yet retains the potential to turn into any kind of differentiated cells. Polycomb group protein such as Suz 12 repress most of the regulatory genes which, activated, are turned to be developmental genes. These protein molecules keep the ESC in an undifferentiated state. Many of the regulator genes silenced by polycomb proteins are also occupied by such ESC transcription factors as Oct 4, Sox 2, and Nanog. Both polycomb and transcription factor proteins seem to cooperate to keep the ESC in an undifferentiated state, pluripotent, and self-renewable. A normal prion protein (PrP) is found throughout the body from blood to the brain. Prion diseases such as mad cow disease (bovine spongiform encephalopathy) are caused when a normal prion protein misfolds to give rise to PrP$^{SC}$ and assault brain tissue. Why has human body kept such a deadly and enigmatic protein? Although our body has preserved the prion protein, prion diseases are of rare occurrence. Deadly prion diseases have been intensively studied, but normal prion problems are not. Very few facts on the benefit of prion proteins have been known so far. It was found that PrP was hugely expressed on the stem cell surface of bone marrow and on the cells of neural progenitor, PrP seems to have some function in cell maturation and facilitate the division of stem cells and their self-renewal. PrP also might help guide the decision of neural progenitor cell to become a neuron.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.1
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• pp.8-15
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• 2008

Saussurea lappa (SL) has been used to reduce abdominal pain and tenesmus in traditional oriental medicine. SL and major compounds of SL, sesquiterpene lactones, have been suggested to possess various biological effects, including anti-tumor, anti-ulcer, anti-inflammatory, anti-viral and cardiotonic activities. Recently, it has been reported that ethanol extracts from roots of SL have antiproliferative effects on gastric cancer cells. To explore the possibility that SL has chemopreventive effects on prostate cancer, we examined whether the hexane extract of SL (HESL) inhibits the growth of LNCaP human prostate cancer cells. Cells were incubated with various concentrations ($0{\sim}4$ mg/L) of HESL in DMEM/F12 containing 5% charcoal stripped fetal bovine serum. HESL substantially decreased viable cell numbers and induced apoptosis of LNCaP cells in dose-dependent manners. HESL increased the levels of cleaved caspase-8, -9, -7 and -3, and poly (ADP-ribose) polymerase. HESL increased the levels of the pro-apoptotic Bak and truncated-Bid proteins whereas it had no effect on the anti-apoptotic Bcl-2, Bcl-xL, or Mcl-1. The present results indicate that HESL inhibits the growth of human prostate cancer cells by inducing apoptosis, which involves the activation of the caspase cascades.