• Journal of Periodontal and Implant Science
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• v.35 no.3
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• pp.549-561
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• 2005

이 연구의 목적은 새로이 제작된 chitosan-poly(L-lactic acid)(PLLA) 다층 다공성 차폐막의 생체적합성 및 골세포활성도 및 골재생능을 평가하는 것이다. 제작된 차폐막을 24 well에 넣고 clonal osteoblast-like cell line(MC3T3-E1)을 접종한 군을 실험군으로, 차폐막을 사용하지 않은 대조군으로 하였다. 배양 1일, 7일 및 14일째에 각 well에서 세포수를 측정하였다. 주사전자현미경을 이용하여 차폐막에 부착된 세포의 형태관찰을 시행하였다. RNA 추출 및 RT-PCR을 실시한 후, agarose gel상에서 전기영동하여 조골세포 표식자인 collagen type I(COL), osteopontin(OP) 및 osteocalcin(OC) mRNA의 발현을 관찰하였다. 제작된 매트릭스의 생체적합성 및 골재생능을 관찰하기 위하여 백서의 두개골에 직경 8mm의 원형 결손부를 형성한 후 차폐막을 이식한 군을 실험군으로, 아무 것도 넣지 않은 군을 대조군으로 하여 4주 경과 후 실험동물을 희생시킨 후 조직학적관찰을 시행하였다. 시간경과에 따른 부착세포수 관찰결과, 배양 14일까지 조골세포의 수가 지속적으로 증가하였고, 주사전자현미경으로 세포의 형태 관찰결과, 배양된 세포들은 중층의 형태로 성장하면서 시간경과에 따라 세포가 응집되는 양상을 나타내었다. 관찰 기간동안 COL, OP, 및 OC mRNA의 발현이 관찰되어 배양 전 기간동안 조골세포의 형질이 잘 유지되고 있음을 알 수 있었다. 백서 두개골 결손부에 이식된 차폐막은 염증반응 없이 주위 조직과 우수한 생체적합성을 나타내었으며, 차폐막을 이식하지 하지 않은 대조군에 비해 높은 신생골 형성을 나타내었다. 이상의 관찰결과로 새로이 제작된 chitosan-PLLA 차폐막은 우수한 생체적합성 및 골재생능을 나타냄을 알 수 있었으며, 향후 이를 골조직 재생 및 치주조직유도재생 분야에 응용될 수 있을 것으로 생각된다.

• Journal of Periodontal and Implant Science
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• v.31 no.2
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• pp.277-285
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• 2001

Periodontal disease is characterized by inflammation and subsequent loss and/or damage to tooth-supporting tissues such as bone, cementum,and periodontal ligament. Periodontal ligament and cementum are the key tissues in the initial process of regeneration following periodontal disease. Therefore, studies on cementoblasts, which form cementum are emphasized. It is still unclear which cells cementoblast differentiate from. This study was conducted under the hypothesis that PDL fibroblast can differentiate into either cementoblast or osteoblast depending on the conditions of surrounding tissue. Clinically, with excessive traction force of orthodontic appliances or excessive occlusion hypercementosis is observed, and this has been confirmed histologically. Consequently, activation of cementoblast can be expected in rats when mechanical stimuli are given to PDL fibroblast. Therefore, the purpose of this article is to prove that PDL fibroblast differentiates into cementoblast in rats under mechanical stimuli using histologic and molecular methods. In this study, twenty rats were given hard diet. Ten of them were sacrificed after 1 week, and the others were sacrificed after two weeks. Slides were made from tooth specimen, and they were studied under the microscope. In addition, PDL fibroblast and cementum from the extracted teeth were analyzed with Northern blotting. In histologic examination, as time passed, PDL fibroblast migrated to the dentin side, differentiated into cementoblast, and formed new cementum. In Northern blotting, it was found that mRNA expression of cementoblast-specific proteins such as BSP, OC, OPN, and type I collagen were more prominent in rats sacrificed after 2 weeks of hard-diet than rats sacrificed after 1 week. From these findings we can conclude that PDL fibroblast can differentiate into cementoblast under mechanical stimuli. We think that 'Rat Models' used in this study will be beneficial to future studies regarding cementoblast.

• Polymer(Korea)
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• v.29 no.5
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• pp.501-507
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• 2005

Small intestinal submucosa (SIS) had been widely used as a biomaterial without immune rejection responses. SIS sponges prepared by crosslinking with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). SIS powders dissolved in $3\%(v/v)$ acetic acid aqueous solution for 48hrs and freeze-dried. EDC solution ($H_2O$ : ethanol = 5 : 95) as a crosslink agent was used in concentration of 100mM. In vitro, rat-BMSCs seeded in SIS sponges and induced the osteogenesis for 28 days. We have characterized the osteogenic potential of rat-BMSCs in SIS sponges by alkaline phosphatase activity(ALP), n assay, SEM and RT-PCR for osteogenic phenotype. In SEM, all morphology of SIS sponges was regular and showed interconnected pore structure. By RT-PCR analysis, we observed type I collagen expression. These results demonstrate osteogenic differentiation of rat-BMSCs. In conclusion, we confirmed that the morphology of surface, cross-section, and side of SIS sponges were highly porous with good interconnections between each pores, which can support the surface of cell growth, proliferation, and differentiation. This result indicates that SIS sponge is useful for osteogenesis of BMSCs.

• Polymer(Korea)
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• v.33 no.1
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• pp.26-32
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• 2009

The aim of this research was to prepare microparticulate systems based on poly (lactide-co-glycolide)(PLGA) for the local release of ipriflavone in order to reduce bone loss. We developed the IP loaded PLGA microspheres using relatively simple oil-in-water(O/W) solvent evaporation method. HPLC was used to perform the in vitro release test of IP and morphology of cell attached on the micro-spheres was investigated using SEM. Cytotoxicity was assayed by cell counting kit-8 (CCK-8) test. Osteogenic differential cells were analyzed by ALP activity. Through RT-PCR analysis, we observed osteocalcin, ALP, and Type I collagen mRNA expression. The release of IP in vitro was more prolonged over 42 days and IP/PLGA microspheres showed the improvement on the cell proliferation, ALP activity and RT-PCR comparing with control (only PLGA). This initial research will be used to direct future work involved in developing this composite injectable bone tissue engineering system.

• Animal Bioscience
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• v.35 no.6
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• pp.902-915
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• 2022

Objective: Diet acidification supplementation is known to influence intestinal morphology, gut microbiota, and on phosphorus (P) utilization of broilers. Alterations in intestinal barrier and microbiota have been associated with systemic inflammation and thus regulating bone turnover. Hence the effect of acidifier addition to drinking water on tibia mass and the linkages between intestinal integrity and bone were studied. Methods: One-d-old male broilers were randomly assigned to normal water (control) or continuous supply of acidified water (2% the blend of 2-hydroxy-4-methylthiobutyric acid, lactic, and phosphoric acid) group with 5 replicates of 10 chicks per replicate for 42 d. Results: Acidification of drinking water improved the ash percentage and calcium content of tibia at 42 d. Broilers receiving acidified water had increased serum P concentration compared to control birds. The acidified group showed improved intestinal barrier, evidenced by increased wall thickness, villus height, the villus height to crypt depth ratio, and upregulated mucin-2 expression in ileum. Broilers receiving drinking water containing mixed organic acids had a higher proportion of Firmicutes and the ratio of Firmicutes and Bacteroidetes, as well as a lower population of Proteobacteria. Meanwhile, the addition of acidifier to drinking water resulted in declined ileal and serum proinflammatory factors level and increased immunoglobulin concentrations in serum. Concerning bone remodeling, acidifier addition was linked to a decrease in serum C-terminal cross-linked telopeptide of type I collagen and tartrate-resistant acid phosphatase reflecting bone resorption, whereas it did not apparently change serum alkaline phosphatase activity that is a bone formation marker. Conclusion: Acidified drinking water increased tibia mineral deposition of broilers, which was probably linked with higher P utilization and decreased bone resorption through improved intestinal integrity and gut microbiota and through decreased systemic inflammation.

• The Journal of Internal Korean Medicine
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• v.44 no.3
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• pp.402-417
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• 2023

Objective: Liver fibrosis is a highly conserved wound-healing response and the final common pathway of chronic inflammatory injury. This study aimed to evaluate the potential anti-fibrotic effect of the combination of Rhei Radix et Rhizoma water extract (RW) and silymarin in a thioacetamide (TAA)-induced liver fibrosis model. Methods: The liver fibrosis mouse model was established through the intraperitoneal injection of TAA (1 week 100 mg/kg, 2-3 weeks 200 mg/kg, 4-8 weeks 400 mg/kg) three times per week for eight weeks. Animal experiments were conducted in five groups; Normal, Control (TAA-induced liver fibrosis mice), Sily (silymarin 50 mg/kg), RSL (RW 50 mg/kg+silymarin 50 mg/kg), and RSH (RW 100 mg/kg+silymarin 50 mg/kg). Biochemical analyses were measured in serum, including aspartate aminotransferase (AST), alanine aminotransferase (ALT), malondialdehyde (MDA), and ammonia levels. Liver inflammatory cytokines and fibrous biomarkers were measured by Western blot analysis, and liver histopathology was evaluated through tissue staining. Results: A significant decrease in the liver function markers AST and ALT and a reduction in ammonia and total bilirubin were observed in the group treated with RSL and RSH. Measurement of reactive oxygen species and MDA revealed a significant decrease in the RSL and RSH administration group compared to the TAA induction group. The expression of extracellular matrix-related proteins, such as transforming growth factor β1, α-smooth muscle actin, and collagen type I alpha 1, was likewise significantly decreased. All drug-administered groups had increased matrix metalloproteinase-9 but a decreasing tissue inhibitor of matrix metalloproteinase-1. RSL and RSH exerted a significant upregulation of NADPH oxidase 2, p22^phox, and p47^phox, which are oxidative stress-related factors. Furthermore, pro-inflammatory proteins such as cyclooxygenase 2 and interleukin-1β were markedly suppressed through the inhibition of nuclear factor kappa B activation. Conclusions: The administration of RW and silymarin suppressed the NADPH oxidase factor protein level and showed a tendency to reduce inflammation-related enzymes. These results suggest that the combined administration of RW and silymarin improves acute liver injury induced by TAA.

• Food Science of Animal Resources
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• v.44 no.1
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• pp.51-73
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• 2024

This study investigated how birth weight differences in piglets affected carcass and muscle fiber properties as well as meat quality at slaughter. Within litters, piglets were grouped according to their birth weight as either normal (NBW; 1.62-1.73 kg) or low (LBW; 1.18-1.29 kg). At 5 weeks of age, NBW piglets were randomly transitioned to control (C) or isocaloric high fat diets derived from non-dairy (HF), while LBW piglets were randomly transitioned to high fat diets derived from non-dairy (HF) or dairy sources (HFHD). Piglets were reared in individual pens under standardized housing and feeding conditions. Live weight was recorded weekly, and pigs were slaughtered at 12 weeks of age. Hot carcass weights, dressing percentages, lean meat yield, and primal cut proportions were determined. The m. longissimus thoracis was collected from the right side of the carcass for measurement of physical and chemical properties of meat and muscle fiber characteristics. Results indicated that LBW pigs compensated for their live weight compared to NBW pigs at 6 weeks of age. The mean muscle fiber diameter of LBW-HFHD group is significantly higher than NBW-C and NBW-HF group, and the type I muscle fiber diameter is significantly higher than NBW-C group. Dairy fat inclusion in LBW pig diet reduced carcass back fat thickness. This increased the calculated lean meat yield to be comparable to that of NBW pigs fed a commercial diet. Incorporating dairy-sourced high-fat into LBW pigs' diets appears to be an effective strategy for producing carcasses equivalent to NBW pigs.

• Journal of Life Science
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• v.32 no.2
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• pp.101-107
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• 2022

In this research, the anti-inflammatory, antioxidant, antiaging, and skin whitening properties of pulp and seed extracts of passion fruit were studied. The result of the primary skin irritation test using a skin-attached patch determined the skin irritation index to be 0.00 for the passion fruit extract. In addition, RAW 264.7 macrophages produce NO by stimulation of lipopolysaccharides, and the application of extracts to this resulted in significantly lower NOs, confirming the excellent anti-inflammatory properties of passion fruit extracts. The 2,2-diphenyl-1-picrylhydrazyl test further confirmed that the passion fruit extract exhibits a good 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate radical scavenging ability of 5.11% and strong antioxidant properties. The presence of collagen type I in the skin is a measure of aging and various skin diseases. The results obtained from the analysis of the activity of human procollagen I alpha 1 confirmed that the passion fruit extract reduces the synthesis of procollagen. In addition, the skin whitening property of the passion fruit extract was confirmed by the melanin inhibition test, and a sample was obtained that contained more than 2% of arbutin, a whitening agent approved by the Ministry of Food and Drug Safety, which is generally present in the form of a white powder and is used as a functional ingredient. This confirms that the whitening efficacy of the passion fruit extract obtained from nature contributes to the development of functional raw materials for cosmetics and food.

• Journal of Periodontal and Implant Science
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• v.35 no.2
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• pp.321-333
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• 2005

1. 목적 in vitro 상에서 법랑기질유도체가 치주인대섬유아세포, 불멸화 조골세포와 백악질 유래세포의 증식과 유전자 발현에 미치는 영향을 알아보고자 하였다. 2. 연구방법 및 재료 <세포증식 연구> 교정을 목적으로 발거한 치아에서 분리, 배양한 치주인대섬유아세포와 백악질유래세포, 그리고 $SaOs_2$ 세포를 이용하였다. 법랑기질유도체가 세포 증식에 미치는 영향을 알아보기 위해, 35 mm Petri dish에 dish 당 $5{\times}10^3$ 개의 세포를 접종하였다. 대조군은 1% 항생제와 10% FBS를 포함한 DMEM 배지를 이용했고, 5mM 초산을 첨가한 군과 첨가하지 않은 두 개의 대조군이 이용되었다. 실험군은 100 ${\mu}g/ml$의 법랑기질유도제를 첨가한 군과 100 ${\mu}g/ml$의 법랑기질유도체와 5 mM의 초산을 첨가한 2개의 실험군이 이용되었다. 각 군은 세 개의 배양접시에 행해졌고, 1, 3, 8일에 세포의 수를 각각 측정하였다. 결과는 repeated measures ANOVA로 통계 처리하였다. <유전자 발현 연구> 각 세포의 형질 특성을 알아보기 위해 RT-PCR을 실시하여 조골세포 분화 표식자와 연관된 Human collagen type I(COL I), human osteopontin(OP), human osteocalcin(OC), human alkaline phosphatase(ALP)와 human bone sialoprotein(BSP)의 mRNA 발현을 실험 1, 3, 8일에 걸쳐, 세 군의 차이를 비교 관찰하였다. 3. 결과 <세포증식 연구> 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포의 증식은 법랑기질유도체에 의해 영향을 받지 않았다. 대조군과 초산이 포함된 대조군 그리고 법랑기질유도체와 초산이 포함된 실험군에서 유의할 만한 세포 수의 차이가 실험 기간 1, 3, 8일에 걸쳐 나타나지 않았다(p<0.05). <유전자 발현 연구> ALP와 COL I은 세 군의 세포에서 모두 발현되었고, 발현 정도는 EMD에 영향을 받지 않았다. OC은 세 군에서 모두 비교적 약하게 발현되었고, 특히 $SaOs_2$ cell과 백악질유래세포에서 약하게 발현되었다. EMD는 OC의 발현정도를 약하게 하였다. OP은 백악질유래세포에서 1, 3, 8일에 걸쳐 EMD 유무에 관련 없이 발현되지 않았다. 그러나 치주인대세포와 $SaOs_2$ cell에서는 강하게 발현되었다. BSP는 치주인대세포와 $SaOs_2$ cell에서 1, 3, 8일에 걸쳐 비교적 고르게 발현되었다. EMD 배지에서 배양된 백악질유래세포는 8일에는 BSP가 발현되지 않았다. 4 결론 이번 실험 결과에 의하면 법랑기질유도체는 치주인대세포, 불멸화 조골세포와 백악질 유래세포의 증식에 있어 유의성 있는 효과를 나타내지 않았다. 그러나, 유전자 발현에 있어서는, 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포 모두에서 OC mRNA의 발현을 억제하는 효과를 나타내었다. EMD는 세포의 증식에는 영향을 미치지 않지만, 유전자 발현에 있어 일부 영향을 미치는 것으로 보인다. 법랑기질유도체가 세포의 증식과 유전자 발현에 미치는 영향은 배양된 세포의 형질특성, 배양환경, 배양일수 등에 따라 달라질 수 있다. 그러므로 법랑기질유도체가 in vitro 상에서 세포에 미치는 영향은 보다 정량화된 연구가 필요하다.

• Journal of Oral Medicine and Pain
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• v.30 no.3
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• pp.287-300
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• 2005

Squamous cell carcinoma (SCC) in head and neck show a variability in the response to chemotherapy, even when it present with similar histological tumor type, grade, and clinical stage. The purpose of present study it to identify predictive bio-marker for the sensitivity or resistance to conventional chemotherapeutic agents, 5-fluorouracil (5-FU) and Cisplatin Oral cancer cell lines were used in present study. MTT assay was performed to evaluate the sensitivity and/or resistance to 5-FU and Cisplatin. And RT-PCR was carried out for evaluation of the mRNA expressions of various genes associated with mutation, inflammation (COX pathway), cell cycle, senescence and extracellular matrix (ECM). The molecules which are correlated with the sensitivity to 5-FU are XPA, XPC, OGG, APEX, COX-2, PPAR, Cyclin E, Cyclin B1, CDC2, hTERT, hTR, TIMP-3, TIMP-4 and HSP47. And the molecules are correlated with the sensitivity to Cisplatin are COX-1, iNOS, eNOS, PCNA, collagen 1 and MMP-9. Taken together, when choosing the appropriate chemotherpeutic agents for patients, considering the molecules which are correlated or reversely correlated is helpful to choose the resonable agents for cancer patients.