• Biomolecules & Therapeutics
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• 제8권2호
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• pp.199-205
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• 2000

On the inter-order protoplast fusants of Lentinus edodes and Ganoderma lucidum was the antitumor activity test performed and the fusant P22 was selected. The hot water extract of the cultured mycelia of P22 were purified by DEAE-cellulose chromatographya and the resulting purified fraction was designated as P22A. It was found to be a proteoglycan whose molecular weight was 47 kDa. When examined for immunopotentiation activity, P22A increased the number of colony forming unit in the bone marrow stem cells to 3-folds. It also potentiated the secretion of nitric oxide in activated macrophages to 2-folds. In humoral immune response, it increased the activities of the alkaline phosphatase in differentiated B cells to 1.6-folds and the number of plaque forming cells to 1.8-folds. In cellular immune response, it restored the depressed response of delayed type hypersensitivity in tumor bearing mice to normal level. These results suggest that P22A have potential to restore the decreased immune activity of the tumor bearing mice to normal level.

• 한국정밀공학회지
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• 제31권5호
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• pp.375-380
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• 2014

To enhance biocompatibility between the orthopedic implant stem and obsteoblast cells, bone-forming cells, micro-size holes are patterned in Ti plate surface. Initially, the house built laser power stabilization system is applied to the laser micro patterning machine to convince repeatable result. Various pulse widths are irradiated Ti plate and relationship between diameters of patterned holes and pulsed width is derived. Effect of multi pulse is observed and optimal pulse number is considered to avoid heat affected zone. After MG-63 osbeoblast cells are cultured, micro patterned Ti plates are compared with control plates. In SEM image, cells are well aligned and aggregation is observed in both 60, and $100{\mu}m$ patterned plates. Finally, free form surface stem model is prepared to test micro hole patterning.

• International Journal of Stem Cells
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• 제16권3호
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• pp.304-314
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• 2023

Background and Objectives: Ear cartilage malformations are commonly encountered problems in reconstructive surgery, since cartilage has low self-regenerating capacity. Malformations that impose psychological and social burden on one's life are currently treated using ear prosthesis, synthetic implants or autologous flaps from rib cartilage. These approaches are challenging because not only they request high surgical expertise, but also they lack flexibility and induce severe donor-site morbidity. Through the last decade, tissue engineering gained attention where it aims at regenerating human tissues or organs in order to restore normal functions. This technique consists of three main elements, cells, growth factors, and above all, a scaffold that supports cells and guides their behavior. Several studies have investigated different scaffolds prepared from both synthetic or natural materials and their effects on cellular differentiation and behavior. Methods and Results: In this study, we investigated a natural scaffold (alginate) as tridimensional hydrogel seeded with progenitors from different origins such as bone marrow, perichondrium and dental pulp. In contact with the scaffold, these cells remained viable and were able to differentiate into chondrocytes when cultured in vitro. Quantitative and qualitative results show the presence of different chondrogenic markers as well as elastic ones for the purpose of ear cartilage, upon different culture conditions. Conclusions: We confirmed that auricular perichondrial cells outperform other cells to produce chondrogenic tissue in normal oxygen levels and we report for the first time the effect of hypoxia on these cells. Our results provide updates for cartilage engineering for future clinical applications.

• 대한치과교정학회지
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• 제23권2호
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• pp.229-248
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• 1993

결합 조직의 대사를 억제하고, 골조직이나 골세포의 골기질 대사 역시 억제하는 것으로 알려져 있는 $Interferon-\gamma(IFN-\gamma)$의 교정치료에의 이용 가능성을 평가하기 위하여, 교정력에 의한 골 개조 과정에서 중심적 역할을 하는 것으로 알려진 치주인대 세포를 primary culture하여 배양된 세포에 $IFN-\gamma$를 투여함으로써 이것이 골조직 기질의 합성능에 미치는 영향에 대하여 관찰하였다. $IFN-\gamma$는 세포의 DNA합성능을 약하게 증가시켰으며, 세포내 DNA 총량에는 영향을 미치지 않았다. 따라서 이 실험에서 사용한 용량의 $IFN-\gamma$는 세포에 독성이 없다고 할 수 있으며, 다른 결합조직 세포에서 나타나는 항증식 효과와는 반대되는 결과였다. $IFN-\gamma$는 비교원성 단백질(NCP)의 합성을 증가시키는 양상을 나타내었으나, 교원질(CDP)의 합성은 감소시키는 경향을 보여, 총단백질에 대한 교원질의 합성비율은 $IFN-\gamma$에 의해 용량 의존적으로 감소하였다. 또한 교원질의 mRNA양은 단백질 합성과 유관하게 $IFN-\gamma$에 의해 억제되었다. 따라서 $IFN-\gamma$는 교원질 합성의 전사과정 혹은 그 이후의 과정에 영향을 미칠 것으로 예상할 수 있다. 한편 Indomethacin의 투여로 $IFN-\gamma$에 의해 억제된 교원질의 합성이 영향을 받지 않았기 때문에 $IFN-\gamma$에 의한 교원질 합성 과정에 prostaglandin이 관련되지 않았음을 알 수 있었다. 반면 fibronectin의 합성은 10 및 100 U/ml의 $IFN-\gamma$ 투여시에는 영향을 받지 않았으나, 1000U/ml의 $IFN-\gamma$투여시에는 유의한 증가를 나타내어, 교원질에서와는 다른 영향을 나타내었다. 또한 mRNA steady state level에서도 $IFN-\gamma$는 교원질의에서와는 달리 fibronectin mRNA 양에는 영향을 미치지 않았다. 즉 $IFN-\gamma$ fibronectin 유전자 발현에 영향을 미치는 부위는 전사나, 전사후 변형단계가 아닌 단백질 합성단계에서 영향을 미침을 보여주었다. 따라서 $IFN-\gamma$ 교원질과 fibronectin합성 조절 영향을 미치는 부위 서로 다름을 알 수 있겠다. Alkaline phospatase의 활성은 10-1000 U/ml의 $IFN-\gamma$를 투여시 약하게 증가시키는 경향을 보였으나 석회화가 일어날 정도로 높게 증가시키지는 못했다. 따라서 $IFN-\gamma$는 골기질의 주성분인 type I 교원질의 합성을 선택적으로 억제하는 기능과 alkaline phosphatase의 활성을 크게 증가시키지 못한 점 등으로 미루어 볼 때, 골개조를 억제하는 방향으로 작용한다고 볼 수 있으며, 교정치료 과정중 골개조를 억제하는 부위에서 사용을 시도해 볼 수 있겠다.

• 한국키틴키토산학회지
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• 제23권4호
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• pp.277-284
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• 2018

본 연구에서는 제주 자생해조류인 감태로부터 제조한 ethyl acetate 분획물의 비만 세포에 대한 항알러지 활성에 관한 연구를 수행하였다. IgE/BSA로 활성화시킨 비만 세포에서 EC-EtoAc는 세포 독성 없이 비만 세포의 탈과립 지표로 사용되는 ${\beta}$-hexosaminidase의 분비 뿐만 아니라 알러지 유발성 케모카인(TARC)과 사이토카인(IL-$1{\beta}$, IL-4, IL-6, IL-10, IL-13, IFN-${\gamma}$ 및 TNF-${\alpha}$)의 mRNA 발현을 억제하였다. 또한, EC-EtoAc는 비만 세포의 표면 리셉터인 $Fc{\varepsilon}RI$의 발현 및 IgE와 $Fc{\varepsilon}RI$의 결합능을 감소시켰다. 이 모든 결과로부터, 감태 유래 분획물인 EC-EtoAc가 활성화된 비만 세포에서 $Fc{\varepsilon}RI$의 발현 및 IgE와 $Fc{\varepsilon}RI$의 결합능을 감소시킴으로써 알러지 유발성 매개 인자들의 분비와 발현 등을 감소시켜 항알러지 효능을 가진다는 것을 알 수 있었다. 또한, 이 연구 결과는 EC-EtoAc가 항알러지 효능을 보이는 천연 소재로서 이용가치가 있다는 것을 제시한다.

• 폴리머
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• 제31권1호
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• pp.14-19
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• 2007

탈미네랄화된 골분(demineralized bone particle, DBP)은 골/연골 형성의 강력한 유도인자로 사용된다. 본 연구에서는 용매 캐스팅/염 추출법을 이용해 함량별 DBP와 PLGA가 하이브리드화된 다공성 지지체를 실제 디스크 형태와 유사하게 제조하였다. 제조된 지지체의 특성을 분석하기 위하여 다공도, 표면 젖음성 및 물 흡수성을 측정하였다. 디스크 세포인 섬유륜 및 수핵 세포는 토끼로부터 분리하여 제조된 지지체에 각각 파종한 후, 지지체를 재조합하여 배양하였다. 지지체에 파종된 디스크 세포의 생존율과 증식률은 MTT(3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide) 분석 방법을 이용하였고, 면역결핍 쥐의 피하에 삽입하여 이들의 디스크 조직 형성 정도를 확인하였다. 피하에 이식된 지지체를 적출하여 육안으로 관찰하고 모폴로지의 변화를 확인한 후, 조직을 파라핀으로 고정시켜 슬라이드를 제조하여 hematoxylin과 eosin 염색을 수행하였다. 천연/합성 하이브리드 담체로서의 DBP/PLGA 담체가 PLGA 단독으로 사용하였을 때와 비교하여 볼 때 디스크 조직의 형성이 우수하였으며, 특히 20, 40%의 DBP가 함유된 지지체가 세포의 성장과, 디스크 조직화에 유리함을 확인하였다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제31권6호
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• pp.478-484
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• 2009

Three-dimensional porous scaffolds play an important role in tissue engineering strategies. They provide a void volume in which vascularization, new tissue formation, and remodeling can occur. Like any grafted materials, the ideal scaffold for bone tissue engineering should be biocompatible without causing an inflammatory response. It should also possess biodegradability, which provides a suitable three-dimensional environment for the cell function together with the capacity for gradual resorption and replacement by host bone tissue. Various scaffolds have already been developed for bone tissue engineering applications, including naturally derived materials, bioceramics, and synthetic polymers. The advantages of biodegradable synthetic polymers include the ability to tailor specific functions. The purpose of this study was to examine the osteogenic activity of periosteal-derived cells in a polydioxanone/pluronic F127 scaffold. Periosteal-derived cells were successfully differentiated into osteoblasts in the polydioxanone/pluronic F127 scaffold. ALP activity showed its peak level at 2 weeks of culture, followed by decreased activity during the culture period. Similar to biochemical data, the level of ALP mRNA in the periosteal-derived cells was also largely elevated at 2 weeks of culture. The level of osteocalcin mRNA was gradually increased during entire culture period. Calcium content was detactable at 1 week and increased in a time-dependent manner up to the entire duration of culture. Our results suggest that polydioxanone/pluronic F127 could be a suitable scaffold of periosteal-derived cells for bone tissue engineering.

• 치위생과학회지
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• 제19권4호
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• pp.254-260
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• 2019

Background: The primary aims of periodontal disease treatment is to remove dental plaque and calculus, the main causes of tooth loss, and restore periodontal tissue destroyed by inflammation. Periodontal disease treatment should also help maintain the alveolar bone, alleviate inflammation, and promote periodontal ligament cell proliferation, which is essential for tissue regeneration. Conventional antibiotics and anti-inflammatories have adverse side effects, especially during long-term use, so there is a need for adjunct treatment agents derived from natural products. The purpose of this study was to investigate whether the herbal flavone baicalein has the osteogenic activity under inflammatory conditions, and assess the involvement of osteoblast immediate early response 3 (IER3) expression. Methods: Human osteoblastic MG-63 cells were cultured with the pro-inflammatory cytokines tumor necrosis factor α and interleukin 1β in the presence and absence of baicalein. Proliferation was assessed using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, and expression of IER3 mRNA was assessed using real-time polymerase chain reaction. The expression of IER3 protein levels and activation of associated signal transduction pathways were assessed using western blotting. Results: Baicalein increased IER3 mRNA and protein expression synergistically. In addition, baicalein reversed the suppression of cell proliferation, and the downregulation of osteogenic transcription factor runt-related transcription factor 2 and osterix induced by pro-inflammatory cytokines. Baicalein also upregulated the phosphorylation of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK 1/2). The upregulation of IER3 by pro-inflammatory cytokines was blocked by pretreatment with inhibitors of AKT, p38, JNK, and ERK 1/2. Conclusion: Baicalein mitigates the deleterious responses of osteoblasts to pro-inflammatory cytokines. Further, IER3 enhanced the effect of baicalein via activation of AKT, p38, JNK, and ERK pathways.

• Archives of Plastic Surgery
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• 제35권3호
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• pp.229-234
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• 2008

Purpose: There are some reports presenting that peripheral blood contain circulating hematopoietic cells as well as, in significantly smaller quantities, mesenchymal stem cells. The purposes of this study is to isolate and characterize circulating mesenchymal progenitor cells with osteogenic potential from human peripheral blood. Methods: Human buffycoat containing mononuclear cells was harvested from peripheral blood of normal persons and isolated using a density gradient centrifugation and serially subcultured in osteogenic media for 1-4 weeks. The proliferation capability, phase-contrast microscopy, transmission electron microscopy, immunophenotype FACS analysis, Alizarin red staining and RT-PCR assays for osteogenic differentiation potential were performed. Results: The phenotype of cultured cells changed from small round or cuboidal cells at passage 1 into large spindle-shaped fibroblastic morphology cells at passage 4. Surface marker expressed CD14, but did not express CD34, CD80, CD83. Strong positive staining was observed for Alizarin reds in osteogenic medium on day 14, Using RT-PCR, the mRNA levels of bone- specific genes, such as ALP, c-bfa-1 and osteocalcin were detected. Conclusion: A new subset of peripheral blood derived progenitor cells described here has the ability to proliferate and differentiate into osteogenic cell lineages in vitro, and to be candidate for regenerative therapy.

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.274-279
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• 2015

Receptor activator of nuclear factor-kappa B ligand (RANKL) is a critical factor in osteoclastogenesis. It makes osteoclasts differentiate and multinucleate in bone remodeling. In the present study, RANKL was expressed as a soluble maltose binding protein (MBP)-fusion protein using the Escherichia coli maltose binding domain tag system (pMAL) expression vector system. The host cell E. coli DH5α was cultured and induced by isopropyl β-_D-1-thiogalactopyranoside for rRANKL expression. Cells were disrupted by sonication to collect soluble MBP-fused rRANKL. The MBP-fusion rRANKL was purified with MBP Trap affinity chromatography and treated with Tobacco Etch Virus nuclear inclusion endopeptidase (TEV protease) to remove the MBP fusion protein. Dialysis was then carried out to remove binding maltose from the cleaved rRANKL solution. The cleaved rRANKL was purified with a second MBP Trap affinity chromatography to separate unsevered MBP-fusion rRANKL and cleaved MBP fusion protein. The purified rRANKL was shown to have biological activity by performing in vitro cell tests. In conclusion, biologically active rRANKL was successfully purified by a simple two-step chromatography purification process with one column.