• 폴리머
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• 제36권4호
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• pp.401-406
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• 2012

KUSA-A1 골조세포로부터 골재생을 유도하기 위하여, 스폰지형(CSS) 및 부직포형(CSNW)의 키토산 지지체를 적용하였다. CSNW의 표면적 및 공극 크기는 CSS에 비해 상대적으로 큰 값을 나타낸 반면, 공극 부피는 CSNW의 경우가 CSS에 비해 작은 값을 보였다. 세포고정 시험 결과는 CSNW의 경우가 더 적합한 결과를 나타내었으며, 이는 지지체의 넓은 표면적에 기인한 것으로 판단되었다. In vivo 실험을 위하여 세포를 각각의 지지체에 투여 후 일주일간 배양하였으며, BALB/C 무모생쥐의 피하조직에 이식하였다. 이식된 지지체는 각각 수술후 1, 4, 6 및 8주에 채취되어 면역학적 염색을 실시하였다. CSS는 수술후 4주에서 6주 사이에 붕괴되기는 하였으나, 조직의 안정성은 CSNW에 비해 우수한 것으로 관찰되었다. 골조직의 생성은 CSNW와 CSS에 대해 각각 4주 및 8주에서 이루어짐을 확인하였다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권6호
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• pp.588-593
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• 2008

There are increasing reports regarding regeneration of the defected tissues using tissue engineering technique. In this technique, multipotential stem cells are essential. There are many potential sources of adult stem cells, such as bone marrow, umbilical cord blood, fat, muscle, dental tissues and skin. Among them, skin is highly accessible and easily obtained with a minimum of donor site complications. Moreover, skin is an abundant adult stem cell sources and has the potential for self-replication and immune privilege. In this study, we isolated skin-derived precursor cells (SKPs) from the ear of adult miniature pigs. In these SKPs, the expression of transcriptional factors, Oct-4, Sox-2, and Nanog were detected by RT-PCR. In vitro osteogenesis and adipogenesis were observed at 3 weeks after transdifferentiations as assayed by positive von Kossa and Oil-red O staining, respectively. In addition, expression of osteocalcin and osteonectin in the osteogenic differentiation medium and $PPAR{\gamma}2$ and aP2 in the adipogenic differentiation medium were detected by RT-PCR. In vitro neurogenesis of porcine SKPs was observed during 24 and 72 hours after treatment of neurogenic differentiation medium. The results of this study suggest that SKPs demonstrate the properties of pluripotence or multipotence and multi-lineage differentiation. This indicates that autogenous SKPs are a reliable and useful source of adult stem cells for regenerative medicine.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2011년도 제41회 하계 정기 학술대회 초록집
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• pp.202-202
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• 2011

The research of 3-dimensional (3-D) scaffold for tissue engineering has been widely investigated as the importance of the 3-D scaffold increased. 3-D scaffold is needed to support for cells to proliferate and maintain their biological functions. Furthermore, its architecture defines the shape of the new bone and cartilage growth. Polycaprolactone (PCL) has been one of the most promising materials for fabricating 3-D scaffold owing to its excellent mechanical property and biocompatibility. However, there are practical problems for using it, in vitro and in vivo; extracellular matrix components and nutrients cannot penetrate into the inner space of scaffold, due to its hydrophobic property, and thus cell seeding and attachment onto the inner surface remain as a challenge. Thus, the surface modification strategy of 3-D PCL scaffold is prerequisite for successful tissue engineering. Herein, we utilized a mussel-inspired approach for surface modification of 3-D PCL scaffold. Modification of 3-D PCL scaffolds was carried out by simple immersion of scaffolds into the dopamine solution and stimulated body fluid, and as a result, hydroxyapatite-immobilized 3-D PCL scaffolds were obtained. After surface modification, the wettability of 3-D PCL scaffold was considerably changed, and infiltration of the pre-osteoblastic cells into the 3-D scaffold followed by the attachment onto the surface was successfully achieved.

• Tissue Engineering and Regenerative Medicine
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• 제15권6호
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• pp.793-801
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• 2018

BACKGROUND: The aim of this study was to evaluate the combined effect of low-level laser treatment (LLLT) and recombinant human bone morphological protein-2 (rhBMP-2) applied to hypoxic-cultured MC3T3-E1 osteoblastic cells and to determine possible signaling pathways underlying differentiation and mineralization of osteoblasts under hypoxia. METHODS: MC3T3-E1 cells were cultured under 1% oxygen tension for 72 h. Cell cultures were divided into four groups: normoxia control, low-level laser (LLL) alone, rhBMP-2 combined with LLLT, and rhBMP-2 under hypoxia. Laser irradiation was applied at 0, 24, and 48 h. Cells were treated with rhBMP-2 at 50 ng/mL. Alkaline phosphatase activity was measured at 3, 7, and 14 days to evaluate osteoblastic differentiation. Cell mineralization was determined with Alizarin red S staining at 7 and 14 days. Western blot assays were performed to evaluate whether p38/protein kinase D (PKD) signaling was involved. RESULTS: The results indicate that LLLT and rhBMP-2 synergistically increased alkaline phosphatase (ALP) activity and mineralization. Western blot analyses showed that expression of type I collagen, runt-related transcription factor 2 (RUNX2), and Osterix (Osx), increased and expression of hypoxia-inducible factor 1-alpha ($HIF-1{\alpha}$), decreased more in the LLLT and rhBMP-2 combined group than in the rhBMP-2 or LLL alone groups. Moreover, LLLT and rhBMP-2 stimulated p38 phosphorylation and rhBMP-2 and LLLT increased Prkd1 phosphorylation. CONCLUSION: Combined treatment with rhBMP-2 and LLL induced differentiation and mineralization of hypoxic-cultured MC3T3-E1 osteoblasts by activating p38/PKD signaling in vitro.

• Tissue Engineering and Regenerative Medicine
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• 제15권6호
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• pp.699-709
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• 2018

BACKGROUND: Diabetes mellitus is a major health concern in current scenario which has been found to affect people of almost all ages. The disease has huge impact on global health; therefore, alternate methods apart from insulin injection are being explored to cure diabetes. Therefore, this review mainly focuses on the current status and therapeutic potential of stem cells mainly mesenchymal stem cells (MSCs) for Type 1 diabetes mellitus in preclinical animal models as well as humans. METHODS: Current treatment for Type 1 diabetes mellitus mainly includes use of insulin which has its own limitations and also the underlying mechanism of diseases is still not explored. Therefore, alternate methods to cure diabetes are being explored. Stem cells are being investigated as an alternative therapy for treatment of various diseases including diabetes. Few preclinical studies have also been conducted using undifferentiated MSCs as well as in vitro MSCs differentiated into ${\beta}$ islet cells. RESULTS: These stem cell transplant studies have highlighted the benefits of MSCs, which have shown promising results. Few human trials using stem cells have also affirmed the potential of these cells in alleviating the symptoms. CONCLUSION: Stem cell transplantation may prove to be a safe and effective treatment for patients with Type 1 diabetes mellitus.

• Journal of Periodontal and Implant Science
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• 제51권3호
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• pp.213-223
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• 2021

Purpose: The atmospheric pressure plasma jet (APPJ) has been introduced as an effective disinfection method for titanium surfaces due to their massive radical generation at low temperatures. Helium (He) has been widely applied as a discharge gas in APPJ due to its bactericidal effects and was proven to be effective in our previous study. This study aimed to evaluate the safety and effects of He-APPJ application at both the cell and tissue levels. Methods: Cellular-level responses were examined using human gingival fibroblasts and osteoblasts (MC3T3-E1 cells). He-APPJ was administered to the cells in the experimental group, while the control group received only He-gas treatment. Immediate cell responses and recovery after He-APPJ treatment were examined in both cell groups. The effect of He-APPJ on osteogenic differentiation was evaluated via an alkaline phosphatase activity assay. In vivo, He-APPJ treatment was administered to rat calvarial bone and the adjacent periosteum, and samples were harvested for histological examination. Results: He-APPJ treatment for 5 minutes induced irreversible effects in both human gingival fibroblasts and osteoblasts in vitro. Immediate cell detachment of human gingival fibroblasts and osteoblasts was shown regardless of treatment time. However, the detached areas in the groups treated for 1 or 3 minutes were completely repopulated within 7 days. Alkaline phosphatase activity was not influenced by 1 or 3 minutes of plasma treatment, but was significantly lower in the 5 minute-treated group (P=0.002). In vivo, He-APPJ treatment was administered to rat calvaria and periosteum for 1 or 3 minutes. No pathogenic changes occurred at 7 days after He-APPJ treatment in the He-APPJ-treated group compared to the control group (He gas only). Conclusions: Direct He-APPJ treatment for up to 3 minutes showed no harmful effects at either the cell or tissue level.

• Macromolecular Research
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• 제17권7호
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• pp.516-521
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• 2009

Techflex$^{(R)}$, a polypropylene-lined, multilayer infusion bag, was studied for its compatibility with diazepam, in comparison to the conventional infusion bag, Safeflex$^{(R)}$, which is comprised of poly(vinyl chloride) (PVC). Diazepam was diluted in 0.9% sodium chloride isotonic solution and stored in the infusion bags for 24 h. To evaluate the sorption of diazepam into the infusion bags during storage, the concentration of the drug remaining in the bag was measured using gas chromatography-mass spectroscopy. The PVC bags exhibited a marked sorption of diazepam, with a drug loss reaching up to 90% of the initial concentration after 24 h of contact, whereas Techflex$^{(R)}$ inhibited the drug sorption, showing approximately 10%, under the same conditions. The differences in the sorption behaviors of the bags are discussed in terms of solubility parameters and crystallinities of the polymers.

• 대한골관절종양학회지
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• 제17권2호
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• pp.65-72
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• 2011

목적: 골종양을 절제한 후 동종골을 이용한 재건술 후 발생한 합병증을 평가하고 그 합병증에 대한 문헌고찰을 하고자 한다. 대상 및 방법: 골종양 절제 후 동종골을 이용한 재건술을 시행한 15예에 대하여 임상적 및 방사선학적 자료를 통해 후향적으로 연구를 시행하였다. 결과: 남자가 8예, 여자가 7예이었으며 평균 나이는 27.1세(1-56세), 평균 추시 기간은 89.5개월(33-146개월)였다. 21예(80.0%)에서 평균 8.35개월(4-12개월)에 방사선학적 골유합 소견을 보였다. Musculoskeletal Tumor Society 점수 평균은 73.5%(46.6-93.0%)였다. 동종골 이식과 관련된 술 후 합병증을 모두 기록하였다. 추시 기간 동안 9예(60.6%)에서 한 가지의 합병증이 발생하였고 3예(20.0%)에서 두 가지 이상의 합병증이 발생하였다. 합병증으로는 감염 3예, 골절 2예, 불유합 2예, 하지 부동 2예, 내반 변형이 2예였다. 합병증이 발생하지 않은 평균 기간은 60.8개월(6-144개월)이었다. 동종골의 평균 생존기간은 80.2개월이었고 5년 생존률은 83.0%였다. 결론: 동종골의 합병증을 줄이기 위하여 동종골을 이용한 재건술시 자가비골을 추가하는 것이 추천된다. 더나아가 조직 공학 기술과 줄기 세포 및 혈소판 풍부 혈장의 적용이 동종골의 재흡수나 불유합 등의 합병증을 줄일 수 있을 것으로 생각된다.

• Nutrition Research and Practice
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• 제11권3호
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• pp.190-197
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• 2017

BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of $50-250{\mu}g/mL$. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to $250{\mu}g/mL$ and were 149% and 129% at $250{\mu}g/mL$ concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of $500{\mu}g/mL$. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.

• Archives of Plastic Surgery
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• 제36권3호
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• pp.247-253
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• 2009

Purpose: Hydroxyapatite(HA) has been widely used due to its chemical similarity to bone and good biocompatibility. HA is composed of macropores and micropores. Too much irregularities of the micropores are ineffective against the adhesion and proliferation of osteoblast. Many efforts have been tried to overcome these drawbacks. HA crystal coating on the irregular surface of HA scaffold, crystallized HA, is one of the method to improve cell adhesion. Meanwhile, the collagen has been incorporated with HA to create composite scaffold that chemically resembles the natural extracellular matrix components of bone. The authors proposed to examine the effect of collagen - coated crystallized HA on the adhesion and proliferation of osteoblast. Method: HA powder containing $10{\mu}m$ pore size was manufactured as 1 cm pellet size. For the making crystallized HA, 0.1 M EDTA solution was used to dissolve HA powder and heated $100^{\circ}C$ for 48 hours. Next, the crystallized HA pellets were coated with collagen (0.1, 0.5, and 1%). The osteoblasts were seeded into HA pellets and incubated for the various times (1, 5, and 9 days). After the indicating days, methylthiazol tetrazolium (MTT) assay was performed for cell proliferation and alkaline phosphatase (ALP) activty was measured for bone formation. Result: In SEM study, the surface of crystallized HA pellet was more regular than HA pellet. MTT assay showed that the proliferation of osteoblasts increased in a collagen dose - dependent and time - dependent manner and had a maximum effect at 1% collagen concentration. ALP activity also increased in a collagen dose - dependent manner and had a highest effect at 1% collagen concentration. Conclusion: These data showed that crystallization and collagen coating of HA was effective for osteoblast proliferation and ALP activity. Therefore, our results suggest that crystallized - HA scaffold with collagen coating is may be a good strategy for tissue engineering application for bone formation.