• 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.

• Journal of Oral Medicine and Pain
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• 제37권2호
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• pp.125-133
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• 2012

측두하악관절 골관절염은 관절조직에 가해지는 과부하에 의해 관절면과 하부 골조직에서 일련의 퇴행성과정과 함께 관절부의 국소적 압통, 하악운동에 의한 관절의 염발음, 하악의 운동범위 제한, 심한 경우에는 과두지지가 없어져 구치부 과다접촉과 전치부 개교합이 나타날 수 있는 퇴행성질환이다. 일반적으로는 임상검사 후 파노라마방사선사진, 측두하악관절 파노라마, 경두개방사선사진을 촬영한 뒤 진단하게 되는데 이들 일반적인 방사선사진들은 질환 초기의 미세한 골변화를 평가하는데 제한적이다. 따라서 본 연구에서는 일반방사선학적 검사에서 골조직의 퇴행성변화가 의심되어 확진을 위해 단층촬영 및 골스캔 검사를 시행한 환자의 영상의학적 검사소견들 간의 일치도와 임상소견과의 관계를 평가하여 골관절염의 진단에서 이들 다양한 검사법들의 한계와 그 유용성을 평가하고 향후 추가연구를 위한 예비자료를 얻기위해 시행하였다. 골관절염의 진단에서 일반방사선사진과 단층촬영, 골스캔 검사의 결과 일치도가 비교적 높게 나타났지만 위음성 가능성이 존재하므로 임상검사소견에 따른 단계적인 영상의학적 검사 시행이 바람직할 것으로 사료된다.

• Journal of Periodontal and Implant Science
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• 제37권4호
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• pp.859-869
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• 2007

Naked DNA and standard vectors have been previously used for gene delivery. Among these, PEI can efficiently condense DNA and has high intrinsic endosomal activities. The aim of this study is to investigate whether the cationic polycation PEI could increase the transfection efficiency of BMP expressing DNA using a vector-loaded collagen sponge model. BMP-2/pcDNA3.1 plasmid was constructed by subcloning human BMP-2 cDNA into the pcDNA3.1 plasmid vector. PEI/DNA complexes were prepared by mixing PEI and BMP-2/pcDNA3.1 and the constructed complexes were loaded into the collagen sponges. In vitro studies, BMSCs were transfected with the PEI/BMP-2/pcDNA3.1 complexes from collgen sponge. The level of secreted BMP-2 and alkaline phosphatase activities of transfected BMSCs were significantly higher in PEI/BMP-2/pcDNA3.1 group than in BMP-2/pcDNA3.1 group (p<0.05). Transfected BMSCs were cultured and mineralization was observed only in cells treated with PEI/BMP-2/pcDNA3.1 complexes. In vivo studies, PEI/BMP-2/pcDNA3.1/collagen, BMP-2/pcDNA3.1/collagen and blank collagen were grafted in skeletal muscle of nude mice. Ectopic bone formation was shown in PEI/BMP-2/pcDNA3.1/collagen grafted mouse 4 weeks postimplantation, while not in BMP-2/pcDNA3.1 grafted tissue. This study suggests that PEI-condensed DNA encoding for BMP-2 is capable of inducing bone formation in ectopic site and might increase the transfection rate of BMP-2/pcDNA3.1. As a non-viral vector, PEI offers the potential in gene therapy for bone engineering.

• Asian Pacific Journal of Cancer Prevention
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• 제16권2호
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• pp.571-578
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• 2015

Prostate cancer is the most common cancer in men. In this study, we investigated immune responses of cytotoxic T lymphocytes (CTLs) against TRAMP-C2 prostate cancer cells after activation by dendritic cells (DCs) loaded with TRAMP-C2 freeze-thaw antigen and/or PEP-3 peptide in vitro. Bone marrow-derived DC from the bone marrow of the C57BL/6 were induced to mature by using the cytokine of rhGM-CSF and rhIL-4, and loaded with either the freeze-thaw antigen or PEP-3 peptide or both of them. Maturation of DCs was detected by flow cytometry. The killing efficiency of the CTLs on TRAMP-C2 cells were detected by flow cytometry, CCK8, colony formation, transwell migration, and wound-healing assay. The levels of the IFN-${\gamma}$, TNF-${\beta}$ and IL-12 were measured by enzyme-linked immunosorbent assay (ELISA). Compared with the unloaded DCs, the loaded DCs had significantly increased expression of several phenotypes related to DC maturation. CTLs activated by DCs loaded with freeze-thaw antigen and PEP-3 peptide had more evident cytotoxicity against TRAMP-C2 cells in vitro. The secretion levels of IFN-${\gamma}$, TNF-${\beta}$ and IL-12, secreted by DCs loaded with antigen and PEP-3 and interaction with T cells, were higher than in the other groups. Our results suggest that the CTLs activated by DCs loaded with TRAMP-C2 freeze-thaw antigen and PEP-3 peptide exert a remarkable killing efficiency against TRAMP-C2 cells in vitro.

• BMB Reports
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• 제43권1호
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• pp.52-56
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• 2010

Silk fibroin, produced by the silkworm Bombyx mori, has been widely studied as a scaffold in tissue engineering. Although it has been shown to be slowly biodegradable, cellular responses to degraded silk fibroin fragments are largely unknown. In this study, silk fibroin was added to MG-63 cell cultures, and changes in gene expression in the MG-63 cells were screened by DNA microarray analysis. Genes showing a significant (2-fold) change were selected and their expression changes confirmed by quantitative RT-PCR and western blotting. DNA microarray results showed that alkaline phosphatase (ALP), collagen type-I alpha-1, fibronectin, and transforming growth factor-${\beta}1$ expressions significantly increased. The effect of degraded silk fibroin on osteoblastogenic gene expression was confirmed by observing up-regulation of ALP activity in MG-63 cells. The finding that small fragments of silk fibroin are able to increase the expression of osteoblastogenic genes suggests that controlled degradation of silk fibroin might accelerate new bone formation.

• 대한치과보철학회지
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• 제29권1호
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• pp.167-213
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• 1991

The successful replacement of missing teeth has been one driving aim behind the emergence of implant dentistry as both a technology and clinical vocation for over four decades. To date, a multitude of dental implant devices had been designed and utilized in the patient population. Most of these devices have been designed without support of the engineering criteria. The long-term success of any dental implant is dependent upon the optimization of stresses which occurs during oral function and parafunction. Although many studies have examined the biologic interactions between dental implants and living tissue, few studies have been reported on the biomechanical aspects of dental implants. The purpose of this study was to analyze the stress distribution of osseointegrated prosthesis on certain conditions, such as amount of load, location of load, length of fixtures, number of fixtures used, arch shape, bone quality, etc. Three dimentional finite element analysis was used for this study. FEM models were created using commercial software(Super SAP. for IBM 16 bit AT computer. All elements were 8-node brick, isoparametric. Mandible and prosthesis was modeled with 780 elements and 1074 nodes. The results were as follows : 1. In case of cantilever extension, there was a compressive stress at the base of the first implant and a tensile stress at the base of the second implant. 2. The stresses were linearly proportional to the amount of load. 3. The stresses were linearly proportional to the length of cantilever. 4. There was a stress concentration at the neck of the implant and bone under horizontal loads.

• Macromolecular Research
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• 제13권4호
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• pp.285-292
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• 2005

We developed alginate beads loaded with transforming growth $factor-{\beta}_{1}(TGF-{\beta}_{1})$ to examine the possible application of the scaffold and cytokine carrier in tissue engineering. In this study, bone marrow stromal cells (BMSCs) and $TGF{\beta}_{1}$ were uniformly encapsulated in the alginate beads and then cultured in vitro. The cell morphology and shape of the alginate beads were observed using inverted microscope, scanning electron microscope (SEM), histological staining and RT-PCR to confirm chondrogenic differentiation. The amount of the $TGF{\beta}_{1}$ released from the $TGF-{\beta}_{1}$ loaded alginate beads was analyzed for 28 days in vitro in a phosphate buffered saline (pH 7.4) at $37^{\circ}C$. We observed the release profile of $TGF-{\beta}_{1}$ from $TGF-{\beta}_{1}$ loaded alginate beads with a sustained release pattern for 35 days. Microscopic observation showed the open cell pore structure and abundant cells with a round morphology in the alginate beads. In addition, histology and RT-PCR results revealed the evidence of chondrogenic differentiation in the beads. In conclusion, these results confirmed that $TGF-{\beta}_{1}$ loaded alginate beads provide excellent conditions for chondrogenic differentiation.

• 한국표면공학회:학술대회논문집
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• 한국표면공학회 2016년도 추계학술대회 논문집
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• pp.195-195
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• 2016

Titanium and its alloys are widely used as implants in orthopedics, dentistry and cardiology due to their outstanding properties, such as high strength, high level of hemocompatibility and enhanced biocompatibility. Hence, recent works showed that the synthesis of new Ti-based alloys for implant application involves more biocompatible metallic alloying element, such as, Nb, Hf, Zr and Mo. In particular, Nb and Hf are one of the most effective Ti ${\beta}-stabilizer$ and reducing the elastic modulus. Plasma electrolyte oxidation (PEO) is known as excellent method in the biocompatibility of biomaterial due to quickly coating time and controlled coating condition. The anodized oxide layer and diameter modulation of Ti alloys can be obtained function of improvement of cell adhesion. Silicon (Si) and magnesium (Mg) has a beneficial effect on bone. Si in particular has been found to be essential for normal bone and cartilage growth and development. In vitro studies have shown that Mg plays very important roles in essential for normal growth and metabolism of skeletal tissue in vertebrates and can be detected as minor constituents in teeth and bone. The aim of this study is to research Si and Mg doped hydroxyapatite film formation by plasma electrolytic oxidation. Ti-29Nb-xHf (x= 0, 3, 7 and 15wt%, mass fraction) alloys were prepared Ti-29Nb-xHf alloys of containing Hf up from 0 wt% to 15 wt% were melted by using a vacuum furnace. Ti-29Nb-xHf alloys were homogenized for 2 hr at $1050^{\circ}C$. Each alloy was anodized in solution containing typically 0.15 M calcium acetate monohydrate + 0.02 M calcium glycerophosphate at room temperature. A direct current power source was used for the process of anodization. Anodized alloys was prepared using 270V~300V anodization voltage at room. A Si and Mg coating was produced by RF-magnetron sputtering system. RF power of 100W was applied to the target for 1h at room temperature. The microstructure, phase and composition of Si and Mg coated oxide surface of Ti-29Nb-xHf alloys were examined by FE-SEM, EDS, and XRD.

• 한국결정성장학회지
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• 제27권5호
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• pp.235-242
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• 2017

기공 제어가 가능한 다공질 인공 지지체를 제조하기 위해 HA 분말에 기공형성제 역할을 하는 PMMA 분말을 첨가하여 TBA를 용매로 한 slurry를 합성한 후 동결주조와 소결을 거쳐 주상형 기공채널이 상호 연결되어 있는 다공질 HA 지지체를 제조하였다. PMMA 분말의 첨가량에 따른 HA 지지체의 결정구조는 XRD로 측정하였고 SEM을 통하여 지지체의 표면 및 내부 단면을 관찰하였는데, 소결과정에서 PMMA의 탈지가 지지체의 내부구조와 HA 분말의 결정성에 영향을 미치는 것으로 결과가 나타났다. 또한 지지체의 물리적 및 기계적 특성을 평가하여 기공형성제의 첨가량을 조절함으로써 기공률 및 기공 크기와 압축 강도의 제어가 가능하였다. 본 연구 결과, HA 지지체가 천연 해면골과 구조 및 특성이 유사하였으며 이를 통해 PMMA 첨가 다공질 HA 지지체가 조직공학용 인공 골지지체로서 자가골을 대체하여 사용이 가능한 것으로 판단된다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.98-100
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• 2003

합성고분자(PET, ePTFE)로 제작된 기존의 혈관 패치는 혈전생성으로 인한 혈관 막힘 현상과 생체 비적합성으로 인한 석회화 현상 때문에 장기간 혈관 기능을 수행할 수 없다. 본 연구에서는 혈관조직을 형성하는 세포들로 분화가 가능한 중간엽줄기세포와 생체적합성 매트릭스를 이용하여 혈관용 패치를 개발하였다. 이식 후 3주에 관찰하였을 때 조직공학적으로 제조된 혈관 패치는 동물 임상시험에서 혈관막힘 현상 없이 혈관기능을 수행하였고 실제 혈관과 유사한 조직으로 재생되었다. 동물모델에서의 장기간 추가 보완 연구를 거친다면 본 연구에서 개발된 혈관 패치는 기존의 재료를 대체하여 많은 혈관질환 치료에 적용이 가능할 것으로 사료된다.