• Journal of Life Science
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• v.33 no.8
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• pp.612-622
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• 2023

T-cell deficiency may occur in various clinical conditions including congenital defects, cell/organ transplantation, HIV infection and aging. In this regard, the development of artificial thymus has recently been attracting much attention. To achieve this aim, the development of techniques for 3D culture of thymic stromal cells is necessary because thymocytes grown only in a 3D thymic microenvironment can be differentiated fully to become mature, immunocompetent T cells; the same cannot be achieved for thymocytes grown in 2D. This study aimed to develop a nanotechnology-based 3D culture technique using polymeric scaffolds for thymic epithelial cells (TECs), the main component of thymic stromal cells. Scanning electron microscopic observation revealed that the pores of both PCL and PCL/PLGA scaffolds were filled with TECs. Interestingly, TECs grown in 3D on polydopamine-coated scaffolds exhibited enhanced cell attachment and proliferation compared to those grown on non-coated scaffolds. In addition, the gene expression of thymopoietic factors was upregulated in TECs cultured in 3D on polydopamine-coated scaffolds compared to those cultured in 2D. Taken together, the results of the present study demonstrate an efficient 3D culture model for TECs using polymeric scaffolds and provide new insights into a novel platform technology that can be applied to develop functional, biocompatible scaffolds for the 3D culture of thymocytes. This will eventually shed light on techniques for the in vitro development of T cells as well as the synthesis of artificial thymus.

• Polymer(Korea)
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• v.36 no.6
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• pp.789-794
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• 2012

Poly(lactic-co-glycolic acid) (PLGA) has been most widely used due to its advantages such as good biodegradability, controllable rate of degradation and metabolizable degradation products. We manufactured composite scaffolds of PLGA scaffold penetrated DBP gel (PLGA/DBP gel) by a simple method, solvent casting/salt leaching prep of PLGA scaffolds and subsequent soaking in DBP gel. Chondrocytes were seeded on the PLGA/DBP gel. The mechanical strength of scaffold, histology (H&E, Safranin-O, Alcian-blue) and immunohistochemistry (collagen type I, collagen type II) were performed to elucidate in vitro and in vivo cartilage-specific extracellular matrices. It was better to keep the characteristic of chondrocytes in the PLGA/DBP gel scaffolds than that PLGA scaffolds. This study suggests that PLGA/DBP gel scaffold may serve as a potential cell delivery vehicle and a structural basis for in vivo tissue engineered cartilage.

• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.213-231
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• 2000

이 연구의 목적은 다공성의 CPP 내부에 쥐의 장골의 골수에서 유래된 세포를 접종하고 3차 원적으로 배양하여 CPP가 골 형성을 위한 조직공학의 지지체로 적용가능한가를 연구하는 것과 Calcium PolyPhosphate(CPP)의 돌연변이 유발성을 검사하는 것이다. 무수 ($Ca(H_2PO_4)$)를 condensation하여 무결정의 ($Ca(PO_3)$)를 얻고 이를 용융하고 냉각시킨 후 분쇄하여 Calcium polyphosphate(CPP) powder를 얻었다. 다공성의 CPP는 5% $SiO_2$를 첨가하여 sponge 형태로 $450-550{\mu}m$ 소공의 크기를 가지는 것과(CPP-45ppi) $200-300{\mu}m$의 소공의 크기를 가지는 것(CCP-60ppi) 2가지로 제작하였다. 각각의 CPP matrices는 $5mm{\times}5mm{\times}1mm$의 블록 형태로 만들었다. 체중 100g 내외의 백서에서 장골(femur, tibia)을 채취하여 백서의 장골 골수 세포를 분리하여 배양한 후 24well에 CPP block을 넣고 CPP block 당 $10^5$개의 배양한 세포를 접종하였다. 배양 1, 7, 14, 및 21 일째에 각 well에서 trypsin EDTA를 이용하여 2회 반복하여 cell을 분리하였고, 원심분리한 후 hemacytometer로 측정하였다. 또, 45ppi와 60ppi, 그 리 고 Tissue Culture Polystyrene(control group)에 접종, 배양된 세포들의 염기성 인산분해효소활성도를 배양 7, 14, 및 21 일째에 각각 측정하였다. 각 기간별로 배양된 세포-CPP 혼합체내에서 세포의 부착 및 증식과 형성된 조직의 3차원적 형태를 관찰하기 위하여 주사전자현미경하에서의 관찰하였다. CPP의 돌연변이 유발성 검사 (mutagenicity test)를 위해 hypoxanthine-guanine phosphoribosyl transferase(HPRT) assay를 하였다. NIH3T3 cell line과 CHO-K1 cell line으로 각각 $1000{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$ 그리고 $1{\mu}g/m{\ell}$의 CPP 농도에서 측정하였다. 통계적 분석을 위해서 모든 측정은 각군당 4개체 이상 시험하였고, 각 측정값은 평균값${\pm}$표준편차로 나타내었다. 각 군간의 통계적 유의성 검정을 위해서 Analysis of variance(ANOVA)를 이용하였고 Tukey의 방법으로 사후분석을 실시하였다. 제작된 CPP matrices 소공들이 서로간에 연결이 잘 되어있는 형태였다. 두 가지로 제조된 CPP(45ppi와 60ppi) 모두에서 세포의 부착이 잘 일어났고, 부착된 세포의 분열도 잘 일어났다. 2 가지의 CPP 모두에서 7, 14, 21일째의 세포 수는 1일째에 비해 유의성 있게 증가하였다(P<0.01). 3차원적 구조인 Calcium PolyPhosphate에서 배양한 세포는 24well dish(tissue culture polystyrene)에서 평면적으로 배양한 대조군의 세포에서 보다 염기성 인산분해효소 (Alkaline Phosphatase)를 유의성 있게 높게 나타냈다. 주사전자현미경에서 세포-CPP 혼합체를 관찰한 결과, CPP block에 세포들이 잘부착되어 있었고, 시간이 지남에 따라 세포가 여러 층을 형성하면서 뭉치는 현상을 보였다. 또, HPRT assay 결과 , Calcium PolyPhosphate는 돌연변이 유발성을 보이지 않았다. 이상의 결과로 볼 때 CPP에는 세포부착이 잘 일어나고, 지지체 상에서 세포의 분열도 활발하게 일어나므로 골조직을 위한 조직공학의 우수한 지지체가 될 수 있을 것으로 사료된다.

• Journal of the KSME
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• v.52 no.12
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• pp.49-53
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• 2012

이 글에서는 조직공학에 사용되는 재료들의 기계적 특성에 대해 간략하게 정리하였다. 당연하게도, 각각의 재료들은 그 기계적/화학적 특성에 맞는 제작방법이 필요하며, 이러한 기계적/화학적 특성들은 타겟 장기/조직이 필요로 하는 그것과 유사해야 한다.

• Polymer(Korea)
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• v.37 no.5
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• pp.632-637
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• 2013

It has been widely accepted that costal cartilage cells (CCs) have more excellent initial proliferation capacity than articular cartilage cells as well as the easiness for isolation and collection. This study demonstrated that CCs might be one of the substitutes for articular cartilage cells by tissue engineered cartilage. Poly(lactic-co-glycolic acid) (PLGA) has been extensively tested and used as scaffold material but it was limited by the low attachment of cells and the induction of inflammatory cells. Base on previous our studies, we confirmed demineralized bone particle (DBP) had the power of the reduction of inflammatory reaction and the stimulation proliferation of cells. We fabricated PLGA scaffold loaded with 10, 20, 40 and 80 wt% DBP and then tested the possibility of the regeneration of cartilage using CCs. Assays of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and scanning electron microscope (SEM) carried out to evaluate the attachment and proliferation of CCs in DBP/PLGA scaffolds. Glycosaminoglycan (sGAG) and collagen contents assay were conducted to confirm the effects of DBP on formation of extracellular matrix. This study demonstrated that DBP/PLGA scaffolds showed significant positive effects on cell growth and proliferation due to the vitality of DBP as well as the possibility of the application of CCs for tissue engineered cartilage.

• Polymer(Korea)
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• v.38 no.2
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• pp.150-155
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• 2014

Polycaprolactone (PCL) is a synthetic biodegradable polymer with excellent mechanical properties. $TiO_2$ (titanium dioxide) has a hydrophilic, high density and excellent biocompatibility. In this work, we produced three-dimensional porous scaffolds with PCL and $TiO_2$ nanoparticles using a salt-leaching method. Physical properties of the scaffolds were analyzed by FE-SEM, FTIR, TGA and compressive strength. Interestingly, the addition of $TiO_2$ nanoparticles decreased the water absorption and swelling ratio of the porous scaffolds. However, the compressive strength was increased by $TiO_2$. CCK-8 assay, which is generally used for the analysis of cell growth, shows that $TiO_2$ nanoparticles have no cytotoxicity. Taken together, we suggest that the PLC/$TiO_2$-scaffold can be used for biomedical applications.

• Polymer(Korea)
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• v.32 no.5
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• pp.458-464
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• 2008

A successful scaffold should have a highly porous structure and good mechanical stability. High porosity and appropriate pore size provide structural matrix for initial cell attachment and proliferation enabling the exchange of nutrients between the scaffold and environment. In this paper the highly porous scaffold of poly(${\varepsilon}$-caprolactone) electrospun nanofibers could be manufactured with an auxiliary electrode and chemical blowing agent (BA) under several processing conditions, such as the concentration of PCL solution, weight percent of a chemical blowing agent, and decomposition time of a chemical blowing agent. To attain stable electrospinnability and blown nanofiber mats having high microporosity and large pore, a processing condition, 8wt% of PCL solution and 0.5wt% of a chemical blowing agent under $100^{\circ}C$ and decomposition time of $2{\sim}3\;s$, was used. The growth characteristic of human dermal fibroblasts cells cultured in the mats showed the good adhesion and proliferation on the blown mat compared to a normal electrospun mat.

• Journal of the Korean Society of Manufacturing Process Engineers
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• v.18 no.2
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• pp.44-51
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• 2019

With the goal of tissue regeneration for organs damaged through an accident or a disease, research on tissue engineering has been conducted to produce 3-D scaffolds that can support the cells in the attachment and growth for the cell proliferation and differentiation. A scaffold requires a suitable pore size and porosity to increase the nutrient circulation or oxygen supply for the attachment and growth of cells. The existing production methods such as solvent-casting particulate leaching, phase separation, and fiber bonding have certain disadvantages. With these methods, it is difficult to obtain a free desired shape. In addition, certain pore sizes and interconnectivities among the pores may not be guaranteed. To solve these problems, this study has fabricated a scaffold with a 3-D shaped nose using Alginate, which is a natural polymer obtained through Fused Deposition Modeling (FDM), one of the CAD/CAM-based Solid Freeform Fabrication (SFF) methods.

• Transactions of the Korean Society of Mechanical Engineers A
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• v.40 no.10
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• pp.865-871
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• 2016

In recent years, traditional scaffold fabrication techniques such as gas foaming, salt leaching, sponge replica, and freeze casting in tissue engineering have significantly limited sufficient mechanical property and cell interaction effect due to only random pores. Fused deposition modeling is the most apposite technology for fabricating the 3D scaffolds using the polymeric materials in tissue engineering application. In this study, 3D slurry mould was fabricated with a blended biphasic calcium phosphate (BCP)/Silica/Alginic acid sodium salt slurry in PCL mould and heated for two hours at $100^{\circ}C$ to harden the blended slurry. 3D dual-pore BCP/Silica scaffold, composed of macro pores interconnected with micro pores, was successfully fabricated by sintering at furnace of $1100^{\circ}C$. Surface morphology and 3D shape of dual-pore BCP/Silica scaffold from scanning electron microscopy were observed. Also, the mechanical properties of 3D BCP/Silica scaffold, according to blending ratio of alginic acid sodium salt, were evaluated through compression test.

• Applied Microscopy
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• v.31 no.4
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• pp.325-331
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• 2001

Integumentary structures of the stone flounder, Karefus bicoloratus were examined by means of the light and transmission electron microscopy. Stratified epidermal layer consists of supporting cells, unicellular glands and granular cells. The epidermal layer could be classified into superficial, intermediated and basal layer by morphology and structure of the supporting cells . The cytoplasm of supporting cells is divided into cortex and medullar part. In the cortex microfilaments are well developed. Mucous cells of unicellular gland were observed in the superficial and intermediated layer of the epidermis. The mucous materials were identified as glycoprotein of neutral and carboxylated mucosubstance by histochemical methods. Club cell has well developed smooth endoplasmic reticula and Golgi complex in the cytoplasm. Granular cells were observed in the intermediated and basal layer, and the cytoplasm is occupied with membrane-bounded granules of electron dense. Three types of pigment cells could be distinguished with electron density of cytoplasmic inclusions. Nerve myelins were observed near the pigment cells.