• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.223-223
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• 2005

• Membrane Journal
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• v.11 no.1
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• pp.29-37
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• 2001

본 논문은 인공 진피와 조직공학용 scaffold로 이용하기 위해 다공성 membrane로서 gelatin-based sponge의 효율성을 연구하였다. 불용성의 다공성 membrane은 1-ethyl-(3-3dimethylaminopropyl)carbodiimide(EDC)로 가교하여 제조하였다. Fourier-transformed infrared (FT-IR) spectroscopy, scanning electron microscopy(SEM) 그리고 Instron analysis로 다공성 membrane의 특성을 조사하였다. 다공성 membrane은 용적당 큰 표면적을 제공하는 micro porous한 구조를 가지고 있다. Gelatin/hyaluronic acid (HA) membrane의 공경크기는 40~200$\mu\textrm{m}$이다. HA의 첨가는 다공성 membrane의 기계적 강도와 세포부착능력에 영향을 미쳤다. Gelatin/HA 다공성 membrane의 압축강도는 collagen과 비슷하며, 세포배양과 인공진피 transplantation에 있어서의 충분한 기계적 강도를 가지고 있다. Fibroblasts를 함유한 진피기질을 제조하기 위해 직경 8mm의 다공성 membran에 4$\times$10(sup)5cells/membrane의 세포밀도로 fibroblast를 배양하였다. GH91 porous membrane에서의 fibroblast 부착성은 GH55 porous membrane에서보다 우수하였다. 삼차원 구조의 gelatin/HA membrane matrix에서의 fibroblast의 배양은 생체내 조건과 유사한 생리적 환경을 제공하였다.

• Proceedings of the KAIS Fall Conference
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• 2011.12a
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• pp.330-333
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• 2011

Allomatrix (Wright Medical Tech, Inc., USA), is a newly designed, injectable putty with a reliable demineralized bone matrix(DBM), derived from human bone. The compound contains 86% DBM and other bone growth factors such as bone morphogenic protein (BMP)-2, BMP-4, insulin-like growth factor (IGF)-1, and transforming growth factor (TGF)-${\beta}1$. It has excellent os-teoinduction abilities. In addition, DBM is known to have osteoconduction capacity as a scaffold due to its collagen matrix. This product contains a powder, which is a mix of DBM and surgical grade calcium sulfate as a carrier. A practitioner can blend the powder with calcium sulfate solution, making a putty-type material which has the advantages of ease of handling, better fixation, and no need for a membrane, because it can function as membrane itself. This study reports the clinical and radiographic results of various guided bone regeneration cases using Allomatrix, demonstrating its strong potential as a graft material.

• Journal of Biomedical Engineering Research
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• v.29 no.1
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• pp.52-58
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• 2008

The object of this study is to investigate the effects of intermittent cyclic stretching on the smooth muscle cells (SMCs) seeded onto aligned multi-layered fibrous scaffold. To make multi-layered fibrous scaffold, polyurethane (PU) and poly(ethylene oxide) (PEO) were electrospun alternatively, then were immersed into distilled water to extract PEO. Various types of scaffolds were fabricated depending on fiber directions, i.e., aligned or randomly oriented. The direction of stretching was either parallel or vertical to the fiber direction for the aligned scaffolds. The stretching was also applied to the randomly aligned scaffolds. The duration of stretching was 2 min with 15 min resting period. During the stretching, the maximum and minimum strain was adjusted to be 10 and 7%, respectively with the frequency of 1 Hz. The bioactivities of cells on the scaffolds were assessed by quantifying DNA, collagen, and glycosaminoglycan (GAG) levels. And the cell morphology was observed by staining F-actin. SMCs under parallel stretching to the fiber direction responded more positively than those in other conditions. From the results, we could explain the morphological effect of a substrate on cellular activities. In addition the synergistic effects of substrate and mechanical stimuli effects were confirmed.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.8
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• pp.618-626
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• 2019

Stem cell therapy is not expected to bestow any therapeutic benefit because of the low engraftment rates after transplantation.Various cell-carrying scaffolds have been developed in order to overcome this problem. When the scaffold is formed by 3-dimensional (3D) printing, it is possible to create various shapes of scaffolds for specific regions of injury. At the same time, scaffolds provide stem cells as therapeutic-agents and mechanically support an injured region. PCL is not only cost effective, but it is also a widely used material for 3D printing. Therefore, rapid and economical technology development can be achieved when PCL is printed and used as a cell carrier. Yet PCL materials do not perform well as cell carriers, and only a few cells survive on the PCL surface. In this study, we tried to determine the conditions that maximize the cell-loading capacity on the PCL surface to overcome this issue. By applying a plasma treated condition and then collagen coating known to improve the cell loading capacity, it was confirmed that the 3% collagen coating after plasma treatment showed the best cell engraftment capacity during 72 hours after cell loading. By applying the spheroid cell culture method and scaffold structure change, which can affect the cell loading ability, the spheroid cell culture methods vastly improved cell engraftment, and the scaffold structure did not affect the cell engraftment properties. We will conduct further experiments using PCL material as a cell carrier and as based the excellent results of this study.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.11
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• pp.1492-1498
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• 2009

In the dermis, fibroblast plays an important role in the turnover of the dermal extracellular matrix. Collagen I and III, which are the most important dermal proteins of the extracellular matrix, function as a stabilizing scaffold of dermal connective tissues, as well as a regulator of differentiation and migration of dermal cells. In this study, we investigated the effect of various nutrients, such as ascorbic acid, silicon, Fe, lysine and proline which function as cofactors or building blocks on collagen synthesis. When the physiological concentrations of ascorbic acid (0-100 ${\mu}M$), silicon (0-50 ${\mu}M$), Fe (0-50 ${\mu}M$), lysine (0-150 ${\mu}M$) and proline (0-300 ${\mu}M$) were treated at HS27 for either 3 or 5 days, 5 day treatment of ascorbic acid at the low concentration (5-10 ${\mu}M$) increased the expression of collagen I and III protein by 115-1300% without increasing cell proliferation. 3 or 5 days treatment of Fe increased the expression of collagen I and III proteins up to 323% in parallel with cell proliferation by 164%. However, cell proliferation and expression of collagen I and III protein in silicon treated HS27 did not differ. Proline and lysine only increased cell proliferation up to 247.9%. Taken together, we demonstrate that the physiological concentrations of ascorbic acid and Fe enhance the expression of collagen I and III protein for treatment of 3 or 5 days.

• Archives of Plastic Surgery
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• v.33 no.1
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• pp.46-52
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• 2006

High-density micromass culture was needed to take three dimensions culture with ASCs(adipose derived stromal cells) and chondrogenesis. However, the synthetic polymer has hydrophobic character and low affinity to cells and other biomolecules. Therefore, the surface modification without changes of physical and chemical properties is necessary for more suitable condition to cells and biomolecules. This study was performed to investigate the effect of surface modification of poly (lactic-co-glycolic acid)(PLGA) scaffold by plasma treatment (P(+)) on the adhesion, proliferation and chondrogenesis of ASCs, and not plasma treatment (P(-)). ASCs were isolated from human subcutaneous adipose tissue obtained by lipectomy and liposuction. At 1 hour 30 minutes and 3days after cell seeding onto the P(-) group and the P(+) group, total DNA amount of attached and proliferated ASCs markedly increased in the P(+) group (p < 0.05). The changes of the actin under confocal microscope were done for evaluation of cellular affinity, at 1 hour 30 minutes, the shape of the cells was spherical form in all group. At 3rd day, the shape of the cells was fiber network form and finely arranged in P(+) group rather than in P(-) group. RT-PCR analysis of cartilage-specific type II collagen and link protein were expressed in 1, 2 weeks of induction. Amount of Glycoaminoglycan (GAG) markedly increased in P(+) group(p < 0.05). In a week, extracellular matrix was not observed in the Alcian blue and Safranin O staining. However in 2 weeks, it was observed that sulfated proteoglycan increased in P(+) group rather than in P(-) group. In conclusion, we recognized that plasma treatment of PLGA scaffold could increase the hydrophilic property of cells, and provide suitable environment for high-density micromass culture to chondrogenesis

• Polymer(Korea)
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• v.37 no.2
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• pp.141-147
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• 2013

Poly(lactide-co-glycolic acid) (PLGA) has been widely used in the drug delivery and tissue engineering applications because of its good mechanical strength and biodegradation profile. However, cell attachment to the scaffold is low compared with that on fibrin although cells can be attached to the polymer surface. In this study, PLGA scaffolds were soaked in cells-fibrin suspension and polymerized with dropping fibrinogen-thrombin solution. Cellular proliferation activity was observed in PLGA/fibrin-seeded costal cartilage cells (CC) on 1, 3, and 7 days using the MTT assay and SEM. The effects of fibrin on the extracellular matrix (ECM) formation were evaluated using CC cell-seeded PLGA/fibrin scaffolds. The PLGA/fibrin scaffolds elicited more production of glycosaminoglycan (GAG) and collagen than the PLGA scaffold. In this study, fibrin incorporated PLGA scaffolds were prepared to evaluate the effects of fibrin on the cell attachment and proliferation in vitro and in vivo. In this result, we confirmed that proliferation of cells in PLGA/fibrin scaffolds were better than in PLGA scaffolds. The PLGA/fibrin scaffolds provide suitable environment for growth and proliferation of costal cartilage cells.

• 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.

• Design & Manufacturing
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• v.15 no.2
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• pp.11-16
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• 2021

Recently, three-dimensional (3D) cell culture systems, which are superior to conventional two-dimensional (2D) vascular systems that mimic the in vivo environment, are being actively studied to reproduce drug responses and cell differentiation in organisms. Conventional two-dimensional cell culture methods (scaffold-based and non-scaffold-based) have a limited cell growth rate because the culture cannot supply the culture medium as consistently as microvessels. To solve this problem, we would like to propose a 3D culture system with an environment similar to living cells by continuously supplying the culture medium to the bottom of the 3D cell support. The 3D culture system is a structure in which microvascular structures are combined under a scaffold (agar, collagen, etc.) where cells can settle and grow. First, we have manufactured molds for the formation of four types of microvessel-mimicking chips: width / height ①100 ㎛ / 100 ㎛, ②100 ㎛ / 50 ㎛, ③ 150 ㎛ / 100 ㎛, and ④ 200 ㎛ / 100 ㎛. By injection molding, four types of microfluidic chips were made with GPPS (general purpose polystyrene), and a 100㎛-thick PDMS (polydimethylsiloxane) film was attached to the top of each microfluidic chip. As a result of observing the flow of the culture medium in the microchannel, it was confirmed that when the aspect ratio (height/width) of the microchannel is 1.5 or more, the fluid flows from the inlet to the outlet without a backflow phenomenon. In addition, the culture efficiency experiments of colorectal cancer cells (SW490) were performed in a 3D culture system in which PDMS films with different pore diameters (1/25/45 ㎛) were combined on a microfluidic chip. As a result, it was found that the cell growth rate increased up to 1.3 times and the cell death rate decreased by 71% as a result of the 3D culture system having a hole membrane with a diameter of 10 ㎛ or more compared to the conventional commercial. Based on the results of this study, it is possible to expand and build various 3D cell culture systems that can maximize cell culture efficiency by cell type by adjusting the shape of the microchannel, the size of the film hole, and the flow rate of the inlet.