• Archives of Plastic Surgery
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• 제34권4호
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• pp.413-419
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• 2007

Purpose: In this study, porous type I collagen scaffolds were cross-linked using dehydrothermal(DHT) treatment and/or 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide(EDC), in the presence and absence of chondroitin-6-sulfate(CS) and cultured autologous chondrocytes(Chondro) for cartilage regeneration. Methods: Cartilage defects were created in the proximal part of the ear of New Zealand rabbits. Four prepared types of scaffolds(n=4) were inserted. The groups included Chondro-Collagen-DHT(Group 1), Chondro- Collagen-DHT-EDC(Group 2), Chondro-CS-Collagen- DHT(Group 3), and Chondro-CS-Collagen-DHT-EDC (Group 4). Histomorphometric analysis and cartilage-specific gene expression of the reconstructed tissues were evaluated 4, 8, and 12 weeks after implantation. Results: EDC cross-linked groups 2 and 4 regenerated more cartilage than other groups. However, calcification was observed in the 4th week after implantation. CS did not increase chondrogenesis in all groups. Cartilage-specific type II collagen mRNA expression increased in the course of time in all groups.Conclusion: EDC cross-linking methods maintain the scaffold and promote extracellular matrix production of chondrocytes.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.48-50
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• 2002

Cartilage defects are common and painful conditions that affect people of all ages. Although many techniques have developed, none of the current available treatment options is satisfactory. Recent advances in biology and materials science have pushed tissue engineering to the forefront of new cartilage repair techniques. The purpose of this study is to determine effective regeneration method for tissue-engineered cartilage. A serum free medium was developed for cartilage tissue engineering. Chondrocyte passage number was found to influence greatly on cartilage tissue formation in vivo. Injectable, biodegradable polymer matrix was developed for chondrocyte transplantation through injection. Transplantation of chondrocytes mixed with the injectable matrices resulted in the cartilage formation in nude mice's subcutaneous sites and rabbit knees. This study may lead to the development of tissue-engineered cartilage appropriate for clinical applications.

• Archives of Plastic Surgery
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• 제33권6호
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• pp.723-731
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• 2006

Purpose: Collagen is the principal structural biomolecule in cartilage extracellular matrix, which makes it a logical target for cartilage engineering. In this study, porous type I collagen scaffolds were cross-linked using dehydrothermal(DHT) treatment and/or 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide(EDC), in the presence and absence of chondroitin-6-sulfate(CS) for cartilage regeneration. Methods: Cartilage defects were created in the proximal part of the ear of New Zealand rabbits. Four types of scaffolds(n=4) were inserted. The types included DHT cross-linked(Group 1), DHT and EDC cross- linked(Group 2), CS added DHT cross-linked(Group 3), and CS added DHT and EDC cross-linked(Group 4). Histomorphometric analysis and cartilage-specific gene expression of the reconstructed tissues were evaluated respectively 4, 8, and 12 weeks after implantation. Results: The largest quantity of regenerated cartilage was found in DHT cross-linked groups 1 and 3 in the 8th week and then decreased in the 12th week, while calcification increased. Calcification was observed from the 8th week and the area increased in the 12th week. Group 4 was treated with EDC cross-linking and CS, and the matrix did not degrade in the 12th week. Cartilage-specific type II collagen mRNA expression increased with time in all groups. Conclusion: CS did not increase chondrogenesis in all groups. EDC cross-linking may prevent chondrocyte infiltration from the perichondrium into the collagen scaffold.

• 대한기계학회:학술대회논문집
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• 대한기계학회 2007년도 춘계학술대회A
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• pp.1265-1270
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• 2007

Conventional methods for fabricating three-dimensional (3-D) scaffolds have substantial limitations. In this paper, we present 3-D scaffolds that can be made repeatedly with the same dimensions using a microstereolithography system. This system allows the fabrication of a pre-designed internal structure, such as pore size and porosity, by stacking photopolymerized materials. The scaffolds must be manufactured in a material that is biocompatible and biodegradable. In this regard, we synthesized liquid photocurable biodegradable TMC/TMP, followed by acrylation at terminal ends. And also, solidification properties of TMC/TMP polymer are to be obtained through experiments. Cell adhesion to scaffolds significantly affects tissue regeneration. As a typical example, we seeded chondrocytes on two types of 3-D scaffold and compared the adhesion results. Based on these results, the scaffold geometry is one of the most important factors in chondrocyte adhesion. These 3-D scaffolds could be key factors for studying cell behavior in complex environments and eventually lead to the optimum design of scaffolds for the regeneration of various tissues, such as cartilage and bone.

• 한국정밀공학회:학술대회논문집
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• 한국정밀공학회 2011년도 춘계학술대회 논문집
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• pp.1605-1606
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• 2011

• 대한정형외과학회지
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• 제54권6호
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• pp.475-477
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• 2019

줄기세포를 임상에 이용하고자 하는 시도는 원래 종래의 치료에 반응하지 않는 치명적인 난치질환을 이용하기 위하여 시작되었다. 근골격계질환들은 그로 인하여 사망하지는 않지만 발생빈도가 높고 임상실험이 비교적 용이한 점이 있어 이 분야의 임상적용이 타분야에 비하여 많이 이루어졌다. 그러나 근골격계에서 줄기세포 치료에 의한 재생치료와 관련되어 시험관 내 연구와 전임상 연구가 많은 것과 비교하면 임상적용에 있어 신뢰할 만한 높은 신뢰도를 가진 연구결과는 드물다. 근골격계에서 골, 연골, 건의 재생을 위하여 줄기세포 치료를 적용할 수 있는데 골의 대규모 결손, 대퇴골두 무혈성괴사, 골연골결손, 골관절염, 회전근개 파열등에서 시도되고 있다. 현재까지는 주로 증례보고나 후향적 논문이 대부분이지만 향후 전향적 무작위 비교연구 등 높은 증거능력을 가진 연구 결과들이 나오기를 기대한다.

• 대한기관식도과학회지
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• 제2권1호
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• pp.46-56
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• 1996

Perichondrium is generally used for reconstruction of airway and successful regeneration of cartilage framework using perichondrium are reported by several authors. It has many advantages for airway reconstruction. It can maintain the stable framework and it has higher flexibility so it's easy to design according to the shape of defects. It resist strongly against infection process. Its airtightness and rapid mucosalization enables to predict good postoperative recovery and results. To investigate the differences in cartilage regeneration between avascular and vascularized perichondrial flap, this study was designed with vascularized flaps composed of vascularized perichondrium and central auricular artery and vein. Morphologic study was performed to determine the fate of the grafted perichondrium at regular intervals using light microscopy with H & E stain. Chondrogenic potential and formation of cartilaginous plate of experimental group was superior than in the control group. Grafted perichondrium was fed by consistent feeding vessel. At 6 weeks, the regenerated cartilage could hardly be distinguished form the normal cartilage framework but in control group, regenerated cartilaginous tissue was generally immature in same period. In conclusion, this study indicated that consistent vasculature to grafed tissue is the essential factor for successful reconstruction of cartilaginous framework.

• 한국잠사곤충학회지
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• 제50권1호
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• pp.15-19
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• 2012

최근 다양한 생체재료를 이용하여 연골재생과 관련한 많은 연구가 진행되고 있다. 실크단백질은 생체적합성이 뛰어나며, 우수한 기계적 강도를 가지고 있는 천연 고분자 물질로서 최근 생체재료로 사용하기위한 연구가 세계적으로 많이 이루어지고 있다. 본 연구는 실크단백질이 연골재생에 효과가 있는지를 확인하기위하여 수행되었다. 우리는 연골세포를 코뼈로부터 분리하고, 3종류의 배지 (DMEM, DMEM/F12, RPMI)와 서로 다른 농도의 ascorbic acid를 사용하여 최적 배양조건을 확립하였다. 그 결과 우리가 분리한 연골세포는 10% FBS와 $100{\mu}M$ ascorbic acid가 함유된 DMEM배지에서 가장 잘 생장하였다. 연골에 대한 실크의 영향을 관찰하기위해서, 실크 피브로인 용액을 제작하고 이를 멸균한것과 멸균하지 않은 것으로 구분하여 연골세포 배양 시 첨가하여 연골분화에 대한 마커인자인 제2형 콜라겐의 발현량을 측정하였다. 멸균하지 않은 실크 피브로인 첨가시 제2형 콜라겐의 발현량이 2.7배 증가하였으나, 멸균된 실크 피브로인의 첨가는 제2형 콜라겐의 발현량을 오히려 감소시켰다. 또한 실크 피브로인은 제10형 콜라겐의 발현을 증가시키는 것을 확인하였다. 이 효과는 특히 연골세포를 3차원 배양할 때 더 컸다. 본 연구결과를 통하여 우리는 연골을 재생하는데 있어서 실크 단백질을 가능성을 보았으며, 향후 연구에서 연골재생과 실크의 관계를 좀 더 정밀하게 파악하고자 한다.

• Nutrition Research and Practice
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• 제13권6호
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• pp.480-487
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• 2019

BACKGROUND/OBJECTIVES: Osteoarthritis (OA) is a major public health issue in Japan and other countries, and foods that prevent or treat OA are in strong demand. Proteins and peptides in chicken meat and bones are known for being rich in functional and nutritional ingredients for the improvement of osteoporosis. We speculated that chicken legs, a food consumed in many regions of the world, may also contain such ingredients. In this study, we aim to (i) evaluate the effect of chicken leg extract (CLE) on the promotion of cartilage matrix production and (ii) identify the active ingredient in CLE that contributes to this function. MATERIALS/METHODS: Artificial CLE digest was prepared, and the acid mucopolysaccharide production-promoting activity of the CLE digest was evaluated by alcian blue staining of ATDC5 cells. CLE was orally administered to rabbits with burr holes in the knee joint of the femur, and the degree of regeneration of cartilage matrix was evaluated. Furthermore, we investigated orally administered CLE-derived peptides in human plasma using LC-MS. From measuring the acid mucopolysaccharide production-promotion activity of these peptides, a molecule considered to be an active ingredient in the CLE digest was identified. RESULTS: CLE digest promoted acid mucopolysaccharide production and facilitated regeneration of cartilage matrix in in vitro and in vivo experiments. Four peptides including phenylalanyl-hydroxyproline (Phe-Hyp) were detected as CLE-derived peptides in human plasma. The effect of CLE was inferred to be due to Phe-Hyp, which was confirmed to be present in the CLE digest. CONCLUSIONS: It was shown that CLE stimulated the production of articular cartilage matrix both in vitro and in vivo, and that CLE could be an effective food for preventing or treating OA. Furthermore, only Phe-Hyp was confirmed as the active compound in the CLE digest, suggesting that the activity of CLE was due to Phe-Hyp.

• Macromolecular Research
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• 제16권6호
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• pp.517-523
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• 2008

A RGD (Arg-Gly-Asp) conjugated chitosan hydrogel was used as a cell-supporting scaffold for articular cartilage regeneration. Thermosensitive chitosan-Pluronic (CP) has potential biomedical applications on account of its biocompatibility and injectability. A RGD-conjugated CP (RGD-CP) copolymer was prepared by coupling the carboxyl group in the peptide with the residual amine group in the CP copolymer. The chemical structure of RGD-CP was characterized by $^1H$ NMR and FT IR. The concentration of conjugated RGD was quantified by amino acid analysis (AAA) and rheology of the RGD-CP hydrogel was investigated. The amount of bound RGD was $0.135{\mu}g$ per 1 mg of CP copolymer. The viscoelastic parameters of RGD-CP hydrogel showed thermo-sensitivity and suitable mechanical strength at body temperature for cell scaffolds (a> 100 kPa storage modulus). The viability of the bovine chondrocyte and the amount of synthesized glycosaminoglycans (GAGs) on the RGD-CP hydrogels were evaluated together with the alginate hydrogels as a control over a 14 day period. Both results showed that the RGD-CP hydrogel was superior to the alginate hydrogel. These results show that conjugating RGD to CP hydro gels improves cell viability and proliferation, including extra cellular matrix (ECM) expression. Therefore, RGD conjugated CP hydrogels are quite suitable for a chondrocyte culture and have potential applications to the tissue engineering of articular cartilage tissue.