[논문]변형된 키토산 알지네이트 겔 poly (L-Lactic-co-ε-Caprolactone) 지지체의 연골 조직 재생 평가

황야원; 최석화; 김근형

doi:10.17555/jvc.2015.06.32.3.224

[국내논문] 변형된 키토산 알지네이트 겔 poly (L-Lactic-co-ε-Caprolactone) 지지체의 연골 조직 재생 평가
Chitosan-alginate Gel Modified Poly (L-Lactic-co-ε-Caprolactone) (PLCL) as a Scaffold for Cartilage Tissue Engineering 원문보기

Journal of veterinary clinics = 한국임상수의학회지, v.32 no.3, 2015년, pp.224 - 230

(충북대학교 수의과대학 수의외과학 교실) , 황야원 (충북대학교 수의과대학 수의외과학 교실) , 최석화 (충북대학교 수의과대학 수의외과학 교실) , 김근형 (충북대학교 수의과대학 수의외과학 교실)

초록
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본 연구는 키토산 알지네이트 수화겔을 사용하여 제작된 연골세포의 3차원 구조를 유지하며 생물학적, 생리학적인 기능을 유지하는데 적합한 poly (L-Lactic-co-${\varepsilon}$-Caprolactone) (PLCL) 지지체의 효과에 대한 연구이다. 체내에서 수화겔은 단독으로 지지체 역할을 하기에는 부하를 견디기에 약하다. 이에 본 연구에서는 연골세포와 유사한 세포, 세포외 기질의 3차원적 구성을 만들기 위해 PLCL 지지체와 수화겔을 사용하여 합성 지지체를 제작하였다. 염화나트륨을 사용한 입자 침출 기법으로 85%의 다공성, $300-500{\mu}m$ 크기의 구멍을 가진 탄성력 높은 지지체를 제작하였다. 소의 연골세포와 키토산 알지네이트 겔 혼합물이 PLCL 지지체에 적용되었고 대조군의 알지네이트와 비교 연구하였다. 키토산 알지네이트 수화겔과 연골세포가 혼합된 경우에 알지네이트 단독 사용에 비해 세포 성숙, 증식, 세포외 기질의 합성, sGAG 생성과 II 형 콜라겐의 발현 등의 효과가 좋은 것으로 확인되었다. 본 연구 결과를 통해 PLCL 지지체에 연골세포와 키토산 알지네이트 겔 혼합물을 적용할 경우 세포 증식과 기질의 합성에 적합한 환경을 만들 수 있으며 연골의 복구와 재생에 효과적으로 사용될 수 있을 것으로 기대된다.

Abstract ▼ AI-Helper

This study was designed in the fabricated poly (L-Lactic-co-${\varepsilon}$-Caprolactone) (PLCL) scaffold using chitosan-alginate hydrogel, which would be more suitable to maintain the biological and physiological functions continuing three dimensional spatial organizations for chondrocytes. As a scaffold, hydrogels alone is weak at endure complex loading within the body. In this study, we made cell hybrid scaffold constructs with poly (L-Lactic-co-${\varepsilon}$-Caprolactone) (PLCL) scaffold and hydrogels to make a three-dimensional composition of cells and extracellular matrix, which would be a mimic of a native cartilage. Using a particle leaching technique with NaCl, we fabricated a highly-elastic scaffold from PLCL with 85% porosity and $300-500{\mu}m$ pore size. A mixture of bovine chondrocytes and chitosan-alginate gel was seeded and compared with alginate as a control on the PLCL scaffold. The cell maturation, proliferation, extracellular matrix synthesis, glycosaminoglycans (sGAG) production and collagen type-II expressions were better in chondrocytes seeded in chitosan-alginate hydrogel than in alginate only. These results indicate that chondrocytes with chitosan-alginate gel on PLCL scaffolds provide an appropriate biomimetic environment for cell proliferation and matrix synthesis, which could successfully be used for cartilage repair and regeneration.

주제어

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문제 정의

Therefore, the objectives of this study were to investigate the use of chitosan-alginate mixed hydrogel that might be useful to enhance the chondrocytes function in creating cartilage tissue onto the fabricated poly (L-Lactic-co-ε-Caprolactone) (PLCL) scaffold for advanced use for the repair of damaged cartilage.

제안 방법

The sections were stained at one week with H&E and Alcian blue staining methods.

데이터처리

The statistical significance in the differences between the mean values among the groups was analyzed by one-way analysis of variance (ANOVA) with Tukey’s test using the SPSS 11.5 statistical software (SPSS Inc., Chicago, USA).

이론/모형

Primary chondrocytes were used for this experiment. Cell number and viability were assessed with a hemocytometer by Trypan blue exclusion assay. More than 95% of the cells were usually viable after isolation.
Schematic diagram to illustrate the fabrication process of PLCL scaffold. PLCL scaffold fabricated by the particulateleaching method.

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