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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2019.1
- 수록면
- 285 - 299 (15page)
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초록· 키워드
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BACKGROUND: This study aimed to observe the cartilaginous matrix production in SRY (sex determining region Y)-box 9 (SOX9)- and/or telomerase reverse transcriptase (TERT)-transfected chondrocytes from monolayer to three-dimensional (3D) culture. METHODS: The genes were transferred into chondrocytes at passage-1 (P1) via lipofection. The post-transfected chondrocytes (SOX9-, TERT- and SOX9/TERT) were analysed at P1, P2 and P3. The non-transfected group was used as control. The 3D culture was established using the chondrocytes seeded in a disc-shaped PLGA/fibrin and PLGA scaffolds. The resulting 3D ‘‘cells-scaffolds’’ constructs were analysed at week-1, -2 and -3. The histoarchitecture was evaluated using haematoxylin and eosin, alcian blue and safranin o stains. The quantitative sulphated glycosaminoglycan (sGAG) content was measured using biochemical assay. The cartilage-specific markers expression were analysed via real-time polymerase chain reaction. RESULTS: All monolayer cultured chondrocytes showed flattened, fibroblast-like appearance throughout passages. Proteoglycan and sGAG were not detected at the pericellular matrix region of the chondrocytes. The sGAG content assay indicated the matrix production depletion in the culture. The cartilage-specific markers, COL2A1 and ACAN, were downregulated. However, the dedifferentiation marker, COL1A1 was upregulated. In 3D ‘‘cells-scaffolds’’ constructs, regardless of transfection groups, chondrocytes seeded in PLGA/fibrin showed a more uniform distribution and produced denser matrix than the PLGA group especially at week-3. Both sGAG and proteoglycan were clearly visualised in the constructs, supported by the increment of sGAG content, quantitatively. Both COL2A1 and ACAN were upregulated in SOX9/TERT-PLGA and SOX9/TERT-PLGA/fibrin respectively. While, COL1A1 was downregulated in SOX9/TERT-PLGA. CONCLUSION: These findings indicated that the SOX9/TERT-transfected chondrocytes incorporation into 3D scaffolds facilitates the cartilage regeneration which is viable structurally and functionally.
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참고문헌 (48)
참고문헌 신청
Langer R / 1993 / Tissue engineering / Science 260:920~926
Abdul Rahman R / 2015 / The potential of 3-dimensional construct engineered from poly(lactic-co-glycolic acid)/fibrin hybrid scaffold seeded with bone marrow mesenchymal stem cells for in vitro cartilage tissue engineering / Tissue Cell 47:420~430
Munirah Sha’ban / 2011 / Sox-9 Transient Transfection Enhances Chondrogenic Expression of Osteoarthritic Human Articular Chondrocytes In Vitro: Preliminary Analysis / 조직공학과 재생의학 8(1):32~41
Frisch J / 2016 / TGF-b gene transfer and overexpression via rAAV vectors stimulates chondrogenic events in human bone marrow aspirates / J Cell Mol Med 20:430~440
Griffin DJ / 2016 / Mechanical properties and structure-function relationships in articular cartilage repaired using IGF-I gene-enhanced chondrocytes / J Orthop Res 34:149~153
Abdul Rahman R / 2015 / The potential of 3-dimensional construct engineered from poly(lactic-co-glycolic acid)/fibrin hybrid scaffold seeded with bone marrow mesenchymal stem cells for in vitro cartilage tissue engineering / Tissue Cell 47:420~430
Munirah Sha’ban / 2011 / Sox-9 Transient Transfection Enhances Chondrogenic Expression of Osteoarthritic Human Articular Chondrocytes In Vitro: Preliminary Analysis / 조직공학과 재생의학 8(1):32~41
Frisch J / 2016 / TGF-b gene transfer and overexpression via rAAV vectors stimulates chondrogenic events in human bone marrow aspirates / J Cell Mol Med 20:430~440
Griffin DJ / 2016 / Mechanical properties and structure-function relationships in articular cartilage repaired using IGF-I gene-enhanced chondrocytes / J Orthop Res 34:149~153
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