Korean Chemical Engineering Research

The Korean Institute of Chemical Engineers (한국화학공학회)
권호 표지
DOI QR코드

DOI QR Code

Enhanced Chondrogenic Differentiation of Human Adipose-derived Stem Cells with Inverse Opal Scaffolds

역오팔 구조 지지체를 이용한 인간 지방 유래 줄기 세포의 연골 분화 촉진

  • Bhang, Suk Ho (Department of Biomedical Engineering, Washington University) ;
  • Yu, Taekyung (Department of Chemical Engineering, College of Engineering, Kyung Hee University)
  • 방석호 (세인트루이스 워싱턴대학교 생물의공학과) ;
  • 유태경 (경희대학교 화학공학과)
  • Received : 2013.10.10
  • Accepted : 2013.11.07
  • Published : 2013.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

In this report, we present an inverse opal scaffold that can enhance the chondrogenic differentiation of human adipose-derived stem cells (hADSCs) without drug, gene, or cytokine supplement. Inverse opal scaffolds based on poly(D,L-lactide-co-glycolide) were formed with uniform $200{\mu}m$ pores. Due to uniform pore sizes and well-controlled interconnectivity of inverse opal scaffold, hADSCs were allowed to distribute homogeneously throughout the scaffolds. As a result, high cell density culture with scaffold was possible. Since the hADSCs cultured in inverse opal scaffolds were subjected to limited supplies of oxygen and nutrients, these cells were naturally preconditioned to a hypoxic environment that stimulated the up-regulation of hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$). As a result, apoptotic activity of hADSCs until 3 weeks after initial cell seeding was significantly reduced and chondrogenic differentiation related molecular signal cascades were up regulated (transforming growth factor-beta, phosphorylated AKT, and phosphorylated p38 expression). In contrast, hADSCs cultured with small and non-uniform porous scaffolds showed significantly increased apoptotic activity with decreased chondrogenic differentiation. Taken together, inverse opal scaffold could potentially be used as an effective tool for improving chondrogenesis using stem cells.

본 연구는 역오팔 지지체를 이용하여 인간지방유래 줄기세포의 연골 분화를 촉진하는 내용을 담고 있다. 비 다공성 구조를 가진 지지체에서 세포를 분화 시도하였을 경우 분화가 잘 촉진되지 않는 것에 비해 200 nm 정도의 균일한 구멍을 가지는 poly(D,L-lactide-co-glycolide)로 구성된 역오팔 지지체는 그 다공성 구조로 인하여 지지체의 내부까지 산소와 유기물의 수송을 가능하게 하여 지지체 내에서 어떤 유전적, 약물적 처리 없이 인간지방유래 줄기세포가 분화가 잘 되게 하는 것을 확인하였다.

Keywords

References

  1. Lee, C. R., Grodzinsky, A. J., Hsu, H. P., Martin, S. D. and Spector, M., "Effects of Harvest and Selected Cartilage Repair Procedures on the Physical and Biochemical Properties of Articular Cartilage in the Canine Knee," J. Orthop. Res., 18, 790(2000). https://doi.org/10.1002/jor.1100180517
  2. Rodriguez, A. M., Elabd, C., Amri, E. Z., Ailhaud, G. and Dani, C., "The Human Adipose Tissue is a Source of Multipotent Stem Cells," Biochimie., 87, 125(2005). https://doi.org/10.1016/j.biochi.2004.11.007
  3. Casteilla, L., Planat-Benard, V., Cousin, B., Silvestre, J. S., Laharrague, P., Charrie`re, G. and Penicaud, L., "Plasticity of Adipose Tissue: A Promising Therapeutic Avenue in the Treatment of Cardiovascular and Blood Diseases," Arch. Mal. Coeur. Vaiss., 98, 922(2005).
  4. Oedayrajsingh-Varma, M., Ham, S. V., Knippenberg, M., Helder, M. N., Klein-Nulend, J., Schouten, T. E., Ritt, M. J. and Milligen, F. J. V., "Adipose Tissue-derived Mesenchymal Stem Cell Yield and Growth Characteristics Are Affected by the Tissue-harvesting Procedure," Cytotherapy, 8, 166(2006). https://doi.org/10.1080/14653240600621125
  5. Yoon, H. H., Bhang, S. H., Shin, J. Y., Shin, J. and Kim B. S., "Enhanced Cartilage Formation via Three-dimensional Cell Engineering of Human Adipose-derived Stem Cells," Tissue Eng., 18, 1949(2012). https://doi.org/10.1089/ten.tea.2011.0647
  6. Afizah, H., Yang, Z., Hui, J. H., Ouyang, H. W. and Lee, E. H., "A Comparison Between the Chondrogenic Potential of Human Bone Marrow Stem Cells (BMSCs) and Adipose-derived Stem Cells (ADSCs) Taken From the Same Donor," Tissue Eng., 13, 659(2007). https://doi.org/10.1089/ten.2006.0118
  7. Park, S. J., Kim, S. H. and Hong, S. K., "Preparation of Microcapsules and Release Behaviors of Biodegradable Poly(L-lactide)/Poly(ethylene glycol) Copolymers," Korean Chem. Eng. Res.(HWAHAK KONGHAK), 41, 497(2003).
  8. Lee, M. J., Seo, D. Y., Lee, H. E. and Choi, G. J., "Therapeutic Effect of Chitosan Modification on Salmon-calcitonin-loaded PLGA Nanoparticles," Korean J. Chem. Eng., 28, 1406(2011). https://doi.org/10.1007/s11814-010-0508-9
  9. Yoo, D. J, "Computer-aided Design and Fabrication of Bio-mimetic Scaffold for Tissue Engineering Using the Triply Periodic Minimal Surface," J. Kor. Soc. Prec. Eng., 28, 834(2011).
  10. Shin, Y. M. and Shin, H. S., "Current Status of Tissue Engineering Scaffolds Using Biodegradable Polymers," Polym. Sci. Tech., 18, 458(2007).
  11. Choi, S. W., Zhang, Y. and Xia, Y., "Three-dimensional Scaffolds for Tissue Engineering: the Importance of Uniformity in Pore Size and Structure," Langmuir, 26, 19001(2010). https://doi.org/10.1021/la104206h
  12. Oh, M. J., Moon, S. K., Kim, S. E. and Choi, S. W., "Preparation of Inverse Opal Scaffolds with Uniform Pore Sizes and Their Effect on Cell Proliferation," Biomaterials Research, 17, 126(2013).
  13. Zhang, Y., Choi, S. W. and Xia, Y., "Modifying the Pores of An Inverse Opal Scaffold with Chitosan Microstructures for Truly Three-dimensional Cell Culture," Macromol. Rapid Commun., 33, 296(2012). https://doi.org/10.1002/marc.201100695
  14. Amarilio, R., Viukov, S. V., Sharir, A., Eshkar-Oren, I., Johnson, R. S. and Zelzer, E., "HIF1alpha Regulation of Sox9 is Necessary to Maintain Differentiation of Hypoxic Prechondrogenic Cells During Early Skeletogenesis," Development, 134, 3917(2007). https://doi.org/10.1242/dev.008441
  15. Murphy, C. L. and Polak, J. M., "Control of Human Articular Chondrocyte Differentiation by Reduced Oxygen Tension," J. Cell Physiol., 199, 451(2004). https://doi.org/10.1002/jcp.10481
  16. Sekiya, I., Tsuji, K., Koopman, P., Watanabe, H., Yamada, Y., Shinomiya, K., Nifuji, A. and Noda, M., "SOX9 Enhances Aggrecan Gene Promoter/Enhancer Activity and is Upregulated by Retinoic Acid in a Cartilage-derived Cell Line TC6," J. Biol. Chem., 275, 10738(2000).
  17. Oh, C. D., Chang, S. H., Yoon, Y. M., Lee, S. J., Lee, Y. S., Kang, S. S. and Chun, J. S., "Opposing Role of Mitogen-activated Protein Kinase Subtypes, erk-1/2 and p38, in the Regulation of Chondrogenesis of Mesenchymes," J. Biol. Chem., 275, 5613(2000). https://doi.org/10.1074/jbc.275.8.5613
  18. Kanichai, M., Ferguson, D., Prendergast, P. J. and Campbell, V. A., "Hypoxia Promotes Chondrogenesis in Rat Mesenchymal Stem Cells: a Role for AKT and Hypoxia-inducible Factor (HIF)-1alpha," J. Cell Physiol., 216, 708(2008). https://doi.org/10.1002/jcp.21446