Document Type : Original Article

Authors

1 Professor, Department of Anatomical Sciences and Molecular Biology, Isfahan University of Medical Sciences, Isfahan, Iran

2 Ph.D. Candidate, Anatomy Department, Medical Faculty, Shaheed Beheshti University of Medical Sciences, Tehran, Iran

3 Associate Professor, Department of Anatomical Sciences and Molecular Biology, Isfahan University of Medical Sciences, Isfahan, Iran

4 Assistant Professor, Nanotechnology and Tissue Engineering Group, Department of Advanced Medical Technology, Isfahan University of Medical Sciences, Isfahan, Iran

5 Assistant Professor. Department of Anatomical Sciences and Molecular Biology, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

Abstract

Background: Recently, cartilage tissue engineering is the best candidate for regeneration of cartilage defects. We evaluated the potential of fibrin and PLGA/fibrin scaffolds in providing a suitable environment for growth and chondrogenic differentiation of human adipose derived stem cells (hADSCs) in the presence of icariin.
Method: The Three-dimensional (3-D) PLGA scaffold was prepared using the solvent casting/salt leaching technique and the hybrid scaffold was fabricated by fibrin. hADSCs were isolated from human adipose tissue. 3-D PLGA/fibrin scaffolds were seeded with cultured hADSCs and analyzed 14 days later, Monolayer culture was used for the control group. The viabilities of cells in different groups were assessed by MTT. The expression of chondrogenic related genes, hypertrophic marker and Fibrotic marker were quantified by RT-PCR.
Results:MTT results show that viability in the control group was significantly higher than those in the Fibrin and PLGA/Fibrin groups. Also viability in the PLGA/Fibrin group affected by icariin was higher than that in Fibrin group.
The results of the real-time PCR showed that SOX9, Agg, Coll 2, and Coll 1 gene expression in the fibrin and PLGA/fibrin groups were significantly higher than those in the control group. Coll 10 gene expression in the fibrin group was higher in comparison to the control group but not significantly. type SOX9, Coll 2 and Coll 1 gene expression in the fibrin group was significantly lower compared to the PLGA/fibrin group.
Conclusions: The study reveals that the corporation of PLGA with fibrin is an effective way to potentially enhance articular cartilage regeneration of hADSCs in the presence of icariin.

Keywords

  1. Samanipour R, Karbasi S, Hashemibeni B. Comparing behavior of chondrocyte cells on different polyhydroxybutyrate scaffold structure for cartilage tissue engineering. Anatomical Sciences 2016; 13(2):105-16.
  2. Peterson L, Minas T, Brittberg M, Nilsson A, Sjögren-Jansson E, Lindahl A. Two- to 9-year outcome after autologous chondrocyte transplantation of the knee. Clin Orthop Relat Res 2000; (374):212-34.
  3. Park JG, Lee JH, Kim JN, Kang JA, Kim KJ, Park KD, et al. Chondrogenic differentiation of human adipose tissue-derived stem cells in functional PLGA scaffolds. Tissue Engineering and Regenerative Medicine 2011; 8(1):47-54.
  4. Valiani A, Hashemibeni B, Esfandiary E, Ansar MM, Kazemi M, Esmaeili N. Study of carbon nano tubes effects on the chondrogenesis of human adipose derived stem cells in alginate scaffold. Int J Prev Med 2014; 5(7):825-34.
  5. De Ugarte DA, Morizono K, Elbarbary A, Alfonso Z, Zuk PA, Zhu M, et al. Comparison of multi-lineage cells from human adipose tissue and bone marrow. Cells Tissues Organs 2003; 174(3):101-9.
  6. Park K, Cho KJ, Kim JJ, Kim IH, Han DK. Functional PLGA scaffolds for chondrogenesis of bone-marrow-derived mesenchymal stem cells. Macromol Biosci 2009; 9(3):221-9.
  7. Pilia M, Guda T, Appleford M. Development of composite scaffolds for load-bearing segmental bone defects. Biomed Res Int 2013; 2013:458253.
  8. Lü JM, Wang X, Marin-Muller C, Wang H, Lin PH, Yao Q, et al. Current advances in research and clinical applications of PLGA-based nanotechnology. Expert Rev Mol Diagn 2009; 9(4):325-41.
  9. Uematsu K, Hattori K, Ishimoto Y, Yamauchi J, Habata T, Takakura Y, et al. Cartilage regeneration using mesenchymal stem cells and a three-dimensional poly-lactic-glycolic acid (PLGA) scaffold. Biomaterials 2005; 26(20):4273-9.
  10. Ahmed TA, Dare EV, Hincke M. Fibrin: a versatile scaffold for tissue engineering applications. Tissue Eng Part B Rev 2008; 14(2):199-215.
  11. Wang W, Li D, Wang MC, Li YL, Gao CY. A hybrid scaffold of poly (lactide-co-glycolide) sponge filled with fibrin gel for cartilage tissue engineering. Chinese Journal of Polymer Science 2011; 29(2):233-240.
  12. Lin Z, Willers C, Xu J, Zheng MH. The chondrocyte: biology and clinical application. Tissue Eng 2006; 12(7):1971-84.
  13. Eyrich D, Wiese H, Maier G, Skodacek D, Appel B, Sarhan H, et al. In vitro and in vivo cartilage engineering using a combination of chondrocyte-seeded long-term stable fibrin gels and polycaprolactone-based polyurethane scaffolds. Tissue Eng 2007; 13(9):2207-18.
  14. Lee CR, Grad S, Gorna K, Gogolewski S, Goessl A, Alini M. Fibrin-polyurethane composites for articular cartilage tissue engineering: a preliminary analysis. Tissue Eng 2005; 11(9-10):1562-73.
  15. Breen A, O'Brien T, Pandit A. Fibrin as a delivery system for therapeutic drugs and biomolecules. Tissue Eng Part B Rev 2009; 15(2):201-14.
  16. Zhang DW, Cheng Y, Wang NL, Zhang JC, Yang MS, Yao XS. Effects of total flavonoids and flavonol glycosides from epimedium koreanum nakai on the proliferation and differentiation of primary osteoblasts. Phytomedicine 2008; 15(1-2):55-61.
  17. Li D, Yuan T, Zhang X, Xiao Y, Wang R, Fan Y, et al. Icariin: a potential promoting compound for cartilage tissue engineering. Osteoarthritis Cartilage 2012; 20(12):1647-56.
  18. Hsieh TP, Sheu SY, Sun JS, Chen MH, Liu MH. Icariin isolated from epimedium pubescens regulates osteoblasts anabolism through BMP-2, SMAD4, and Cbfa1 expression. Phytomedicine 2010; 17(6):414-23.
  19. Zhang X, Guo Y, Li DX, Wang R, Fan HS, Xiao YM, et al. The effect of loading icariin on biocompatibility and bioactivity of porous β-TCP ceramic. J Mater Sci Mater Med 2011; 22(2):371-9.
  20. Tavakoli E, Mehdikhani-Nahrkhalaji M, Hashemi-Beni B, Zargar-Kharazi A, Kharaziha M. Preparation, characterization and mechanical assessment of poly (lactide-co-glycolide)/hyaluronic acid/fibrin/bioactive glass nano-composite scaffolds for cartilage tissue engineering applications. Procedia Materials Science 2015; 11:124-30.
  21. Yang SH, Wu CC, Shih TTF, Chen PQ, Lin FH. Three‐dimensional culture of human nucleus pulposus cells in fibrin clot: comparisons on cellular proliferation and matrix synthesis with cells in alginate. Artif Organs 2008; 32(1):70-3.
  22. Mardani M, Hashemibeni B, Ansar MM, Zarkesh Esfahani SH, Kazemi M, Goharian V, et al. Comparison between chondrogenic markers of differentiated chondrocytes from adipose derived stem cells and articular chondrocytes in vitro. Iran J Basic Med Sci 2013; 16(6):763-73.
  23. Sha'ban M, Kim SH, Idrus RB, Khang G. Fibrin and poly (lactic-co-glycolic acid) hybrid scaffold promotes early chondrogenesis of articular chondrocytes: an in vitro study. J Orthop Surg Res 2008; (3):17.
  24. Esfandiary E, Valiani A, Hashemibeni B, Moradi I, Narimani M. The evaluation of toxicity of carbon nanotubes on the human adipose-derived-stem cells in-vitro. Adv Biomed Res 2014; 3:40.
  25. Mardani M, Roshankhah S, Hashemibeni B, Salahshoor M, Naghsh E, Esfandiari E. Induction of chondrogenic differentiation of human adipose-derived stem cells by low frequency electric field. Adv Biomed Res 2016; 5:97.
  26. Ansar MM, Esfandiariy E, Mardani M, Hashemibeni B, Zarkesh-Esfahani SH, Hatef M, et al. A comparative study of aggrecan synthesis between natural articular chondrocytes and differentiated chondrocytes from adipose derived stem cells in 3D culture. Adv Biomed Res 2012; 1:24.
  27. Eyrich D, Brandl F, Appel B, Wiese H, Maier G, Wenzel M, et al. Long-term stable fibrin gels for cartilage engineering. Biomaterials 2007; 28(1):55-65.
  28. Weisel JW. Fibrinogen and fibrin. Adv Protein Chem 2005; 70:247-99.
  29. Gentile P, Chiono V, Carmagnola I, Hatton PV. An overview of poly (lactic-co-glycolic) acid (PLGA)-based biomaterials for bone tissue engineering. Int J Mol Sci 2014; 15(3):3640-59.
  30. Zhao H, Ma L, Gao C, Shen J. A composite scaffold of PLGA microspheres/fibrin gel for cartilage tissue engineering: fabrication, physical properties, and cell responsiveness. J Biomed Mater Res B Appl Biomater 2009; 88(1):240-9.
  31. Zhu Y, Gao C, Liu X, He T, Shen J. Immobilization of biomacromolecules onto aminolyzed poly (L-lactic acid) toward acceleration of endothelium regeneration. Tissue Eng 2004; 10(1-2):53-61.
  32. Weisel JW. The mechanical properties of fibrin for basic scientists and clinicians. Biophys Chem 2004; 112(2-3):267-76.
  33. Cheng NC, Estes BT, Young TH, Guilak F. Engineered cartilage using primary chondrocytes cultured in a porous cartilage-derived matrix. Regen Med 2011; 6(1):81-93.