Comparison of the efficacy of Piascledine and transforming growth factor β1 on chondrogenic differentiation of human adipose-derived stem cells in fibrin and fibrin-alginate scaffolds

Document Type : Original Article


1 Department of Anatomical Sciences, Medical School, Isfahan University of Medical Sciences, Isfahan, Iran

2 Department of Molecular Biology, Medical School, Isfahan University of Medical Sciences, Isfahan, Iran


Objective(s):The aim of this study was to compare the chondrogenic induction potential of Piascledine and TGF-β1 on adipose-derived stem cells (ADSCs) in fibrin and fibrin-alginate scaffolds. 
Materials and Methods: Human subcutaneous adipose tissues were harvested from three patients who were scheduled to undergo liposuction. Isolated ADSCs were proliferated in a culture medium. Then, the cells were seeded in fibrin or fibrin-alginate scaffolds and cultured for 14 days in a chondrogenic medium containing Piascledine, TGF-β1, or both. The rate of cell proliferation and survival was evaluated by using MTT [3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide] assay and the rate of the expression of type II collagen, aggrecan, and type X collagen genes was evaluated by real-time polymerase chain reaction (real-time PCR) method.
Results: The MTT results showed that Piascledine is able to enhance the proliferation and survival of ADSCs in fibrin scaffolds in comparison to other groups (P<0.05). Real-time PCR evaluation revealed that the expression of type II collagen was higher in TGF- β1groups, but the expression of aggrecan was higher in TGF-β1 alone or along with Piascledine in fibrin-alginate scaffolds. Furthermore, the expression of type X collagen was lower in Piascledine alone or along with TGF-β1 in fibrin scaffold.
Conclusion: Piascledine can enhance the proliferation and differentiation of ADSCs in fibrin scaffolds.


Main Subjects

1. Hardingham T, Tew S, Murdoch A. Tissue engineering: Chondrocytes and cartilage. Arthritis Res 2002; 4: 63-8.
 2. Mitchell N, Shepard N. The resurfacing of adult rabbit articular cartilage by multiple perforations through the subchondral bone. J Bone Joint Surg Am 1976; 58: 230-3.
3. Mardani M, Hashemibeni B, Ansar MM, Esfahani SH, Kazemi M, Goharian V, Esmaeili N, Esfandiary E. Comparison between chondrogenic markers of differentiated chondrocytes from adipose derived stem cells and articular chondrocytes in vitro. Iranian journal of basic medical sciences. 2013; 16(6):763.
4. Colter DC, Sekiya I, Prockop DJ. Identification of a subpopulation of rapidly self-renewing and multipotential adult stem cells in colonies of human marrow stromal cells. Proc Natl Acad Sci USA 2001; 98: 7841-45. 
5. Hahemibeni B, Razavi Sh, Esfandiary E, Karbasi S, Mardani M, Nasresfahani M. Induction of chondrogenic differentiation of human adipose‑derived stem cells with TGF‑β3 in pellet culture system. Iran J Basic Med Sci 2008; 11(1): 10-17. 
6. Giovannini S, Diaz-Romero J, Aigner T, Heini P, Mainil-Varlet P, Nesic D. Micromass co-culture of human articular chondrocytes and human bone marrow mesenchymal stem cells to investigate stable neocartilage tissue formation in vitro. Eur Cell Mater. 2010; 20:59.
7. Langer R, Vacanti JP. Tissue engineering. Science 1993; 260(5110): 920-6.
8. Henrotin YE, Labasse AH, Jaspar JM, De Groote DD, Zheng SX, Guillou GB, Reginster JY. Effects of three avocado/soybean unsaponifiable mixtures on metalloproteinases, cytokines and prostaglandin E 2 production by human articular chondrocytes. Clinical rheumatology. 1998; 17(1):31-9.
9. Mauviel A, Daireaux M, Hartmann DJ, Galera P, Loyau G, Pujol JP. Effects of unsaponifiable extracts of avocado/soy beans (PIAS) on the production of collagen by cultures of synoviocytes, articular chondrocytes and skin fibroblasts. Rev Rhum Mal Osteoartic 1989; 56(2): 207-11.  [In French].      
10. Hunter DJ. Pharmacologic therapy for osteoarthritis—the era of disease modification. Nat Rev Rheumatol 2010; 7(1): 13-22.
11. Breen A, Dockery P, O'Brien T, Pandit A. Fibrin scaffold promotes adenoviral gene transfer and controlled vector delivery. J of Biomed Mater Res A 2009;89(4):876-84.
12. Chen G, Ushida T, Tateishi T. A biodegradable hybrid sponge nested with collagen microsponges. J Biomed Mater Res 2000; 51(2): 273-79.
13. Lahiji A, Sohrabi A, Hangerford DS, Frondoza CG. Chitosan supports the expression of extracellular matrix proteins in human osteoblasts and chondrocytes. J Biomed Mater Res 2000; 51(4): 586-95.
14. Zhou H, Xu HH. The fast release of stem cells from alginate-fibrin microbeads in injectable scaffolds for bone tissue engineering. Biomaterials 2011; 32(30): 7503-13.
15. Buser Z, Liu J, Thorne KJ, Coughlin D, Lotz JC. Inflammatory response of intervertebral disc cells is reduced by fibrin sealant scaffold in vitro. J Tissue Eng Regen Med 2014;8(1):77-84.
16. Shikanov A, Xu M, Woodruff TK, Shea LD. Interpenetrating fibrin-alginate matrices for in vitro ovarian follicle development. Biomaterials 2009; 30(29): 5476-85.
17. Morris VJ. Gelation of polysaccharides. In: Mitchell JR, Ledward DA, editors. Functional properties of food macromolecules. New York: Elsevier; 1986: 121-8. 
18. Hashemibeni B, Razavi S, Esfandiary E, Karbasi S, Mardani M, Sadeghi F, Esfahani MN, Nadali F, Shafiezade H. Effect of Transforming Growth Factor-ß3 and Bone Morphogenetic Protein-6 Growth Factors on Chondrogenic Differentiation of Adipose-Derived Stem Cells in Alginate Scaffold. J Isfahan Med Sch  2010; 28(112). [In Persian].
19.Yang SH, Wu CC, Shih TT, 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.       
20. 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.
21.  Rowley JA, Madlambayan G, Mooney DJ. Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials 1999; 20(1): 45-53.       
22. Wang ZY, Zhang QZ, Konno M, Saito S. Sol-gel transition of alginate solution by the addition of various divalent cations: 13C-nmr spectroscopic study. Biopolymers 1993; 33(4): 703-11.
23. 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. Advanced biomedical research. 2014; 3(1):40.
24. Yan J, Dong L, Zhang B, Qi N. Effects of extremely low-frequency magnetic field on growth and differentiation of human mesenchymal stem cells. Electromagnetic biology and medicine 2010; 29(4):165-76.
25. Creecy CM, O'Neill CF, Arulanandam BP, Sylvia VL, Navara CS, Bizios R. Mesenchymal stem cell osteodifferentiation in response to alternating electric current. Tissue Engineering Part A  2012; 19(3-4):467-74.
26. Grimaud E, Heymann D, Rédini F. Recent advances in TGF-β effects on chondrocyte metabolism: potential therapeutic roles of TGF-β in cartilage disorders. Cytokine & growth factor reviews 2002 Jun 30; 13(3):241-57
27. Christiansen BA, Bhatti S, Goudarzi R, Emami S. Management of osteoarthritis with avocado/soybean unsaponifiables. Cartilage 2015; 6(1):30-44.
28. Didehvar H, Golshan-Iranpour F, Valiani A, Hashemibeni B, Esmaeeli M. Comparing the effects of transforming growth factor beta1 (TGF-ß1) and piascledine on the expression of collagen II, X and aggrecan genes in chondrogenesis of human adipose-derived stem cells in fibrin alginate composite scaffold. J Isfahan Med Sch 2016; 34(373): 157-65. [In Persian].
29. Esmaeily M, Hashemibeni B, Valiani A, Amirpour N, Purmollaabbasi B, Kazemi M. Effect of Piasclidine on induction of chondrogenesis by human adipose-derived stem cells in fibrin scaffold. J Isfahan Med Sch 2016; 33(357): 1862-70. [In Persian].
30. Lamaud E, Robert AM, Wepierre J. Biochemical effects of unsaponifiable lipidic components of avocado and soya bean administered percutaneously on the connective tissue components of hairless rat skin. Int J Cosmet Sci 1979; 1(4):213-9. 
31. Henrotin YE, Sanchez C, Deberg MA, Piccardi N, Guillou GB, Msika P, Reginster JY. Avocado/soybean unsaponifiables increase aggrecan synthesis and reduce catabolic and proinflammatory mediator production by human osteoarthritic chondrocytes. Journal of rheumatology 2003; 30(8):1825-34.
32. Altinel L, Saritas ZK, Kose KC, Pamuk K, Aksoy Y, Serteser M. Treatment with unsaponifiable extracts of avocado and soybean increases TGF-beta1 and TGF-beta2 levels in canine joint fluid. Tohoku J Exp Med 2007; 211(2):181-6.
33. Lippiello L, Nardo JV, Harlan R, Chiou T. Metabolic effects of avocado/soy unsaponifiables on articular chondrocytes. Evid Based Complement Alternat Med  2008; 5(2):191-7.
34. Kucharz EJ. Application of avocado/soybean unsaponifiable mixtures (piascledine) in treatment of patients with osteoarthritis. Ortopedia, traumatologia, rehabilitacja 2003; 5(2):248-51.
35. Hunziker EB. Growth-factor-induced healing of partial-thickness defects in adult articular cartilage. Osteoarthritis Cartilage 2001; 9(1): 22-32.
36. George M, Abraham TE. Polyionic hydrocolloids for the intestinal delivery of protein drugs: alginate and chitosan--a review. J Control Release 2006; 114(1):1-14.
37. Rabie A, Esfandiari E, Fesharaki M, Sanaie M ,Aminmansur B, Hashemibeni B. Access to a three dimensional osteoblasts culture originating human calvaria in Iran. J Isfahan Med Sch 2010; 27(102): 777-87. [In Persian].
38. Harrison P, Wilbourn B, Debili N, Vainchenker W, Breton-Gorius J, Lawrie AS, et al. Uptake of plasma fibrinogen into the alpha granules of human megakaryocytes and platelets. J Clin Invest 1989; 84(4): 1320-4.
39. Le Nihouannen D, Guehennec LL, Rouillon T, Pilet P, Bilban M, Layrolle P, et al. Micro-architecture of calcium phosphate granules and fibrin glue composites for bone tissue engineering. Biomaterials 2006; 27(13): 2716-22.
40. Dragoo JL, Carlson G, McCormick F, Khan-Farooqi H, Zhu M, Zuk PA, et al. Healing full-thickness cartilage defects using adipose-derived stem cells. Tissue Eng 2007; 13(7): 1615-21.
41. Wei Y, Hu Y, Hao W, Han Y, Meng G, Zhang D, et al. A novel injectable scaffold for cartilage tissue engineering using adipose-derived adult stem cells. J Orthop Res 2008; 26(1): 27-33.
42. Girandon L, Kregar-Velikonja N, Bozikov K, Barlic A. In vitro models for adipose tissue engineering with adipose-derived stem cells using different scaffolds of natural origin. Folia Biol (Praha) 2011; 57(2):47-56.
43. Zhao L, Weir MD, Xu HH. An injectable calcium phosphate-alginate hydrogel-umbilical cord mesenchymal stem cell paste for bone tissue engineering. Biomaterials 2010; 31(25): 6502-10.
44. Chien CS, Ho HO, Liang YC, Ko PH, Sheu MT, Chen CH. Incorporation of exudates of human platelet-rich fibrin gel in biodegradable fibrin scaffolds for tissue engineering of cartilage. J Biomed Mater Res B Appl Biomater 2012; 100(4): 948-55.
45. Stevens MM, Qanadilo HF, Langer R, Prasad S, V. A rapid-curing alginate gel system: utility in periosteum-derived cartilage tissue engineering. Biomaterials 2004; 25(5): 887-94.
46. Stevens MM, Marini RP, Martin I, Langer R, Prasad S, V. FGF-2 enhances TGF-beta1-induced periosteal chondrogenesis. J Orthop Res 2004; 22(5): 1114-9.
47. Ma HL, Hung SC, Lin SY, Chen YL, Lo WH. Chondrogenesis of human mesenchymal stem cells encapsulated in alginate beads. J Biomed Mater Res 2003; 64(2): 273-81.