Comparison of TGF-β3 and avocado/soybean unsaponifiable on chondrogenesis of human adipose-derived stem cells on poly (lactic-co-glycolic) acid/ hyaluronic acid hybrid scaffold

Document Type : Original Article

Authors

1 Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

2 Department of Anatomy, Faculty of Medicine, Hormozgan University of Medical Sciences, Bandar Abbas, Iran

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

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

5 Fertility and Infertility Research Center, Hormozgan University of Medical Sciences, Bandar Abbas, Iran

Abstract

Objective(s): Avocado/soybean unsaponifible (ASU) possesses properties including chondroprotective, anticatabolic, and anabolic. The goal behind this research was to detect the effect of ASU and TGF-β3 on the chondrogenesis of human adipose-derived stem cells (hADSCs) on poly (lactic-co-glycolic) acid (PLGA)/ hyaluronic acid (PLGA/HA) hybrid scaffold.
Materials and Methods: First hADSCs were seeded in PLGA/Hyaluronic acid scaffold and cultured in chondrogenic media. These cells were assigned into 4 groups: control, TGFβ-3, ASU, and TGFβ-3+ASU. The viability was assessed separately by MTT. Real-time PCR was used to quantify the expression of chondrogenic specific genes [Sox9, collagen type II (ColII), Aggrecan (AGG)] and collagen type X (ColX). Moreover, Western blotting was employed to evaluate protein expression levels of collagens type II and X.
Results: These findings indicated a significant increase in the proliferation and survival of hADSCs differentiated cells by ASU compared with the control group (P=0.008). Real-time PCR results revealed significant differences in the expression of AGG, SOX9, ColII, and ColX genes in the control group when compared with other groups (ASU, TGF-β3, and TGF-β3+ASU). ColII protein production significantly dropped in the TGF-β3 group in comparison with the TGF-β3+ASU group (0.000). The ColII (P=0.002) and ColX (P=0.002) protein production proved significantly higher in the TGF-β3+ASU group compared with the ASU group.
Conclusion: Using the synergist form TGFβ-3, ASU induces chondrogenesis in hADSCs in PLGA/HA composite scaffold. This can be deduced with reduction of special markers of hyaline cartilage in comparison with ASU and decreased hypertrophic marker compared with TGF-β3.

Keywords

References

1.    Bhosale AM, Richardson JB. Articular cartilage: structure, injuries and review of management. Br Med Bull 2008; 87:77-95.
2.    Hashemibeni B, Mardani M, Bahrami M, Valiani A, Mehr MS, Pourentezari M. Comparison of fibrin and PLGA/fibrin scaffolds for chondrogenesis of human adipose derived stem cells by icariin. J Kerman Univ Med Sci 2020; 27:14-23.
3.    Hashemibeni B, Mardani M, Valiani A, Pourentezari M, Anvari M, Yadegari M, et al. Effects of avocado/soybean on the chondrogenesis of human adipose-derived stem cells cultured on polylactic-co-glycolic acid/fibrin hybrid scaffold. Appl Biotechnol Rep 2019; 6:145-150.
4.    Oliva J, Florentino A, Bardag-Gorce F, Niihara Y. Engineering, differentiation and harvesting of human adipose-derived stem cell multilayer cell sheets. Regen Med 2019; 14:151-163.
5.    Zhang J, Dong S, Sivak WN, Sun HB, Chang P. stem cells in cartilage diseases and repair. Stem Cells Int 2018; 2018:1-2.
6.    Yamagata K, Nakayamada S, Tanaka Y. Use of mesenchymal stem cells seeded on the scaffold in articular cartilage repair. Inflamm Regen 2018; 38:4-12.
7.    Dicker KT, Gurski LA, Pradhan-Bhatt S, Witt RL, Farach-Carson MC, Jia X. Hyaluronan: a simple polysaccharide with diverse biological functions. Acta Biomater 2014; 10:1558-1570.
8.    Rippe M, Cosenza V, Auzély-Velty R. Design of soft nanocarriers combining hyaluronic acid with another functional polymer for cancer therapy and other biomedical applications. Pharmaceutics 2019; 11:338-363.
9.    Yoo HS, Lee EA, Yoon JJ, Park TG. Hyaluronic acid modified biodegradable scaffolds for cartilage tissue engineering. Biomaterials. Biomaterials 2005; 26:1925-1933.
10.    Gupta RC, Lall R, Srivastava A, Sinha A. Hyaluronic acid: molecular mechanisms and therapeutic trajectory. Front vet sci 2019; 6:192-214.
11.    Lindenhayn K, Perka C, Spitzer RS, Heilmann HH, Pommerening K, Mennicke J, et al. Retention of hyaluronic acid in alginate beads: aspects for in vitro cartilage engineering. J Biomed Mater Res 1999; 44:149-155.
12.    Chen MJ, Whiteley JP, Please CP, Schwab A, Ehlicke F, Waters SL, et al. Inducing chondrogenesis in MSC/chondrocyte co-cultures using exogenous TGF-β: a mathematical model. J Theor Biol 2018; 439:1-13.
13.    Tayalia P, Mooney DJ. Controlled growth factor delivery for tissue engineering. Adv Mater 2009; 21:3269-3285.
14.    Heldin C-H, Miyazono K, Ten Dijke P. TGF-β signalling from cell membrane to nucleus through SMAD proteins. Nature 1997; 390:465-471.
15.    Coricor G, Serra R. TGF-β regulates phosphorylation and stabilization of Sox9 protein in chondrocytes through p38 and Smad dependent mechanisms. Sci Rep 2016; 6:38616-38627.
16.     Bi W, Deng JM, Zhang Z, Behringer RR, De Crombrugghe B. Sox9 is required for cartilage formation. Nat Genet 1999; 22:85-89.
17.    Lefebvre V, Behringer R, De Crombrugghe B. L-Sox5, Sox6 and Sox9 control essential steps of the chondrocyte differentiation pathway. Osteoarthr Cartil 2001; 9:69-75.
18.    Lee HL, Yu B, Deng P, Wang CY, Hong C. transforming growth factor‐β‐induced kdm4b promotes chondrogenic differentiation of human mesenchymal stem cells. Stem Cells 2016; 34:711-719.
19.    Christiansen BA, Bhatti S, Goudarzi R, Emami S. Management of osteoarthritis with avocado/soybean unsaponifiables. Cartilage 2015; 6:30-44.
20.    Ownby SL, Fortuno LV, Au AY, Grzanna MW, Rashmir-Raven AM, Frondoza CG. Expression of pro-inflammatory mediators is inhibited by an avocado/soybean unsaponifiables and epigallocatechin gallate combination. J Inflamm 2014; 11:8-15.
21.    Hashemibeni B, Valiani A, Esmaeli M, Kazemi M, Aliakbari M, Iranpour FG. 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. Iran J Basic Med Sci 2018; 21:212-218.
22.    Munirah S, Kim S, Ruszymah BH, Khang G. The use of fibrin and poly (lactic-co-glycolic acid) hybrid scaffold for articular cartilage tissue engineering: an in vivo analysis. Eur Cell Mater  2008; 21:41-52.
23.    Song JE, Kim MJ, Yoon H, Yoo H, Lee YJ, Kim HN, et al. Effect of hyaluronic acid (HA) in a HA/PLGA scaffold on annulus fibrosus regeneration: In vivo tests. Macromol Res 2013; 21:1075-1082.
24.     Hashemibeni B, Razavi S, 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. IJBMS 2008; 11:10-17.
25.    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; 3:40-52.
26.    Hashemibeni B, Jafary F, Esmaeil N, Goharian V, Feizi G, Heidari F, et al. Comparison of phenotypic characterization between differentiated osteoblasts from stem cells and calvaria osteoblasts in vitro. Int J Prev Med 2013; 4:180-186.
27.    Gugjoo M, Amarpal G, Aithal H, Kinjavdekar P. Cartilage tissue engineering: role of mesenchymal stem cells along with growth factors & scaffolds. Indian J Med Res 2016; 144:339-347.
28.     Lammi M, Piltti J, Prittinen J, Qu C. Challenges in fabrication of tissue-engineered cartilage with correct cellular colonization and extracellular matrix assembly. Int J Mol Sci 2018; 19:2700-2720.
29.    Cui JH, Park SR, Park K, Choi BH, Min B-h. Preconditioning of mesenchymal stem cells with low-intensity ultrasound for cartilage formation in vivo. Tissue Eng 2007; 13:351-360.
30.     Chung C, Burdick JA. Engineering cartilage tissue. Adv Drug Deliv Rev 2008; 60:243-262.
31.    Valiani A, Izadi M, Bahramian H, Esfandiari E, Hashemibeni B. Comparison between the effect of kartogenin and TGFβ3 on chondrogenesis of human adipose-derived stem cells in fibrin scaffold. Bratisl Lek Listy 2017; 118:591-597.
32.    Tuli R, Tuli S, Nandi S, Huang X, Manner PA, Hozack WJ, et al. Transforming growth factor-β-mediated chondrogenesis of human mesenchymal progenitor cells involves N-cadherin and mitogen-activated protein kinase and Wnt signaling cross-talk. J Biol Chem 2003; 278:41227-41236.
33.    Yoon ST, Kim KS, Li J, Park JS, Akamaru T, Elmer WA, et al. The effect of bone morphogenetic protein-2 on rat intervertebral disc cells in vitro. Spine. 2003; 28:1773-1780.
34.    Boumediene K, Felisaz N, Bogdanowicz P, Galera P, Pujol JP. Avocado/soya unsaponifiables enhance the expression of transforming growth factor β1 and β2 in cultured articular chondrocytes. Arthritis Rheumatol 1999; 42:148-156.
35.    Izadi M, Valiani A, Bahramian H, Esfandiari E, Hashemibeni B. Which factor is better for cartilage tissue engineering from human adipose-derived stem cells? kartogenin or avocado soybean unsaponifiable.  Pharmacophore 2018; 9:140-148.  
36.    Henrotin YE, Sanchez C, Deberg MA, Piccardi N, Guillou GB, Msika P, et al. Avocado/soybean unsaponifiables increase aggrecan synthesis and reduce catabolic and proinflammatory mediator production by human osteoarthritic chondrocytes.
Iranian Journal of Basic Medical Sciences
Volume 24, Issue 1
January 2021
Pages 24-29

Files

Share

How to cite

Statistics