Icariin promotes stable chondrogenic differentiation of bone marrow mesenchymal stem cells in self‑assembling peptide nanofiber hydrogel scaffolds

Wang,Zhicong; Li,Kaihua; Sun,Huijun; Wang,Ji; Fu,Zhuodong; Liu,Mozhen

doi:10.3892/mmr.2018.8913

Icariin promotes stable chondrogenic differentiation of bone marrow mesenchymal stem cells in self‑assembling peptide nanofiber hydrogel scaffolds

Authors:
- Zhicong Wang
- Kaihua Li
- Huijun Sun
- Ji Wang
- Zhuodong Fu
- Mozhen Liu
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Affiliations: Department of Orthopedic Surgery, People's Hospital of Deyang City, Deyang, Sichuan 618000, P.R. China, Department of Orthopedic Surgery, General Hospital of Fengfeng Group, Handan, Hebei 056200, P.R. China, Department of Clinical Pharmacology, Dalian Medical University, Dalian, Liaoning 116044, P.R. China, Department of Orthopedic Surgery, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China
Published online on: April 23, 2018 https://doi.org/10.3892/mmr.2018.8913
Pages: 8237-8243

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Abstract

Icariin, a traditional Chinese medicine, has previously been demonstrated to promote chondrogenesis of bone marrow mesenchymal stem cells (BMSCs) in traditional 2D cell culture. The present study investigated whether icariin has the potential to promote stable chondrogenic differentiation of BMSCs without hypertrophy in a 3D microenvironment. BMSCs were cultivated in a self‑assembling peptide nanofiber hydrogel scaffold in chondrogenic medium for 3 weeks. Icariin was added to the medium throughout the culture period at concentrations of 1x10‑6 M. Chondrogenic differentiation markers, including collagen II and SRY‑type high mobility group box 9 (SOX9) were detected by immunofluorescence, reverse transcription‑quantitative polymerase chain reaction and toluidine blue staining. Hypertrophic differentiation was further assessed by detecting collagen X and collagen I gene expression levels and alkaline phosphatase activity. The results demonstrated that icariin significantly enhanced cartilage extracellular matrix synthesis and gene expression levels of collagen II and SOX9, and additionally promoted more chondrocyte‑like rounded morphology in BMSCs. Furthermore, chondrogenic medium led to hypertrophic differentiation via upregulation of collagen X and collagen I gene expression levels and alkaline phosphatase activity, which was not potentiated by icariin. In conclusion, these results suggested that icariin treatment may promote chondrogenic differentiation of BMSCs, and inhibit the side effect of growth factor activity, thus preventing further hypertrophic differentiation. Therefore, icariin may be a potential compound for cartilage tissue engineering.

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1	Lubis AM and Lubis VK: Adult bone marrow stem cells in cartilage therapy. Acta Med Indones. 44:62–68. 2012.PubMed/NCBI
2	Giovannini S, Diaz-Romero J, Aigner T, Heini P, Mainil-Varlet P and 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. 20:245–259. 2010. View Article : Google Scholar : PubMed/NCBI
3	Aung A, Gupta G, Majid G and Varghese S: Osteoarthritic chondrocyte-secreted morphogens induce chondrogenic differentiation of human mesenchymal stem cells. Arthritis Rheum. 63:148–158. 2011. View Article : Google Scholar : PubMed/NCBI
4	Pelttari K, Winter A, Steck E, Goetzke K, Hennig T, Ochs BG, Aigner T and Richter W: Premature induction of hypertrophy during in vitro chondrogenesis of human mesenchymal stem cells correlates with calcification and vascular invasion after ectopic transplantation in SCID mice. Arthritis Rheum. 54:3254–3266. 2006. View Article : Google Scholar : PubMed/NCBI
5	Mueller MB, Fischer M, Zellner J, Berner A, Dienstknecht T, Kujat R, Prantl L, Nerlich M, Tuan RS and Angele P: Effect of parathyroid hormone-related protein in an in vitro hypertrophy model for mesenchymal stem cell chondrogenesis. Int Orthop. 37:945–951. 2013. View Article : Google Scholar : PubMed/NCBI
6	Zhang L, Zhang X, Li KF, Li DX, Xiao YM, Fan YJ and Zhang XD: Icariin promotes extracellular matrix synthesis and gene expression of chondrocytes in vitro. Phytother Res. 26:1385–1392. 2012. View Article : Google Scholar : PubMed/NCBI
7	Li D, Yuan T and Zhang X, Xiao Y, Wang R, Fan Y and Zhang X: Icariin: A potential promoting compound for cartilage tissue engineering. Osteoarthritis Cartilage. 20:1647–1656. 2012. View Article : Google Scholar : PubMed/NCBI
8	Fan JJ, Cao LG, Wu T, Wang DX, Jin D, Jiang S, Zhang ZY, Bi L and Pei GX: The dose-effect of icariin on the proliferation and osteogenic differentiation of human bone mesenchymal stem cells. Molecules. 16:10123–10133. 2011. View Article : Google Scholar : PubMed/NCBI
9	Liu MH, Sun JS, Tsai SW, Sheu SY and Chen MH: Icariin protects murine chondrocytes from lipopolysaccharide-induced inflammatory responses and extracellular matrix degradation. Nutr Res. 30:57–65. 2010. View Article : Google Scholar : PubMed/NCBI
10	Sun P, Liu Y, Deng X, Yu C, Dai N, Yuan X, Chen L, Yu S, Si W, Wang X, et al: An inhibitor of cathepsin K, icariin suppresses cartilage and bone degradation in mice of collagen-induced arthritis. Phytomedicine. 20:975–979. 2013. View Article : Google Scholar : PubMed/NCBI
11	Wang ZC, Sun HJ, Li KH, Fu C and Liu MZ: Icariin promotes directed chondrogenic differentiation of bone marrow mesenchymal stem cells but not hypertrophy in vitro. Exp Ther Med. 8:1528–1534. 2014. View Article : Google Scholar : PubMed/NCBI
12	Li J, Mareddy S, Tan DM, Crawford R, Long X, Miao X and Xiao Y: A minimal common osteochondrocytic differentiation medium for the osteogenic and chondrogenic differentiation of bone marrow stromal cells in the construction of osteochondral graft. Tissue Eng Part A. 15:2481–2490. 2009. View Article : Google Scholar : PubMed/NCBI
13	Caron MM, Emans PJ, Coolsen MM, Voss L, Surtel DA, Cremers A, van Rhijn LW and Welting TJ: Redifferentiation of dedifferentiated human articular chondrocytes: Comparison of 2D and 3D cultures. Osteoarthritis Cartilage. 20:1170–1178. 2012. View Article : Google Scholar : PubMed/NCBI
14	Kisiday J, Jin M, Kurz B, Hung H, Semino C, Zhang S and Grodzinsky AJ: Self-assembling peptide hydrogel fosters chondrocyte extracellular matrix production and cell division: Implications for cartilage tissue repair. Proc Natl Acad Sci USA. 99:9996–10001. 2002. View Article : Google Scholar : PubMed/NCBI
15	Kopesky PW, Vanderploeg EJ, Sandy JS, Kurz B and Grodzinsky AJ: Self-assembling peptide hydrogels modulate in vitro chondrogenesis of bovine bone marrow stromal cells. Tissue Eng Part A. 16:465–477. 2010. View Article : Google Scholar : PubMed/NCBI
16	Wang B, Sun C, Shao Z, Yang S, Che B, Wu Q and Liu J: Designer self-assembling Peptide nanofiber scaffolds containing link protein N-terminal peptide induce chondrogenesis of rabbit bone marrow stem cells. Biomed Res Int. 2014:4219542014. View Article : Google Scholar : PubMed/NCBI
17	Erickson IE, Huang AH, Chung C, Li RT, Burdick JA and Mauck RL: Differential maturation and structure-function relationships in mesenchymal stem cell- and chondrocyte-seeded hydrogels. Tissue Eng Part A. 15:1041–1052. 2009. View Article : Google Scholar : PubMed/NCBI
18	Kisiday JD, Kopesky PW, Evans CH, Grodzinsky AJ, McIlwraith CW and Frisbie DD: Evaluation of adult equine bone marrow- and adipose-derived progenitor cell chondrogenesis in hydrogel cultures. J Orthop Res. 26:322–331. 2008. View Article : Google Scholar : PubMed/NCBI
19	Miller RE, Grodzinsky AJ, Vanderploeg EJ, Lee C, Ferris DJ, Barrett MF, Kisiday JD and Frisbie DD: Effect of self-assembling peptide, chondrogenic factors, and bone marrow-derived stromal cells on osteochondral repair. Osteoarthritis Cartilage. 18:1608–1619. 2010. View Article : Google Scholar : PubMed/NCBI
20	Miller RE, Grodzinsky AJ, Barrett MF, Hung HH, Frank EH, Werpy NM, McIlwraith CW and Frisbie DD: Effects of the combination of microfracture and self-assembling Peptide filling on the repair of a clinically relevant trochlear defect in an equine model. J Bone Joint Surg Am. 96:1601–1609. 2014. View Article : Google Scholar : PubMed/NCBI
21	He B, Yuan X, Zhou A, Zhang H and Jiang D: Designer functionalised self-assembling peptide nanofibre scaffolds for cartilage tissue engineering. Expert Rev Mol Med. 16:e122014. View Article : Google Scholar : PubMed/NCBI
22	Cooke ME, Allon AA, Cheng T, Kuo AC, Kim HT, Vail TP, Marcucio RS, Schneider RA, Lotz JC and Alliston T: Structured three-dimensional co-culture of mesenchymal stem cells with chondrocytes promotes chondrogenic differentiation without hypertrophy. Osteoarthritis Cartilage. 19:1210–1218. 2011. View Article : Google Scholar : PubMed/NCBI
23	Yuan T, He L, Yang J, Zhang L, Xiao Y, Fan Y and Zhang X: Conjugated icariin promotes tissue-engineered cartilage formation in hyaluronic acid/collagen hydrogel. Process Biochemistry. 50:2242–2250. 2015. View Article : Google Scholar
24	He L, Yang J, Lu J, Xiao Y, Fan Y and Zhang X: Preparation and characterization of a novel hyaluronic acid-icariin conjugate hydrogel. Materials Lett. 136:41–44. 2014. View Article : Google Scholar
25	Yan H, Zhou Z, Huang T, Peng C, Liu Q, Zhou H, Zeng W, Liu L, Ou V, He S and Huang H: Controlled release in vitro of icariin from gelatin/hyaluronic acid composite microspheres. Polymer Bulletin. 73:1055–1066. 2016. View Article : Google Scholar
26	Ichinose S, Muneta T, Koga H, Segawa Y, Tagami M, Tsuji K and Sekiya I: Morphological differences during in vitro chondrogenesis of bone marrow-, synovium-MSCs, and chondrocytes. Lab Invest. 90:210–221. 2010. View Article : Google Scholar : PubMed/NCBI
27	Ichinose S, Tagami M, Muneta T, Mukohyama H and Sekiya I: Comparative sequential morphological analyses during in vitro chondrogenesis and osteogenesis of mesenchymal stem cells embedded in collagen gels. Med Mol Morphol. 46:24–33. 2013. View Article : Google Scholar : PubMed/NCBI