Chondrocytes cultured in silk-based biomaterials maintain function and cell morphology

Abstract

Objective:

To characterize the morphology of chondrocytes and the expression and secretion of active collagen II by these cells cultured within a regenerated silk fibroin film. Silk fibroin film cytocompatibility and the effect of silk fibroin on chondrocytes in vitro were also evaluated.

Methods:

Chondrocytes were transfected with a lentivirus containing a green fluorescent protein marker and cultured within a regenerated silk fibroin film. Effects on chondrocyte adhesion, growth, and expression of functional collagen II were assessed in vitro by analysis with immunofluorescent histochemistry and laser scanning confocal microscopy.

Results:

The results of this study showed that the regenerated silk fibroin film had no cytotoxic effect on chondrocytes. The regenerated silk fibroin film facilitated the adhesion of chondrocytes with typical morphology. Chondrocytes cultured within silk fibroin films exhibited the expression of collagen II in vitro.

Conclusion:

Regenerated silk fibroin film was found to be an excellent biomaterial with good cytocompatibility for chondrocytes, because these cells remained functional and maintained normal cell morphology when cultured in silk-based biomaterials. These results suggest that silk-based chondrocyte biomaterial complexes may provide a feasible and functional biomaterial for repairing clinical cartilage defects.

Keywords

Regenerated silk fibroin film biomaterials cytocompatibility chondrocytes

Get full access to this article

View all access options for this article.

References

Little

Bawolin

Chen

Mechanical properties of natural cartilage and tissue-engineered constructs. Tissue Eng Part B Rev 2011; 17(4): 213–227.

Sancho-Tello

Forriol

Gastaldi

et al . Time evolution of in vivo articular cartilage repair induced by bone marrow stimulation and scaffold implantation in rabbits. Int J Artif Organs 2015; 38(4): 210–223.

Wang

Yuan

Guo

Q-Y

et al . Mesenchymal stem cells for treating articular cartilage defects and osteoarthritis. Cell Transplant 2015; 24: 1661–1678.

Musumeci

Castrogiovanni

Leonardi

et al . New perspectives for articular cartilage repair treatment through tissue engineering: a contemporary review. World J Orthop 2014; 5(2): 80–88.

Mobasheri

Kalamegam

Musumeci

et al . Chondrocyte and mesenchymal stem cell-based therapies for cartilage repair in osteoarthritis and related orthopaedic conditions. Maturitas 2014; 78(3): 188–198.

Musumeci

Lo Furno

Loreto

et al . Mesenchymal stem cells from adipose tissue which have been differentiated into chondrocytes in three-dimensional culture express lubricin. Exp Biol Med 2011; 236: 1333–1341.

Bartlett

Skinner

Gooding

et al . Autologous chondrocyte implantation versus matrix-induced autologous chondrocyte implantation for osteochondral defects of the knee: a prospective, randomised study. J Bone Joint Surg Br 2005; 87(5): 640–645.

Vikingsson

Gallego Ferrer

Gómez-Tejedor

et al . An “in vitro” experimental model to predict the mechanical behavior of macroporous scaffolds implanted in articular cartilage. J Mech Behav Biomed Mater 2014; 32: 125–131.

Kushibe

Nezu

Nishizaki

et al . Tracheal allotransplantation maintaining cartilage viability with long-term cryopreserved allografts. Ann Thorac Surg 2001; 71: 1666–1669.

10.

Yildirim

Haliloglu

Sapci

et al . Tracheal reconstruction with porous high-density polyethylene tracheal prosthesis. Ann Otol Rhinol Laryngol 2000; 109: 981–987.

11.

Freed

Grande

Lingbin

et al . Joint resurfacing using allograft chondrocytes and synthetic biodegradable polymer scaffolds. J Biomed Mater Res 1994; 28: 891–889.

12.

Mori

Tsukada

New silk protein: modification of silk protein by gene engineering for production of biomaterials. J Biotechnol 2000; 74(2): 95–103.

13.

Horan

Antle

Collette

et al . In vitro degradation of silk fibroin. Biomaterials 2005; 26(17): 3385–3393.

14.

Meinel

Hofmann

Karageorgiou

et al . The inflammatory responses to silk films in vitro and in vivo. Biomaterials 2005; 26(2): 147–155.

15.

Demers

Griffin

De Vroey

et al . Harmonization of animal care and use guidance. Science 2006; 312(5): 700–701.

16.

Jiang

Wen

et al . Construction of a functional silk-based biomaterial complex with immortalized chondrocytes in vivo. J Biomed Mater Res A 2014; 102(4): 1071–1078.

17.

Ogiso

Minoura

Enzymatic degradation behavior of porous silk fibroin sheets. Biomaterials 2003; 24: 357–365.

18.

Altman

Diaz

Jakuba

et al . Silk-based biomaterials. Biomaterials 2003; 24: 401–416.

19.

Zhou

Confalonieri

Medina

et al . Fine organization of Bombyx mori fibroin heavy chain gene. Nucleic Acids Res 2000; 28: 2413–2419.

20.

Motta

Migliaresi

Faccioni

et al . Fibroin hydrogels for biomedical applications: preparation, characterization and in vitro cell culture studies. J Biomater Sci Polym Ed 2004; 15: 851–864.

21.

Liu

Zhang

et al . Study on blood compatibility of silk fibroin membrane. Chin J Hemorheol 2002; 12: 266–269.

22.

Liu

Miao

Sheng

et al . Cytocompatibility of regenerated silk fibroin film: a medical biomaterial applicable to wound healing. J Zhejiang Univ Sci B 2010; 11(1): 10–16.

23.

Ouyang

Wang

et al . In vivo histocompatibility evaluation of polyurethane membrane modified by superfine silk-fibroin powder. J Huazhong Univ Sci Technolog Med Sci 2009; 29(4): 508–511.

24.

Wang

Wenk

et al . Silk coatings on PLGA and alginate microspheres for protein delivery. Biomaterials 2007; 28(28): 4161–4169.

25.

Unger

Michael

Kirsten

et al . Growth of human cells on a non-woven silk fibroin net: a potential for use in tissue engineering. Biomaterials 2004; 25(6): 1069–1075.

26.

Altman

Horan

et al . Silk matrix for tissue engineered anterior cruciate ligaments. Biomaterials 2002; 23(20): 4131–4141.

27.

Minoura

Aiba

Gotoh

et al . Attachment and growth of fibroblast cells on silk fibroin. Biochem Biophys Res Comm 1995; 208(2): 511–516.

28.

Inounye

Kurokawa

Nishikawa

et al . Use of Bombyx mori silk fibroin as a substratum for cultivation of animal cells. J Biochem Biophys Methods 1998; 37: 159–164.

29.

Chiarin

Petrini

Bozzini

et al . Silk fibroin/poly(carbonate)-urethane as a substrate for cell growth: in vitro interactions with human cells. Biomaterials 2003; 24: 789–799.

30.

Gotoh

Tsukada

Minoura

et al . Synthesis of poly(ethylene glycol)-silk fibroin conjugates and surface interaction between L-929 cells and the conjugates. Biomaterials 1997; 18(3): 267–271.

31.

Setzen

Williams

EF.

Tissue response to suture materials implanted subcutaneously in a rabbit model. Plast Reconstr Surg 1997; 100: 1788–1795.

32.

Mandal

Kundu

SC.

Cell proliferation and migration in silk fibroin 3D scaffolds. Biomaterials 2009; 30(15): 2956–2965.

33.

Fang

Chen

Yang

et al . In vitro and in vivo research on using Antheraea pernyi silk fibroin as tissue engineering tendon scaffolds. Mater Sci Eng C 2009; 29: 1527–1534.

34.

Wang

Rudym

Walsh

et al . In vivo degradation of three-dimensional silk fibroin scaffolds. Biomaterials 2008; 29: 3415–3428.