Cultivation of Intervertebral Disc Cells in Medium Fortified with Growth Factors Improved In Vitro Chondrogenesis

Various chondrogenic growth factors have been shown to promote proliferation, matrix production and early chondrogenesis of intervertebral disc (IVD). The objective of this study is to reconstruct IVD via tissue engineering technique and to evaluate in-vitro chon-drogenesis of chondrocytes cultured in basic growth medium with chondrogenic growth factors (FDGFs) and without chondrogenic growth factors (FD). The annulus fibrosus (AF) and nucleus pulposus (NP) were aseptically dissected from the lumbar discs of sheep and the tissues enzymatically digested and cultured in-vitro. Passage 1 cells were used to form in-vitro construct with autologous fibrin as the biomaterial. All constructs were stabi-lized for 14 days in their respective growth media. Histological and sulphated-glycosaminoglycan (sGAG) analysis were performed at the end of the experiment. The AF cells showed greater growth kinetic profile and higher viability in FDGFs while the NP cells showed no differences for both medium. The construct of AF and NP in FDGFs showed a bigger size construct compared to FD. The qualities of both constructs were simi-lar as evidence by Safranin O staining in both groups. In conclusion, FDGFs increase the viability and quantity of cells for AF and not in NP. However, FDGFs supplementation improved in-vitro constructs formation for both cells type.

[1]  Soon Hee Kim,et al.  The use of fibrin and poly(lactic-co-glycolic acid) hybrid scaffold for articular cartilage tissue engineering: an in vivo analysis. , 2008, European cells & materials.

[2]  M. Kneževič,et al.  Evaluation of rabbit auricular chondrocyte isolation and growth parameters in cell culture , 2007, Cell biology international.

[3]  J. Blaker,et al.  Three-dimensional culture of annulus fibrosus cells within PDLLA/Bioglass composite foam scaffolds: assessment of cell attachment, proliferation and extracellular matrix production. , 2007, Biomaterials.

[4]  S Holm,et al.  Nutrition of the intervertebral disk. An in vivo study of solute transport. , 1977, Clinical orthopaedics and related research.

[5]  Qiong Wu,et al.  Evaluation of three-dimensional scaffolds prepared from poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) for growth of allogeneic chondrocytes for cartilage repair in rabbits. , 2008, Biomaterials.

[6]  H. An,et al.  Growth factors and the intervertebral disc. , 2004, The spine journal : official journal of the North American Spine Society.

[7]  J. Gough,et al.  In vitro studies of annulus fibrosus disc cell attachment, differentiation and matrix production on PDLLA/45S5 Bioglass composite films. , 2006, Biomaterials.

[8]  C. K. Hui,et al.  The Re-Expression of Collagen Type 2, Aggrecan and Sox 9 in Tissue-Engineered Human Articular Cartilage , 2005 .

[9]  F. Strutz,et al.  TGF-beta 1 induces proliferation in human renal fibroblasts via induction of basic fibroblast growth factor (FGF-2). , 2001, Kidney international.

[10]  Charles A Vacanti,et al.  Tissue-Engineered Composites of Anulus Fibrosus and Nucleus Pulposus for Intervertebral Disc Replacement , 2004, Spine.

[11]  C. Laurencin,et al.  Biphasic scaffold for annulus fibrosus tissue regeneration. , 2008, Biomaterials.

[12]  K. Chua,et al.  Pediatric auricular chondrocytes gene expression analysis in monolayer culture and engineered elastic cartilage. , 2007, International journal of pediatric otorhinolaryngology.

[13]  K. Chua,et al.  Basic fibroblast growth factor with human serum supplementation: enhancement of human chondrocyte proliferation and promotion of cartilage regeneration. , 2007, Singapore medical journal.

[14]  Sally Roberts,et al.  Degeneration of the intervertebral disc , 2003, Arthritis research & therapy.

[15]  J. Hoyland,et al.  Degeneration of intervertebral discs: current understanding of cellular and molecular events, and implications for novel therapies , 2001, Expert Reviews in Molecular Medicine.

[16]  K. Chua,et al.  Insulin-transferrin-selenium prevent human chondrocyte dedifferentiation and promote the formation of high quality tissue engineered human hyaline cartilage. , 2005, European cells & materials.

[17]  Munirah Sha'ban,et al.  Fibrin promotes proliferation and matrix production of intervertebral disc cells cultured in three-dimensional poly(lactic-co-glycolic acid) scaffold , 2008, Journal of biomaterials science. Polymer edition.