Abstract
Ligament-bone interface (enthesis) is a complex structure which comprises of ligament, fibrocartilage and bone. The fibrocartilage transformation adds significant insertional strength to the interface and makes it highly resistant to avulsion forces. Many ACL grafts cannot generate native interfacial region, leading to their failure. Co-culture has proved to be an effective way to generate new tissues in tissue engineering. Studies have found important signaling molecules in transduction pathway of chondrogenesis to be transmitted via gap junctions. We hypothesized that stem cells cocultured between ligament and bone cells would enable transmission of chondrogenic factors from bone/ligament cells to bone marrow stem cells (BMSCs) via gap junctions, resulting in their differentiation into fibrocartilage. To test this hypothesis, we studied to establish effective co-culture system. In this study, two set of co-culture (BMSCs and ligament cells; BMSCs and bone cells) were established. Confocal microscopy showed efficient dye transfer from bone/ligament cells into BMSCs. This was further confirmed and quantified by FACS, which showed a gradual temporal increase in the percentage of BMSCs acquiring Calcein. RT-PCR analysis showed that the bone cells-BMSC and the ligament cells-BMSC co-culture systems expressed higher amounts of collagen type-2, as compared to the various monocultures. The results proved the establishment of effective co-culture. The findings provide important information for the development of a more promising ligamentfibrocartilagebone graft.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Gao, J., K. Messner (1996) Quantitative comparison of soft tissuebone interface at chondral ligament insertions in the rabbit knee joint. J Anat 188 (Pt2): p.367–373
Tadokoro, K., et al. (2004) Evaluation of hamstring strength and tendon regrowth after harvesting for anterior cruciate ligament reconstruction. Am J Sports Med 32(7): p. 1644–1650
Takeda, Y., et al. (2006) Hamstring muscle function after tendon harvest for anterior cruciate ligament reconstruction: evaluation with T2 relaxation time of magnetic resonance imaging. Am J Sports Med 34(2): p.281–288
Roe, J., et al. (2005) A 7-year follow-up of patellar tendon and hamstring tendon grafts for arthroscopic anterior cruciate ligament reconstruction: differences and similarities. Am J Sports Med 34(9): p.1337–1345
Moffat K.L., Lu HH, et al. (2006) Charaterization of the Mechanical Properties and Mineral Distribution of the Anterior Cruciate Ligament-to-Bone Insertion Site. EMBS Annual International Conference. 2366–2369.
Cooper, R. R., Misol, S. (1970) Tendon and ligament insertion. Alight and electron microscopic study. J. Bone Joint Surg. Am. 52,1.
Fujioka H, Thakur R, Wang GJ, et al. (1998) Comparison of surgically attached and non-attached repair of the rat Achilles tendon-bone interface. Cellular organization and type X collagen expression. Connect Tissue Res 37:205–218.
Hendriks, J. et al. (2007) Co-culture in cartilage tissue engineering. J Tissue Eng Regen Med 1:170–178.
Zhang, W et al. (1999) Direct Gap Junction Communication between Malignant Glioma Cells and Astrocytes. Cancer Research 59, 1994–2003 April 15.
Turhani, D., et al. (2005) In vitro study of adherent mandibular osteoblast-like cells on carrier materials. Int J Oral Maxillofac Surg. 34(5): p. 543–550.
Nagineni, C.N., et al. (1992) Characterization of the intrinsic properties of the anterior cruciate and medial collateral ligament cells: an in vitro cell culture study. J Orthop Res, 10(4): p. 465–475.
Sahoo, S., et al. (2006) Characterization of a novel polymeric scaffold for potential application in tendon/ligament tissue engineering. Tissue Eng. 12(1): p. 91–99.
Czyz, J., et al. (2000) Gap-junctional coupling measured by flow cytometry. Exp Cell Res. 255(1): p. 40–46.
Li, X et al. (2005) Modulation of chondrocytic properties of fatderived mesenchymal cells in co-cultures with nucleus pulposus. Connect Tissue Res. 46(2):75–82.
Visconti C.S. et al. (1996) Biochemical analysis of collagens at the ligament-bone interface reveals presence of cartilage-specific collagens. Arch Biochem Biophys. Apr 1;328(1):135–142
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 International Federation of Medical and Biological Engineering
About this paper
Cite this paper
He, P.F., Sahoo, S., Goh, J.C., Toh, S.L. (2009). Establishing a Coculture System for Ligament-Bone Interface Tissue Engineering. In: Lim, C.T., Goh, J.C.H. (eds) 13th International Conference on Biomedical Engineering. IFMBE Proceedings, vol 23. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-92841-6_375
Download citation
DOI: https://doi.org/10.1007/978-3-540-92841-6_375
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-540-92840-9
Online ISBN: 978-3-540-92841-6
eBook Packages: EngineeringEngineering (R0)