Hostname: page-component-78c5997874-4rdpn Total loading time: 0 Render date: 2024-11-13T09:58:09.376Z Has data issue: false hasContentIssue false
  • English
  • Français

Collagen-Chondroitin Sulphate-Hydroxyapatite Porous Composites: A Histochemical and Electron Microscopy Approach

Published online by Cambridge University Press:  26 January 2010

Otilia Zarnescu ,
Oana Craciunescu  and
Lucia Moldovan
Otilia Zarnescu*
Affiliation:
Department of Animal Biology, Faculty of Biology, University of Bucharest, Splaiul Independentei 91-95, R-050095, Bucharest, Romania
Oana Craciunescu
Affiliation:
Department of Cellular and Molecular Biology, National Institute of Research and Development for Biological Sciences, Splaiul Independentei 296, R-060031, Bucharest, Romania
Lucia Moldovan
Affiliation:
Department of Cellular and Molecular Biology, National Institute of Research and Development for Biological Sciences, Splaiul Independentei 296, R-060031, Bucharest, Romania
*
Corresponding author. E-mail: otilia@bio.unibuc.ro
Get access

Abstract

In this study the structure of collagen-chondroitin sulphate-hydroxyapatite porous composites is investigated by histochemical (Von Kossa staining), immunohistochemical, and transmission electron microscopy. Examination of composites on picrosirius red stained sections showed that polarization colors of collagen were generally in the range of orange-red. Immunofluorescence data indicate that chondroitin sulphate was either chemically incorporated into the bulk structure of collagen scaffolds or attached on surfaces of collagen bundles. Depending on the ratio between collagen:chondroitin sulphate:hydroxyapatite, von Kossa histochemical staining showed a progressive loading of collagen-chondroitin sulphate bundles with hydroxyapatite. Transmission electron microscopy studies have shown that composites contain mostly collagen fibrils aggregated with random orientation with very few collagen fibers showing the 67-nm banding pattern. Hydroxyapatite deposits of various sizes occurred among the collagen fibrils.

Keywords

compositecollagenhydroxyapatitechondroitin sulphatehistochemistryimmunohistochemistryTEM
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 16 , Issue 2 , April 2010 , pp. 137 - 142
Copyright
Copyright © Microscopy Society of America 2010

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Burger, M., Sherman, B.S. & Sobel, A.E. (1962). Observations of the influence of chondroitin sulphate on the rate of bone repair. J Bone Joint Surg 44B, 675–687.Google Scholar
Dayan, D., Hiss, Y., Hirshberg, A., Bubis, J.J. & Wolman, M. (1989). Are the polarization colors of picrosirius red-stained collagen determined only by the diameter of the fibers? Histochemistry 93, 27–29.CrossRefGoogle ScholarPubMed
Declercq, H., Van Der Vreken, N., De Maeyer, E., Verbeeck, R., Schacht, E., De Ridder, L. & Cornelissen, M. (2004). Isolation, proliferation and differentiation of osteoblastic cells to study cell/biomaterial interactions: Comparison of different isolation techniques and source. Biomaterials 25, 757–768.CrossRefGoogle ScholarPubMed
Glimcher, M.J. (1992). The nature of the mineral component of bone and the mechanism of calcification. In Metabolic Bone Disease and Clinically Related Disorders, Coe, F.L. & Favus, M.J. (Eds.), pp. 265–286. New York: Raven Press Ltd.Google Scholar
Holbrook, K.A. & Smith, L.T. (1993). Morphology of connective tissue: Structure of the skin and tendon. In Connective Tissue and Its Heritable Disorders—Molecular, Genetic and Medical Aspects, Royce, P.M. & Steinmann, B. (Eds.), pp. 51–71. New York: Wiley-Liss.Google Scholar
Junqueira, L.C., Cossermelli, W. & Brentani, R. (1978). Differential staining of collagens type I, II and III by Sirius Red and polarization microscopy. Arch Histol Jpn 41, 267–274.Google Scholar
Kamakura, S., Sasaki, K., Honda, Y., Anada, T., Matsui, K., Echigo, S. & Suzuki, O. (2007). Dehydrothermal treatment of collagen influences on bone regeneration by octacalcium phosphate (OCP) collagen composites. J Tissue Eng Regen Med 1, 450–456.CrossRefGoogle Scholar
Kikuchi, M., Ikoma, T., Itoh, S., Matsumoto, H.N., Koyama, Y., Takakuda, K., Shinomiya, K. & Tanaka, J. (2004). Biomimetic synthesis of bone-like nanocomposites using the self organization mechanism of hydroxyapatite and collagen. Compos Sci Technol 64, 819–825.Google Scholar
Liu, C., Han, Z. & Czernuszka, J.T. (2009). Gradient collagen/nanohydroxyapatite composite scaffold: Development and characterization. Acta Biomater 5, 661–669.Google Scholar
Moldovan, L., Craciunescu, O., Oprita, E.I., Balan, M. & Zarnescu, O. (2009). Collagen-chondroitin sulfate-hydroxyapatite porous composites: Preparation, characterization and in vitro biocompatibility testing. Roum Biotechnol Lett 14, 4462–4469.Google Scholar
Moldovan, L., Oancea, O., Zarnescu, O. & Barboi, G. (1996). In vitro biocompatibility evaluation of collagen-based sponges. Rev Roum Biol Biol Anim 4, 59–64.Google Scholar
Murugan, R. & Ramakrishna, S. (2005). Development of nanocomposites for bone grafting. Comp Sci & Tech 65, 2385–2406.Google Scholar
Rhee, S.H. & Tanaka, J. (2002). Self-assembly phenomenon of hydroxyapatite nanocrystals on chondroitin sulfate. J Mater Sci Mater Med 13, 597–600.CrossRefGoogle ScholarPubMed
Supová, M. (2009). Problem of hydroxyapatite dispersion in polymer matrices: A review. J Mater Sci Mater Med 20, 1201–213.CrossRefGoogle ScholarPubMed
Suvorova, E.I., Klechkovskaya, V.V., Komarov, V.F., Severin, A.V., Melikhov, I.V. & Buffat, P.A. (2006). Electron microscopy of biomaterials based on hydroxyapatite. Crystallogr Rep 51, 881–887.CrossRefGoogle Scholar
Weiner, S. & Traub, W. (1986). Organization of hydroxyapatite crystals within collagen fibrils. FEBS Lett 206, 262–266.Google Scholar
Xie, J., Baumann, M.J. & McCabe, L.R. (2004). Osteoblasts respond to hydroxyapatite surfaces with immediate changes in gene expression. J Biomed Mater Res A 71, 108–117.CrossRefGoogle ScholarPubMed
Yunoki, S., Marukawa, E., Ikoma, T., Sotome, S., Fan, H., Zhang, X., Shinomiya, K. & Tanaka, J. (2007). Effect of collagen fibril formation on bioresorbability of hydroxyapatite/collagen composites. J Mater Sci Mater Med 18, 2179–2183.Google Scholar
Zhang, S.M., Cui, F.Z., Liao, S.S., Zhu, Y. & Han, L. (2003). Synthesis and biocompatibility of porous nano-hydroxyapatite/collagen/alginate composite. J Mater Sci Mater Med 14, 641–645.CrossRefGoogle ScholarPubMed