Type II collagen-chondroitin sulfate-hyaluronan scaffold cross-linked by genipin for cartilage tissue engineering

https://doi.org/10.1016/j.jbiosc.2008.09.020Get rights and content

Abstract

Owing to of the limited repair capacity of articular cartilage, it is essential to develop tissue-engineered cartilage for patients suffering from joint disease. Chondroitin sulfate (CS) and hyaluronan (HA) are the components of the cartilage extracellular matrix (ECM) and are known to influence the proliferation and differentiation of chondrocytes. Scaffolds composed of type-II collagen, CS, and HA may create an environment that can preserve the normal phenotype of cells to promote regeneration of cartilage-like constructs. In this investigation, we prepared and characterized 3-dimensional type-II collagen scaffolds both with and without HA and CS. Porous composite scaffolds fabricated by freeze-drying showed interconnected pores with mean diameters of 140 ± 30 µm and porosities of 92–95% after cross-linking with genipin. After a 14-day in vitro culture, morphologically round chondrocytes were found to be uniformly distributed throughout the sponges. Expression of genes of aggrecan, type-II collagen and cartilage oligomeric matrix protein (COMP) was statistically and significantly increased on scaffolds with CS and HA than those without CS and HA. Furthermore, there was a markedly greater accumulation of proteoglycans (PGs) on the scaffolds with CS and HA.

Section snippets

Collagen II sponge fabrication and cross-linking

COL II made from bovine trachea was prepared using previously defined protocols (19). A sample of 100 mg lyophilized collagen thus made was homogenized in 0.5 M acetic acid at 4 °C to give a final COL II concentration of 1% (wt/vol). The well-mixed slurry was poured into molds and then frozen and lyophilized at − 20 °C and − 58 °C, respectively. The fabricated COL II sponges were immersed overnight in 70% alcohol to remove acetic acid before cross-linking. CS with MW of ca. 100,000 from shark

Three-dimensional composite scaffold fabrication

The scaffolds used for tissue engineering must have high porosity with a large surface area/volume ratio to provide a greater space for cells and to allow the production of new ECM. In this study, porous COL II sponges were fabricated, both with and without CS and HA, using high purity type-II atelocollagen and cross-linker genipin to mimic the native ECM of articular cartilage. The best cross-linker for primary amino groups is the naturally existing reagent genipin, which has been reported to

Acknowledgments

This research is supported by the Ministry of Economic Affairs, Taiwan, under the Technology Development Program for Academia grant 91-EC-17-A-17-S1-0009.

References (26)

  • SungH.W. et al.

    In vitro surface characterization of a biological patch fixed with a naturally occurring cross-linking agent

    Biomaterials

    (2000)
  • BassleerC.T. et al.

    Effects of chondroitin sulphate and interleukin-1 on human articular chondrocytes cultivated in clusters

    Osteoarthr. Cartil.

    (1998)
  • BanuN. et al.

    Markedly different effects of hyaluronic acid and chondroitin sulfate-A on the differentiation of human articular chondrocytes in micromass and 3-D honeycomb rotation cultures

    J. Biomed. Mater. Res.

    (2007)
  • Cited by (111)

    • Biomaterials and tissue engineering approaches using glycosaminoglycans for tissue repair: Lessons learned from the native extracellular matrix

      2023, Acta Biomaterialia
      Citation Excerpt :

      reported sponges containing gelatin-CS-HA in poly (lactic-co-glycolic acid) (PLGA) improved chondrogenic differentiation of MSCs and MSC/TGF-β3 immobilized sponges regenerated chondral defects in a rabbit model [172]. Similarly collagen-based HA and CS containing hydrogels also showed improved chondrogenic activity [173]. Chitosan and HA copolymers have also been extensively studied for cartilage tissue engineering [174].

    • In the quest of the optimal chondrichthyan for the development of collagen sponges for articular cartilage

      2021, Journal of Science: Advanced Materials and Devices
      Citation Excerpt :

      These observations not only confirm that the pore size and porosity were suitable for cell and ECM migration within the three-dimensional conformation of the scaffolds, but also indicate the chondrogenic potential of the scaffolds. Again these observations are in agreement with previous work in the field, where Jellyfish (R. esculentum) collagen sponges with pore size of 40–200 μm (at the centre of the scaffold, the pore diameter was 74 ± 18 μm and at superficial zones, the pore diameter was 50 ± 20 μm) [14] and bovine collagen type II sponges with pore size of 140 ± 30 μm [50] maintained chondrocyte phenotype and Jellyfish (R. esculentum) collagen sponges with porosity of 98.2 ± 0.4% induced chondrogenic differentiation of human mesenchymal stem cells [16]. With respect to gene analysis, in general all collagen scaffolds showed a higher chondro-inductive potential compared to TCP, as evidenced by downregulated expression of COL1A1 and COL3A1 (except the LSD scaffold) and upregulated expression of COL10A1, SOX9, COMP and ACAN (except the CR scaffold).

    View all citing articles on Scopus
    View full text