Scaffold-free cartilage tissue engineering with a small population of human nasoseptal chondrocytes

Laryngoscope. 2017 Mar;127(3):E91-E99. doi: 10.1002/lary.26396. Epub 2016 Nov 15.

Abstract

Objective: Cartilage tissue engineering is a promising approach to provide suitable materials for nasal reconstruction; however, it typically requires large numbers of cells. We have previously shown that a small number of chondrocytes cultivated within a continuous flow bioreactor can elicit substantial tissue growth, but translation to human chondrocytes is not trivial. Here, we aimed to demonstrate the application of the bioreactor to generate large-sized tissues from a small population of primary human nasoseptal chondrocytes.

Study design: Experimental study.

Methods: Chondrocytes were cultured in the bioreactor using different medium compositions, with varying amounts of serum and with or without growth factors. Resulting engineered tissues were analyzed for physical properties, biochemical composition, tissue microstructure, and protein localization.

Results: Bioreactor-cultivated constructs grown with serum and growth factors (basic fibroblast growth factor and transforming growth factor beta 2) had greater thickness, as well as DNA and glycosaminoglycan (GAG) contents, compared to low serum and no growth factor controls. These constructs also showed the most intense proteoglycan and collagen II staining.

Conclusion: The combination of bioreactor conditions, serum, and growth factors allowed the generation of large, thick scaffold-free human cartilaginous tissues that resembled the native nasoseptal cartilage. There also may be implications for patient selection in future clinical applications of these engineered tissues because their GAG content decreased with donor age.

Level of evidence: NA. Laryngoscope, 127:E91-E99, 2017.

Keywords: Cartilage tissue engineering; continuous flow bioreactor; growth factors; human nasoseptal cartilage; primary cells; scaffold-free.

MeSH terms

  • Biomechanical Phenomena
  • Cell Culture Techniques
  • Cells, Cultured
  • Chondrocytes / cytology*
  • Chondrocytes / pathology
  • Fibroblast Growth Factor 2 / administration & dosage
  • Humans
  • Immunohistochemistry
  • Nasal Septum / cytology
  • Nasal Surgical Procedures / methods
  • Plastic Surgery Procedures / methods
  • Receptors, Transforming Growth Factor beta / administration & dosage
  • Tensile Strength*
  • Tissue Engineering / methods*
  • Tissue Scaffolds
  • Tissue and Organ Harvesting

Substances

  • Receptors, Transforming Growth Factor beta
  • Fibroblast Growth Factor 2