title: [Experimental study on tissue engineered cartilage complex three-dimensional nano-scaffold with collagen type II and hyaluronic acid in vitro].
abstract: OBJECTIVE
  To explore the possibility of constructing tissue engineered cartilage complex three-dimensional nano-scaffold with collagen type II and hyaluronic acid (HA) by electrospinning.
  
  
  METHODS
  The three-dimensional porous nano-scaffolds were prepared by electrospinning techniques with collagen type II and HA (8 : 1, W : W), which was dissolved in mixed solvent of 3-trifluoroethanol and water (1 : 1, V : V). The morphology were observed by light microscope and scanning electron microscope (SEM). And the porosity, water absorption rate, contact angle, and degradation rate were detected. Chondrocytes were harvested from 1-week-old Japanese white rabbit, which was disgested by 0.25% trypsin 30 minutes and 1% collagenase overlight. The passage 2 chondrocytes were seeded on the nano-scaffold. The cell adhesion and proliferation were evaluated by cell counting kit 8 (CCK-8). The cell-scaffold composites were cultured for 2 weeks in vitro, and the biological morphology and extracelluar matrix (ECM) secretion were observed by histological analysis.
  
  
  RESULTS
  The optimal electrospinning condition of nano-scaffold was 10% electrospinning solution concentration, 10 cm receiver distance, 5 mL/h spinning injection speed. The scaffold had uniform diameter and good porosity through the light microscope and SEM. The diameter was 300-600 nm, and the porosity was 89.5% +/- 25.0%. The contact angle was (35.6 +/- 3.4) degrees, and the water absorption was 1 120% +/- 34% at 24 hours, which indicated excellent hydrophilicity. The degradation rate was 42.24% +/- 1.51% at 48 days. CCK-8 results showed that the adhesive rate of cells with scaffold was 169.14% +/- 11.26% at 12 hours, and the cell survival rate was 126.03% +/- 4.54% at 7 days. The histological and immunohistochemical staining results showed that the chondrocytes could grow well on the scaffold and secreted ECM. And the similar cartilage lacuma structure could be found at 2 weeks after co-culture, which suggested that hyaline cartilage formed.
  
  
  CONCLUSIONS
  The collage type II and HA complex three-dimensional nano-scaffold has good physicochemical properties and excellent biocompatibility, so it can be used as a tissue engineered cartilage
authors.affiliation: Research Institute of Tissue Engineering and Stem Cell, Nanchong Central Hospital, Second Clinical College of North Sichuan Medical College, Nanchong Sichuan 637000, PR China.
authors.forename: Zelong
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Research Institute of Tissue Engineering and Stem Cell, Nanchong Central Hospital, Second Clinical College of North Sichuan Medical College, Nanchong Sichuan 637000, PR China.
authors.forename: Zhu
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Research Institute of Tissue Engineering and Stem Cell, Nanchong Central Hospital, Second Clinical College of North Sichuan Medical College, Nanchong Sichuan 637000, PR China.
authors.forename: Kang
authors.initials: K
authors.lastname: Liu
authors.affiliation: Research Institute of Tissue Engineering and Stem Cell, Nanchong Central Hospital, Second Clinical College of North Sichuan Medical College, Nanchong Sichuan 637000, PR China.
authors.forename: Yiguang
authors.initials: Y
authors.lastname: Bai
authors.affiliation: Department of Plastic and Aesthetic Surgery, Nanchong Central Hospital, Second Clinical College of North Sichuan Medical College, Nanchong Sichuan 637000, PR China.
authors.forename: Ting
authors.initials: T
authors.lastname: Jiang
authors.affiliation: Department of Spinal Surgery, Affiliated Hospital of Luzhou Medical College, PR China.
authors.forename: Daxiong
authors.initials: D
authors.lastname: Feng
authors.affiliation: Research Institute of Tissue Engineering and Stem Cell, Nanchong Central Hospital, Second Clinical College of North Sichuan Medical College, Nanchong Sichuan 637000, PR China.
authors.forename: Gang
authors.initials: G
authors.lastname: Feng
mesh_terms: D000818:Animals; D001672:Biocompatible Materials; D002356:Cartilage / Q000166:cytology*; D002448:Cell Adhesion; D049109:Cell Proliferation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology*; D024043:Collagen Type II / Q000737:chemistry*; D005109:Extracellular Matrix / Q000737:chemistry; D005240:Feasibility Studies; D006820:Hyaluronic Acid / Q000737:chemistry*; D008422:Materials Testing; D049329:Nanostructures; D016062:Porosity; D011817:Rabbits; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D001672:Biocompatible Materials; D024043:Collagen Type II; D006820:Hyaluronic Acid