1.Preparation and evaluation of an articular cartilage extracellular matrix/human umbilical cord Wharton gel composite scaffold
Tongguang XIAO ; Chunxiang HAO ; Xiaoguang JING ; Xuejian LIU ; Ganggang GUO ; Jianhua YANG ; Quanyi GUO
Chinese Journal of Tissue Engineering Research 2017;21(22):3470-3475
BACKGROUND:Soft tissue engineering mainly includes seed cells,scaffolds,cytokines and bioreactors,among which,the scaffolds are the key link in the construction of tissue-engineered cartilage.OBJECTIVE:To prepare an articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffold,and to evaluate its physicochemical properties and biocompatibility.METHODS:The articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffold was prepared by freeze thawing drying method using porcine articular cartilage extracellular matrix and human umbilical cord Wharton glue as raw materials.The porosity,water absorption,tissue composition and longitudinal compressive elastic modulus of the scaffold were measured and histologically stained.Rabbit chondrocytes were co-cultured with the articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffold for 7 days.Then,scanning electron microscopy,live-dead cell staining and hematoxylin-eosin staining were performed.In addition,rabbit chondrocytes were cultured in the extract of the articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffold and cell culture medium for 6 days,respectively;and MTT assay was used to detect cell proliferation.RESULTS AND CONCLUSION:The articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffolds had a cross-section of uniform porous network structure and a vertical cross-section of the vertical tubular structure,and the pore wall was densely covered with cartilage fibers.The composite porous scaffold was positive for hematoxylin-eosin staining,safranin O staining and toluidine blue staining,and contained collagen and glycosaminoglycan ingredients.The water absorption,porosity and longitudinal compressive elastic modulus of the scaffolds were (17.418 8±0.909 0)%,(81.495 1±6.621 0)% and (2.833 3±0.456 4) kPa,respectively.After 7 days of co-culture,rabbit chondrocytes adhered to the scaffold and proliferated,and further grew into the pores of the scaffold.Moreover,the scaffold was non-toxic to the rabbit chondrocytes.To conclude,the physiochemical properties and biochemical components of articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffolds are similar to those of natural cartilage,and the scaffold has good biocompatibility.