Collagen II/hyaluronan/chondroitin-6-sulfate tri-copolymer scaffold for nucleus pulposus tissue engineering.
This study aims to investigate the bioactivity of collagen II/hyaluronan/chondroitin-6-sulfate tri-copolymer as bionic scaffold for nucleus pulposus (NP) tissue engineering. Collagen II (C II) (pH 1-2) was mixed with hyaluronan (HyA) and lyophilized to prepare C II/HyA matrices. Chondroitin 6-sulfate (6-CS) was covalently attached to the C II/HyA matrices using 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). Then, cells were expanded from rabbit NP and seeded in the tri-copolymer scaffold. Cell-scaffold hybrids were maintained for up to 28 days in culture. Cell viability/proliferation, extracellular matrix (ECM)-related gene expression, and the content of sulfated glycosaminoglycans (s-GAG) were evaluated. Our results are as following: when cultured for 28 days, the cell-scaffold hybrids maintained active cell viability/proliferation and exhibited a significantly increased s-GAG content. In addition, rabbit NP cells cultured in the scaffold demonstrated a significantly higher level of C II and aggrecan gene expression and a significantly lower level of Collagen I (C I) gene expression when compared with that of monolayer cells. Histological studies and scanning electron microscopy (SEM) further indicated newly secreted ECM deposits in the scaffolds. In conclusion, the C II/HyA-CS scaffold may be an alternative material for NP tissue engineering due to its satisfactory bioactivity, and it deserves further in vivo investigation.
Department of Orthopedics, Xinqiao Hospital, The Third Military Medical University, Chongqing 400037, People's Republic of China.
Bo
B
Huang
Chang-Qing
CQ
Li
Yue
Y
Zhou
Gang
G
Luo
Chuan-Zhi
CZ
Zhang
D000818:Animals; D001672:Biocompatible Materials; D006690:Bisbenzimidazole; D049109:Cell Proliferation; D002470:Cell Survival; D002809:Chondroitin Sulfates / Q000737:chemistry*; D024043:Collagen Type II / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D004396:Coloring Agents; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D005109:Extracellular Matrix / Q000737:chemistry; D015870:Gene Expression / Q000187:drug effects; D006820:Hyaluronic Acid / Q000737:chemistry*; D007150:Immunohistochemistry; D008751:Methylene Blue / Q000031:analogs & derivatives; D008855:Microscopy, Electron, Scanning; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012844:Sincalide / Q000378:metabolism; D054457:Tissue Scaffolds / Q000737:chemistry*
D001672:Biocompatible Materials; D024043:Collagen Type II; D004396:Coloring Agents; C435946:dimethylmethylene blue; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates; D004247:DNA; D006690:Bisbenzimidazole; D012844:Sincalide; D008751:Methylene Blue