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.
Huang
Bo
B
Department of Orthopedics, Xinqiao Hospital, The Third Military Medical University, Chongqing 400037, People's Republic of China.
Li
Chang-Qing
CQ
Zhou
Yue
Y
Luo
Gang
G
Zhang
Chuan-Zhi
CZ
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