Biocompatibility of NGF-grafted GTG membranes for peripheral nerve repair using cultured Schwann cells.
We previously developed a biodegradable composite with potentially good biocompatibility composed by tricalcium phosphate and gluataraldehyde cross-linking gelatin (GTG) with good mechanical property feasible for surgical manipulation. The purpose of this study was to evaluate the feasibility of immobilizing nerve growth factor (NGF) onto the composite (GTG) with carbodiimide (GEN composite). Cultured Schwann cells were seeded onto the GTG and GEN composites. For comparison, GTG membrane soaked in NGF solution without carbodiimide (GN composite) as cross-linking agent was also used to culture Schwann cells. Cell morphology was observed by a scanning electron microscope. Cell survival, cytotoxicity and cellular metabolism on the NGF-grafted GTG membrane were assessed quantitatively in terms of cell protein content, leakage of cytosolic lactate dehydrogenase (LDH) activity and by the well-established MTT assay, respectively. The result of LDH study did not show significant difference among GTG, NGF-modified GTG and control group. This indicated that GTG composite, whether cross-linking with NGF or not, has little cytotoxic effect. Comparing the protein content and MTT assay among GEN, GN composite and control group, the data confirmed more attachment of Schwann cells on GEN composite. Although GTG cross-linking with NGF did not promote Schwann cell proliferation, the techniques we used in this study provided a method to fabricate a novel biomaterial incorporation of Schwann cells and covalently immobilized NGF.
Chen
Pei-Ru
PR
Institute of Biomedical Engineering, College of Medicine, College of Engineering, National Taiwan University, No 1, Sec. 1, Jen-Ai Road, Tiapei, Taiwan.
Chen
Ming-Hong
MH
Sun
Jui-Sheng
JS
Chen
Mei-Hsiu
MH
Tsai
Chien-Chen
CC
Lin
Feng-Huei
FH
D000818:Animals; D000831:Animals, Newborn; D002130:Calcium Phosphates / Q000737:chemistry; D002478:Cells, Cultured; D005780:Gelatin / Q000031:analogs & derivatives / Q000737:chemistry; D005976:Glutaral / Q000737:chemistry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008422:Materials Testing; D008567:Membranes, Artificial; D008855:Microscopy, Electron, Scanning; D020932:Nerve Growth Factor / Q000737:chemistry*; D009416:Nerve Regeneration; D059348:Peripheral Nerve Injuries; D010525:Peripheral Nerves / Q000503:physiopathology; D011506:Proteins / Q000032:analysis; D051381:Rats; D011917:Rats, Inbred Lew; D012583:Schwann Cells / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D013385:Succinate Dehydrogenase / Q000378:metabolism; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
D002130:Calcium Phosphates; D008567:Membranes, Artificial; D011506:Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D005780:Gelatin; D020932:Nerve Growth Factor; D007770:L-Lactate Dehydrogenase; D013385:Succinate Dehydrogenase; C022616:thiazolyl blue; C018392:tricalcium phosphate; D005976:Glutaral