Release characteristics and bioactivity of gelatin-tricalcium phosphate membranes covalently immobilized with nerve growth factors
Resource
Biomaterials 26 (33): 6579-6587
Journal
Biomaterials
Journal Volume
26
Journal Issue
33
Pages
6579-6587
Date Issued
2005
Date
2005
Author(s)
Abstract
The gelatin-tricalcium phosphate membranes were cross-linking with low concentration glutaraldehyde solution (GTG). This material has good mechanical property, biocompatibility, and is feasible for surgical manipulation. For axonal regeneration, nerve growth factors (NGF) were immobilized onto the composite (GTG) with carbodiimide. The purpose of this study was to evaluate the release characteristics and bioactivity of NGF after covalent immobilization onto the GTG membranes (GEN). NGF immobilized onto and released from the composite was quantified using ELISA method. PC 12 cells were cultured on the GTG and GEN composites. Cell survival, cytotoxicity, and cellular activity were evaluated by total protein content, LDH activity, and MTT assay respectively. Neurite outgrowth assay was used to evaluate the biological activity of NGF released from GEN composite. From ELISA measurement, the releasing curve for NGF showing two distinctive parts with different slopes indicated that NGF were released from the composite in diffusion-controlled mechanism and degradation-controlled mechanism respectively. While culturing with PC 12 cells, LDH leakage results implied that whether GTG composite cross-linked with NGF or not showed little cytotoxicity. The total protein content and cellular activity of PC 12 cells were lower on GTG and GEN membranes than control group. However, 56%±3.98 of PC 12 cells showed significant neurite outgrowth on GEN membranes which was statistically higher than GTG without NGF immobilization. In addition, sustained release of bioactive NGF for two months had been demonstrated by neurite outgrowth assay. From these experiments, it can be concluded that the technique used in the present study is capable of immobilizing NGF onto GTG membranes covalently and remaining the bioactivity of NGF. Therefore, GEN composite can be materials for sustained release of bioactive NGF and a candidate for future therapeutic application in nerve repair. © 2005 Elsevier Ltd. All rights reserved.
Subjects
Biodegradable nerve conduit; Nerve growth factor; Neurite outgrowth; NGF release; PC 12 cell
Other Subjects
Biocompatibility; Cells; Degradation; Diffusion; Mechanical properties; Neurology; Toxicity; Axonal regeneration; Gelatin-tricalcium phosphate membranes; Nerve growth factors (NGF); Surgical manipulation; Biological membranes; calcium phosphate; gelatin; glutaraldehyde; lactate dehydrogenase; nerve growth factor; analytic method; animal cell; animal cell culture; article; cell strain; cell survival; comparative study; composite material; controlled study; cross linking; cytotoxicity; degradation; diffusion; enzyme activity; enzyme linked immunosorbent assay; immobilization; nerve fiber growth; nonhuman; priority journal; protein content; rat; statistical significance; sustained drug release; Animals; Biocompatible Materials; Calcium Phosphates; Carbodiimides; Cell Membrane; Cell Proliferation; Cell Size; Cell Survival; Cells, Cultured; Cross-Linking Reagents; Diffusion; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Gelatin; Kinetics; L-Lactate Dehydrogenase; Models, Chemical; Nerve Growth Factors; Neurites; Neurons; PC12 Cells; Rats; Temperature; Tetrazolium Salts; Thiazoles; Time Factors
Type
journal article
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