Release characteristics and bioactivity of gelatin-tricalcium phosphate membranes covalently immobilized with nerve growth factors

Pei Ru Chen, Ming Hong Chen, Feng Huei Lin, Wen Yu Su

Research output: Contribution to journalArticle

56 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)6579-6587
Number of pages9
JournalBiomaterials
Volume26
Issue number33
DOIs
Publication statusPublished - Nov 1 2005
Externally publishedYes

Fingerprint

Nerve Growth Factors
Gelatin
Bioactivity
Phosphates
Membranes
Composite materials
Assays
Cytotoxicity
Immobilization
tricalcium phosphate
Enzyme-Linked Immunosorbent Assay
Proteins
Carbodiimides
Glutaral
Biocompatibility
Regeneration
Cultured Cells
Cell Survival
Repair
Cells

Keywords

  • Biodegradable nerve conduit
  • Nerve growth factor
  • Neurite outgrowth
  • NGF release
  • PC 12 cell

ASJC Scopus subject areas

  • Bioengineering
  • Ceramics and Composites
  • Biophysics
  • Biomaterials
  • Mechanics of Materials

Cite this

Release characteristics and bioactivity of gelatin-tricalcium phosphate membranes covalently immobilized with nerve growth factors. / Chen, Pei Ru; Chen, Ming Hong; Lin, Feng Huei; Su, Wen Yu.

In: Biomaterials, Vol. 26, No. 33, 01.11.2005, p. 6579-6587.

Research output: Contribution to journalArticle

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