Controlled release of nerve growth factor from heparin-conjugated fibrin gel within the nerve growth factor-delivering implant
이진용, 김성민, 김명진, 이종호,
소속 상세정보
이진용 ( Lee Jin-Yong ) - Korea University Guro Hospital Department of Oral and Maxillofacial Surgery
김성민 ( Kim Soung-Min ) - Seoul National University School of Dentistry Department of Oral and Maxillofacial Surgery
김명진 ( Kim Myung-Jin ) - Seoul National University School of Dentistry Department of Oral and Maxillofacial Surgery
이종호 ( Lee Jong-Ho ) - Seoul National University School of Dentistry Department of Oral and Maxillofacial Surgery
KMID : 0355620140400010003
Abstract
Objectives: Although nerve growth factor (NGF) could promote the functional regeneration of an injured peripheral nerve, it is very difficult for NGF to sustain the therapeutic dose in the defect due to its short half-life. In this study, we loaded the NGF-bound heparin-conjugated fibrin (HCF) gel in the NGF-delivering implants and analyzed the time-dependent release of NGF and its bioactivity to evaluate the clinical effectiveness.
Materials and Methods: NGF solution was made of 1.0 mg of NGF and 1.0 mL of phosphate buffered saline (PBS). Experimental group A consisted of three implants, in which 0.25 μL of NGF solution, 0.75 μL of HCF, 1.0 μL of fibrinogen and 2.0 μL of thrombin was injected via apex hole with micropipette and gelated, were put into the centrifuge tube. Three implants of experimental group B were prepared with the mixture of 0.5 μL of NGF solution, 0.5 μL HCF, 1.0 μL of fibrinogen and 2.0 μL of thrombin. These six centrifuge tubes were filled with 1.0 mL of PBS and stirred in the water-filled beaker at 50 rpm. At 1, 3, 5, 7, 10, and 14 days, 1.0 mL of solution in each tubes was collected and preserved at -20oC with adding same amount of fresh PBS. Enzyme-linked immunosorbent assay (ELISA) was done to determine in vitro release profile of NGF and its bioactivity was evaluated with neural differentiation of pheochromocytoma (PC12) cells.
Results: The average concentration of released NGF in the group A and B increased for the first 5 days and then gradually decreased. Almost all of NGF was released during 10 days. Released NGF from two groups could promote neural differentiation and neurite outgrowth of PC12 cells and these bioactivity was maintained over 14 days.
Conclusion: Controlled release system using NGF-HCF gel via NGF-delivering implant could be an another vehicle of delivering NGF to promote the nerve regeneration of dental implant related nerve damage .
키워드
Nerve growth factor-delivering implant; Nerve growth factor; Controlled release; Nerve regeneration
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