Dermal fibroblasts genetically engineered to release nerve growth factor
| Autor: | Priscilla Chauvin, Charles W. Patrick, Björn G. Stark, Gregory R. D. Evans, Bei Zheng, Mathias Schmidt, Zhen Fan, Sonja Herman |
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| Rok vydání: | 2002 |
| Předmět: |
Time Factors
Tissue replacement Schwann cell Transfection Rats Sprague-Dawley Dermis Nerve Growth Factor Medicine Animals Fibroblast Skin business.industry Genetically engineered Anatomy Fibroblasts medicine.disease In vitro Cell biology Rats medicine.anatomical_structure Nerve growth factor Peripheral neuropathy Surgery Schwann Cells business Genetic Engineering |
| Zdroj: | Annals of plastic surgery. 47(6) |
| ISSN: | 0148-7043 |
| Popis: | Current surgical strategies for repair of critical nerves involves the transfer of normal donor nerve from an uninjured body location. One possible alternative to autogenous tissue replacement is the development of engineered constructs to replace those elements necessary for axonal proliferation. Delivery of growth factors is one strategy to enhance synthetic nerve constructs. Thus, this study focused on the delivery of nerve growth factor (NGF) by genetic engineering to begin approaching the microenvironment dictated, in part, by Schwann's cells. Rat dermal fibroblasts (DFBs) were modified genetically to release rat NGF. The reporter gene LacZ was used to assess the optimum nonviral transfection method commercially available before NGF transfection. FuGENE6 provided the optimum transfection efficiency (24% maximum, 20.1 +/- 1.9% 5-day average) as measured by beta-galactosidase catalytic activity. NGF release from transfected DFBs was assessed over a 3-day period. Compared with control (no transfection) DFBs and DFBs transfected with vector alone, DFBs transfected with an expression vector encoding rat beta-NGF demonstrated significantly (p0.05) higher levels of NGF, with a 3-day maximum of 111 pg NGF per milliliter. When normalized to cell number, NGF-transfected DFBs released 1.2 pg NGF per milliliter/10(3) cells. The NGF-transfected DFBs demonstrated a maximal NGF release rate at day 1 (1.2 ng NGF/10(6) cells per day), followed by a markedly lower, sustained release rate at days 2 and 3 (0.44 ng NGF/10(6) cells per day and 0.48 ng NGF/10(6) cells per day respectively). The release rate curves for control and vector-transfected DFBs also exhibited a maximal NGF release rate at day 1, but were followed by a decreasing release rate, potentially representing in vitro degradation of NGF present in fetal bovine serum. Although not first with the development of growth factor delivery through fibroblasts, these findings suggest that rat DFBs can be modified genetically to act like Schwann's cells to deliver NGF. |
| Databáze: | OpenAIRE |
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