Effect of nerve growth factor on biological characteristics of scar fibroblasts / 中国组织工程研究

ABSTRACT

BACKGROUND: Nerve growth factor is secreted and synthetized by a variety of cells, such as inflammatory calls and repairing calls, its biological effects are diverse and closely related to the process of wound repair, but its mechanism is not yet clear.OBJECTIVE: To observe the influence of nerve growth factor on the biological characteristics of scar fibroblasts.METHODS: Eight clinical surgical resection specimens, including 5 face and neck hyperplastic scar or keloid specimens, did not receive any treatment; three were prepuce specimens following circumcision (normal tissue). By use of tissue block method, the scar and normal skin fibroblasts were cultured, followed by digestion passage. The scar tissue and normal tissue flbroblasts at 3-6passages in the logarithmic phase were seeded in 96-well plate and divided into the experimental group (scar flbroblest group) and the control group (normal skin fibroblasts group), with two parallel holes in each group were added with 3,33, 0.33 mg/L nerve growth factor, 50 μL. Inverted microscope was used to observe fibroblast morphology. At 24, 48, 72 hours after culture, the absorbanca value was measured using MTT. Fibroblast DNA content and cell apoptosis were determined by flow cytometry.RESULTS AND CONCLUSION: The fibroblasts were adherent cells, the scar and normal skin tissues were shown to cell free out of tissue block and gradual expansion at 4-6 days after incubation. Compared with normal skin fibroblasts, the pathological scar fibroblasts became larger, irregular shape and arrangement. MTT results showed that nerve growth factor could promote the normal and hypertrophic scar fibroblasts growth, which becomes more apparent. Flow cytometry results showed that by adding nerve growth factor, the percentage of scar fibroblasts at proliferating S-G_2-M phase was higher than that in the control;group; with a Iower level of apoptosis. It is indicated that nerve growth factor plays an obviously promoting role on normal and scar skin fibroblasts growth and proliferation, especially on the scar skin.