摘要
背景:神经生长因子由多种细胞分泌合成,如炎性细胞、修复细胞等,其生物学作用呈多样性,与创伤修复过程密切相关,但其作用机制尚不明了。目的:观察神经生长因子对瘢痕成纤维细胞生物学特征的影响。方法:8份临床手术切除标本,其中5份为面颈部增生性瘢痕或瘢痕疙瘩标本,未接受过任何治疗;3份为包皮环切术切除的包皮标本(正常组织)。采用组织块法培养瘢痕及正常皮肤成纤维细胞,消化法传代。取3～6代对数生长期瘢痕组织及正常组织成纤维细胞,接种于96孔培养板中,设置实验组(瘢痕成纤维细胞组)和对照组(正常皮肤成纤维细胞组),每组设两个平行孔,分别加入3.33,0.33mg/L神经生长因子各50μL。采用倒置显微镜观察成纤维细胞形态。培养24,48,72h后,MTT测定各孔的吸光度。流式细胞术分析成纤维细胞DNA含量及细胞凋亡情况。结果与结论:成纤维细胞为贴壁生长型细胞,瘢痕和正常皮肤组织块在接种后4～6d可见细胞从组织块中游出,逐渐向周围扩展开。与正常皮肤成纤维细胞相比,病理性瘢痕成纤维细胞体积较大,形态和排列不规则。MTT结果显示,神经生长因子可促进正常及瘢痕成纤维细胞的增长,后者更为明显。流式细胞术测定结果显示,加入神经生长因子后,瘢痕成纤维细胞增殖期S-G2-M细胞百分数高于对照组,且凋亡水平降低。提示神经生长因子对正常及瘢痕皮肤成纤维细胞的生长趋势与增殖活性有明显的促进作用,对瘢痕皮肤的作用更明显。
BACKGROUND： Nerve growth factor is secreted and synthetized by a variety of cells, such as inflammatory cells and repairing cells, 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 fibroblasts at 3-6 passages in the logarithmic phase were seeded in 96-well plate and divided into the experimental group （scar fibroblast 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 absorbance 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-G2-M phase was higher than that in the control group, with a lower 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.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第7期1208-1212,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
云南省自然科学基金资助项目(2005C0072M)
云南省社会发展攻关项目(2007CA006)~~
