预损伤嗅黏膜对自体嗅鞘细胞体外培养的作用

Effects of pre-damage olfactory mucosa on in vitro culture of autologous olfactory ensheathing cells

目的:比较预损伤嗅黏膜与正常嗅黏膜来源的嗅鞘细胞生长特性和生物学活性的差异。方法:雄性成年SD大鼠10只,随机分为嗅黏膜损伤组、正常组,5只/组。嗅黏膜损伤组大鼠麻醉后采用自制弯头针,深入鼻腔内直至鼻中隔后1/3处,针尖抵住鼻中隔后切割数下至出血,填塞酒精棉条压迫止血。正常组大鼠不做任何处理。伤后7d取两组大鼠鼻中隔,将双侧鼻黏膜后1/3从鼻中隔刮下,胰酶消化后体外分离培养嗅黏膜嗅鞘细胞,调整浓度至1×109L-1,采用改良NASH差速贴壁法进行纯化。结果:嗅鞘细胞纯度达70%时,正常组需14d,嗅黏膜损伤组需12d。正常组培养7d后细胞进入对数期,嗅黏膜损伤组培养5d后细胞进入对数期,两组嗅鞘细胞进入平台期后生长趋于缓慢,数量维持在106左右。在细胞生长周期的相同时间点,嗅黏膜损伤组嗅鞘细胞活性大于正常组(t=19.0013,P<0.001),两组嗅鞘细胞分泌的神经营养因子3质量浓度基本相似(P>0.05)。结论:预损伤处理嗅黏膜能够加快嗅黏膜嗅鞘细胞的增殖分化,且活性较高。

OBJECTIVE： To compare the differences in growth and biological activity of olfactory ensheathing cells from injured olfactory mucosa and normal olfactory mucosa. METHODS： Ten male adult Sprague Dawley rats were randomly assigned into olfactory mucosa injury and normal olfactory mucosa groups, 5 rats for each group. In the olfactory mucosa injury group, a self-made bending head acupuncture was put into the nasal cavity till reached the 1/3 posterior to nasal septum. Incision was made till bleeding and pressed using alcohol sliver. Rats in the normal group were left intact. At 7 days following surgery, the rat nasal septum was obtained in two groups. 1/3 of bilateral mucosa was removed from the nasal septum. Olfactory mucosa olfactory ensheathing cells were incubated utilizing trypsin in vitro at 1 ×10^9 L^-1. Cells were purified by modified NASH differential adherence. RESULTS： It took 14 days and 12 days respectively in the normal group and the olfactory mucosa injury group to reach 70% purity of olfactory ensheathing cells. Cells in the normal group entered the logarithmic phase at 7 days; Cells in the olfactory mucosa injury group entered the logarithmic phase at 5 days. Cells from both groups grew slowly, at about 106. At the same time points, cell activity was higher in the olfactory mucosa injury group compared with the normal group （t=19.001 3, P 〈 0.001）. Neurotrophic factor-3 mass concentration was similar between both groups （P〉 0.05）. CONCLUSION： Pretreatment of olfactory mucosa can accelerate the proliferation and differentiation of olfactory mucosa olfactory ensheathing cells, with a high activity.