保存人鼻中隔软骨活性的3种体外方法比较  

Comparison of human nasal septum cartilage preservation using three different methods in vitro

作　　者：杨宪法[1] 赵大庆[2] 崔鹏程[1] 徐凯[2] 

机构地区：[1]解放军第四军医大学唐都医院耳鼻喉科,陕西省西安市710038 [2]解放军第四军医大学西京医院耳鼻咽喉-头颈外科,陕西省西安市710032

出　　处：《中国组织工程研究与临床康复》2008年第42期8235-8238,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：陕西省科技攻关项目(2005K11-G4(3))~~

摘　　要：背景:软骨损伤后难以自身修复,以活性软骨进行修复效果较好,但活性软骨的可靠来源尚未根本解决。目的:以3种不同方法体外保存成人鼻中隔软骨块的活性比较。设计、时间及地点:单一样本观察,实验于2007-06/2008-03在解放军第四军医大学唐都医院耳鼻喉科实验室及西京医院全军耳鼻咽喉-头颈外科中心实验室完成。材料:材料为临床常规手术中切除的成人正常鼻中隔软骨20块,按国务院2006年《医疗机构管理条例》规定,已与患者签订知情同意书。方法:将所取软骨,块剪成大小约15mm×10mm×2mm软骨块,分别置于4,-20,-80℃冷冻及RPMI-1640液于37℃培养保存,于保存24h,7,14,21d和30d时取样品。主要观察指标:以苏木精-伊红、AB/PAS及Masson三色染色和锥虫蓝排斥试验检测软骨活性。结果:用RPMI-1640培养液保存的成人鼻中隔软骨,在整个保存期中均能保持较好活性,软骨细胞活性率保持在80%左右。以-80℃冷冻保存7d及-20℃保存14d时,软骨基本失去活性,当4℃保存在21d时,软骨可保持较好活性,保存30d时,软骨活性明显降低。结论:与低温保存方法相比,用组织培养法保存成人软骨块能较好地保持其活性,有望为临床治疗提供一种取材方便、新型有效的软骨修复材料。BACKGROUND： Injured cartilage cannot be self-repaired, and vital cartilage can be used to repair injured cartilage with a good outcome. However, reliable source of vital cartilage does still not be solved. OBJECTIVE： To compare the activity of human adult nasal septum cartilage preserved in vitro using three different methods. DESIGN, TIME AND SETTING： The single sample observation was performed at the Laboratory of Department of Otorhinolaryngology, Tangdu Hospital, Fourth Military Medical University of Chinese PLA, and Central Laboratory of Department of Ear, Nose, Throat, Head and Neck, Xijing Hospital, Fourth Military Medical University of Chinese PLA from June 2007 to March 2008. MATERIALS： Twenty normal human adult anasal septum cartilages were obtained from clinical septectomy. Patients had signed the informed consent according to the rules of Medical Establishment Management made by the State Council in 2006. METHODS： The size of cartilage fragments was about 15 mm×10 mm × 2 mm. The fragments were separately preserved at 4, -20 and -80℃ and in the RPMI-1640 medium at 37℃. Samples were collected at 24 hours, 7, 14, 21 and 30 days after preservation. MAIN OUTCOME MEASURES： Cartilage activity was detected using hematoxylin-eosin, AB/PAS and Masson 3 staining, and trypan blue staining exclusion test. RESULTS： Cartilage preserved in the RPMI-1640 medium retained its viability during the whole storage time. The viability ratio of cartilage samples were about 80%. Cartilages kept at -80 ℃ for 7 days and at -20 ℃ for 14 days showed a quick decreased viability. Cartilage preserved at 4 ℃ retained its viability at 21 days, but lost its viability when preserved for 30 days. CONCLUSION： Compared with hypothermia preservation, tissue culture method could successfully keep the viability of cartilages. It may offer a new effective easily obtained cartilage repair material for clinical treatment.

关 键 词：鼻中隔 软骨 组织培养技术 低温保存 组织构建 

分 类 号：R622[医药卫生—整形外科]