摘要
背景:选择适合的生物支架与骨髓间充质干细胞作为新型的种子细胞复合能否构建出理想的组织工程化软骨?目的:观察将扩增的兔骨髓间充质干细胞接种于聚羟基乙酸构建人工软骨的可行性。设计:单一样本观察。单位:泸州医学院附属医院耳鼻咽喉头颈外科。材料:实验于2004-10/2005-10在泸州医学院完成。选用8只日本大耳兔,雌雄不拘,清洁级,二三个月龄,体质量1.5～2.0kg,常温常湿环境饲养。聚羟基乙酸由美国Albany公司生产。方法:分离、获取、扩增实验兔骨髓间充质干细胞,将聚羟基乙酸剪成1cm×1cm×1cm大小,并以多聚赖氨酸包被,通过将扩增的骨髓间充质干细胞接种于聚羟基乙酸上,按4mL/cm3多点播散的方式均匀地接种在预湿的聚羟基乙酸支架上,体外诱导培养3周,作为实验组。将不加入骨髓间充质干细胞的聚羟基乙酸支架作为对照组。将实验组和对照组的标本分别植入4只自体兔腹腔内,进行体内培养6～12周后,分别取出实验组与对照组的标本,对其进行大体观察。同时以100g/L中性甲醛固定,行5μm切片,采用苏木精-伊红染色观察标本的组织形态学特点,进行阿新蓝染色以观察酸性粘多糖形成,进行甲苯胺蓝染色以观察异染性基质的形成,进行免疫组织化学染色检测Ⅱ型胶原蛋白的表达。主要观察指标:两组支架植入实验兔体内后6,12周时标本大体观察结果、不同染色观察结果及Ⅱ型胶原蛋白的表达。结果:纳入8只实验兔均进入结果分析。①支架植入实验兔体内后6及12周标本大体观察结果:支架植入兔腹腔6周后,实验组标本均仍被腹腔大网膜包裹,剥去大网膜后,有3个标本成淡黄色,光滑,质地中,外形与植入前基本一致,1个标本呈灰黑色,质地很软,标本不能成形。12周后,实验组标本的外形仍与植入前基本一致,呈灰白色,光滑,但质地明显较硬。而第6,12周的对照组的标
BACKGROUND： Whether choosing a suitable biological scaffold compounding with mesenchymal stem cells （MSCs） can construct an ideal tissue-engineering cartilage or not should be researched further.
OBJECTIVE： To investigate the feasibility of constructing artificial cartilage by using amplified rabbit MSCs which were inoculated on poly-glycolic acid （PGA）.
DESIGN： Single sample observation.
SETTING ： Department of Otolaryngology-Head ＆ Neck Surgery, Affiliated Hospital of Sichuan Luzhou Medical College.
MATERIALS ： The experiment was carried out in the Luzhou Medical College from October 2004 to October 2005. A total of 8 Japanese large ear rabbits, of both genders, clean grade, aged from 2 to 3 months, weighing 1.5-2.0 kg, were fed in normal temperature and humidity. Poly-glycolic acid was provided by Albany Company, USA.
METHODS： Rabbit MSCs were separated, obtained and amplified. In addition, poly-glycolic acid was sheared into pieces with the size of 1 cm × 1 cm × 1 cm and embedded with poly-L-lysine. Amplified MSCs were inoculated on the surface of poly-glycolic acid, and then, they were averagely grown on pre-wet scaffold of poly-glycolic acid according to 4 mL/cm^2 multi-points spreading style and cultured in vitro for 3 weeks. After the operations mentioned above, samples were regarded as the experimental group. Scaffolds of poly-glycolic acid without MSCs were considered as the control group. Samples in both experimental group and control group were transplanted into abdominal cavity of 4 rabbits, respectively, cultured in vivo for 6-12 weeks, taken out and observed generally. Meanwhile, the samples were fixed with 100 g/L neutrality formaldehyde, cut into sections with the thickness of 5 μm, stained with haematine-eosin （HE） and observed their histomorphological characteristics. Moreover, the samples were stained with alcian blue for observation of glycosaminoglycans formation, with toluidine blue for observation of metachromasia matrix formation, and with immunohistoch
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第14期2761-2764,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
四川省科技厅科研基金资助(2006J13163)~~
