三种组织工程鼓膜支架材料的生物相容性

Biocompatibility of three kinds of tissue engineered tympanic membrane

背景:自体组织修复鼓膜穿孔存在增加手术创伤、取材有限等缺点,构建组织工程鼓膜是鼓膜穿孔治疗的发展方向。目的:构建3种不同材料的组织工程鼓膜支架材料,并评价其生物相容性。设计、时间及地点:随机对照动物实验,细胞学体内观察,于2004-08/2005-02在解放军第四军医大学组织工程中心完成。材料:将健康小猪的硬脑膜、真皮及人羊膜进行脱细胞处理。方法:将预先培养的豚鼠鼓膜成纤维细胞种植于3种材料表面,观察成纤维细胞的生长状态;18只豚鼠随机分为3组,于右侧背皮下分别埋植制备的脱细胞硬脑膜、真皮及人羊膜。主要观察指标:3种脱细胞生物支架材料的显微结构及组织相容性。结果:3种脱细胞材料均为网状结构,无任何细胞及细胞残核存在;豚鼠鼓膜成纤维细胞能够很好地贴附于3种脱细胞材料生长,无细胞毒性;将3种组织材料分别埋植于豚鼠皮下后,无埋植材料排出现象,1周时植入材料周围有较明显炎症反应,其中以脱细胞真皮周围组织反应较轻,4周时炎症反应消失。结论:通过脱细胞方法可以成功建立具有良好生物相容性的组织工程鼓膜支架材料;3种材料中以脱细胞猪真皮的生物相容性最佳。

BACKGROUND： Repairing ear-drum perforations with autologous has the disadvantage of sources limited and adding more operative trauma, Constructing tissue engineered tympanic membrane maybe one of the best choices for treating tympanic membrane perforation in future. OBJECTIVE： To construct three different kinds of tympanic membrane by tissue engineering technique and compare the biocompatibility. DESIGN, TIME AND SETTING： The randomized animal control experiment of in vivo cytology observation was performed in Tissue Engineering Center of the Fourth Military Medical University of Chinese PLA between August 2004 and February 2005. MATERIALS： The original cells were removed from healthy pig cerebral dura mater, dermis and human amnion. METHODS： Cultured tympanic membrane fibroblast cells of guinea pig were seeded on the surface of three materials, then observed the growth state. Eighteen guinea pigs were randomly divided into 3 groups, and embed in acellular cerebral dura mater, dermis and human amnion at right side of dorsal cutaneous, respectively. MAIN OUTCOME MEASURES： Microstructure and histocompatibility of three scaffold materials were observed. RESULTS： Three scaffolds were reticular formation without residual nuclear. Fibroblasts could attached and extended on these scaffolds well, there was no cytotoxicity. Though there was obvious inflammatory reaction 1 week after embedding materials into subcutaneous tissues, it disappeared at 4 weeks. Guinea pigs showed no evidence of rejection or extrution. CONCLUSION： Tissue engineered tympanic membrane materials can be constructed by decellularization methods. These scaffold materials have good biocompatibility, especially the derma of the pig.