三种组织工程鼓膜的构建

目的 构建3种不同材料的组织工程鼓膜,并对其进行生物学特性检测. 方法 体重 30 kg 健康雄性小白猪1只,制备脱细胞硬脑膜及脱细胞真皮,并从牛皮中提取液态胶原,将第3代豚鼠鼓膜成纤维细胞 1×106个/mL,各 0.2 mL 种于3种材料表面进行培养,倒置相差显微镜下观察成纤维细胞的生长状态.培养1周后,将3种材料分别进行扫描电镜及HE染色观察,测定材料厚度及其张力. 结果 成纤维细胞在3种材料表面生长良好,呈束状平行排列,其中以胶原与脱细胞真皮表面成纤维细胞生长情况较佳.HE 染色及扫描电镜下可见成纤维细胞呈单层于3种材料表面连续生长.胶原膜、脱细胞真皮及脱细胞硬脑膜的平均厚度分别为 9.4、10.0 及 10.4 μm.张力强度测定,复合成纤维细胞的胶原组织工程鼓膜张力 (1.417±0.030) N/mm2,复合成纤维细胞的脱细胞真皮张力为 (24.500±2.040)N/mm2,复合成纤维细胞的脱细胞硬脑膜张力为(53.300±2.600)N/mm2;复合成纤维细胞的脱细胞真皮张力与正常豚鼠鼓膜张力26.700 N/mm2最接近. 结论 脱细胞猪真皮为支架复合成纤维细胞构建的组织工程鼓膜,其张力与厚度均与正常豚鼠鼓膜相接近,是构建组织工程鼓膜的理想材料.

RECONSTRUCTION OF THREE DIFFERENT KINDS OF TYMPANIC MEMBRANE IN VITRO BY TISSUE ENGINEERING TECHNIQUE

OBJECTIVE: To reconstruct three different kinds of tympanic membrane in vitro by tissue engineering technique, and to examine their histological structures and mechanical properties. METHODS: The skin and dura of pig (weight 30 kg) were processed with high satuated saline and enzymes to make extracellular matrix. Meanwhile, fibroblasts (1 x 10(6)/mL, 0.2 mL) were seeded on the surface of these two scaffolds and collagen. The composite tissues were cultured in vitro for 1 week and examined in histological structure and mechanical properties. RESULTS: Fibroblasts cultured were spindle-shaped and could grow and attach to these scaffolds with a arrangement of sarciniform and parallel. The reconstructed tissue of ECM and collagen appeared to integrate well and had better bio-compatibility. The mean thickness of the collagen, the skin and the dura (all covered with fibroblasts) were 9.4, 10.0 and 10.4 microm respectively. The tension of the collagen was (1.417+/-0.030) N/mm2, of the acellular dermal matrix was (24.500+/-2.040) N/mm2 (being close to the tension of normal tympanic membrane, 26.700 N/mm2), of the acellular dura was (53.300+/-2.600) N/mm2. CONCLUSION: The results suggest that the tension and the thinkness of acellular dermal matrix is similar to the normal tympanic membrane of guinea pig, it is an ideal material for tympanoplasty.