构建人工涎腺样组织的脱细胞气管基质支架

背景：目前还没有找到理想的构建组织工程化人工涎腺的支架材料，具有良女，中空样结构的脱细胞气管基质材料有着良好的可应用的发展前景。 目的：将下颌下腺细胞与家免及SD大鼠的脱细胞气管基质共培养，评价其生物相容性。 设计、时间及地点：随机对照动物实验，于2003—02／2005—05在四川大学华西医院的卫生部移植与免疫莺点实验室完成。 材料：用改良去污和酶交联法，去除SD大鼠和家兔气管的细胞，制备脱细胞气管基质。 方法；SD大鼠18只随机分为2组，一组颊部皮下植入SD大鼠的脱细胞气管，另一组植入家兔的脱细胞气管。将传至第3代的下颌下腺细胞接种到2种脱细胞气管和PGA膜上培养，对照组进行无材料12孔板单纯细胞接种，进行细胞／支架材料复合体培养。 主要观察指标：光镜、扫描电镜观察脱细胞气管基质的组织结构和内壁的拓扑结构。于1，4，12周，对埋置在颊部皮下脱细胞气管周围的组织进行炎症反应评估。在培养的1～7d，每天对各组细胞进行计数，并分别于培养第1，3，5，7天，以MTT法测定各组细胞的A值及其上清液中的淀粉酶活性。 结果：两种动物气管中的细胞被完全去除。在颊部皮下埋置的两种脱细胞气管周围的组织未见明显炎症反应。生长在2种脱细胞气管上的下颌下腺细胞数明显多于PGA膜（P〈0．05），其上清液中淀粉酶活性及细胞的A值也高于PGA膜（P〈005）。 结论：自制的脱细胞气管基质保持了中空状，并有良好的组织相容性和细胞相容性。

Biocompatibility of acellular tracheae as scaffold for artificial salivary gland organoid

BACKGROUND: An important initial step in developing a tissue engineering artificial salivary gland organoid is to find an ideal scaffold. To find other new biomaterials should to be further studied.OBJECTIVE: To obtain an acellular matrix from tracheae of rabbits and SD rats, and to investigate its biocompatibility as a primary step toward developing a tissue engineering artificial salivary gland organoid.DESIGN, TIME AND SETTING: A randomized controlled animal study, which was performed at the Key Laboratory of Transplantation Engineering and Immunology of Ministry of Health, West China Hospital of Sichuan University from February 2003 to May 2005.MATERIALS: A modified detergent and enzyme link extraction procedure was performed to remove cells from SD rats and rabbits tracheae. The histology, topography of inner-surface and biocompatibility were studied on both acellular tracheae.METHODS: Eighteen SD rats were randomly divided into 2 groups. One group was planted acellular trachea of SD rats. Other group consisted of acellular trachea of rabbits. On the third generations, submandibular gland cells were inoculated on two acellular tracheae and cultured on PGA membrane; while, cells in the control group were inoculated on 12-well culture plate, and cell/scaffold complex was cultured at the same time.MAIN OUTCOME MEASURES: The structure and topography of inner-surface of the acellular tracheae matrixes were observed both by light and scanning electron microscopy. The inflammatory response of the tissue around acellular tracheae implanted under the skin of cheek was evaluated at 1, 4, 12 weeks. The numbers of cells grown on the acellular tracheae and PGA film were counted at 1, 2, 3, 4, 5, 6, 7 days. At 1, 3, 5, 7 days, the mean values of metabolic activity test and the amylase activities of supernatants of the cells/scaffold complexes were examined.RESULTS: The cells were completely removed from both tracheae. No evident inflammatory response was found in tissues around two kinds of acellular tracheae implanted under the skin of cheek. The number of submandibular gland cells (SSG) grown on the two kinds of naturally derived biomaterials was much more than grown on PGA (P<0.05). The mean values of metabolic activity test and the amylase activities of supernatants containing cell/acellular matrixes were much higher than that of cell/PGA (P<0.05).CONCLUSION: The acelhilar tracheae matrixes made by our laboratory can be used as scaffold in the study of tissue engineering artificial salivary gland organoid.