大鼠颌下腺脱细胞支架的制备研究

doi:10.13701/j.cnki.kqyxyj.2020.10.014

口腔医学研究 ›› 2020, Vol. 36 ›› Issue (10): 948-952.DOI: 10.13701/j.cnki.kqyxyj.2020.10.014

大鼠颌下腺脱细胞支架的制备研究

戴太强¹, 张琳琳², 安莹², 许方方¹, 邵小夕¹, 刘彦普¹, 刘斌^3*

1.军事口腔医学国家重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔疾病临床医学研究中心,第四军医大学口腔医院颌面外科陕西西安 710032;
2.军事口腔医学国家重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔生物工程技术研究中心,第四军医大学口腔医院牙周病科陕西西安 710032;
3.军事口腔医学国家重点实验室,国家口腔疾病临床医学研究中心,第四军医大学口腔医院实验中心陕西西安 710032

收稿日期:2020-03-24 出版日期:2020-10-28 发布日期:2020-10-19
通讯作者: *刘斌,E-mail:kqyljd_liu@126.com
作者简介:戴太强(1989~ ),男,山东德州人,博士,主治医师,研究方向:涎腺组织工程再生。

Preparation of Decellularized Scaffolds from Submandibular Glands of Rats

DAI Taiqiang¹, ZHANG Linlin², AN Ying², XU Fangfang¹, SHAO Xiaoxi¹, LIU Yanpu¹, LIU Bin^3*

1. State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shanxi Clinical Research Center for Oral Diseases, Department of Oral and Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University, Xi'an 710032, China;
2. State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shanxi Engineering Research Center for Dental Materials and Advanced Manufacture, Department of Periodontology, School of Stomatology, The Fourth Military Medical University, Xi'an 710032, China;
3. State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases, Laboratory Animal Center, School of Stomatology, The Fourth Military Medical University, Xi'an 710032, China

Received:2020-03-24 Online:2020-10-28 Published:2020-10-19

摘要/Abstract

摘要： 目的: 探究利用灌注法制备大鼠颌下腺脱细胞支架的可行性,为组织工程涎腺的构建提供支架材料支持。方法: 建立大鼠颌下腺灌注系统,利用脱细胞制剂的组合制备大鼠颌下腺脱细胞支架, HE染色及扫描电镜观察组织结构;导管及静脉铸型观察管道系统完整性;Masson染色、免疫荧光染色及胶原定量检测细胞外基质成分存留情况,DNA定量检测有无DNA残留。结果: 经静脉及导管插管,可以建立大鼠颌下腺的灌注系统,并成功制备大鼠颌下腺脱细胞支架,其基本保持了颌下腺原有的组织结构,并存留有细胞外基质中的Ⅰ、Ⅳ型胶原、纤维连接蛋白、层粘连蛋白。同时,DNA定量显示,其残留小于50 μg/g冻干脱细胞支架;导管及静脉铸型显示,管道系统基本完整。结论: 利用灌注法首次成功制备大鼠颌下腺脱细胞支架,这将为组织工程涎腺的构建提供更加理想的支架材料支持。

关键词: 颌下腺, 脱细胞支架, 组织工程再生

Abstract: Objective: To explore the method for preparation of decellularized scaffold (DS) from submandibular glands (SMGs) of rats and provide scaffolds for tissue engineering regeneration of salivary glands. Methods: Perfusion system of SMGs was built by ductal and venous cannula. Decellularized SMGs scaffolds were produced by freeze-thaw cycle and sequence perfusion with SDS, Triton X-100, and DNase I through the cannula of vein. HE staining and SEM were performed to observe the general and microstructure of the DS, while Masson staining and immunofluorescence staining were performed to assess extracellular matrix retention. At the same time, the casts of duct and vein for SMGs and DS were compared to analyze the retention of ductal and venous system. Finally, the retention of collagen and DNA was detected by quantitative analysis. Results: Perfusion system was constructed by ductal and venous cannula and DS was produced successfully by perfusion method. DS held the original structure and kept retention of collagen type Ⅰ, Ⅳ, fibronectin, and laminin in extracellular matrix. DNA quantification showed that less than 50 μg/g DNA remained on dry tissue. The casts suggested relatively intact ductal and venous system after decellularization. Conclusion: Decellularized salivary glands scaffolds can be successfully prepared by perfusion method and it will provide scaffolds for tissue engineering construction of salivary glands.

Key words: submandibular glands, decellularized scaffolds, tissue engineering regeneration

戴太强, 张琳琳, 安莹, 许方方, 邵小夕, 刘彦普, 刘斌. 大鼠颌下腺脱细胞支架的制备研究[J]. 口腔医学研究, 2020, 36(10): 948-952.

DAI Taiqiang, ZHANG Linlin, AN Ying, XU Fangfang, SHAO Xiaoxi, LIU Yanpu, LIU Bin. Preparation of Decellularized Scaffolds from Submandibular Glands of Rats[J]. Journal of Oral Science Research, 2020, 36(10): 948-952.

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大鼠颌下腺脱细胞支架的制备研究

Preparation of Decellularized Scaffolds from Submandibular Glands of Rats

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