The preparation for decellularized scaffolds in the rat uterine and its evaluation
GUO Fang1,3, YANG Jie1, CHEN Jingjing1, LIU Li1, CHEN Yunzhi2, ZHENG Feiyun3
1.Department of Gynecology, the Third Clinical College of Wenzhou Medical University, Wenzhou People’s Hospital, Wenzhou, 325000; 2.Department of General Surgery, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015; 3.Department of Gynecology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015
GUO Fang,YANG Jie,CHEN Jingjing, et al. The preparation for decellularized scaffolds in the rat uterine and its evaluation[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2018, 48(3): 162-167.
Abstract：Objective: To explore the feasible strategy for the preparation of natural uterine decellularized scaffolds and to evaluate it systematically. Methods: Female SD rats were used, detergents including TritonX-100 and sodium dodecyl sulfate were perfused into the uterus through the uterine artery after physical freezing and thawing and enzymatic hydrolysis. After decellularization, scaffolds were evaluated by general observation, methylene blue staining, HE staining, quantitative analysis of genomic DNA, cytokine content determination, transmission and scanning electron microscopy (TEM and SEM), collagen detection and identification. Results: After the perfusion, the transparent uterine scaffold was obtained, HE staining and TEM showed that there was no cell residual in it. DNA content was confirmed to be (45.6±7.3) ng/mg, less than 5% of the normal uterus (P<0.01). Methylene blue staining and SEM showed that vascular network and spatial structure were intact. Collagen content increased from (0.57±0.12) μg/mg in fresh uterine tissue to (0.88±0.10) μg/mg in scaffolds (P<0.05), combined with IHC study, indicating that extracellular matrix components of the scaffolds were completely preserved. In addition, ELISA results showed that the cytokines EGF, bFGF and TGF-β were still retained in the decellularized scaffolds (66%, 85% and 54%, respectively), indicating that they still had some biological activity. Conclusion: The decellularized uterine scaffold could be successfully prepared with its physical and chemical properties preserved, and it may be used as an alternative platform for uterine tissue engineering study.
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