Department of Veterinary Science, Islamic Azad University Shabestar Branch, 5381637181, Shabestar, Iran.
Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
BMC Res Notes. 2022 Nov 8;15(1):346. doi: 10.1186/s13104-022-06233-7.
Recently, the decellularization technique is introduced as one of the tissue engineering procedures for the treatment of various deficiencies. Here, we aimed to assess the dynamic activity of CCs and HUVECs within decellularized bovine ovarian tissue transplanted subcutaneously in rats. Ovarian tissue was decellularized using a cocktail consisting of different chemicals, and the efficiency of decellularization was assessed using hematoxylin-eosin and DAPI staining. The cell survival was evaluated using an LDH leakage assay. Thereafter, decellularized samples were recellularized using HUVECs and CCs, encapsulated inside alginate (1.2%)-gelatin, (1%) hydrogel, and transplanted subcutaneously to rats. The existence of CD31- and estrogen-positive cells was assessed using immunohistochemistry staining.
Bright-field imaging and DAPI staining revealed the lack of nuclei with naive matrix structure in ovarian tissue subjected to decellularization protocol. SEM imaging revealed a normal matrix in decellularized ovaries. LDH assay showed a lack of cytotoxicity for CCs after 7-days compared to the control group. Immunohistochemistry staining showed both CD31- and estrogen-positive cells in CCs + HUVECs compared to the CCs group. CD31 cells appeared with flattened morphology aligned with matrix fibers. The existence of estrogen and CD31 positive cells showed the efficiency of decellularized ovarian tissue to restore cellular function and activity.
最近，脱细胞技术被引入组织工程程序之一，用于治疗各种缺陷。在这里，我们旨在评估 CCs 和 HUVECs 在牛卵巢组织脱细胞后移植到大鼠皮下的动态活性。使用包含不同化学物质的鸡尾酒对卵巢组织进行脱细胞处理，并使用苏木精-伊红和 DAPI 染色评估脱细胞效率。使用 LDH 漏出测定评估细胞存活率。此后，使用 HUVECs 和 CCs 对脱细胞样本进行再细胞化，将其包裹在藻酸盐（1.2%）-明胶、（1%）水凝胶中，并移植到大鼠皮下。使用免疫组织化学染色评估 CD31 和雌激素阳性细胞的存在。
明场成像和 DAPI 染色显示，在经历脱细胞方案的卵巢组织中缺乏细胞核和原始基质结构。SEM 成像显示脱细胞卵巢中的正常基质。LDH 测定显示，与对照组相比，CCs 在 7 天后缺乏细胞毒性。与 CCs 组相比，CCs+HUVECs 中均显示 CD31 和雌激素阳性细胞。CD31 细胞呈现出与基质纤维对齐的扁平形态。雌激素和 CD31 阳性细胞的存在表明脱细胞卵巢组织恢复细胞功能和活性的效率。
