Abstract：
Decellularization is an innovative method to create natural scaffolds by removing all cellular materials while preserving the composition and three-dimensional ultrastructure of the extracellular matrix (ECM). The obtention of decellularized reproductive organs in cats might facilitate the development of assisted reproductive techniques not only in this species but also in other felids. The aim was to compare the efficiency of three decellularization protocols on reproductive organs (ovary, oviduct, and uterine horn) in domestic cats. The decellularization protocol involved 0.1% sodium dodecyl sulfate and 1%Triton X-100. Protocol 1 (P1) entailed 2-cycles of decellularization using these detergents. Protocol 2 (P2) was like P1 but included 3-cycles. Protocol 3 (P3) was similar to P2, with the addition of deoxyribonuclease incubation. Reproductive organs from nine cats were separated into two sides. One side served as the control (non-decellularized organ) while the contralateral side was the treated group (decellularized organ). The treated organs were subdivided into 3 groups (n = 3 per group) for each protocol. Both control and treated samples were analyzed for DNA content, histology (nuclear and ECM (collagen, elastin, and glycosaminoglycans (GAGs)) density), ultrastructure by electron microscopy, and cytotoxicity. The results of the study showed that P3 was the only protocol that displayed no nucleus residue and significantly reduced DNA content in decellularized samples (in all the studied organs) compared to the control (P < 0.05). The ECM content in the ovaries remained similar across all protocols compared with controls (P > 0.05). However, elastic fibers and GAGs decreased in decellularized oviducts (P < 0.05), while collagen levels remained unchanged (P > 0.05). Regarding the uterus, the ECM content decreased in decellularized uterine horns from P3 (P < 0.05). Electron microscopy revealed that the microarchitecture of the decellularized samples was maintained compared to controls. The decellularized tissues, upon being washed for 24 h, showed cytocompatibility following co-incubation with sperm. In conclusion, when comparing different decellularization methods, P3 proved to be the most efficient in removing nuclear material from reproductive organs compared to P1 and P2. P3 demonstrated its success in decellularizing ovarian samples by significantly decreasing DNA content while maintaining ECM components and tissue microarchitecture. However, P3 was less effective in maintaining ECM contents in decellularized oviducts and uterine horns.
摘要：
去细胞化是通过去除所有细胞材料同时保留细胞外基质（ECM）的组成和三维超微结构来创建天然支架的创新方法。在猫中获得脱细胞生殖器官可能不仅在该物种中而且在其他猫科动物中都促进了辅助生殖技术的发展。目的是比较三种去细胞化方案对生殖器官（卵巢，输卵管,和子宫角）在家猫中。脱细胞方案涉及0.1%十二烷基硫酸钠和1%TritonX-100。方案1(P1)需要使用这些洗涤剂进行2个循环的去细胞化。方案2(P2)类似于P1,但包括3个循环。方案3(P3)类似于P2,添加脱氧核糖核酸酶孵育。将9只猫的生殖器官分成两侧。一侧用作对照(非脱细胞器官),而对侧是治疗组(脱细胞器官)。对于每个方案,将处理的器官细分为3组(每组n=3)。分析对照和处理过的样品的DNA含量，组织学(核和ECM(胶原蛋白，弹性蛋白,和糖胺聚糖(GAG)密度)，超微结构通过电子显微镜，和细胞毒性。研究结果表明，P3是唯一的方案，与对照组相比，在脱细胞样品（在所有研究的器官中）中没有显示出核残基并显着降低了DNA含量（P<0.05）。与对照相比,卵巢中的ECM含量在所有方案中保持相似(P>0.05)。然而,去细胞输卵管中的弹性纤维和GAGs减少（P<0.05），而胶原蛋白水平保持不变（P>0.05）。关于子宫,从P3开始脱细胞子宫角中ECM含量降低（P<0.05）。电子显微镜显示,与对照相比,脱细胞样品的微结构得以维持。去细胞化的组织,清洗24小时后，与精子共孵育后显示细胞相容性。总之,当比较不同的去细胞化方法时，与P1和P2相比，P3被证明是最有效的从生殖器官中去除核材料。P3通过显着降低DNA含量同时保持ECM成分和组织微结构，证明了其在使卵巢样品脱细胞化方面的成功。然而,P3在维持去细胞化输卵管和子宫角中的ECM含量方面效果较差。
