Almeida Gustavo Henrique Doná Rodrigues, da Silva Raquel Souza, Gibin Mariana Sversut, Gonzaga Victória Hellen de Souza, Dos Santos Henrique, Igleisa Rebeca Piatniczka, Fernandes Leticia Alves, Fernandes Iorrane Couto, Nesiyama Thais Naomi Gonçalves, Sato Francielle, Baesso Mauro Luciano, Hernandes Luzmarina, Rinaldi Jaqueline de Carvalho, Meirelles Flávio Vieira, Astolfi-Ferreira Claudete S, Ferreira Antonio José Piantino, Carreira Ana Claudia Oliveira
Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo 03828-000, Brazil.
Department of Physics, State University of Maringá, Maringá 87020-900, Brazil.
Biomimetics (Basel). 2024 Jun 24;9(7):382. doi: 10.3390/biomimetics9070382.
The uterine tube extracellular matrix is a key component that regulates tubal tissue physiology, and it has a region-specific structural distribution, which is directly associated to its functions. Considering this, the application of biological matrices in culture systems is an interesting strategy to develop biomimetic tubal microenvironments and enhance their complexity. However, there are no established protocols to produce tubal biological matrices that consider the organ morphophysiology for such applications. Therefore, this study aimed to establish region-specific protocols to obtain decellularized scaffolds derived from porcine infundibulum, ampulla, and isthmus to provide suitable sources of biomaterials for tissue-engineering approaches. Porcine uterine tubes were decellularized in solutions of 0.1% SDS and 0.5% Triton X-100. The decellularization efficiency was evaluated by DAPI staining and DNA quantification. We analyzed the ECM composition and structure by optical and scanning electronic microscopy, FTIR, and Raman spectroscopy. DNA and DAPI assays validated the decellularization, presenting a significative reduction in cellular content. Structural and spectroscopy analyses revealed that the produced scaffolds remained well structured and with the ECM composition preserved. YS and HEK293 cells were used to attest cytocompatibility, allowing high cell viability rates and successful interaction with the scaffolds. These results suggest that such matrices are applicable for future biotechnological approaches in the reproductive field.
输卵管细胞外基质是调节输卵管组织生理学的关键组成部分，其具有区域特异性的结构分布，这与其功能直接相关。考虑到这一点，在培养系统中应用生物基质是开发仿生输卵管微环境并提高其复杂性的一种有趣策略。然而，目前尚无既定方案可用于制备考虑到此类应用的器官形态生理学的输卵管生物基质。因此，本研究旨在建立区域特异性方案，以获得源自猪输卵管漏斗部、壶腹部和峡部的脱细胞支架，为组织工程方法提供合适的生物材料来源。将猪输卵管在0.1%十二烷基硫酸钠和0.5% Triton X-100溶液中进行脱细胞处理。通过DAPI染色和DNA定量评估脱细胞效率。我们通过光学显微镜、扫描电子显微镜、傅里叶变换红外光谱和拉曼光谱分析了细胞外基质的组成和结构。DNA和DAPI检测验证了脱细胞效果，显示细胞含量显著降低。结构和光谱分析表明，所制备的支架结构良好，细胞外基质组成得以保留。使用永生化输卵管上皮细胞（YS）和人胚肾293细胞（HEK293）来证明细胞相容性，结果显示细胞活力率高且与支架成功相互作用。这些结果表明，此类基质适用于生殖领域未来的生物技术应用。
