Dos Santos Ana Caroline, de Andrade Livia Maria Barbosa, Candelária Raí André Querino, de Carvalho Juliana Casanovas, Valbão Maria Carolina Miglino, Barreto Rodrigo da Silva Nunes, de Faria Marcelo Domingues, Buchaim Rogerio Leone, Buchaim Daniela Vieira, Miglino Maria Angelica
Graduate Program in Anatomy of Domestic and Wild Animals, Faculty of Veterinary Medicine and Animal Science, University of São Paulo (FMVZ/USP), São Paulo 05508-270, Brazil.
Medical School, University of Marília (UNIMAR), Marília 17525-902, Brazil.
Bioengineering (Basel). 2025 Jan 9;12(1):52. doi: 10.3390/bioengineering12010052.
The shortage of tissues and damaged organs led to the development of tissue engineering. Biological scaffolds, created from the extracellular matrix (ECM) of organs and tissues, have emerged as a promising solution for transplants. The ECM of decellularized auricular cartilage is a potential tool for producing ideal scaffolds for the recellularization and implantation of new tissue in damaged areas. In order to be classified as an ideal scaffold, it must be acellular, preserving its proteins and physical characteristics necessary for cell adhesion. This study aimed to develop a decellularization protocol for pig ear cartilage and evaluate the integrity of the ECM. Four tests were performed using different methods and protocols, with four pig ears from which the skin and subcutaneous tissue were removed, leaving only the cartilage. The most efficient protocol was the combination of trypsin with a sodium hydroxide solution (0.2 N) and SDS (1%) without altering the ECM conformation or the collagen architecture. In conclusion, it was observed that auricular cartilage is difficult to decellularize, influenced by material size, exposure time, and the composition of the solution. Freezing and thawing did not affect the procedure. The sample thickness significantly impacted the decellularization time.
组织和受损器官的短缺促使了组织工程的发展。由器官和组织的细胞外基质（ECM）制成的生物支架已成为移植的一种有前景的解决方案。脱细胞耳软骨的ECM是为受损区域新组织的再细胞化和植入生产理想支架的潜在工具。为了被归类为理想支架，它必须是无细胞的，保留其细胞粘附所需的蛋白质和物理特性。本研究旨在开发一种猪耳软骨脱细胞方案并评估ECM的完整性。使用不同方法和方案进行了四项测试，用了四只猪耳，去除了皮肤和皮下组织，仅留下软骨。最有效的方案是胰蛋白酶与氢氧化钠溶液（0.2N）和SDS（1%）的组合，且不改变ECM构象或胶原结构。总之，观察到耳软骨难以脱细胞，受材料大小、暴露时间和溶液组成影响。冻融不影响该过程。样品厚度显著影响脱细胞时间。
