Decellularization and in vitro characterization of porcine small intestine scaffolds for complex wound treatments

Abstract

Introduction: complicated skin injuries have become a global health problem, being difficult to treat due to the body’s limited healing process. Many studies aim to enhance traditional treatments for skin injuries, which have many disadvantages. Therefore, wound healing research is aiming towards tissue engineering options, such as decellularized matrix, which have shown great healing and biocompatibility competencies. Objectives: to obtain and characterize the properties of a decellularized biological matrix derived from the small intestine of animals. Methods: porcine small intestine was prepared and decellularized using four different methods: Triton X-100, sodium dodecyl sulfate (SDS) and sodium deoxycholate (SDC) for one or two cycles of 6 hours or 24 hours, and peracetic acid for one cycle of 2 hours. The remaining DNA was quantified with Nanodrop and electrophoresis characterization. Histology stains and Scanning Electron Microscopy (SEM) were performed to assess surface structure and integrity. Resistance assays were conducted to measure mechanical strength. Finally, degradability assays with different buffers were performed. Results: no differences between the decellularization protocols regarding remaining DNA were found, making protocols of one cycle of six hours more efficient. With the least remaining DNA content and better structure perseveration, TX-100 could be considered as the best protocol. No statistically difference between protocols and native tissue were found during the mechanical analysis. Biodegradability assays showed the expected degradability properties of the produced matrix. Conclusions: promising results were achieved to obtain decellularized biological matrices that could serve as a treatment for complicated skin wounds. More in vitro and molecular studies should be carried out in future studies to further characterize these scaffolds.

Keywords

• bioactive scaffold
• decellularization
• porcine small intestine
• skin wound
• tissue engineering