Development of a Cell-Based Biosensor for Residual Detergent Detection in Decellularized Scaffolds

Area of Science:

• Biomaterials Engineering
• Tissue Engineering
• Cellular and Molecular Biology

Background:

• Ex vivo organ engineering using decellularized scaffolds and stem cells offers a promising alternative to organ transplantation.
• A key challenge is ensuring scaffold cytocompatibility by accurately detecting residual detergents.
• Current evaluation methods lack the required precision and specificity.

Purpose of the Study:

• To develop a highly sensitive and specific cell-based biosensor (CBB) for detecting residual detergents in decellularized tissues.
• To establish a reliable method for assessing scaffold cytocompatibility in regenerative medicine applications.

Main Methods:

• Engineered a CBB utilizing the NF-κB inducible promoter linked to a reporter gene (secreted alkaline phosphatase - SEAP).
• Cloned promoter sections containing NF-κB binding sequences upstream of the SEAP gene in HEK293 cells.
• Validated biosensor sensitivity, specificity, and stability using quantitative PCR (qPCR) and SEAP activity assays after exposure to sodium dodecyl sulfate (SDS).

Main Results:

• The developed CBB detected minute amounts of SDS detergent as low as 3.467 pM.
• Optimal biosensor performance was observed 8 hours post-detergent exposure, with stable function across high cell passage numbers.
• A threshold SEAP activity of 4.36 U/L indicated a reduction in transplanted cell viability below 70% in decellularized tissues.

Conclusions:

• A novel CBB was successfully developed for sensitive and efficient detection of residual detergents like SDS.
• This biosensor provides a precise tool for evaluating the cytocompatibility of decellularized scaffolds.
• The findings advance the safety and efficacy of ex vivo organ engineering for transplantation.