Tissue decellularization

Natural extracellular matrix (ECM) obtained from decellularized tissues has a wide range of applications for regenerative medicine in both preclinical and clinical settings. Numerous products are currently utilized for applications in wound care, pericardial reconstruction, heart valve replacement, and ophthalmology. These materials are used as inductive templates for tissue and organ reconstruction, by providing a native and complex microenvironment for cell growth and tissue remodeling.

However, the ability to promote regeneration is critically dependent upon the methods used to prepare the scaffold material. Every decellularization method may alter the composition and the micro-architecture of ECM, causing variations of the biological activity and mechanical integrity of the decellularized material. CHP specifically detects denatured collagen in the decellularized tissue/organ, enabling assessment of the effect of decellularization upon ECM at the molecular level.

CHP staining basement membrane complex of a porcine urinary bladder

 

Staining with fluorescent F-CHP directly reveals different contents of denatured collagen in the basement membrane complex of a porcine urinary bladder decellularized with different detergents (1% SDS, 8 mM CHAPS, 4% SD, 3% Triton X-100). Scale bar: 50 μm. Images adapted from ref [1].

With simple image analysis, the CHP stain allows quantitative comparison of molecular collagen denaturation among ECM materials decellularized with different protocols, which is not feasible for current methods like convention histological stain, SEM or TEM.[1] CHP is a powerful tool for optimization and standardization of the decellularization technology. 

 

Reference

  1. Molecular assessment of collagen denaturation in decellularized tissues using a collagen hybridizing peptide. Acta Biomaterialia (2017) (open access) more info