Characterization of Decellularized Implants for Extracellular Matrix Integrity and Immune Response Elicitation

Abstract

Biological scaffold or implant is a popular choice for the preparation of tissue-engineered organs and has the potential to address donor shortage in the clinics. However, biological scaffolds prepared by physical or chemical agents cause damage to the extracellular matrix by potentially inducing immune responses after implantation. The current study explores an alternative route for the preparation of acellular scaffolds and explores the fate of the prepared scaffolds in a milieu of immune cells following implantation without using immunosuppressant. Using the syngeneic (Lewis male-Lewis female) and allogeneic (Brown Norway male-Lewis female) models and different tissue routes (subcutaneous vs omentum) for transplantation, normal blood vascular scaffolds were implanted which was converted to acellular vascular scaffolds by in vivo natural decellularization at the end of 2 months of observation. We also prepared chemically decellularized acellular scaffolds from normal untreated blood vascular scaffolds using a cocktail of chemicals which was also similarly placed in subcutaneous and omentum sites. Here, we applied in-depth quantitative proteomics along with histology and image analysis to comprehensively describe and compare the proteome of the natural and chemically decellularized scaffold. Our data confirm that site-specific advantages exist in modulating the ECM and regulating the immune responses (macrophage and T cells) following implantation, which possibly led to the production of an acellular scaffold (natural decellularization) under in vivo conditions. The current approach opens up the possibility to create tailor-made acellular scaffolds to build functional blood vessels. In addition, the identification of different tissue sites facilitates differential immune response against the scaffolds. This study provides a rich resource aimed toward an enhanced mechanistic understanding to study immune responses under similar settings in the field of transplantation and regenerative medicine.

Angiogenesis and Re-endothelialization in decellularized scaffolds: Recent advances and current challenges in tissue engineering

“…Banerjee D et al implanted the acellular and cellular aorta scaffolds in the omentum and subcutaneous position. It was observed that in contrast to omental implants, the subcutaneous site implantation minimized subsequent immune responses ( Banerjee et al, 2022 ). The change of ECM microstructure during the decellularization process is the third factor.…”

Macrophage response mediated by extracellular matrix: recent progress

“…These nuances might be elaborated by the combination of techniques in the transcriptional and post-transcriptional level (e.g. proteomics [59]), which will be elucidated in section 3.5. What's more, since the reproducibility, scalability, cost, and homology [60], mice are one of the most commonly used models in scRNA-seq.…”

“…Applying the combination of protein and genomic tools, Banerjee et al [59] explore the fate of the chemical decellularized or normal ECM scaffolds at different body routes (subcutaneous and omentum) using genetically different donor and recipient rats (syngeneic and allogeneic). Alpha actin (Acta2) or alpha-smooth muscle actin (a-SMA), which is associated with fibrous capsule was not upregulated in any of the biomaterials, showing the good biocompatibility of ECM scaffold.…”

Polymer–Metal Bilayer with Alkoxy Groups for Antibacterial Improvement

“…However, the main problems with metal prostheses were their bacterial attachment and colonization [ 2 ]. A bacteria-contaminated implants can cause, for example, acute cellular rejection [ 3 ]. As an alternative, in the second half of the 20th century, polymers were introduced into medicine and pharmacy.…”