Biochemical and immunomodulatory insights of extracellular matrix from decellularized human whole cervix: recellularization and in vivo ECM remodelling interplay.
Atul Kumar OjhaRagavi RajasekaranArmaan Kunwar HansdaPriyanka ChoudhuryAsmita BiswasSunita SharmaP Pal ChaudhuryNantu DograRitobrata GoswamiKoel ChaudhurySantanu DharaPublished in: Biofabrication (2024)
Extracellular matrix (ECM) rich whole organ bio-scaffolds, preserving structural integrity and essential growth factors, has potential towards regeneration and reconstruction. Women with cervical anomalies or trauma can benefit from clinical cervicovaginal repair using constructs rich in site specific ECM. In this study, complete human cervix decellularization was achieved using a modified perfusion-based stir bench top decellularization method. This was followed by physico-chemical processes including perfusion of ionic agents, enzymatic treatment and washing using detergent solutions for a duration of 10-12 days. Histopathological analysis, as well as DNA quantification confirmed the efficacy of the decellularization process. Tissue ultrastructure integrity was preserved and the same was validated via scanning electron microscopy (SEM) and transmission electron microscopy (TEM) studies. Biochemical analysis and structural characterizations like FTIR, Raman spectroscopy of decellularised tissues demonstrated preservation of important proteins, crucial growth factors, collagen, and glycosaminoglycans (GAGs ). In vitro studies, using THP-1and HUVEC cells, demonstrated macrophage polarisation from M1 to M2 and vascular functional genes enhancement, respectively, when treated with decellularized cervical matrix. human decellularised cervical matrix (hdCM) () grafts were recellularized with site specific human cervical epithelial cells and HUVEC, showing non-cytotoxic cell viability and enhanced proliferation. Futhermore, hdCM scaffolds showed immunomodulatory effects in vivo on small rodent model via upregulation of M2 macrophage genes as compared to decellularised rat cervical matrix scaffolds (DRC). hdCM scaffolds underwent neo-vascularization followed by ECM remodelling with enhanced tissue integration.
Keyphrases
- extracellular matrix
- electron microscopy
- endothelial cells
- tissue engineering
- induced pluripotent stem cells
- pluripotent stem cells
- raman spectroscopy
- stem cells
- adipose tissue
- cell proliferation
- preterm birth
- gene expression
- oxidative stress
- genome wide
- cell death
- long non coding rna
- risk assessment
- high resolution
- smoking cessation
- combination therapy
- wound healing