Bioinspired Collagen/κ-Carrageenan 3D Matrix for In Vitro Modeling of Vascular Calcification

Area of Science:

• Biomaterials Science
• Vascular Biology
• Regenerative Medicine

Background:

• Pathological soft tissue calcification, especially in vascular structures, is a key factor in cardiovascular diseases, leading to vascular stiffening and dysfunction.
• The cellular mechanisms driving the transdifferentiation of vascular smooth muscle cells into osteochondroblast-like cells, crucial for calcification, are not fully understood.
• Existing models lack the complexity to accurately replicate the vascular extracellular matrix (ECM) environment.

Purpose of the Study:

• To develop a bioinspired 3D scaffold mimicking the vascular ECM for studying pathological calcification.
• To investigate the transdifferentiation of mouse vascular smooth muscle cells (MOVAS) into a mineralizing phenotype within this novel scaffold.
• To explore the role of scaffold composition, specifically type I collagen (Col) and κ-carrageenan (κ-Carr), in directing cellular responses and calcification.

Main Methods:

• Fabrication of a 3D scaffold using type I collagen (Col) and κ-carrageenan (κ-Carr).
• Culture of MOVAS cells on the scaffold under osteogenic conditions.
• Assessment of cell behavior (adhesion, spreading) and osteogenic differentiation (marker expression, alkaline phosphatase activity).
• Characterization of mineral deposition using spectroscopic and thermogravimetric analyses.

Main Results:

• The 3D scaffold successfully supported MOVAS cell adhesion, spreading, and osteogenic transdifferentiation.
• Overexpression of osteogenic markers (TNAP, RUNX2) and increased alkaline phosphatase activity confirmed MOVAS differentiation.
• Formation of carbonated apatite minerals with a calcium-deficient apatite structure indicated controlled mineral deposition.
• κ-carrageenan incorporation enhanced the calcification process, highlighting the importance of biochemical cues.

Conclusions:

• The developed bioinspired 3D scaffold effectively replicates the vascular ECM, providing a physiologically relevant model for studying pathological calcification.
• This model facilitates the investigation of MOVAS transdifferentiation and mineralization processes.
• The findings suggest potential for developing regenerative biomaterials and therapeutic strategies to prevent vascular calcification in cardiovascular diseases.