Electrospun Decellularized Lung Matrix Scaffold for Airway Smooth Muscle Culture

Abstract

Chronic respiratory disease affects many people worldwide with little known about the intricate mechanisms driving the pathology, making it difficult to develop novel therapies. Improving the understanding of airway smooth muscle and extracellular matrix (ECM) interactions is key to developing treatments for this leading cause of death. With currently no relevant or controllable in vivo or in vitro models to investigate cell–ECM interactions in the small airways, the development of a biomimetic in vitro model with cell attachment, signaling, and organization is needed. The goal of this study was to create a biologically and structurally relevant in vitro model of small airway smooth muscle. In order to achieve this goal, a scaffold was engineered from synthetic poly-l-lactic acid (PLLA) and decellularized pig lung ECM (PLECM). PLECM scaffolds have improved physical characteristics over synthetic scaffolds, by exhibiting a significant decrease in the elastic modulus and an increase in hydrophilicity. Histological staining and SDS-PAGE showed that essential proteins or protein fragments found in natural ECM were present after processing. Human bronchial smooth muscle cells (HBSMCs) seeded onto PLECM 3D scaffolds formed confluent layers and maintained a contractile phenotype, as demonstrated by the organized arrangement of actin filaments within the cell and expected contractile protein expression of calponin 1. HBSMCs cultured on electrospun PLECM scaffold also increased alpha-1 type 1 collagen compared to those cultured on PLLA scaffolds. In summary, this research demonstrates that a PLLA/PLECM composite electrospun mat is a promising tool to produce an in vitro model of the airway with the potential for a better understanding of bronchiole smooth muscle behavior in diseased or normal states.

Fabrication and Characterization of Electrospun Decellularized Muscle-Derived Scaffolds

“…Previous studies with electrospinning dECM have relied on blending with carrier polymers. [27][28][29][30][31][32] We have identified key steps that are crucial for the successful formation of stable, electrospun dECM scaffolds. By homogenizing the dECM into a uniform slurry, any remaining connective tissue could be effectively removed.…”

“…Recent studies have reported dECM blended with synthetic polymers to create hybrid scaffolds with enhanced biochemical properties compared with traditional synthetic materials. [27][28][29][30][31][32] However, dECM alone has been shown to be difficult to electrospin and produce stable scaffolds for analysis. To our knowledge, there are no current studies in the literature of successful electrospinning of muscle dECM without a carrier polymer.…”

Evolution of Bioengineered Lung Models: Recent Advances and Challenges in Tissue Mimicry for Studying the Role of Mechanical Forces in Cell Biology

“…Furthermore, it has been shown that decellularized extracellular matrix (dECM) containing essential proteins for cell attachment and proliferation, could be a good candidate to improve the biocompatibility of copolymers . In a recent study, hybrid electrospun scaffold of poly‐L‐lactic acid and dECM (derived from pig lung) for in vitro ASM model showed that dECM improved physical characteristics (e.g., wettability) …”

Co-culture cell-derived extracellular matrix loaded electrospun microfibrous scaffolds for bone tissue engineering

“…[38][39][40][41] These scaffolds have shown superior mechanical properties and maintenance of bioactivity in various tissue engineering applications. [42][43][44] Different approaches have been applied to bone tissue engineering applications. Gibson and co-workers incorporated decellularized ECM nanoparticles from bone into a biosynthetic nanofiber composite scaffold 45 while Jeon and colleagues have cultured pre-osteoblasts on electrospun PCL scaffolds and decellularized it to obtain decellularized cell-derived ECM scaffolds.…”