Bio-functional Collagen Matrix Scaffold Composition Differentially Promotes Paracrine Activity In Human Induced Pluripotent Stem Cell Derived Vascular Smooth Muscle Cells
Kaiti Duan, B.S.1, Biraja Dash, PhD2, Henry C. Hsia, MD2.
1Frank H. Netter MD School of Medicine, Hamden, CT, USA, 2Yale School of Medicine, New Haven, CT, USA.
Purpose: Induced pluripotent stem cell-derived vascular smooth muscle cells (iPSC-VSMC) have the potential to treat chronic wounds by secreting proangiogenic factor vascular endothelial growth factor (VEGF) (1). However, little is known how the extracellular matrix (ECM) composition may impact the cellsí paracrine secretion profile. In this study, our objective was to understand the effects of ECM density and functionalities on the secretory profile of human iPSC-VSMCs.
Methods: Type-I collagen was used as the material for the scaffolds and were incorporated with functional biomolecules: hyaluronic acid (HA), fibronectin, and laminin. This allowed fabrication of collagen scaffolds with different functionalities. The functionalities were used in combination with three different densities of type-I collagen (1.25mg/ml, 2.5mg/ml, and 5mg/ml) to study iPSC-VSMC viability and paracrine secretion profile. Several pro-angiogenetic factors included VEGF, SDF, PDGF, bFGF, angiopoietin 1, IL-8, and TGF-beta were investigated. The anti-inflammatory factor IL-10 was also evaluated.
Result: Human iPSC-VSMCs embedded in collagen scaffolds containing functional biomolecules showed an increase in cell viability across all the collagen densities compared to the collagen scaffolds without functional biomolecules. The greatest significant level of cell proliferation was display by the 1.25mg/ml scaffolds (P-value=0.0001). Enhanced VEGF was observed in all three of the functionalized scaffolds at 1.25mg/ml collagen density (P value=0.0086) and also in the 2.5mg/ml collagen density scaffold containing HA (p value=0.0026), whereas no significant difference in VEGF level was found among 5mg/ml functionalized scaffolds. Interestingly, fibronectin-functionalized 5mg/ml collagen scaffold exhibited substantially elevated bFGF secretion (P value=0.0049). There was also a positive correlation between increasing amounts of fibronectin with increasing bFGF paracrine secretion (P value=0.0001). Furthermore, our data showed respective upregulation of IL-8 and IL-10 secretion in fibronectin-functionalized collagen scaffolds at the 1.25mg/ml and 2.5mg/ml collagen densities.
Conclusion: These results suggested that functionalization along with density differentially regulate paracrine function of human iPSC-VSMCs. Future studies in elucidating the underlying signaling pathways and mouse model experiments will further deepen our understanding of the interaction between ECM and human iPSC-VSMCs. This will ultimately optimize human iPSC-VSMCs as a viable therapeutic agent for chronic wound healing.
Kuzuya M, Satake S, Esaki T, Yamada K, Hayashi T, Naito M, et al. Induction of angiogenesis by smooth muscle cell-derived factor: Possible role in neovascularization in atherosclerotic plaque. Journal of Cellular Physiology. 1995;164(3):658-67.
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