Christopher B Sylvester1,2, Aarthi Pugazenthi3, K Jane Grande-Allen1, Ravi K Ghanta4. 1. Deparment of Bioengineering, Rice University, Houston, TX, USA. 2. Medical Scientist Training Program, Baylor College of Medicine, Houston, TX, USA. 3. Michael E. DeBakey Department of Surgery, Baylor College of Medicine, One Baylor Plaza MS390, Houston, TX, 77030, USA. 4. Michael E. DeBakey Department of Surgery, Baylor College of Medicine, One Baylor Plaza MS390, Houston, TX, 77030, USA. ravi.ghanta@bcm.edu.
Abstract
PURPOSE: Cellular therapy with mesenchymal stem cells (MSCs) shows promise for restoring function after myocardial infarction (MI). However, cellular therapy has yet to be clinically translated, in part because of difficulty in studying how MSCs interact with the post-MI scar microenvironment. This study aimed to design an in vitro model to study MSC behavior in the post-MI scar stiffness microenvironment. METHODS: Using poly(ethylene glycol)-acrylate (PEG) conjugated to bioactive peptides, rat MSCs were encapsulated in hydrogels of varying stiffnesses and crosslinking densities. Cell viability was assessed through 14 days using calcein and ethidium homodimer staining. To simulate post-MI pro-fibrotic signaling, transforming growth factor-beta (TGFβ) was added to selected cultures. Immunofluorescence and qRT-PCR were used to assess changes in cardiac transdifferentiation or paracrine secretion, two proposed methods of MSCs in cellular therapy. RESULTS: Bioactivated PEG hydrogels with stiffnesses between 1.6 and 151.0 kPa were prepared. Rat MSCs demonstrated up to 71.6% viability after 3 days of encapsulated culture, and survived within the hydrogels up to 14 days. Encapsulation decreased MSC expression of cardiac troponin T and most growth factors, except interleukin-6. Meanwhile, TGFβ caused increased cardiac troponin T expression but decreased secreted factor expression. Varying hydrogel stiffness did not have an effect on cardiac troponin T or secreted factor expression. CONCLUSIONS: These findings suggest that a 3D microenvironment hinders two key mechanisms by which MSCs could improve cardiac function after post-MI scar formation, namely cardiac transdifferentiation and secreted factor production. Future studies incorporating MSCs other cell types should broaden understanding of the post-MI scar microenvironment.
PURPOSE: Cellular therapy with mesenchymal stem cells (MSCs) shows promise for restoring function after myocardial infarction (MI). However, cellular therapy has yet to be clinically translated, in part because of difficulty in studying how MSCs interact with the post-MI scar microenvironment. This study aimed to design an in vitro model to study MSC behavior in the post-MI scar stiffness microenvironment. METHODS: Using poly(ethylene glycol)-acrylate (PEG) conjugated to bioactive peptides, rat MSCs were encapsulated in hydrogels of varying stiffnesses and crosslinking densities. Cell viability was assessed through 14 days using calcein and ethidium homodimer staining. To simulate post-MI pro-fibrotic signaling, transforming growth factor-beta (TGFβ) was added to selected cultures. Immunofluorescence and qRT-PCR were used to assess changes in cardiac transdifferentiation or paracrine secretion, two proposed methods of MSCs in cellular therapy. RESULTS: Bioactivated PEG hydrogels with stiffnesses between 1.6 and 151.0 kPa were prepared. Rat MSCs demonstrated up to 71.6% viability after 3 days of encapsulated culture, and survived within the hydrogels up to 14 days. Encapsulation decreased MSC expression of cardiac troponin T and most growth factors, except interleukin-6. Meanwhile, TGFβ caused increased cardiac troponin T expression but decreased secreted factor expression. Varying hydrogel stiffness did not have an effect on cardiac troponin T or secreted factor expression. CONCLUSIONS: These findings suggest that a 3D microenvironment hinders two key mechanisms by which MSCs could improve cardiac function after post-MI scar formation, namely cardiac transdifferentiation and secreted factor production. Future studies incorporating MSCs other cell types should broaden understanding of the post-MI scar microenvironment.