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A Model for Studying the Biomechanical Effects of Varying Ratios of Collagen Types I and III on Cardiomyocytes

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Abstract

Purpose

To develop a novel model composed solely of Col I and Col III with the lower and upper limits set to include the ratios of Col I and Col III at 3:1 and 9:1 in which the structural and mechanical behavior of the resident CM can be studied. Further, the progression of fibrosis due to change in ratios of Col I:Col III was tested.

Methods

Collagen gels with varying Col I:Col III ratios to represent a healthy (3:1) and diseased myocardial tissue were prepared by manually casting them in wells. Absorbance assay was performed to confirm the gelation of the gels. Rheometric analysis was performed on each of the collagen gels prepared to determine the varying stiffnesses and rheological parameters of the gels made with varying ratios of Col I:Col III. Second Harmonic Generation (SHG) was performed to observe the 3D characterization of the collagen samples. Scanning Electron microscopy was used for acquiring cross sectional images of the lyophilized collagen gels. AC16 CM (human) cell lines were cultured in the prepared gels to study cell morphology and behavior as a result of the varying collagen ratios. Cellular proliferation was studied by performing a Cell Trace Violet Assay and the applied force on each cell was measured by means of Finite Element Analysis (FEA) on CM from each sample.

Results

Second harmonic generation microscopy used to image Col I, displayed a decrease in acquired image intensity with an increase in the non-second harmonic Col III in 3:1 gels. SEM showed a fiber-rich structure in the 3:1 gels with well-distributed pores unlike the 9:1 gels or the 1:0 controls. Rheological analysis showed a decrease in substrate stiffness with an increase of Col III, in comparison with other cases. CM cultured within 3:1 gels exhibited an elongated rod-like morphology with an average end-to-end length of 86 ± 28.8 µm characteristic of healthy CM, accompanied by higher cell growth in comparison with other cases. Finite element analysis used to estimate the forces exerted on CM cultured in the 3:1 gels, showed that the forces were well dispersed, and not concentrated within the center of cells, in comparison with other cases.

Conclusion

This study model can be adopted to simulate various biomechanical environments in which cells crosstalk with the Collagen-matrix in diseased pathologies to generate insights on strategies for prevention of fibrosis.

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Acknowledgments

We acknowledge the technical assistance received from Dr. Armando Varela for kindly assisting us with the bright field microscopy and Dr. David Roberson for his help with the SEM.

Author contributions

BR and BJ conceptualized the study and were responsible for the experimentation, data collection, curation and analysis. SAK helped in selected experiments, writing and editing of the manuscript. SCA and LJS performed the rheological analysis for this manuscript. MD helped with the cell culture experiments. SM and RC performed the Cell Force Estimation Analysis. AMR and CL performed the SHG experiments for this manuscript. All authors participated in the writing and editing of the manuscript.

Funding

The Joddar Lab (IMSTEL) acknowledges NIH BUILD Pilot 8UL1GM118970-02, the NSF PREM (DMR 1205302), the NSF-MRI Grant # DMR-1828268 and NIH 1SC2HL134642-01. CL acknowledges the NSF MRI Grant #DBI 1429708 and both CL and RC acknowledges the NSF-PREM program (#DMR-1827745).

Conflict of interest

The authors declare no conflicts of interest.

Consent for publication

All authors have consented to this manuscript and their role and order of authorship.

Statement of Human and Animal Rights

No human or animal experimentation was executed as a part of this study.

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Authors and Affiliations

  1. Inspired Materials & Stem-Cell Based Tissue Engineering Laboratory (IMSTEL), El Paso, USA

    Brian Roman, Shweta Anil Kumar, Monica Delgado, Sabastian Moncayo & Binata Joddar

  2. Department of Metallurgical, Materials and Biomedical Engineering, University of Texas at El Paso, 500 W University Avenue, El Paso, TX, 79968, USA

    Brian Roman, Shweta Anil Kumar, Monica Delgado & Sabastian Moncayo

  3. Department of Biomedical Engineering, The University of Texas at Austin, Austin, TX, 78712, USA

    Shane C. Allen & Laura J. Suggs

  4. Department of Physics, The University of Texas at El Paso, 500 W University Avenue, El Paso, TX, 79968, USA

    Andres M. Reyes & Chunqiang Li

  5. Department of Mechanical Engineering, The University of Texas at El Paso, 500 W University Avenue, El Paso, TX, 79968, USA

    Ramana Chintalapalle

  6. Border Biomedical Research Center, University of Texas at El Paso, 500 W University Avenue, El Paso, TX, 79968, USA

    Chunqiang Li & Binata Joddar

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Correspondence to Binata Joddar.

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Roman, B., Kumar, S.A., Allen, S.C. et al. A Model for Studying the Biomechanical Effects of Varying Ratios of Collagen Types I and III on Cardiomyocytes. Cardiovasc Eng Tech 12, 311–324 (2021). https://doi.org/10.1007/s13239-020-00514-7

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