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Fabrication of Decellularized Amnion and Chorion Scaffolds to Develop Bioengineered Cell-Laden Constructs

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Abstract

Introduction

Human mesenchymal stem cells (hMSCs) holds great promise for managing several clinical conditions. However, the low engraftment efficiency and obscurity to harvest these cells without compromising the cellular viability, structural and functional properties from the culture niche still remain major obstacles for preparing intact regenerative constructs. Although few studies have demonstrate different methods for generating cell-liberated amniotic scaffolds, a common method for producing completely cell-liberated amnion (D-HAM) and chorion (D-HCM) scaffolds and their cytocompatibility with hMSCs yet to be demonstrated.

Methods

A common process was developed for preparing D-HAM and D-HCM scaffolds for assessing hMSCs engraftment efficiency, proliferation and molecular shifts to generate cell-laden biological discs. The structural and functional integrity of D-HAM and D-HCM was evaluated using different parameters. The compatibility and proliferation efficiency of hMSCs with D-HAM and D-HCM was evaluated.

Results

Histological analysis revealed completely nucleic acid-free D-HAM and D-HCM scaffolds with intact extracellular matrix, mechanical and biological properties almost similar to the native membranes. Human MSCs were able to adhere and engraft on D-HCM better than D-HAM and expanded faster. Ultrastructural observations, crystal violet staining and expression studies showed better structural and functional integrity of hMSCs on D-HCM than D-HAM and control conditions.

Conclusion

A common, simple and reliable process of decellularization can generate large number of cell-liberated amniotic scaffolds in lesser time. D-HCM has better efficiency for hMSCs engraftment and proliferation and can be utilized for preparing suitable cell-laden constructs for tissue engineering applications.

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Acknowledgments

This study was supported by the grant received from Indian Council of Medical Research (ICMR) in form of Research Associate Fellowship (RA, No. 5/3/8/3/ITR-F/2019-ITR) to Sandeep Kumar Vishwakarma. All authors thank to Owaisi Hospital and Research Centre for material supply.

Conflict of interest

Chandrakala Lakkireddy, Nagarapu Raju, Shaik Iqbal Ahmed, Avinash Bardia, Mazharuddin Ali Khan, Sandhya Annamaneni, and Aleem Ahmed Khan declare that they have no conflict of interest.

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  1. Chandrakala Lakkireddy and Sandeep Kumar Vishwakarma share equal first authorship for this manuscript.

Authors and Affiliations

  1. Central Laboratory for Stem Cell Research & Translational Medicine, Centre for Liver Research and Diagnostics, Deccan College of Medical Sciences, Kanchanbagh, Hyderabad, Telangana, 500058, India

    Chandrakala Lakkireddy, Sandeep Kumar Vishwakarma, Nagarapu Raju, Shaik Iqbal Ahmed, Avinash Bardia & Aleem Ahmed Khan

  2. Department of Orthopedics, OHRC, Deccan College of Medical Sciences, Kanchanbagh, Hyderabad, Telangana, India

    Mazharuddin Ali Khan

  3. Department of Genetics, Osmania University, Hyderabad, Telangana, India

    Sandhya Annamaneni

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  1. Chandrakala Lakkireddy
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Lakkireddy, C., Vishwakarma, S.K., Raju, N. et al. Fabrication of Decellularized Amnion and Chorion Scaffolds to Develop Bioengineered Cell-Laden Constructs. Cel. Mol. Bioeng. 15, 137–150 (2022). https://doi.org/10.1007/s12195-021-00707-7

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