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Single-pot biofabrication of living fibers for tissue engineering applications

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Abstract

In this work, morphology, viability, and metabolism of the amniotic mesenchymal stem cells conditioned with different citric acid (CA)/media ratios were investigated using rhodamine-phalloidin/4′,6-diamidino-2-phenylindole staining, live/dead assay, proliferating cell nuclear antigen, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL assay). The cells cultured in 25:75 CA/media displayed well spread actin filaments with a prominent nucleus and evidenced optimum viability. The gelation kinetics of chitosan solution in CA/media (25:75) was monitored via dynamic time sweep analysis on a rheometer. The chemical cross-linking of chitosan with CA was confirmed by nuclear magnetic resonance studies. Subsequently, chitosan solution was extruded in CA/media bath containing cells under benign conditions to form cell-laden fibers (living fibers). The prelabeled cells imaged immediately after fiber formation confirmed the attachment of the cells on the fibers. This approach has several advantages including instantaneous gelation, tunable mechanical properties, and adjustable biodegradability that can provide a platform technology for creating viable three dimensional (3D) building blocks for tissue engineering applications.

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ACKNOWLEDGMENTS

Fellowship from Council of Scientific and Industrial Research (CSIR) and Department of Science and Technology (DST), Govt. of India to P.G. and A.P.R. is acknowledged. Financial support from CSIR, DST, Department of Biotechnology, and Defense Research and Development Organization, (Govt. of India) are acknowledged. Dr. Asit Baran Panda is acknowledged for his help in NMR studies.

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  1. These authors contributed equally to this work.

Authors and Affiliations

  1. Biomaterials and Tissue Engineering Laboratory, School of Medical Science and Technology, Indian Institute of Technology Kharagpur, Kharagpur, 721302, India

    Paulomi Ghosh, Arun Prabhu Rameshbabu, Bhuvaneshwaran Subramanian & Santanu Dhara

  2. CSIR-Indian Institute of Chemical Biology, Kolkata, 700032, West Bengal, India

    Paulomi Ghosh

  3. Polymer Chemistry Laboratory, Department of Applied Chemistry, Indian School of Mines, Dhanbad, 826004, India

    Dipankar Das, Sagar Pal & Sudip Kumar Ghosh

  4. Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur, 721302, India

    Sintu Kumar Samanta, Sagar Pal & Sudip Kumar Ghosh

  5. Department of Gynaecology, Midnapore Medical College, Paschim Medinipur, 721101, India

    Sabyasachi Roy

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  1. Paulomi Ghosh
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  2. Arun Prabhu Rameshbabu
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  3. Dipankar Das
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  4. Bhuvaneshwaran Subramanian
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  6. Sabyasachi Roy
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  7. Sagar Pal
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  8. Sudip Kumar Ghosh
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  9. Santanu Dhara
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Correspondence to Paulomi Ghosh or Santanu Dhara.

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Ghosh, P., Rameshbabu, A.P., Das, D. et al. Single-pot biofabrication of living fibers for tissue engineering applications. Journal of Materials Research 33, 2019–2028 (2018). https://doi.org/10.1557/jmr.2018.143

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