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Secretome Analysis of Human Nasal Fibroblast Identifies Proteins That Promote Wound Healing

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Cell Biology and Translational Medicine, Volume 21

Part of the book series: Advances in Experimental Medicine and Biology ((CBTMED,volume 1450))

Abstract

Conditioned medium from cultured fibroblast cells is recognized to promote wound healing and growth through the secretion of enzymes, extracellular matrix proteins, and various growth factors and cytokines. The objective of this study was to profile the secreted proteins present in nasal fibroblast conditioned medium (NFCM). Nasal fibroblasts isolated from human nasal turbinates were cultured for 72 h in Defined Keratinocytes Serum Free Medium (DKSFM) or serum-free F12: Dulbecco’s Modified Eagle’s Medium (DMEM) to collect conditioned medium, denoted as NFCM_DKSFM and NFCM_FD, respectively. SDS-PAGE was performed to detect the presence of protein bands, followed by MALDI-TOF and mass spectrometry analysis. SignalP, SecretomeP, and TMHMM were used to identify the secreted proteins in conditioned media. PANTHER Classification System was performed to categorize the protein according to protein class, whereas STRING 10 was carried out to evaluate the predicted proteins interactions. SDS-PAGE results showed the presence of various protein with molecular weight ranging from ~10 kDa to ~260 kDa. Four protein bands were identified using MALDI-TOF. The analyses identified 104, 83, and 7 secreted proteins in NFCM_FD, NFCM_DKSFM, and DKSFM, respectively. Four protein classes involved in wound healing were identified, namely calcium-binding proteins, cell adhesion molecules, extracellular matrix proteins, and signaling molecules. STRING10 protein prediction successfully identified various pathways regulated by secretory proteins in NFCM. In conclusion, this study successfully profiled the secreted proteins of nasal fibroblasts and these proteins are predicted to play important roles in RECs wound healing through various pathways.

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Acknowledgments

The authors would like to thank Dr. Nur Atiqah Haizum Abdullah from the Centre for Tissue Engineering and Regenerative Medicine, UKM, for providing guidance on bioinformatics analysis. Protein identification by MALDI-TOF was carried out at Proteomics International Pty Ltd., Perth, Australia. Triple TOF LC-MS analysis was performed at the Australian Proteome Analysis Facility, Sydney, Australia, a NCRIS (National Collaborative Research Infrastructure Strategy)-enabled Bioplatforms Australia infrastructure.

Authors Contribution

All authors participation are as follows: conceptualization: RCM, RBHI, and YL; methodology: RCM and YL; software: WIWI and YL; validation: RCM, WIWI, YL; formal analysis: RCM, WIWI, YL; investigation: RCM, WIWI, YL; resources: RBHI, ABS, YL; data curation: RCM, WIWI, YL; writing – original draft preparation: RCM, WIWI, YL; writing – review and editing: RCM, RBHI, ABS, WIWI, YL; visualization: RCM and YL; supervision: RBHI, ABS, and YL; project administration: RCM, RBHI, YL; funding acquisition: RBHI, ABS, YL.

Declaration of Conflicting Interests

The author(s) declare no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Ethical Approval

This study has been approved by the Universiti Kebangsaan Malaysia Research and Ethics Committee (approval code: FRGS/1/2016/SKK08/UKM/03/1).

Funding

This study was supported by the University Kebangsaan Malaysia research grant from the Ministry of Higher Education Malaysia [grant number: DPP-2014-121, AP-2013-015].

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Authors and Affiliations

  1. Centre for Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia

    Rohaina Che Man, Ruszymah Binti Hj Idrus & Yogeswaran Lokanathan

  2. Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia

    Ruszymah Binti Hj Idrus

  3. Medical Biotechnology Laboratory, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia

    Wan Izlina Wan Ibrahim

  4. Ear, Nose & Throat Consultant Clinic, Ampang Puteri Specialist Hospital, Selangor, Malaysia

    Aminuddin Bin Saim

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  1. Rohaina Che Man
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  5. Yogeswaran Lokanathan
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Correspondence to Yogeswaran Lokanathan .

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  1. Ottawa Hospital, Ottawa, ON, Canada

    Kursad Turksen

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Man, R.C., Idrus, R.B.H., Ibrahim, W.I.W., Saim, A.B., Lokanathan, Y. (2023). Secretome Analysis of Human Nasal Fibroblast Identifies Proteins That Promote Wound Healing. In: Turksen, K. (eds) Cell Biology and Translational Medicine, Volume 21. Advances in Experimental Medicine and Biology(), vol 1450. Springer, Cham. https://doi.org/10.1007/5584_2023_777

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