Skip to main content

Advertisement

SpringerLink
Log in

Biomarkers and heterogeneous fibroblast phenotype associated with incisional hernia

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Development of incisional hernia (IH) is multifactorial but inflammation and abdominal wall ECM (extracellular matrix) disorganization are key pathological events. We investigated if the differential expression of fibroblast biomarkers reflects the cellular milieu and the dysregulated ECM in IH tissues. Expression of fibroblast biomarkers, including connective tissue growth factor, alpha-smooth muscle actin (α-SMA), CD34 (cluster of differentiation 34), cadherin-11 and fibroblast specific protein 1 (FSP1), was examined by histology and immunofluorescence in the hernial-fascial ring/neck tissue (HRT) and hernia sack tissue (HST) harvested from the patients undergoing hernia surgery and compared with normal fascia (FT) and peritoneum (PT) harvested from brain-dead healthy subjects undergoing organ procurement for transplantation. The H&E staining revealed alterations in tissue architecture, fibroblast morphology, and ECM organization in the IH tissues compared to control. The biomarker for undifferentiated fibroblasts, CD34, was significantly higher in HST and decreased in HRT than the respective FT and PT controls. Also, the findings revealed an increased level of CTGF (connective tissue growth factor) with decrease in α-SMA in both HRT and HST compared to the controls. In addition, an increased level of FSP1 (fibroblast specific protein 1) and cadherin-11 in HRT with decreased level in HST were observed relative to the respective controls (FT and PT). Hence, these findings support the heterogeneity of fibroblast population at the laparotomy site that could contribute to the development of IH. Understanding the mechanisms causing the phenotype switch of these fibroblasts would open novel strategies to prevent the development of IH following laparotomy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Data Availability

There is no restrictions on sharing a de-identified data set (without any identifying or sensitive patient information) on the experiments performed on the human tissues collected and processed in an un-identifiable manner. Such data are now included as Supporting Information Excel file. This file contains: (i) the values behind the means, standard deviations and other measures reported, and (ii) the values used to build graphs.

References

  1. Reistrup H, Zetner DB, Andresen K, Rosenberg J (2018) [Prevention of incisional hernia]. Ugeskr Laeger 180(34):V02180094

  2. Deng Y, Ren J, Chen G et al (2017) Injectable in situ cross-linking chitosan-hyaluronic acid based hydrogels for abdominal tissue regeneration. Sci Rep. https://doi.org/10.1038/s41598-017-02962-z

    Article  PubMed  PubMed Central  Google Scholar 

  3. Harrison B, Sanniec K, Janis JE (2016) Collagenopathies—Implications for Abdominal Wall Reconstruction: A Systematic Review. Plast Reconstr Surg - Glob Open 4:e1036. https://doi.org/10.1097/GOX.0000000000001036

    Article  PubMed  PubMed Central  Google Scholar 

  4. Guillen-Marti J, Diaz R, Quiles MT et al (2009) MMPs/TIMPs and inflammatory signalling de-regulation in human incisional hernia tissues. J Cell Mol Med 13:4432–4443. https://doi.org/10.1111/j.1582-4934.2008.00637.x

    Article  CAS  PubMed  Google Scholar 

  5. Henriksen NA, Yadete DH, Sorensen LT et al (2011) Connective tissue alteration in abdominal wall hernia. Br J Surg 98:210–219. https://doi.org/10.1002/bjs.7339

    Article  CAS  PubMed  Google Scholar 

  6. Yahchouchy-Chouillard E, Aura T, Picone O et al (2003) Incisional Hernias. Dig Surg 20:3–9. https://doi.org/10.1159/000068850

    Article  PubMed  Google Scholar 

  7. Ireton JE, Unger JG, Rohrich RJ (2013) The role of wound healing and its everyday application in plastic surgery: a practical perspective and systematic review. Plast Reconstr Surg Glob Open. https://doi.org/10.1097/GOX.0b013e31828ff9f4

    Article  PubMed  PubMed Central  Google Scholar 

  8. Rath AM, Chevrel JP (1998) The healing of laparotomies: review of the literature: Part 1. Physiologic and pathologic aspects Hernia 2:145–149. https://doi.org/10.1007/BF01250034

    Article  Google Scholar 

  9. Diaz R, Quiles MT, Guillem-Marti J et al (2011) Apoptosis-like cell death induction and aberrant fibroblast properties in human incisional hernia fascia. Am J Pathol 178:2641–2653. https://doi.org/10.1016/j.ajpath.2011.02.044

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Dobaczewski M, Gonzalez-Quesada C, Frangogiannis NG (2010) The extracellular matrix as a modulator of the inflammatory and reparative response following myocardial infarction. J Mol Cell Cardiol 48:504–511. https://doi.org/10.1016/j.yjmcc.2009.07.015

    Article  CAS  PubMed  Google Scholar 

  11. Frangogiannis NG (2016) Fibroblast—extracellular matrix interactions in tissue fibrosis. Curr Pathobiol Rep 4:11–18. https://doi.org/10.1007/s40139-016-0099-1

    Article  PubMed  PubMed Central  Google Scholar 

  12. Li B, Wang JH-C (2011) Fibroblasts and myofibroblasts in wound healing: Force generation and measurement. J Tissue Viability 20:108–120. https://doi.org/10.1016/j.jtv.2009.11.004

    Article  PubMed  Google Scholar 

  13. Thankam FG, Palanikumar G, Fitzgibbons RJ, Agrawal DK (2019) Molecular mechanisms and potential therapeutic targets in incisional hernia. J Surg Res 236:134–143. https://doi.org/10.1016/j.jss.2018.11.037

    Article  CAS  PubMed  Google Scholar 

  14. Henshaw FR, Boughton P, Lo L et al (2015) topically applied connective tissue growth factor/CCN2 improves diabetic preclinical cutaneous wound healing: potential role for CTGF in human diabetic foot ulcer healing. J Diabetes Res 2015:1–10. https://doi.org/10.1155/2015/236238

    Article  Google Scholar 

  15. Alfaro MP, Deskins DL, Wallus M et al (2013) A physiological role for connective tissue growth factor in early wound healing. Lab Invest 93:81–95. https://doi.org/10.1038/labinvest.2012.162

    Article  CAS  PubMed  Google Scholar 

  16. Thankam FG, Boosani CS, Dilisio MF et al (2016) MicroRNAs associated with shoulder tendon matrisome disorganization in glenohumeral arthritis. PLoS ONE 11:e0168077. https://doi.org/10.1371/journal.pone.0168077

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Thankam FG, Dilisio MF, Dietz NE, Agrawal DK (2016) TREM-1, HMGB1 and RAGE in the shoulder tendon: dual mechanisms for inflammation based on the coincidence of glenohumeral arthritis. PLoS ONE 11:e0165492. https://doi.org/10.1371/journal.pone.0165492

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Xing L, Culbertson EJ, Wen Y, Franz MG (2013) Early laparotomy wound failure as the mechanism for incisional hernia formation. J Surg Res 182:e35–e42. https://doi.org/10.1016/j.jss.2012.09.009

    Article  PubMed  Google Scholar 

  19. Franz MG (2008) The biology of hernia formation. Surg Clin North Am 88:1–15. https://doi.org/10.1016/j.suc.2007.10.007

    Article  PubMed  PubMed Central  Google Scholar 

  20. Benjamin M, Hillen B (2003) Mechanical influences on cells, tissues and organs ? ?mechanical morphogenesis? Eur J Morphol 41:3–7. https://doi.org/10.1076/ejom.41.1.3.28102

    Article  CAS  PubMed  Google Scholar 

  21. Ravikanth M, Manjunath K, Ramachandran C et al (2011) Heterogenecity of fibroblasts. J Oral Maxillofac Pathol 15:247. https://doi.org/10.4103/0973-029X.84516

    Article  PubMed  PubMed Central  Google Scholar 

  22. Lekic PC, Pender N, McCulloch CA (1997) Is fibroblast heterogeneity relevant to the health, diseases, and treatments of periodontal tissues? Crit Rev Oral Biol Med Off Publ Am Assoc Oral Biol 8:253–268

    Article  CAS  Google Scholar 

  23. Kong P, Christia P, Saxena A et al (2013) Lack of specificity of fibroblast-specific protein 1 in cardiac remodeling and fibrosis. Am J Physiol-Heart Circ Physiol 305:H1363–H1372. https://doi.org/10.1152/ajpheart.00395.2013

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Strutz F, Okada H, Lo CW et al (1995) Identification and characterization of a fibroblast marker: FSP1. J Cell Biol 130:393–405

    Article  CAS  Google Scholar 

  25. Zhang R, Gao Y, Zhao X et al (2018) FSP1-positive fibroblasts are adipogenic niche and regulate adipose homeostasis. PLOS Biol 16:e2001493. https://doi.org/10.1371/journal.pbio.2001493

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Sudheer Shenoy P, Bose B (2017) Identification, isolation, quantification and systems approach towards CD34, a biomarker present in the progenitor/stem cells from diverse lineages. Methods 131:147–156. https://doi.org/10.1016/j.ymeth.2017.06.035

    Article  CAS  PubMed  Google Scholar 

  27. Silverman JS, Tamsen A (1998) CD34 and factor XIIIa-positive microvascular dendritic cells and the family of fibrohistiocytic mesenchymal tumors. Am J Dermatopathol 20:533–536

    Article  CAS  Google Scholar 

  28. San Martin R, Barron DA, Tuxhorn JA et al (2014) Recruitment of CD34(+) fibroblasts in tumor-associated reactive stroma: the reactive microvasculature hypothesis. Am J Pathol 184:1860–1870. https://doi.org/10.1016/j.ajpath.2014.02.021

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Van De Water L, Varney S, Tomasek JJ (2013) Mechanoregulation of the Myofibroblast in Wound Contraction, Scarring, and Fibrosis: Opportunities for New Therapeutic Intervention. Adv Wound Care 2:122–141. https://doi.org/10.1089/wound.2012.0393

    Article  Google Scholar 

  30. Micallef L, Vedrenne N, Billet F et al (2012) The myofibroblast, multiple origins for major roles in normal and pathological tissue repair. Fibrogenesis Tissue Repair 5:S5. https://doi.org/10.1186/1755-1536-5-S1-S5

    Article  PubMed  PubMed Central  Google Scholar 

  31. Yang Z, Sun Z, Liu H et al (2015) Connective tissue growth factor stimulates the proliferation, migration and differentiation of lung fibroblasts during paraquat-induced pulmonary fibrosis. Mol Med Rep 12:1091–1097. https://doi.org/10.3892/mmr.2015.3537

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Tsai C-C, Wu S-B, Chang P-C, Wei Y-H (2015) Alteration of Connective Tissue Growth Factor (CTGF) expression in orbital fibroblasts from patients with graves’ ophthalmopathy. PLoS ONE 10:e0143514. https://doi.org/10.1371/journal.pone.0143514

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Lee CH, Shah B, Moioli EK, Mao JJ (2010) CTGF directs fibroblast differentiation from human mesenchymal stem/stromal cells and defines connective tissue healing in a rodent injury model. J Clin Invest 120:3340–3349. https://doi.org/10.1172/JCI43230

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Chang SK, Noss EH, Chen M et al (2011) Cadherin-11 regulates fibroblast inflammation. Proc Natl Acad Sci 108:8402–8407. https://doi.org/10.1073/pnas.1019437108

    Article  PubMed  PubMed Central  Google Scholar 

  35. Schroer A, Bersi M, Clark C et al (2019) Cadherin-11 blockade reduces inflammation driven fibrotic remodeling and improves outcomes after myocardial infarction: compiled supplmental methods and figures. Biorxiv. https://doi.org/10.1101/533000

    Article  Google Scholar 

  36. Row S, Liu Y, Alimperti S et al (2016) Cadherin-11 is a novel regulator of extracellular matrix synthesis and tissue mechanics. J Cell Sci 129:2950–2961. https://doi.org/10.1242/jcs.183772

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors would like to sincerely acknowledge the staffs of Live on Nebraska (Formerly called NORS (Nebraska Organ Recovery System) for procuring control specimen in this study.

Funding

This research work was supported by NIH-NHLBI grants R01HL147662 and R01HL144125 of DK Agrawal and CU Surgery research funds to Dr. Fitzgibbons. The content of this original article is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

Author information

Authors and Affiliations

  1. Department of Translational Research, Western University of Health Sciences, 309 E. Second Street, Pomona, CA, 91766, USA

    Finosh G. Thankam & Devendra K. Agrawal

  2. Departments of Clinical & Translational Science and Surgery, Creighton University School of Medicine, Omaha, NE, 68178, USA

    Finosh G. Thankam, Nicholas K. Larsen, Ann Varghese, Thao-Nguyen Bui, Matthew Reilly, Robert J. Fitzgibbons & Devendra K. Agrawal

Authors
  1. Finosh G. Thankam
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nicholas K. Larsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ann Varghese
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Thao-Nguyen Bui
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Matthew Reilly
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Robert J. Fitzgibbons
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Devendra K. Agrawal
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Conception and design: FGT, NKL, RJF, DKA; Contributed reagents/ materials/analysis tool: RJF, DKA; Conducting surgeries, analysis, and interpretation of the data: RJF, NKL, FGT, AV, T-NB, MR, DKA; Drafting of the article: FGT, NKL, DKA; Critical revision and editing of the article for important intellectual content: FGT, NKL, RJF, DKA; Final approval of the submitted article: FGT, NKL, AV, T-NB, MR, RJF, DKA.

Corresponding author

Correspondence to Devendra K. Agrawal.

Ethics declarations

Conflict of interest

The authors report no proprietary or commercial interest in any product mentioned or concept discussed in this article. The authors declare no conflict of interest.

Ethical approval

Institutional Review Board (IRB) of Creighton University approved the study under the IRB protocol (2nd May 2018 to 2nd April 2019) of Robert J. Fitzgibbons, MD, a co-author of this article.

Consent to participate

Patients ≥ 19 years of age, of either sex, who were undergoing repair of their incisional hernia were recruited in the study and informed consents and HIPPA forms were obtained and saved in the office of Robert J. Fitzgibbons, MD. The de-identified tissues were sent to the laboratory for further experiments.

Consent for publication

There is no identifiable images or other personal or clinical details of study participants in the data presented. As the corresponding authors, I verify that all authors significantly contributed to various aspect of the study, have read the manuscript and consented to submit for publication in the Molecular and cellular biochemistry.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 1952 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Thankam, F.G., Larsen, N.K., Varghese, A. et al. Biomarkers and heterogeneous fibroblast phenotype associated with incisional hernia. Mol Cell Biochem 476, 3353–3363 (2021). https://doi.org/10.1007/s11010-021-04166-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-021-04166-6

Keywords

Search

Navigation