Skip to main content

Advertisement

SpringerLink
Log in

Second-harmonic generation (SHG) microscopy and hepatic venous pressure gradient-based validation of a novel histological staging system for alcoholic hepatitis

  • Original Article
  • Published:
Virchows Archiv Aims and scope Submit manuscript

Abstract

Alcoholic hepatitis (AH) lacks specific histological staging. A novel fibrosis staging that encompasses perisinusoidal fibrosis and cirrhosis sub-stages, substantiated by Hepatic venous pressure gradient (HVPG) and automated fibrosis quantification, is imperative. To correlate novel histological staging system of AH with second-harmonic generation microscopy (SHG)-based q-fibrosis, HVPG, and activated hepatic stellate cells (HSCs). Liver biopsies of AH (n = 175) were staged semi-quantitatively as F0, F1, F2, F3A and F3B and Laennec substages of cirrhosis 4A, 4B and 4C. Stages were correlated with SHG q-fibrosis parameters, HVPG and HSCs. Mean age 41.2 ± 9.4 years, 96.6% males, bilirubin 20.58 ± 8.0 mg/dl and Maddrey’s discriminant function 78.9 ± 36.7 displayed advanced fibrosis in 98.6%. With increasing histological stages, an increase in q-fibrosis indices and mean HVPG (p < 0.0001) were recorded; stage 4C showed the most significant difference from other stages (p < 0.000). Stages 3A and 3B were comparable with the stages 4A and 4B, respectively, for q-fibrosis (p = 1) and HVPG (p = 1). HSCs (> 30%) were significantly higher in stage 3 (75%) compared with 4 (49%) and 2 (59%), p = 0.018. Overall agreement for histological staging was excellent for all stages (0.82). SHG quantified fibrosis and HVPG corroborates the novel histological staging of AH. Expansive PCF matches with collagen content and clinical severity to early sub-stages of cirrhosis. This highlights the need for an accurate quantification and inclusion of PCF as a separate stage. SHG-based quantification can be a useful adjunct to histological fibrosis staging systems.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

Explore related subjects

Discover the latest articles, news and stories from top researchers in related subjects.

Availability of data and material

Not applicable

Code availability

Not applicable

References

  1. Sarin SK, Choudhury A, Sharma MK et al (2019) Acute-on-chronic liver failure: consensus recommendations of the Asian Pacific association for the study of the liver (APASL): an update. Hepatol Int 13(4):353–390

    Article  PubMed  Google Scholar 

  2. Bataller R, Gao B (2015) Liver fibrosis in alcoholic liver disease. Semin Liver Dis 35(2):146–156

    Article  CAS  PubMed  Google Scholar 

  3. Shasthry SM, Rastogi A, Bihari C, Vijayaraghavan R, Arora V, Sharma MK, Sarin SK (2018) Histological activity score on baseline liver biopsy can predict non-response to steroids in patients with severe alcoholic hepatitis. Virchows Arch 472(4):667–675

    Article  CAS  PubMed  Google Scholar 

  4. Gao B, Bataller R (2011) Alcoholic liver disease: pathogenesis and new therapeutic targets. Gastroenterology 141:1572–1585

    Article  CAS  PubMed  Google Scholar 

  5. Lackner C, Tiniakos D (2019) Fibrosis and alcohol-related liver disease. J Hepatol 70(2):294–304

    Article  PubMed  Google Scholar 

  6. Scheuer PJ (1991) Classification of chronic viral hepatitis: a need for reassessment. J Hepatol 13:372–374

    Article  CAS  PubMed  Google Scholar 

  7. Bedossa P, Poynard T (1996) An algorithm for the grading of activity in chronic hepatitis C. The METAVIR Cooperative Study Group. Hepatology 24:289–293

    Article  CAS  PubMed  Google Scholar 

  8. Ishak K, Baptista A, Bianchi L, Callea F, de Groote J, Gudat F, Denk H, Desmet V, Korb G, MacSween RNM, Phillips MJ, Portmann BG, Poulsen H, Scheuer PJ, Schmid M, Thaler H (1995) Histological grading and staging of chronic hepatitis. J Hepatol 22:696–699

    Article  CAS  PubMed  Google Scholar 

  9. Kleiner DE, Brunt EM, Van Natta M et al (2005) Design and validation of a histological scoring system for nonalcoholic fatty liver disease. Hepatology 41:1313–1321

    Article  PubMed  Google Scholar 

  10. Shoreibah M, Raff E, Bloomer J, Kakati D, Rasheed K, Kuo YF, Singal AK (2016) Alcoholic liver disease presents at advanced stage and progresses faster compared to non-alcoholic fatty liver disease. Ann Hepatol 15:183–189

    PubMed  Google Scholar 

  11. Tiniakos DG, Anstee QM, Burt AD (2017) Fatty liver disease. In: Burt AD, Ferrell LD, Hubscher SG (eds) Mac Sweens Pathology of the Liver. Elsevier, Philadelphia, pp 308–371

    Google Scholar 

  12. Szabo G (2018) More than meets the eye: Severe alcoholic hepatitis can present as acute-on-chronic liver failure. J Hepatol 69(2):269–271

    Article  PubMed  Google Scholar 

  13. Nakano M, Fukusato T (2005) Histological study on comparison between NASH and ALD. Hepatol Res 33:110–115

    Article  PubMed  Google Scholar 

  14. Suk KT, Kim DJ (2015) Staging of liver fibrosis or cirrhosis: The role of hepatic venous pressure gradient measurement. World J Hepatol 7(3):607–615

    Article  PubMed  PubMed Central  Google Scholar 

  15. Restellini S, Goossens N, Clément S, Lanthier N, Negro F, Rubbia-Brandt L, Spahr L (2018) Collagen proportionate area correlates to hepatic venous pressure gradient in non-abstinent cirrhotic patients with alcoholic liver disease. World J Hepatol 10(1):73–81

    Article  PubMed  PubMed Central  Google Scholar 

  16. Garcia-Tsao G, Friedman S, Iredale J, Pinzani M (2010) Now there are many (stages) where before there was one: In search of a pathophysiological classification of cirrhosis. Hepatology 51:1445–1449

    Article  PubMed  Google Scholar 

  17. Rastogi A, Maiwall R, Bihari C, Ahuja A, Kumar A, Singh T, Wani ZA, Sarin SK (2013) Cirrhosis histology and Laennec staging system correlate with high portal pressure. Histopathology 62(5):731–741

    Article  PubMed  Google Scholar 

  18. Kumar M, Sakhuja P, Kumar A et al (2008) Histological subclassification of cirrhosis based on histological-haemodynamic correlation. Aliment Pharmacol Ther 27(9):771–779

    Article  CAS  PubMed  Google Scholar 

  19. Kim SU, Oh HJ, Wanless IR, Lee S, Han KH, Park YN (2012) The Laennec staging system for histological subclassification of cirrhosis is useful for stratification of prognosis in patients with liver cirrhosis. J Hepatol 57:556–563

    Article  PubMed  Google Scholar 

  20. Almpanis Z, Demonakou M, Tiniakos D (2016) Evaluation of liver fibrosis: “Something old, something new”. Ann Gastroenterol 29(4):445–453

    PubMed  PubMed Central  Google Scholar 

  21. Wang Y, Wong GL, He FP et al (2020) Quantifying and monitoring fibrosis in non-alcoholic fatty liver disease using dual-photon microscopy. Gut 69(6):1116–1126

    Article  PubMed  Google Scholar 

  22. Sun Y, Zhou J, Wu X, Chen Y, Piao H, Lu L, Ding H, Nan Y, Jiang W, Wang T, Liu H, Ou X, Wee A, Theise ND, Jia J, You H (2018) Quantitative assessment of liver fibrosis (qFibrosis) reveals precise outcomes in Ishak “stable” patients on anti-HBV therapy. Sci Rep 8(1):2989

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Xu S, Wang Y, Tai DCS, Wang S, Cheng CL, Peng Q, Yan J, Chen Y, Sun J, Liang X, Zhu Y, Rajapakse JC, Welsch RE, So PTC, Wee A, Hou J, Yu H (2014) qFibrosis: a fully-quantitative innovative method incorporating histological features to facilitate accurate fibrosis scoring in animal model and chronic hepatitis B patients. J Hepatol 61:260–269

    Article  PubMed  PubMed Central  Google Scholar 

  24. Wang Y, Vincent R, Yang J, Asgharpour A, Liang X, Idowu MO, Contos MJ, Daitya K, Siddiqui MS, Mirshahi F, Sanyal AJ (2017) Dual-photon microscopy-based quantification of fibrosis-related parameters (q-FP) to model disease progression in steatohepatitis. Hepatology 65:1891–1903

    Article  CAS  PubMed  Google Scholar 

  25. Sun W, Chang S, Tai DC et al (2008) Non-linear optical microscopy: Use of second harmonic generation and two-photon microscopy for automated quantitative liver fibrosis studies. J Biomed Opt 13(6):064010

    Article  PubMed  CAS  Google Scholar 

  26. Guilbert T, Odin C, Grand L et al (2010) A robust collagen scoring method for human liver fibrosis by second harmonic microscopy. Opt Express 18:25794–25807

    Article  CAS  PubMed  Google Scholar 

  27. Higashi T, Friedman SL, Hoshida Y (2017) Hepatic stellate cells as key target in liver fibrosis. Adv Drug Deliv Rev 121:27–42

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Thompson KJ, McKillop IH, Schrum LW (2011) Targeting collagen expression in alcoholic liver disease. World J Gastroenterol 17(20):2473–2481

    Article  PubMed  PubMed Central  Google Scholar 

  29. Friedman SL (2008) Mechanisms of hepatic fibrogenesis. Gastroenterology 134:1655–1669

    Article  CAS  PubMed  Google Scholar 

  30. Singal AK, Louvet A, Shah VH, Kamath PS (2018) Grand Rounds: Alcoholic Hepatitis. J Hepatol 69(2):534–543. https://doi.org/10.1016/j.jhep.2018.05.001

    Article  PubMed  PubMed Central  Google Scholar 

  31. Crabb DW, Bataller R, Chalasani NP, Kamath PS, Lucey M, Mathurin P, McClain C, McCullough A, Mitchell MC, Morgan TR, Nagy L, Radaeva S, Sanyal A, Shah V, Szabo G, NIAAA Alcoholic Hepatitis Consortia (2016) Standard definitions and common data elements for clinical trials in patients with alcoholic hepatitis: recommendation from the NIAAA alcoholic hepatitis consortia. Gastroenterology 150:785–790

    Article  PubMed  Google Scholar 

  32. Dominguez M, Rincon D, Abraldes JG et al (2008) A new scoring system for prognostic stratification of patients with alcoholic hepatitis. Am J Gastroenterol 103:2747–2756

    Article  PubMed  Google Scholar 

  33. Colmenero J, Bataller R, Sancho-Bru P et al (2007) Hepatic expression of candidate genes in patients with alcoholic hepatitis: correlation with disease severity. Gastroenterology 132:687–697

    Article  CAS  PubMed  Google Scholar 

  34. Altamirano J, Miquel R, Katoonizadeh A et al (2014) A histologic scoring system for prognosis of patients with alcoholic hepatitis. Gastroenterology 146(5):1231–9.e96

    Article  PubMed  Google Scholar 

  35. Savolainen V, Perola M, Lalu K, Penttilä A, Virtanen I, Karhunen PJ (1995) Early perivenular fibrogenesis--precirrhotic lesions among moderate alcohol consumers and chronic alcoholics. J Hepatol 23:524–531

    Article  CAS  PubMed  Google Scholar 

  36. Michalak S, Rousselet MC, Bedossa P, Pilette C, Chappard D, Oberti F, Gallois Y, Calès P (2003) Respective roles of porto-septal fibrosis and centrilobular fibrosis in alcoholic liver disease. J Pathol 201:55–62

    Article  PubMed  Google Scholar 

  37. Hall A, Germani G, Isgrò G, Burroughs AK, Dhillon AP (2012) Fibrosis distribution in explanted cirrhotic livers. Histopathology 60:270–277

    Article  PubMed  Google Scholar 

  38. Sanyal AJ, Bosch J, Blei A, Arroyo V (2008) Portal hypertension and its complications. Gastroenterology 134:1715–1728

    Article  PubMed  Google Scholar 

  39. Schmitt-Graff A, Kruger S, Bochard F et al (1991) Modulationof alpha smooth muscle actin and desmin expression in perisinusoidal cells of normal and diseased human livers. Am J Pathol 138:1233–1242

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Rastogi A, Bihari C, Maiwall R, Ahuja A, Sharma MK, Kumar A, Sarin SK (2012) Hepatic stellate cells are involved in the pathogenesis of acute-on-chronic liver failure (ACLF). Virchows Arch 461(4):393–398

    Article  CAS  PubMed  Google Scholar 

  41. Seitz HK, Bataller R, Cortez-Pinto H et al (2018) Alcoholic liver disease. Nat Rev Dis Primers 4(1):18

    Article  PubMed  Google Scholar 

  42. Sun Y, Zhou J, Wang L, Wu X, Chen Y, Piao H, Lu L, Jiang W, Xu Y, Feng B, Nan Y, Xie W, Chen G, Zheng H, Li H, Ding H, Liu H, Lv F, Shao C, Wang T, Ou X, Wang B, Chen S, Wee A, Theise ND, You H, Jia J (2017) New Classification of Liver Biopsy Assessment for Fibrosis in Chronic Hepatitis B Patients Before and After Treatment. Hepatology 65:1438–1450

    Article  CAS  PubMed  Google Scholar 

  43. Yip WW, Burt AD (2006) Alcoholic liver disease. Semin Diagn Pathol 23(3-4):149–160

    Article  PubMed  Google Scholar 

  44. Standish RA, Cholongitas E, Dhillon A, Burroughs AK, Dhillon AP (2006) An appraisal of the histopathological assessment of liver fibrosis. Gut 55(4):569–578

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Rojkind M, Ponce-Noyola P (1982) The extracellular matrix of the liver. Coll Relat Res 2:151–175

    Article  CAS  PubMed  Google Scholar 

  46. Feldman G (1995) Critical analysis of the methods used to morphologically quantify hepatic fibrosis. J Hepatol 22(suppl 2):49–52

    Google Scholar 

  47. Chang PE, Goh GBB, Leow WQ, Shen L, Lim KH, Tan CK (2018) Second harmonic generation microscopy provides accurate automated staging of liver fibrosis in patients with non-alcoholic fatty liver disease. PLoS One 13(6):e0199166

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  48. Bedossa P (2010) Harmony in liver fibrosis. J Hepatol 52(3):313–314

    Article  PubMed  Google Scholar 

  49. Gailhouste L, Le Grand Y, Odin C et al (2010) Fibrillar collagen scoring by second harmonic microscopy: a new tool in the assessment of liver fibrosis. J Hepatol 52(3):398–406

    Article  CAS  PubMed  Google Scholar 

  50. Pirhonen J, Arola J, Sädevirta S, Luukkonen P, Karppinen SM, Pihlajaniemi T, Isomäki A, Hukkanen M, Yki-Järvinen H, Ikonen E (2016) Continuous Grading of Early Fibrosis in NAFLD Using Label-Free Imaging: A Proof-of-Concept Study. PLoS One 11(1):e0147804

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  51. Kumar M, Kumar A, Hissar S, Jain P, Rastogi A, Kumar D, Sakhuja P, Sarin SK (2008) Hepatic venous pressure gradient as a predictor of fibrosis in chronic liver disease because of hepatitis B virus. Liver Int 28(5):690–698

    Article  PubMed  Google Scholar 

  52. Arriazu E, Ruiz de Galarreta M, Cubero FJ, Varela-Rey M, Pérez de Obanos MP, Leung TM, Lopategi A, Benedicto A, Abraham-Enachescu I, Nieto N (2014) Extracellular matrix and liver disease. Antioxid Redox Signal 21(7):1078–1097

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Liver & Biliary Sciences, D-1 Vasant Kunj, Delhi, 110070, India

    Archana Rastogi, Nayana Patil, Rakhi Maiwall, Chhagan Bihari, Ananda Soshee & Shiv K. Sarin

Authors
  1. Archana Rastogi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nayana Patil
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rakhi Maiwall
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chhagan Bihari
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ananda Soshee
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shiv K. Sarin
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors contributed to the study conception and design. Data collection and analysis were performed by Archana Rastogi, Nayana Patil, Rakhi maiwall, Ananda Soshee and Chhagan Bihari. Data interpretation and compilation were performed by Archana Rastogi, Nayana Patil, Rakhi maiwall and Ananda Soshee. The first draft of the manuscript was written by Archana Rastogi and Nayana Patil, and all authors commented on the versions of the manuscript. Final draft was critically assessed for important intellectual content by Shiv K Sarin. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Archana Rastogi.

Ethics declarations

Ethics approval

The study protocol was approved by the Institutional Ethics Committee/ Institutional Review Board (IEC/2019/71/MA06) dated 02.11.2019.

Consent to participate

Not applicable

Consent for publication

Not applicable

Conflicts of interest

None

Additional information

Publisher’s note

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

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Rastogi, A., Patil, N., Maiwall, R. et al. Second-harmonic generation (SHG) microscopy and hepatic venous pressure gradient-based validation of a novel histological staging system for alcoholic hepatitis. Virchows Arch 479, 493–506 (2021). https://doi.org/10.1007/s00428-021-03089-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00428-021-03089-3

Keywords

Search

Navigation