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Clin Myofibroblasts hepatitis Brelated cirrhosis...
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J Clin Pathol 1992;45:446-448 Myofibroblasts in hepatitis B related cirrhosis and hepatocellular carcinoma KY Chau, MA Lily, P C Wu, W L Yau Abstract Peritoneal liver biopsy specimens from eight patients with hepatitis B associated cirrhosis, complicated by hepatocellular carcinoma, were studied for identifica- tion and localisation of myofibroblasts. The avidin-biotin peroxidase complex technique was used on paraffin wax sec- tions, using monoclonal antibodies for actin and desmin, and ultrastructural examination was performed. Myofibro- blasts were found in seven of the eight cirrhotic specimens and in all eight tumour specimens. They were identified in the fibrotic areas by the immunohisto- chemical technique, but ultrastructural examination disclosed their presence in the perisinusoidal space and between tumour cells. Myofibroblasts are transformed fibroblasts with contractile properties. They are found in normal tissue, repair tissue, and a variety of benign, borderline, and malignant fibrous or fibrohistiocytic tumours. Their prevalence and role in the desmoplastic reaction of tumours were examined recently."2 Department of Pathology, Queen Mary Hospital, University of Hong Kong, Hong Kong K Y Chau M A Lily PCWu W L Yau Correspondence to: Dr K Y Chau Accepted for publication 18 September 1991 Methods Immunohistochemical and ultrastructural studies were performed on biopsy specimens from eight patients with hepatitis B associated cirrhosis, complicated by hepatocellular car- cinoma. The biopsy specimens were taken both from cirrhotic areas and tumours using a peritoneoscope. Each specimen was fixed in 10% formalin immediately after biopsy. They were then divided into two portions and processed for histological and ultrastructural studies. These patients were either seropositive for hepatitis B surface antigen or their liver biopsy specimens showed ground-glass hepa- tocytes which stained positively for aldehyde, fuchsin, or both. No other causes for cirrhosis were identified. An avidin-biotin peroxidase complex tech- nique with monoclonal antibodies specific for actin (HHF 35; Biogenex and desmin D 33; Dakopatts) were used to identify myofibroblasts in paraffin wax sections. Both myofibroblasts and smooth muscle cells express actin' and, also weakly, desmin. Elec- tron microscopic examination was also per- formed on all the specimens to identify myofibroblasts. They were characterised ultrastructurally by peripherally located cyto- plasmic actin filaments with dense bodies, in an otherwise typical fibroblast containing prominent rough endoplasmic reticulum, Golgi apparatus, and an indented nucleus. Results Histologically, besides perivascular smooth muscle cells, myofibroblasts expressing actin and, weakly, desmin, were identified. Five out of the eight cirrhotic liver specimens showed occasional numbers of these cells in the fibrous septa; none of these cells was identifiable in the perisinusoidal space using immunohisto- chemistry. In seven of the eight tumour specimens scanty to moderate numbers of these cells were seen in the fibrous tissue surround- ing or separating tumour masses, close to the margins of the tumour islands (fig 1). They were moderate in number in one case and occasional or scanty in six cases. The negative one contained only tumour cells arranged in thick trabeculae showing no fibrous stroma. Positive staining cells were not identified within the tumour masses in any of the cases. Ultrastructurally, myofibroblasts were found in five of the eight cirrhotic specimens, including two immunohistochemically nega- tive cases. They were also found in all eight tumour specimens (fig 2) including the one which showed no fibrous stroma at light microscopy. They could be seen in the peri- sinusoidal space and between tumour cells as well as in fibrotic regions. Discussion Using both modalities of immunohisto- chemical and ultrastructural methods, myofibroblasts were identified in hepatitis B associated cirrhosis and hepatocellular carcin- oma. They were found in seven of the eight cirrhotic specimens and in all eight tumour specimens. They were identified in the fibrotic areas using an immunohistochemical tech- nique, but ultrastructural examination dis- closed their presence in the perisinusoidal space and between tumour cells. The presence of myofibroblasts has been observed in alcoholic cirrhosis.4 They have not been closely studied in other types of cirrhosis or liver disease. Their role in cirrhosis remains to be elucidated, but may be indicative of the basic reparative process that occurs in all types of cirrhosis. In experimentally induced cirr- hosis there was evidence that myofibroblasts were derived from the fat storing cells (Ito 446 on 29 May 2018 by guest. Protected by copyright. http://jcp.bmj.com/ J Clin Pathol: first published as 10.1136/jcp.45.5.446 on 1 May 1992. Downloaded from
Transcript of Clin Myofibroblasts hepatitis Brelated cirrhosis...
J Clin Pathol 1992;45:446-448
Myofibroblasts in hepatitis B related cirrhosis andhepatocellular carcinoma
KY Chau, MA Lily, P C Wu,W L Yau
AbstractPeritoneal liver biopsy specimens fromeight patients with hepatitis B associatedcirrhosis, complicated by hepatocellularcarcinoma, were studied for identifica-tion and localisation of myofibroblasts.The avidin-biotin peroxidase complextechnique was used on paraffin wax sec-tions, using monoclonal antibodies foractin and desmin, and ultrastructuralexamination was performed. Myofibro-blasts were found in seven of the eightcirrhotic specimens and in all eighttumour specimens. They were identifiedin the fibrotic areas by the immunohisto-chemical technique, but ultrastructuralexamination disclosed their presence inthe perisinusoidal space and betweentumour cells.
Myofibroblasts are transformed fibroblastswith contractile properties. They are found innormal tissue, repair tissue, and a variety ofbenign, borderline, and malignant fibrous orfibrohistiocytic tumours. Their prevalence androle in the desmoplastic reaction of tumourswere examined recently."2
Department ofPathology, QueenMary Hospital,University ofHongKong, Hong KongK Y ChauMA LilyPCWuW L YauCorrespondence to:Dr K Y ChauAccepted for publication18 September 1991
MethodsImmunohistochemical and ultrastructuralstudies were performed on biopsy specimensfrom eight patients with hepatitis B associatedcirrhosis, complicated by hepatocellular car-cinoma. The biopsy specimens were taken bothfrom cirrhotic areas and tumours using aperitoneoscope. Each specimen was fixed in10% formalin immediately after biopsy. Theywere then divided into two portions andprocessed for histological and ultrastructuralstudies. These patients were either seropositivefor hepatitis B surface antigen or their liverbiopsy specimens showed ground-glass hepa-tocytes which stained positively for aldehyde,fuchsin, or both. No other causes for cirrhosiswere identified.An avidin-biotin peroxidase complex tech-
nique with monoclonal antibodies specificfor actin (HHF 35; Biogenex and desminD 33; Dakopatts) were used to identifymyofibroblasts in paraffin wax sections. Bothmyofibroblasts and smooth muscle cellsexpress actin' and, also weakly, desmin. Elec-tron microscopic examination was also per-formed on all the specimens to identifymyofibroblasts. They were characterisedultrastructurally by peripherally located cyto-
plasmic actin filaments with dense bodies,in an otherwise typical fibroblast containingprominent rough endoplasmic reticulum,Golgi apparatus, and an indented nucleus.
ResultsHistologically, besides perivascular smoothmuscle cells, myofibroblasts expressing actinand, weakly, desmin, were identified. Five outof the eight cirrhotic liver specimens showedoccasional numbers of these cells in the fibroussepta; none of these cells was identifiable in theperisinusoidal space using immunohisto-chemistry. In seven of the eight tumourspecimens scanty to moderate numbers ofthesecells were seen in the fibrous tissue surround-ing or separating tumour masses, close to themargins of the tumour islands (fig 1). Theywere moderate in number in one case andoccasional or scanty in six cases. The negativeone contained only tumour cells arranged inthick trabeculae showing no fibrous stroma.Positive staining cells were not identifiedwithin the tumour masses in any of the cases.
Ultrastructurally, myofibroblasts werefound in five of the eight cirrhotic specimens,including two immunohistochemically nega-tive cases. They were also found in all eighttumour specimens (fig 2) including the onewhich showed no fibrous stroma at lightmicroscopy. They could be seen in the peri-sinusoidal space and between tumour cells aswell as in fibrotic regions.
DiscussionUsing both modalities of immunohisto-chemical and ultrastructural methods,myofibroblasts were identified in hepatitis Bassociated cirrhosis and hepatocellular carcin-oma. They were found in seven of the eightcirrhotic specimens and in all eight tumourspecimens. They were identified in the fibroticareas using an immunohistochemical tech-nique, but ultrastructural examination dis-closed their presence in the perisinusoidalspace and between tumour cells.The presence of myofibroblasts has been
observed in alcoholic cirrhosis.4 They have notbeen closely studied in other types of cirrhosisor liver disease. Their role in cirrhosis remainsto be elucidated, but may be indicative of thebasic reparative process that occurs in all typesof cirrhosis. In experimentally induced cirr-hosis there was evidence that myofibroblastswere derived from the fat storing cells (Ito
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Myofibroblasts in hepatitis B related cirrhosis and hepatocellular carcinoma 447
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Chau, Lily, Wu, Yau448
cells), working in concert with activatedKupffer cells, platelets, and regeneratinghepatocytes.'A desmoplastic response in hepatocellular
carcinoma is not particularly prominent andhas not been reported to be related to prog-nosis. On the other hand, the encapsulatedhepatocellular carcinoma and the fibrolamellarcarcinoma, which show a distinct stromal reac-tion, are known to carry a better prognosis.67Evidence suggests that the myofibroblastshave a role in limiting tumour expansion andpreventing metastasis by virtue of theirphysical effect and anticollagenolytic proper-ties.89 In a recent report it was suggested thatthe ability of the hepatocellular carcinoma cellsto digest the extracellular matrix was relatedto tumour aggression.10 The presence ofmyofibroblasts in hepatocellular carcinoma istherefore interesting and warrants furtherinvestigation.
1 Ahmed A. The myofibroblast in breast disease. Pathol Annu1990;2:237-86.
2 Arteaga CL, Tandon AK, Hoff DD. Transforming growthfactor beta: Potential antocrine growth inhibitor of oes-trogen receptor negative human breast cancer cells. Can-cer Res 1987;48:3898-904.
3 Tsukada T, McNutt MA, Ross R. HHF35, a muscle actinspecific monoclonal antibody. Am J Pathol 1987;127:389-402.
4 Rudolph R, McClure WJ, Woodward M. Contractilefibroblasts in chronic alcoholic cirrhosis. Gastroenterology1979;76:704-9.
5 Gressner AM, Bachem MG. Cellular sources of non-collagenous matrix proteins: role of fat-storing cells infibrogenesis. Semin Liver Dis 1990;10:30-46.
6 BermanMM, Libbey NP, Foster JH. Hepatocellular carcin-oma of polygonal cell type with fibrous stroma-an atypicalvariant with a favourable prognosis. Cancer 1980;46:1448-55.
7 Hsu HC, Wu TT, Wu MZ: Tumour invasiveness andprognosis in resected hepatocellular carcinoma: clinicaland pathogenetic implications. Cancer 1988;61:2095-9.
8 Barsky SH, Gopalakrishna R. Increase invasion and spon-taneous metastasis ofBL6 melanoma with inhibition ofthedesmoplastic response in C57 BL/6 mice. Cancer Res1987;47: 1663-7.
9 Barsky SH, Nelson LL, Levy VA. Tumour desmoplasiainhibits angiogenesis. Lancet 1987;ii: 1336-7.
10 Grigioni WF, Garbisa S, D'Errico A. Evaluation ofhepatocellular carcinoma aggressiveness by a panel ofextracellular matrix antigens. Am J Pathol 1991;138:647-54.
J Clin Pathol 1992;45:448-449
Use of Romanowsky type (Diff-3) stain fordetecting Helicobacter pylori in smears and tissuesections
A M Zaitoun
AbstractA Romanowsky type (Diff-3) stain wasused for identifying Helicobacter pyloriin gastric biopsy specimens from 50patients with ulcer and non-ulcer dyspep-sia. Air dried smears were prepared fromfresh biopsy tissue and histological sec-tions were prepared from paraffin waxprocessed tissue. The Diff-3 technique isaccomplished in five steps and takesabout 30 seconds. Results using the Diff-3stain correlated 100% with those using theGiemsa stain. The Diff-3 stain is reliable,simple, rapid, easy and clean, and smearsprepared from fresh biopsy tissue can beexamined and an immediate reportgiven. The method is recommended forthe identification of H pylon in smearsprepared from fresh tissue as well as insections prepared from processed tissue.
Many invasive and non-invasive techniques arecurrently, used for research into the naturalhistory of Helicobacter pylori. Recent interesthas focused on finding a simple and quick testfor the detection ofH pylori in gastric biopsyspecimens.'
Histological detection of H pylori in gastricbiopsy specimens can be achieved using severaltechniques, including the Warthin-Starrysilver stain,2 Giemsa,' half-Gram,4 Gimenez,5Cresyl fast violet,6 Brown-Hopps7 andimmunohistochemical methods.8
MethodsGastric biopsy specimens from 50 patients withulcer and non-ulcer dyspepsia were studied byhistological and cytological techniques for thepresence of Hpylori. Gastric biopsy specimensreceived in this laboratory include one antralbiopsy used for direct smear, urease test, andculture, and a second biopsy specimen for thepreparation of histological sections. After grin-ding a biopsy fragment in a sterile grinder airdried smears are prepared and stained by Diff-3 (available from Gainland Chemical Com-pany, Factory Road, Sandycroft, Deeside,Clwyd, Wales). Ggstric biopsy specimens forhistopathological examination are processed informalin, and sections 4 um thick are cut,dewaxed, and stained by the Diff-3 and Giemsastains.
PathologyDepartment, AlQassimi Hospital, POBox 3500 Sharjah,United Arab EmiratesAM ZitounCorrespondence to:Dr A M ZaitounAccepted for publication23 September 1991
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