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Heart 1998;80:509-516 ( November )
Cardiovascular
Medicine, University of Sheffield, Clinical Sciences Centre, Northern
General Hospital, Herries Road, Sheffield S5 7AU, UK
Correspondence to: Dr Malik.
Accepted for publication 22 May 1998
Background
Study of the vascular response to stent
implantation has been hampered by difficulties in sectioning metal and
tissue without distortion of the tissue stent interface. The metal is
often removed before histochemical processing, causing a loss of
arterial architecture. Histological and immunohistochemical sections
should be 5 µm with an intact tissue stent interface.
ObjectivesTo identify the most suitable cutting
and grinding equipment, embedding resin, and slides for producing thin
sections of stented arteries with the stent wires in situ for
histological, immunohistochemical, and transmission electron
microscopic (TEM) analyses.
Methods20 balloon stainless steel stents were
implanted in the coronary arteries of 10 pigs. Twenty eight days later
the stented arterial segments were excised, formalin fixed, embedded in
five different resins (Epon 812, LR white, T9100, T8100, and JB4), and
sectioned with two different high speed saws and a grinder for
histological, immunohistochemical, and TEM analyses. Five stented human
arteries were obtained at necropsy and processed using the best of the
reported methods.
ResultsThe Isomet precision saw and
grinder/polisher unit reliably produced 5 µm sections with most
embedding resins; minimum section thickness with the horizontal saw was
400 µm. Resin T8100, a glycol methacrylate, enabled satisfactory
sectioning, grinding, and histological (toluidine blue, haematoxylin
and eosin, and trichromatic and polychromatic stains) and
immunohistochemical analyses (
smooth muscle actin, von
Willebrand factor, vimentin, proliferating cell nuclear antigen,
and CD68 (mac 387)). T9100 and T8100 embedded stented sections were
suitable for ultrastructural examination with TEM. Stented human
arterial sections showed preserved arterial architecture with the
struts in situ.
ConclusionThis study identified the optimal
methods for embedding, sawing, grinding, and slide mounting of stented
arteries to achieve 5 µm sections with an intact tissue metal
interface, excellent surface qualities, histological and
immunohistochemical staining properties, and suitability for TEM
examination. The technique is applicable to experimental and clinical specimens.
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