Microstructure of gallstones.

Background 
To develop a method for systematic classification of gallbladder stones, analyze the clinical characteristics of each type of stone and provide a theoretical basis for the study of the formation mechanism of different types of gallbladder stones.


Methodology 
A total of 807 consecutive patients with gallbladder stones were enrolled and their gallstones were studied. The material composition of gallbladder stones was analyzed using Fourier Transform Infrared spectroscopy and the distribution and microstructure of material components was observed with Scanning Electron Microscopy. The composition and distribution of elements were analyzed by an X-ray energy spectrometer. Gallbladder stones were classified accordingly, and then, gender, age, medical history and BMI of patients with each type of stone were analyzed.


Principal Findings 
Gallbladder stones were classified into 8 types and more than ten subtypes, including cholesterol stones (297), pigment stones (217), calcium carbonate stones (139), phosphate stones (12), calcium stearate stones (9), protein stones (3), cystine stones (1) and mixed stones (129). Mixed stones were those stones with two or more than two kinds of material components and the content of each component was similar. A total of 11 subtypes of mixed stones were found in this study. Patients with cholesterol stones were mainly female between the ages of 30 and 50, with higher BMI and shorter medical history than patients with pigment stones (P<0.05), however, patients with pigment, calcium carbonate, phosphate stones were mainly male between the ages of 40 and 60.


Conclusion 
The systematic classification of gallbladder stones indicates that different types of stones have different characteristics in terms of the microstructure, elemental composition and distribution, providing an important basis for the mechanistic study of gallbladder stones.

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The Systematic Classification of Gallbladder Stones

A great number of biliary calculi have been studied systematically, mainly by X-ray diffraction methods, with the intention of establishing the crystal forms in which different components separate initially. In order to see whether any phase transitions take place, we have studied fresh calculi immediately after operation as well as calculi removed at operation five years ago. Crystals of cholesterol monohydrate occur in nearly all biliary calculi and this is probably the only form in which cholesterol crystallizes in vivo. A crystalline calcium-bilirubin compound was found for the first time in human calculi; it has earlier been described as occurring in bovine biliary calculi. Calcium carbonate usually occurs as vaterite, but also aragonite and calcite seem to crystallize in vivo. Sodium chloride, though only in small amounts has been found for the first time in biliary calculi.

Crystalline components of biliary calculi.

Several kinds of recently advanced microanalytical techniques — like electron probe x-ray microanalysis, laser Raman microprobe, and Fourier transform-infrared spectroscopy combined with a microsampling method — have been used for the study of the microstructure of gallstones. The organic and inorganic constituents of three kinds of gallstones are characterized on a microscopic scale. The microstructure of a cholesterol-bilirubin gallstone with a layered structure has been studied with particular emphasis. Small white particles contained in both a bilirubin gallstone and a cholesterol-bilirubin mixed gallstone have been found to consist of calcium salt of a fatty acid, probably calcium palmitate. The results obtained here will be very useful for understanding the mechanism of the gallstone-formation process.

Raman microprobe and Fourier transform-infrared microsampling studies of the microstructure of gallstones

Gallstone Disease in Japan: A Report of 812 Cases

Infrared absorption spectra of over 500 pieces of gall stone were measured and analysis of gall stones by infrared spectra was established. This is the best method now known for the analyses of gall stones. Spectra were newly obtained for a group of calculi containing stearic acid, calcium stearate, triglyceride, and polysaccharide. Almost all of the gall stones are composed of a combination of such substances and cholesterol, calcium salt of bilirubin, calcium carbonate, calcium phosphate, and proteins, in various ratios and in variety of modes. A new classification has been proposed based on these chief components. It was also found that there is a vast difference in gall-stone components between urban and rural areas. Cholesterol, which is found in metropolitan area, is rare in rural villages and in its stead ratio of calcium stearate to calcium salt of bilirubin increases.

Emiko Kobayashi

Papers

Medical and Biochemical Application of Infrared Absorption Spectra. IV. Studies on Gall Stones by Infrared Spectra and their New Classification