Abstract
BackgroundSialolithiasis or salivary gland stones are associated with high clinical morbidity. The advances in the treatment of sialolithiasis has been limited, however, by our understanding of their composition. More specifically, there is little information regarding the formation and composition of the protein matrix, the role of mineralogical deposition, or the contributions of cell epithelium and secretions from the salivary glands. A better understanding of these stone characteristics could pave the way for future non-invasive treatment strategies.MethodsTwenty-nine high-quality ductal stone samples were analyzed. The preparation included successive washings to avoid contamination from saliva and blood. The sialoliths were macerated in liquid nitrogen and the maceration was subjected to a sequential, four-step, protein extraction. The four fractions were pooled together, and a standardized aliquot was subjected to tandem liquid chromatography mass spectrometry (LCMS). The data output was subjected to a basic descriptive statistical analysis for parametric confirmation and a subsequent G.O.-KEGG data base functional analysis and classification for biological interpretation.ResultsThe LC–MS output detected 6934 proteins, 824 of which were unique for individual stones. An example of our sialolith protein data is available via ProteomeXchange with the identifier PXD012422. More important, the sialoliths averaged 53% homology with bone-forming proteins that served as a standard comparison, which favorably compared with 62% homology identified among all sialolith sample proteins. The non-homologous protein fraction had a highly variable protein identity. The G.O.-KEGG functional analysis indicated that extracellular exosomes are a primary cellular component in sialolithiasis. Light and electron microscopy also confirmed the presence of exosomal-like features and the presence of intracellular microcrystals.ConclusionSialolith formation presents similarities with the hyperoxaluria that forms kidney stones, which suggests the possibility of a common origin. Further verification of a common origin could fundamentally change the way in which lithiasis is studied and treated.
Highlights
Sialolithiasis or salivary gland stones are associated with high clinical morbidity
Sample and control protein data processing and statistical analysis Because very little is known about the mechanism(s) underlying protein deposition during salivary stone formation, we devised a series of algorithms for analyzing the stones from 29 patients
Based upon the normalized data, which corrected for the differences in protein number, the basic descriptive statistical analysis showed a minimum to maximum of 0.02 to 0.07, mean of 0.0345, median of 0.0310, and mode between 0.025 and 0.035 on the X axis
Summary
Sialolithiasis or salivary gland stones are associated with high clinical morbidity. A variety of anomalous stones or calculi occur with a relatively high frequency in certain organs These formations give rise to a medical condition termed “lithiasis”. Busso et al Clin Proteom (2020) 17:12 and can decrease organ function They cause a high degree of clinical morbidity (pain, swelling, recurrent infections, and organ dysfunction), which may vary depending on the organ affected and the location, number, and invasiveness of stone formation. Sialolithiasis has an incidence of 450 cases per 100,000 individuals/year (mostly treated surgically) (cf L2, L3 and L4).3,4 This implies that the 3250 cases receiving treatment generate costs of approximately $65 million to the healthcare system. We wanted to explore new methodologies for examining these fractions and comparing them with stone formations in other organs
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