Capillary electrophoretic profiling of in‐bone tryptic digests of proteins as a potential tool for the detection of inflammatory states in oral surgery

Abstract

The commonly used histological assessment of pathological states of alveolar bone tissues in oral surgery needs laborious and time-consuming processing by an experienced histologist. Therefore, a simpler and faster methodology is required in this field. Following this demand, this paper reports a straightforward approach using the tryptic cleavage of proteins directly in bone without its demineralization, followed by the capillary electrophoresis-ultraviolet detection profiling of the yielded protein digest. Cleavage-derived peptides were separated by capillary electrophoresis in acidic background electrolytes, pH 2.01-2.54. The best resolution of peptide fragments with the highest peak capacity was achieved in the background electrolyte composed of 55 mM H 3 PO 4 , 14 mM tris(hydroxymethyl)aminomethan, pH 2.01. The differences in the obtained capillary electrophoresis-ultraviolet detection profiles with characteristic patterns for particular bone samples were subsequently discriminated by linear discriminant analysis over principal components. This approach was first verified on porcine bone tissues as model samples; jawbone and calf bone tissues could be discriminated with an accuracy of 100%. Subsequently, the method was capable of differentiating unequivocally between human healthy and inflammatory alveolar bone tissues obtained from oral surgery. This procedure seems to be promising as complement or even an alternative to the traditional histological discrimination between healthy and inflammatory bone tissues in oral surgery.

Recent developments in capillary and microchip electroseparations of peptides (2019–mid 2021)

“…Direct in‐bone tryptic cleavage of proteins without demineralization, followed by the CE‐UV profiling of the yielded peptide fragments has been proposed as a complement to histological evaluation of inflammatory bone tissues in oral surgery using bare FS capillary and BGE composed of 55 mM H 3 PO 4 , 14 mM Tris, pH 2.01 [262]. CE‐UV profiles with characteristic patterns for particular bone samples were discriminated by linear discriminant analysis over principal components.…”

“…(C) A principal component (PC) analysis score plot for the baseline‐adjusted and registered CE–UV profiles of all technical replicates of human jawbone samples taken from healthy and inflammatory tissues. Reprinted with permission from [262].…”

“…(B) The baseline-adjusted and registered CE-UV profiles of tryptic digests of proteins of the healthy and inflammatory tissues of the human jawbone and the relative loadings of the first two principal components. (C) A principal component (PC) analysis score plot for the baselineadjusted and registered CE-UV profiles of all technical replicates of human jawbone samples taken from healthy and inflammatory tissues.Reprinted with permission from[262].…”

Comparison of proteomic approaches used for the detection of potential biomarkers of Alzheimer's disease in blood plasma

“…The resulting spectra were processed with mMass [26]. For PCA, MS‐alone and multiMS‐toolbox [24, 27] were used with default values and normalization by total ion count, and subsequently, for confirmation of the PCA results, by more normalization method [28] using linear registration in both m/z and intensity values.…”

In‐bone protein digestion followed by LC‐MS/MS peptide analysis as a new way towards the routine proteomic characterization of human maxillary and mandibular bone tissue in oral surgery

“…Recently, we have developed a new method for boneprotein identification based on the direct in-bone tryptic cleavage of proteins followed by the LC-MS/MS separation and identification of the released protein-peptide fragments using porcine alveolar bones as model samples [24]. A similar approach, in-bone tryptic protein digestion followed by the CE profiling of the released peptides, has been successfully applied for the differentiation of human healthy and inflammatory alveolar bone tissue obtained from oral surgery [25].…”