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In-bone digestion as a way to characterization of human maxillary and mandibular bone tissues

ABSTRACT: The project is based on in-bone digestion of proteins by trypsin directly in human maxillary and mandibular healthy and pathological bone samples without the prior reduction and alkylation of the disulfide bonds. The released peptides were then separated and identified by LC-MS/MS method.

INSTRUMENT(S): maXis

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Alveolar Bone, Bone Of Jaw

SUBMITTER: Jiri Santrucek

LAB HEAD: Radovan Hynek

PROVIDER: PXD026775 | Pride | 2021-09-07

REPOSITORIES: Pride

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Publications

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.

Hynek Radovan R Michalus Iva I Cejnar Pavel P Šantrůček Jiří J Seidlová Sabina S Kučková Štěpánka Š Sázelová Petra P Kašička Václav V

Electrophoresis 20210912 23

Proteomic characterization of alveolar bones in oral surgery represents an analytical challenge due to their insoluble character. The implementation of a straightforward technique could lead to the routine use of proteomics in this field. This work thus developed a simple technique for the characterization of bone tissue for human maxillary and mandibular bones. It is based on the direct in-bone tryptic digestion of proteins in both healthy and pathological human maxillary and mandibular bone sa ...[more]

PMID: 34453862

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Low-invasive sampling method for taxonomic for the identification of archaeological and paleontological bone by proteomics of their collagens

Project description:Bone collagen is an important organic material for isotopic measurement, radiocarbon and paleoproteomic analyzes, to provide information on diet, dating, taxonomic identification. Current paleoproteomics methods are destructive and require from a few milligrams to several tenths of milligrams of bone for analysis. In many cultures, bones are raw materials for artefact which are conserved in museum which hampers to damage these precious objects during sampling. Here, we describe a minimal sampling method that identifies collagen, taxonomy and post-translational modifications from Holocene and Upper Pleistocene bones dated to 130,000 and 5,000 years ago using dermatological skin tape-discs for sampling. The sampled bone micro-powder were digested following our highly optimized eFASP protocol, then analyzed by MALDI FTICR MS and LC-MS/MS for identifying the genus taxa of the bones. We show that this low-invasive sampling does not deteriorate the bones and achieves results similar to those obtained by destructive sampling. Moreover, this sampling method can be performed at archaeological sites or in museums.

2024-07-25 | PXD044039 | Pride

Protein sequencing of human bone tissue at different ages

Project description:Bone is a dynamic organ in which bone formation mediated by osteoblasts balances against bone resorption mediated by osteoclasts to maintain bone homeostasis. With age, this balance gradually tilts toward bone resorption, leading to bone loss and osteoporosis. At the present study, the bone specimens from children (group A), middle-aged patients (group B), and older individuals (group C) were subjected to proteomics analysis by LC-MS/MS.

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The role of Type VI collagen in alveolar bone

Project description:In this study, an α2(VI) deficient mouse (Col6α2-KO) model was used to examine the role of Type VI collagen in oral tissues. To examine bone properties, µCT was employed, and bone volume and bone mineral density (BMD) was measured in oral tissues. To further investigate its molecular basis, proteome analysis was performed using protein extracted from alveolar bone. In addition, alveolar bone loss progression was evaluated with a periodontitis induced model. µCT analysis showed the Col6α2- KO mice had less volume of alveolar bone, dentin and dental pulp, while the width of periodontal ligament (PDL) was greater than WT. The BMD in alveolar bone and dentin were elevated in Col6α2-KO mice compared with WT. Our proteome analysis showed significant changes in proteins related to ECM organization and elevation of proteins associated with biomineralization in the Col6α2-KO mice. In induced periodontitis, Col6α2-KO mice had greater alveolar bone loss compared to WT. In conclusion, Type VI collagen has role in controlling biomineralization in alveolar bone and that changes in the ECM of alveolar bone could be associated with greater bone loss from periodontitis.

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Homo sapiens expanded into the Northern European Plains by 45,000 years ago

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Gene expression changes in femur bone induced by skeletal muscle unloading

Project description:Gene expression changes in femur bone induced by acute skeletal muscle unloading, which leads to physiological changes including muscle atrophy, weakness and results in bone remodelling and subsequent loss of bone mass, was investigated by hind-limb suspension (HLS) treatment of Male ICR mice (28–32 g body wt; Harlan, Indianapolis, IN). AgilentTM Whole Mouse Genome Oligo Microarrays were utilised to examine the effects of HLS on mRNA expression profiles of the femur bone in the hindlimbs of freely ambulating control and 24h HLS treated mice. Five independent biological replicates of this experiment were carried out. Five independent biological replicates of this experiment (Control and HLS) were carried out.

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