Proteomic Analysis of Gingival Tissue and Alveolar Bone during Alveolar Bone Healing

Abstract

Bone tissue regeneration is orchestrated by the surrounding supporting tissues and involves the build-up of osteogenic cells, which orchestrate remodeling/healing through the expression of numerous mediators and signaling molecules. Periodontal regeneration models have proven useful for studying the interaction and communication between alveolar bone and supporting soft tissue. We applied a quantitative proteomic approach to analyze and compare proteins with altered expression in gingival soft tissue and alveolar bone following tooth extraction. For target identification and validation, hard and soft tissue were extracted from mini-pigs at the indicated times after tooth extraction. From triplicate experiments, 56 proteins in soft tissue and 27 proteins in alveolar bone were found to be differentially expressed before and after tooth extraction. The expression of 21 of those proteins was altered in both soft tissue and bone. Comparison of the activated networks in soft tissue and alveolar bone highlighted their distinct responsibilities in bone and tissue healing. Moreover, we found that there is crosstalk between identified proteins in soft tissue and alveolar bone with respect to cellular assembly, organization, and communication. Among these proteins, we examined in detail the expression patterns and associated networks of ATP5B and fibronectin 1. ATP5B is involved in nucleic acid metabolism, small molecule biochemistry, and neurological disease, and fibronectin 1 is involved in cellular assembly, organization, and maintenance. Collectively, our findings indicate that bone regeneration is accompanied by a profound interaction among networks regulating cellular resources, and they provide novel insight into the molecular mechanisms involved in the healing of periodontal tissue after tooth extraction. Molecular & Cellular Proteomics

Pressure Cycling Technology Assisted Mass Spectrometric Quantification of Gingival Tissue Reveals Proteome Dynamics during the Initiation and Progression of Inflammatory Periodontal Disease

“…The entirety of proteomics findings has suggested that affected gingival tissue during progression contained a defense response that was regulated by many proteins and through various pathways and networks. Although we could not find any earlier data regarding the global quantitative proteome of mouse gingiva, the number of identified or quantified proteins in this study was considerably higher than that reported earlier in the human gingival tissue or the gingiva of mini pigs . The mouse genome reportedly encodes 48 709 genes, of which about one half are protein coding (22 018 genes) .…”

Pilot GWAS of caries in African-Americans shows genetic heterogeneity

“…KLF11 Involved in tooth development, specifically odontoblast differentiation [45] . PDIA6 Differentially expressed in soft tissue and bone after tooth extraction [46]. rs3403492179,933,4124.790.8332.20E-071.0690.5440.000184rs64348732197,504,8263.9550.8184.19E-06−0.029730.34427.11E-06 PGAP1 Mouse gene knockout results in severe facial abnormalities, including lack of mouth, tongue, and mandible [47].…”

“…FOXF2 also encodes a protein located near tooth germ cells during tooth development [140]. The rs1003652 (p-value 4.54 × 10 − 6 ) locus includes several genes that are differentially expressed between various dental, bone, or gingival tissues ( GRHL1, PDIA6 ) [44, 46], and one involved in odontoblast development ( KLF11 ) [45].…”

Differential expression of immunologic proteins in gingiva after socket preservation in mini pigs

“…Differences between the groups were significant in CCR9, CCL5, CXCL2, IL7, and TNF-α. The cytokine and chemokine responses seen at 3 h are known as the early immune response, which is triggered by bleeding following tooth extraction 32 . At 1 week post-extraction, levels of the inflammatory chemokines CCL3, CCL5, and CXCL2 and the chemokine receptors CCR9 and CCR10 had declined in the SP group, as compared to the EO group (however, CCR9 showed no statistical significance.).…”

“…For peptide analysis, gingival tissue was homogenized in buffer containing 1% Triton X-100, 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 2.5 mM sodium pyrophosphate, 1 mM β-glycerolphosphate, 1 mM sodium orthovanadate, 25 mM sodium fluoride, 1 µg/ml leupeptin and 1 mM PMSF. Proteins extracted from gingiva after tooth extraction were analyzed using LC-MS/MS as described previously 32 . To search via ProteinLynx GlobalServer search version 2.3.3 (Waters Corporation, Milford, MA, USA), the parent ion tolerance was set at 100 ppm, and the fragment ion tolerance was set at 0.2 Da.…”