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. 2026 Sep 23;14:1943293. doi: 10.3389/fbioe.2026.1943293

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Copyright © 2026 Raimondi, Costa, Zappini, Piccinini, Gallo, Cuscino, Carcione, Pulsatelli, Cattini, De Luca, Bellavia, Toscano and Giavaresi.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Five grouped bar graphs labeled A to E show fold changes in protein expression (TNFα, Fibronectin, Procollagen 1α, TGF-β1, TGF-β2) in BM and OM at 7 days (blue) and 14 days (red), highlighting statistically significant differences. Panel F presents a heatmap of protein expression across time points and medium conditions, while panel G displays a horizontal bar chart indicating KEGG pathway enrichment with associated p-values. — Analysis of protein released from hMSC cells maintained in basal (BM) or osteogenic medium (OM) for 7 anda 14 days on Ti-AM-Bru and Ti-AM Titanium alloy. BioPlex analysis of supernatant soluble factors (A) TNFa, (B) Fibronectin, (C) ProCollagen a1, (D) TGFb1, (E) TGFb2. Data are expressed as fold of induction (FOI) Ti- Bru vs Ti-AM. A two-way ANOVA and Tukey's multiple comparison test were used to evaluate the effect of Ti-Bru in different colture medium compared to Ti-AM or the effects of treatments during the experimental times (1 symbol, p<0.05; 2 symbols, p < 0.005 and 3 symbols, p < 0.0005). All experiments were triplicated, with the data expressed as Mean ± SD. (F) Heat map rapresentation of media of FOI (Ti-Bru vs Ti-AM) of expression of soluble factors released by hMSCs maintained in BM or OM medium for 7 and 14 days. All graphs and statistical analysis are created using https://BioRender.com. (G) Enrich analysis of protein released. Bar chart of top enriched terms from KEGG 2021 Human gene set library. The top 10 enriched terms for the input gene set are displayed based on the -log10(p-value), with the actual p-value shown next to each term. The term at the top has the most significant overlap with the input query gene set.