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Proteomic analysis of nanoscale-engineered silk fibroin treated Sprague-Dawley BMSC

ABSTRACT: Inspired by the native biomineralization mechanisms of the bone matrix, we developed a growth factor-free, nanoscale-engineered silk fibroin (NESF) hydrogel to enhance in situ biomineralization and promote effective endogenous bone regeneration. By modulating SF nanofiber length to enrich acidic groups, the hydrogel acquired an increased negative surface charge, providing initial physical cues that effectively attracted inorganic ions and promoted Ca2+-PO43- nucleation, resulting in a 6.09-fold enhancement in biomineralization. These intrinsic chemical modifications further boosted the biological activity of the NESF hydrogel by promoting the adhesion and proliferation of BMSCs, and facilitated osteogenic differentiation by elevating intracellular Ca2+ influx and activating the MAPK/ERK signaling pathway.

ORGANISM(S): Rattus Norvegicus

SUBMITTER: Jiaxiang Li

PROVIDER: PXD073431 | iProX | Thu Jan 22 00:00:00 GMT 2026

REPOSITORIES: iProX

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Project description:Background: Ectopic vascular calcifications represent a major clinical problem associated with cardiovascular disease and mortality. However, the mechanisms underlying pathological vascular calcifications are largely unknown hampering the development of therapies to tackle this life threatening medical condition. Results: In order to gain insight into the genes and mechanisms driving this pathological calcification process we analyzed the transcriptional profile of calcifying vascular smooth muscle cells (C-VSMCs). These profiles were compared to differentiating osteoblasts, cells that constitute their physiological calcification counterparts in the body. Overall the transcriptional program of C-VSMC and osteoblasts did not overlap. Several genes, some of them relevant for bone formation, were distinctly modulated by C-VSMCs which did not necessarily lose their smooth muscle cell markers while calcifying. Bioinformatics gene clustering and correlation analysis disclosed limited bone-related mechanisms being shared by two cell types. Extracellular matrix (ECM) and biomineralization genes represented common denominators between pathological vascular and physiological bone calcifications. These genes constitute the strongest link between these cells and represent potential drivers for their shared end-point phenotype. Conclusions: the analyses support the hypothesis that VSMC trans-differentiate into C-VSMCs keeping their own identity while using mechanisms that osteoblasts use to mineralize. The data provide novel insights into groups of genes and biological processes shared in MSC and VSMC osteogenic differentiation. The distinct gene regulation between C-VSMC and osteoblasts might hold clues to find cell-specific pathway modulations, opening the possibility to tackle undesired vascular calcifications without disturbing physiologic bone formation and vice versa. Total RNA obtained from hMSC and hVSMC cultured in osteogenic differentiation medium supplemented with 1.8 mM Ca2+ for 0, 2, 8, 12 or 25 days respectively. For each timepoint 3 replicates were used, with exception for day 0 where 4 replicates were collected.

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Proteomic analysis of nanoscale-engineered silk fibroin treated Sprague-Dawley BMSC

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