Dataset Information

Unraveling Performance: Proteomic Insights into Photo-Crosslinkable Biopolymer Properties

ABSTRACT: Rational design of gelatin methacryloyl (GelMA) hydrogels remains limited by insufficient control over their molecular architecture, despite their broad use in bioactive scaffolds. Especially for applications involving tissue regeneration, a delicate balance is needed between mechanical stability and the ability of cells to remodel their microenvironment. In this study, we present a novel integrative workflow combining bottom-up proteomic analysis, 3D molecular modeling, and structural characterization to elucidate how chemical and physical modifications influence GelMA network formation. A 3D model was built from proteomics data identifying site-specific methacryloyl modifications on gelatin-derived COL1α1 and COL1α2 chains. Increasing methacrylic anhydride equivalents (2.5 vs. 0.5 eq) raised the degree of substitution (GelMA 96 vs. GelMA 34) and the frequency of modification, while preserving the number of modified lysines and arginines. Circular dichroism and X-ray diffraction revealed that highly substituted GelMA showed increased β-strand formation at the expense of helices and turns. Upon photo-crosslinking, networks with different crosslink densities were created and, for the first time, covalent crosslinks were identified and localized using proteomic analysis. This structural mapping demonstrated diverse crosslinking motifs and highlighted how enzyme specificity influences detection. The resulting insights connect GelMA chemistry to its crosslinking behavior and secondary structure, enabling more precise tuning of material properties. This work contributes a foundational strategy for decoding bioactive hydrogel networks and supports improved design of GelMA scaffolds for applications such as osteogenesis.

INSTRUMENT(S):

ORGANISM(S): Bos Taurus (bovine)

SUBMITTER: BRAY FABRICE

LAB HEAD: fabrice bray

PROVIDER: PXD068290 | Pride | 2026-01-13

REPOSITORIES: Pride

ACCESS DATA

Similar Datasets

Project description:Given the clinical need for osteoregenerative materials incorporating controlled biomimetic and biophysical cues, a novel norbornene-modified gelatin was developed with a degree of substitution of 169% compared to the number of amines present in pristine gelatin type B. It is, to the best of our knowledge, the highest substitution degree reported to date for norbornene-functionalised gelatin. Thiol-ene crosslinking exploiting thiolated gelatin as cell-interactive crosslinker resulted in networks with high (>95%) norbornene conversion. Comparing the number of physical crosslinks present, the degree of hydrolytic degradation upon modification, the network density as well as the chemical crosslinking type, the novel thiol-ene network was benchmarked against conventional gelatin-based systems in terms of the effect of biophysical cues, more specifically visco-elastic and topographical properties, affecting osteogenesis both in 2D and in 3D via a biofabrication strategy. The novel thiol-ene network outperformed conventional gelatin-based networks in terms of osteogenesis, as evidenced in 2D dental pulp stem cell seeding assays, resulting from the presentation of both a local (substrate elasticity, 25-40 kPa) and a bulk (compressive modulus, 25-45 kPa) osteogenic substrate modulus in combination with adequate fibrillar cell adhesion spacing to optimally transfer traction forces from the fibrillar ECM (as evidenced by mesh size determination with the rubber-elasticity theory) and resulting in a 1.5- and 1.7-fold increase in alkaline phosphatase activity and calcium production respectively as osteogenic markers (compared to the gold standard gelatin methacryloyl (GelMA)). Dental pulp stem cell encapsulation in extrusion-based biofabricated 3D constructs also showed a favorable response towards the novel thiol-ene network thanks to the combination of its RGD mobility (as observed from proteomic analysis), stress relaxation and substrate rigidity (bulk compressive modulus of 11-30 kPa) to enable a 1.5- and 7-fold increase in alkaline phosphatase activity and calcium production respectively as osteogenic markers (compared to the gold standard GelMA).

			Action	DRS
	1CH.msf	Msf
	1CH.raw	Raw
	1G.msf	Msf
	1G.raw	Raw
	1T.msf	Msf

Dataset Information

Unraveling Performance: Proteomic Insights into Photo-Crosslinkable Biopolymer Properties

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets