Review positions crosslinking as a multidimensional regulator of regenerative biomaterials

Crosslinking is more than a stabilizing step in biopolymer scaffolds: a review presents it as a primary design variable that shapes the environment experienced by cells in tissue engineering and regenerative medicine. By controlling network formation, crosslinking can simultaneously influence mechanics, viscoelasticity, swelling, molecular transport, degradation, ligand accessibility and cell–material interactions. The article is a research review rather than a new experimental study, and it deliberately excludes clinical trials and patient data.

The review considers collagen and gelatin, hyaluronic acid, silk fibroin, chitosan, alginate, fibrin and related matrices. That perspective shifts attention from a scaffold’s nominal composition alone to the physical and biological properties produced by crosslinking. It also supports treating crosslinking as a multidimensional regulator rather than a single measure of scaffold stability.

Different crosslinking methods used in these materials include covalent, photoinduced, enzymatic, ionic, physical and dynamic covalent approaches. The review organizes these approaches around the distinct biological microenvironments they can generate. This comparison matters because the choice of network chemistry is linked to several outcomes at once, including how a material behaves mechanically, how molecules move through it and how its structure changes over time.

A material therefore cannot be characterized adequately through one property when several coupled characteristics may influence its biological performance. Crosslinking is also closely connected to how a scaffold is fabricated. The review examines extrusion bioprinting, electrospinning, freeze-drying, injectable hydrogel formation and porous-scaffold fabrication, emphasizing the coupling between processing routes and network formation.

It further argues that the timing of network creation can be as important as the nominal polymer composition. The central analytical point is that a material’s processing history must be considered when relating its chemistry to its eventual biological environment. Preclinical studies summarized in the review indicate that regenerative performance cannot be inferred from stiffness alone.

Stress relaxation, degradation, transport, interfacial chemistry, immune modulation and preservation of matrix-remodeling capacity are identified as additional considerations. The cited findings are preclinical and therefore do not establish clinical benefit or demonstrate outcomes in patients. The review proposes a mechanistically informed evidence chain to connect the design of a material with its observed behavior.

The sequence begins with material definition, proceeds to properties measured in the hydrated state and then to cellular responses. It extends from those responses to ex vivo or in vivo behavior and, finally, to tissue-specific regenerative outcomes. Reproducibility is another major focus of the proposed validation approach.

The review highlights sterilization, storage and aging-related property drift as factors that may affect a material after fabrication. It calls for controls capable of distinguishing genuine network effects from formulation confounders. Such controls are essential when comparing materials, because an observed difference cannot be assigned to network design if other changes in processing or handling are not adequately addressed.

The review’s final conclusion is that crosslinking should be designed and reported as a multidimensional regulator of the regenerative microenvironment. Its effects on network mechanics, viscoelastic behavior, swelling, transport, degradation, ligand accessibility and cell interactions should be evaluated together with the processing route used to create the material. For regenerative biomaterials, the review argues, crosslinking is not merely a manufacturing operation or a shorthand for stability; it is a central variable linking material chemistry, fabrication and preclinical biological validation.

Subject of Research: Crosslinking, processing and preclinical validation of biopolymer scaffolds for tissue engineering and regenerative medicine

Article Title: Crosslinking-controlled biopolymer biomaterials for tissue engineering and regenerative medicine: network chemistry, processing, and mechanistically informed validation

Article References: Bellucci, S. (2026). Crosslinking-controlled biopolymer biomaterials for tissue engineering and regenerative medicine: network chemistry, processing, and mechanistically informed validation. Frontiers in Bioengineering and Biotechnology, 14, Article 1962375. https://doi.org/10.3389/fbioe.2026.1962375

Image Credits: AI Generated

DOI: 10.3389/fbioe.2026.1962375

Keywords: crosslinking, biopolymer scaffolds, tissue engineering, regenerative medicine, hydrogels, biofabrication, mechanobiology, matrix remodeling, biomaterials, network chemistry, Crosslinking-controlled, biopolymer