A modified hydrogel production protocol to decrease cellular content

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Dermatology

Background:

• Decellularized extracellular matrices (dECMs) are promising biomaterials for tissue regeneration.
• Porcine skin is a potential source for dECM due to its structural similarity to human skin.
• Optimizing decellularization and subsequent hydrogel formulation is crucial for successful clinical translation.

Purpose of the Study:

• To evaluate the efficacy of decellularization processes on porcine-derived skin matrix.
• To determine the optimal hydrogel concentration for minimizing cytotoxicity and maximizing fibroblast proliferation.
• To characterize the resulting hydrogel for potential use in regenerative medicine.

Main Methods:

• Multiple decellularization cycles were applied to porcine skin.
• Histological analysis, DNA quantification, and flow cytometry were used to assess decellularization effectiveness.
• Fibroblast cultures were used to evaluate cytotoxicity and cell proliferation at various hydrogel concentrations.

Main Results:

• Fourth decellularization cycle yielded the lowest DNA concentration (< 50 ng/mg).
• Histological and hematoxylin and eosin staining confirmed the absence of cellular components and revealed a heterogeneous collagen fiber structure.
• Optimal hydrogel concentration ranged from 3% to 25%, with no significant difference in cell response at 24 hours versus 7 days.

Conclusions:

• The decellularization process effectively removed cellular and DNA components from the porcine skin matrix.
• A hydrogel concentration range of 3% to 25% is suitable for supporting fibroblast culture.
• This optimized porcine-derived decellularized skin matrix hydrogel shows potential for tissue engineering applications.