Decellularization and genipin crosslinking of amniotic membrane suitable for tissue engineering applications

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Regenerative Medicine

Background:

• Human amniotic membrane (HAM) shows promise as a scaffold for tissue engineering.
• Enhancing the biostability and biocompatibility of HAM is crucial for its effective application.
• Current limitations hinder the widespread use of native HAM scaffolds.

Purpose of the Study:

• To characterize genipin-crosslinked human amniotic membrane (clAM) as a bioscaffold.
• To evaluate the impact of genipin crosslinking on HAM's degradation, swelling, optical clarity, ultrastructure, and mechanical strength.
• To assess the biocompatibility of clAM with fibroblasts for tissue engineering.

Main Methods:

• Human amniotic membranes (HAM) were processed into native (nAM), decellularized (dAM), and genipin-crosslinked (clAM) groups.
• Decellularization was performed using thermolysin and sodium hydroxide.
• Genipin crosslinking was applied at 0.5% and 1.0% concentrations.
• In vitro degradation, swelling, optical clarity, ultrastructure, mechanical strength, and fibroblast attachment were analyzed.

Main Results:

• Genipin-crosslinked HAM (clAM) exhibited the slowest degradation rate, remaining intact after 30 days in collagenase solution.
• Decellularized HAM (dAM) showed the highest swelling percentage, while retaining collagen content similar to nAM.
• Cell attachment was higher on dAM and 0.5% clAM compared to nAM and 1.0% clAM.
• While dAM had the highest mechanical strength, the difference was not statistically significant.

Conclusions:

• Genipin-crosslinked amniotic membrane (clAM) offers superior biostability and biocompatibility compared to native (nAM) and decellularized (dAM) forms.
• The optimal crosslinking concentration (0.5%) supports good cell attachment.
• clAM possesses favorable characteristics, including swelling, structural integrity, and extracellular matrix content, making it a promising scaffold for diverse tissue engineering applications.