Decellularized human amniotic member hydrogel promotes limbal stem cells proliferation

Area of Science:

• Biomaterials Science
• Ophthalmology
• Stem Cell Biology

Background:

• Allogeneic cultured limbal epithelial stem cell transplantation shows variable success for limbal stem cell deficiency.
• Insufficient stem cell quantity or function are key limitations.

Purpose of the Study:

• To engineer a decellularized amniotic membrane hydrogel (dAM-gel) for improved limbal stem cell transplantation.
• To assess the dAM-gel's properties and its effect on limbal stem cell behavior.

Main Methods:

• Amniotic membrane was decellularized using a freeze-thaw protocol to create dAM-gel.
• Collagen, glycosaminoglycan, and DNA content were analyzed.
• Gelation properties and limbal stem cell migration/proliferation were evaluated.
• Agrin concentration and Yap1-cyclin D1 pathway involvement were investigated.

Main Results:

• The dAM-gel exhibited reduced collagen and glycosaminoglycan content but near-complete DNA removal, minimizing immunogenicity.
• Robust gelation was achieved at 14 mg/ml within approximately 28 minutes.
• dAM-gel significantly enhanced limbal stem cell migration and proliferation, particularly p63+ cells.
• Agrin in dAM-gel was found to promote stem cell proliferation and maintain stemness via the Yap1-cyclin D1 pathway.

Conclusions:

• Decellularized amniotic membrane hydrogel (dAM-gel) is a promising biomaterial for limbal stem cell deficiency treatment.
• dAM-gel supports limbal stem cell proliferation and preserves their stem cell phenotype.
• This approach offers a potential strategy to improve the efficacy of limbal stem cell transplantation.