3D microfibrous scaffolds reinforced hyaluronic acid hydrogel for the conjunctival defects repair and ocular surface reconstruction

The conjunctiva is vital for eye movement and ocular surface protection, but severe conjunctival defects can occur due to diseases or injuries. Effective reconstruction of functional conjunctiva remains a significant challenge. In this work, we developed 3D microfibrous scaffolds using 3D near-field direct-writing electrospinning. These scaffolds were then infused with hyaluronic acid hydrogel containing nerve growth factor, with or without conjunctival epithelial stem cells (CjESCs), to serve as an advanced conjunctival substitute for ocular surface repair. The engineered composite substrate can promote cell growth and proliferation in vitro, indicating its effectiveness in supporting cellular regeneration. Furthermore, using p75 as a stem cell marker, the CjESCs were enriched and inoculated onto the substrate, thereby creating a conjunctival substitute. Upon transplanting the substitute into a rabbit model with large conjunctival defects, favorable outcomes of conjunctival repair were achieved, featuring a stratified conjunctival epithelium rich in goblet cells. RNA sequencing revealed that the substitute transplantation effectively repaired the defects by exerting anti-inflammatory effects and regulating the pluripotency of stem cells. Overall, our research developed a promising tissue-engineered substitute designed for functional conjunctival reconstruction, which is anticipated to offer a therapeutic option for individuals suffering from refractory conjunctival disorders.