Human pluripotent stem cells (hPSCs) hold great promise for the treatment of various human diseases. However, their therapeutic benefits and mechanisms for treating corneal endothelial dysfunction remain undefined. Here, we developed a therapeutic regimen consisting of the combination of hPSC-derived corneal endothelial precursors (CEPs) with nicotinamide (NAM) for effective treatment of corneal endothelial dysfunction. In rabbit and nonhuman primate models, intracameral injection of CEPs and NAM achieved long-term recovery of corneal clarity and thickness, similar with the therapeutic outcome of cultured human corneal endothelial cells (CECs). The transplanted human CEPs exhibited structural and functional integration with host resident CECs. However, the long-term recovery relied on the stimulation of endogenous endothelial regeneration in rabbits, but predominantly on the replacing function of transplanted cells during the 3-year follow-up in nonhuman primates, which resemble human corneal endothelium with limited regenerative capacity. Mechanistically, NAM ensured in vivo proper maturation of transplanted CEPs into functional CECs by preventing premature senescence and endothelial-mesenchymal transition within the TGF-β–enriched aqueous humor. Together, we provide compelling experimental evidence and mechanistic insights of simultaneous delivery of CEPs and NAM as a potential approach for treating corneal endothelial dysfunction.

The therapeutic effects of allogeneic cultivated limbal epithelial transplantation (CLET) for symblephara at different degrees caused by ocular burns were evaluated in this study. A series of interventional cases were involved in this retrospective study. Eighty eyes (80 patients) with symblephara underwent CLET and the success rates of surgical treatment as well as corneal conditions and risk factors for recurrent symblepharon were analyzed. The average age of patients was 32.4 ± 13.7 years (ranged from 4 to 60 years). The average follow-up time was 26.4 ± 13.6 months (ranged from 12 to 60 months). Symblepharon cases were caused by chemical burns (36 eyes) or thermal burns (44 eyes). The first surgical intervention achieved complete success in 40 eyes (50%), partial success in 25 eyes (31.3%), and failure in 15 eyes (18.8%). The rate of complete success was 85.0% in eyes with grade I/II symblephara, 51.5% in eyes with grade III eyes and 22.2% in eyes grade IV symblephara (P = 0.001). The treatment was completely successful in 23.1% of eyes with moderate or severe preoperative inflammatory action and 63.0% of eyes with mild or no inflammation (P = 0.000). The corneal conditions were improved in 43 eyes (53.8%), of which 21 eyes had improved visual acuity. The recurrence of symblepharon after the first CLET was positively correlated with symblepharon length (P = 0.003), preoperative inflammatory activity (P = 0.016) as well as postoperative cicatricial entropion and trichiasis (P = 0.038). CLET was effective on the recovery of anatomically deep fornixes in eyes caused by symblephara and corneal surface condition could be improved simultaneously. The success of surgical treatment was dependent on the effective control of inflammation and timely management of eyelid abnormalities.

Corneal endothelial dysfunction is a major cause of corneal blindness and is mainly treated by corneal transplantation. However, the global shortage of donor cornea hampers its application. Intracameral injection of cultured primary corneal endothelial cells (CECs) was recently confirmed in clinical trials. However, abnormal adhesion of the grafted CECs affects the application of this strategy. In this study, we explored if laminin 511 (LN511) improves the therapeutic function of the intracameral CECs injection for corneal endothelial dysfunction. To mimic the late-stage of corneal endothelial diseases, intense scraping was developed to remove CECs and extracellular matrix of the posterior Descemet's membrane (DM) without DM removal in rabbits. Then, Dulbecco's phosphate-buffered saline (DPBS) and LN511 were intracamerally injected as the control and intervention groups, respectively. We found that the injected LN511 could settle and form a coating on the posterior surface of DM. After CECs transplantation, corneal clarity of rabbits in the LN511 group was rapidly recovered within 7 days, whereas the corneal recovery took 14 days in the DPBS group. Corneal thickness of LN511 group decreased to 413.3±20.8μm 7 days after operation, which was significantly lower than 1086.3±78.6μm of DPBS group (p<0.01). Moreover, for the grafted CECs, LN511 promoted the rapid adhesion, tight junction formation, and expression of Na+/K+-ATPase and ZO-1. In vitro analysis revealed that the functions of LN511 on the cultured human CECs mechanistically depended on the cell density and the nuclear-cytoplasmic translocation of the Yes-associated protein. Our study demonstrated that LN511 precoating promoted the adhesion of the transplanted CECs and enhanced the functional regeneration of the corneal endothelium. Thus, our data suggested that the strategy of LN511 precoating and CECs intracameral injection could be a potential method for the therapy of corneal endothelial dysfunction.