Intrastromal versus subconjunctival injection of mesenchymal stem/stromal cells for promoting corneal repair.

PURPOSE: Different approaches to delivery of mesenchymal stem/stromal cells (MSCs) for ameliorating corneal injuries have been investigated. This study was aimed to compare the efficacy of intrastromal and subconjunctival injection of human bone marrow-derived MSCs (hBM-MSCs) in a corneal epithelial injury model. METHODS: Twenty-four C57BL/6J mice underwent total corneal and limbal epithelial debridement. Then, the mice were divided into three different groups: (1) intrastromal hBM-MSCs injection, (2) subconjunctival hBM-MSCs injection, and (3) injection of frozen medium as a control. Mice were monitored by slit lamp and underwent anterior segment optical coherence tomography (ASOCT). Following euthanasia, the corneas were further evaluated by histology and immunostaining. RESULTS: hBM-MSC injection successfully healed epithelial defects regardless of the delivery route (P < 0.001). However, intrastromal injection was superior to subconjunctival injection in reducing defect area (P = 0.001). Intrastromal injection of hBM-MSCs also significantly reduced corneal opacity and neovascularization and improved ASOCT parameters compared to subconjunctival injection or no treatment (P < 0.001, P = 0.003, and P < 0.001, respectively). Although both of the treatment groups were positive for CK12 and had reduced levels of MUC5AC compared to the control, CK12 staining was stronger in the intrastromal group compared to the subconjunctival group. Also, persistency of MSCs was confirmed by in vivo (up to 2 weeks) and in vitro assessments (up to 4 weeks). CONCLUSIONS: Although the injection of hBM-MSC using both intrastromal and subconjunctival methods improve wound healing and reduce neovascularization and opacity, the intrastromal approach is superior in terms of corneal healing.

Cellular senescence implication in mustard keratopathy.

Mustard agents are vesicants that were used in warfare multiple times. They are potent alkylating agents that activate cellular pathways of apoptosis, increase oxidative stress, and induce inflammation. Eyes are particularly susceptible to mustard exposure with a wide range of ocular surface damage. Three main categories of mustard-related eye injuries are acute, chronic, and delayed-onset manifestations. Mustard keratopathy (MK) is a known complication characterized by corneal opacification, ulceration, thinning, and neovascularization that can lead to severe vision loss and discomfort. Recently, a few reports demonstrated the role of senescence induction as a new pathological mechanism in mustard-related injuries that could affect wound healing. We ran the first murine model of delayed-onset MK and nitrogen mustard-induced senescence, evaluating the pathological signs of senescence in the cornea using beta-galactosidase staining. Our results suggest that nitrogen mustard exposure causes senescence in the corneal cells, which could be the underlying mechanism for chronic and late-onset ocular surface damage. We also found a significant correlation between the percentage of positive beta-galactosidase staining and the degree of fibrosis in the cornea. This provides valuable insight into the possible role of anti-senescence drugs in the near future for accelerating corneal healing and restricting fibrosis in patients with mustard keratopathy.

Physiological Health and Survival of Captive-Reared and Released Juvenile Blanding's Turtles.

AbstractConservation translocations are important in maintaining viable wildlife populations of vulnerable species within their indigenous ranges. To be effective, population restoration efforts (e.g., head start programs) must consider the species' life history, regional ecology, and physiology and the health status of wild and translocated populations. The decline of Blanding's turtles (Emydoidea blandingii) has prompted the initiation of head start programs, but the health and short-term survival of head-started juveniles released to the wild is largely unknown. From May to October 2016 and 2017, we radio tracked captive-reared, recently released juvenile Blanding's turtles and monitored their survivorship and monthly physiological health. We aimed to (1) compare physiological metrics of juveniles before and after release from captivity and between head-started cohorts, (2) identify seasonal trends in physiological metrics of recently released juveniles, (3) compare physiological metrics of recently released and formerly released juveniles, and (4) identify predictors of juvenile survivorship after release from captivity. Juvenile short-term survival was low compared with other studies. Most physiological metrics did not change after release from captivity, negating significant juvenile stress before or after release. Physiological metrics for recently released cohorts varied seasonally, suggesting that these juveniles were likely in good health. Some physiological metrics differed between recently released and formerly released juveniles, demonstrating a potential postrelease acclimatization period. Finally, no physiological metrics significantly predicted survival, but surviving juveniles had a higher percentage of fat. In all, juvenile deaths were not due to poor turtle health but rather to predation from human-subsidized mesocarnivores. Therefore, head-started juvenile Blanding's turtles released in suburban areas may benefit from antipredator training and mesocarnivore control at release sites.