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      Self-Retained Cryopreserved Amniotic Membrane for the Management of Corneal Ulcers

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          Abstract

          Purpose

          To evaluate the clinical outcomes of self-retained cryopreserved amniotic membrane (cAM) for the treatment of corneal ulcers.

          Methods

          This was a single-center, retrospective review of consecutive patients with non-healing corneal ulcers that underwent treatment with self-retained cAM (PROKERA ® Slim). The primary outcome measure was time to complete corneal epithelialization. Ocular discomfort, corneal staining, corneal signs, and visual acuity were assessed at 1 week, 1 month, 3 months, and 6 months. Complications, adverse events, and ulcer recurrence were also recorded.

          Results

          A total of 13 eyes (13 patients) with recalcitrant corneal ulcers were included for analysis, 9 (69%) of which progressed from neurotrophic keratitis (NK). Prior to cAM application, patients used conventional treatments such as artificial tears (n = 11), antibiotics (n = 11), ointment (n = 11), steroids (n = 6), and antivirals (n = 3). Self-retained cAMs (n = 1.5 ± 0.8) were placed for 6.8 ± 3.4 days, during which time antibiotics were continued. Four cases (31%) were subsequently treated with bandage contact lens (n = 3) and tarsorrhaphy (n = 1). All corneal ulcers healed in a median of 14 days (range: 4–43). This was accompanied by a significant improvement in ocular discomfort, corneal staining, and corneal signs at 1 week, 1 month, 3 months, and 6 months ( P<.05). Recurrence was noted in one case. No adverse events were observed.

          Conclusion

          Self-retained cAM may be a valuable, in-office treatment option for healing recalcitrant corneal ulcers of various etiologies, especially those with underlying NK. Further prospective, controlled studies are warranted.

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          Most cited references24

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          Neurotrophic keratopathy

          Neurotrophic Keratopathy (NK) refers to a condition where corneal epitheliopathy leading to frank epithelial defect with or without stromal ulceration (melting) is associated with reduced or absent corneal sensations. Sensory nerves serve nociceptor and trophic functions, which can be affected independently or simultaneously. Loss of trophic function and consequent epithelial breakdown exposes the stroma making it susceptible to enzymatic degradation. Nerve pathology can range from attrition to aberrant re-generation with corresponding symptoms from anaesthesia to hyperaesthesia/allodynia. Many systemic and ocular conditions, including surgery and preserved medications can lead to NK. NK can be mild (epithelium and tear film changes), moderate (non-healing epithelial defect) or severe (stromal melting and perforation). Moderate and severe NK can profoundly affect vision and adversely impact on the quality of life. Medical management with lubricating agents from artificial tears to serum/plasma drops, anti-inflammatory agents, antibiotics and anti-proteases all provide non-specific relief, which may be temporary. Contact lenses, punctal plugs, lid closure with botulinum toxin and surgical interventions like tarsorrhaphy, conjunctival flaps and amniotic membrane provide greater success but often at the cost of obscuring sight. Corneal surgery in a dry ocular surface with reduced sensation is at high risk of failure. The recent advent of biologicals such as biopolymers mimicking heparan sulfate; coenzyme Q10 and antisense oligonucleotide that suppress connexin 43 expression, all offer promise. Recombinant nerve growth factor (cenegermin), recently approved for human use targets the nerve pathology and has the potential of addressing the underlying deficit and becoming a specific therapy for NK.
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            Substance P promotes diabetic corneal epithelial wound healing through molecular mechanisms mediated via the neurokinin-1 receptor.

            Substance P (SP) is a neuropeptide, predominantly released from sensory nerve fibers, with a potentially protective role in diabetic corneal epithelial wound healing. However, the molecular mechanism remains unclear. We investigated the protective mechanism of SP against hyperglycemia-induced corneal epithelial wound healing defects, using type 1 diabetic mice and high glucose-treated corneal epithelial cells. Hyperglycemia induced delayed corneal epithelial wound healing, accompanied by attenuated corneal sensation, mitochondrial dysfunction, and impairments of Akt, epidermal growth factor receptor (EGFR), and Sirt1 activation, as well as decreased reactive oxygen species (ROS) scavenging capacity. However, SP application promoted epithelial wound healing, recovery of corneal sensation, improvement of mitochondrial function, and reactivation of Akt, EGFR, and Sirt1, as well as increased ROS scavenging capacity, in both diabetic mouse corneal epithelium and high glucose-treated corneal epithelial cells. The promotion of SP on diabetic corneal epithelial healing was completely abolished by a neurokinin-1 (NK-1) receptor antagonist. Moreover, the subconjunctival injection of NK-1 receptor antagonist also caused diabetic corneal pathological changes in normal mice. In conclusion, the results suggest that SP-NK-1 receptor signaling plays a critical role in the maintenance of corneal epithelium homeostasis, and that SP signaling through the NK-1 receptor contributes to the promotion of diabetic corneal epithelial wound healing by rescued activation of Akt, EGFR, and Sirt1, improvement of mitochondrial function, and increased ROS scavenging capacity.
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              Antiinflammatory effects of amniotic membrane transplantation in ocular surface disorders.

              To determine whether the sequestration of inflammatory cells plays a role in the antiinflammatory effects of amniotic membrane transplantation to the ocular surface. Amniotic membrane grafts were prepared from placental tissue procured from mothers undergoing planned Cesarean sections. A detailed explanation was given to all donors, and a written consent was obtained before processing. Amniotic membrane tissue was dissected into 3- x 3-cm segments, rinsed in phosphate buffered saline, and stored in dimethyl sulfoxide solutions at -80 degrees C until use. In a clinical series, amniotic membrane patches of the ocular surface were performed in 20 eyes of 20 patients with persistent corneal epithelial defects, or as a prophylactic measure after corneal limbal transplantation. Amniotic membrane patches were harvested after a 1-week observation period and were subjected to histopathologic examinations by hematoxylin and eosin staining. Inflammatory cells trapped within the amniotic membrane were labeled by immunocytochemistry using anti-CD14, CD4, CD8, and CD20 antibodies. TUNEL (TdT-mediated dUTP nick end labeling) staining was done to observe cells undergoing apoptosis. The T cell line Molt 4 was co-cultured with amniotic membrane in vitro to observe adhesion of T cells to amniotic membrane. Various degrees of inflammatory cell infiltration were observed in all clinical samples of amniotic membrane patches. Most of the inflammatory cells stained positively with anti-CD14 antibodies, indicating that these cells were of monocyte/macrophage lineage. Subsets of T cells included both CD4(+) and CD8(+) cells, whereas CD20(+) cells were sparse. TUNEL assays revealed that trapped inflammatory cells exhibited characteristics of cells undergoing apoptosis. Molt 4 invaded within amniotic membrane in an in vitro assay, which was not inhibited by blocking antibodies to beta1 and beta2 integrins. Amniotic membrane attracts and traps inflammatory cells infiltrating the ocular surface, which may explain some of the antiinflammatory properties of the fetal tissue.
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                Author and article information

                Journal
                Clin Ophthalmol
                Clin Ophthalmol
                OPTH
                clinop
                Clinical Ophthalmology (Auckland, N.Z.)
                Dove
                1177-5467
                1177-5483
                26 May 2020
                2020
                : 14
                : 1437-1443
                Affiliations
                [1 ]BostonSight , Needham, MA, USA
                [2 ]Ocular Surface Center and TissueTech Inc , Miami, FL, USA
                [3 ]Department of Ophthalmology, Florida International University , Miami, FL, USA
                [4 ]Department of Biochemistry and Molecular Biology, University of Miami , Miami, FL, USA
                Author notes
                Correspondence: Daniel Brocks BostonSight , 464 Hillside Ave Suite 205, Needham, MA02494, USATel +1 781 726 7337 Email danielbrocks@yahoo.com
                Author information
                http://orcid.org/0000-0003-0788-4374
                Article
                253750
                10.2147/OPTH.S253750
                7266945
                32581504
                91c4af26-0cf2-409a-8939-66348ffa1512
                © 2020 Brocks et al.

                This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License ( http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms ( https://www.dovepress.com/terms.php).

                History
                Page count
                Figures: 1, Tables: 2, References: 38, Pages: 7
                Categories
                Original Research

                Ophthalmology & Optometry
                amniotic membrane,corneal ulcer,neurotrophic keratitis,ocular surface disease

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