Search

Joana Roque,1 Fernando Trancoso Vaz,1 Rita Basto,1 Susana Henriques,1 Ana Sofia Lopes,1 Diana Silva,1 Jorge Santos,2 Graça Pires,1 Maria Lisboa,1 Isabel Prieto1 1Ophthalmology Department, Prof. Doutor Fernando Fonseca Hospital, Amadora, Lisbon, Portugal; 2Department of Mathematics ECT, CIMA IIFA Évora University, Évora, PortugalCorrespondence: Joana RoqueOphthalmology Department, Prof. Doutor Fernando Fonseca Hospital, Amadora, Lisbon, IC19 2720-276, PortugalTel +351932828099Email joana.roque@hff.min-saude.ptPurpose: Amniotic membrane transplantation (AMT) has shown promising results as an antifibrotic agent in trabeculectomy. We aimed to evaluate the additional effect of AMT in MMC-augmented trabeculectomy.Patients and Methods: This retrospective study analyzed the results of the first 12 postoperative months of glaucomatous eyes submitted to Moorfields Safer Surgery Trabeculectomy with MMC alone (non-AMT group) compared to MMC and AMT (AMT group). Both groups were compared in terms of intraocular pressure (IOP), number of antihypertensive medications and need for surgical reinterventions. Absolute and relative success rates 12 months after surgery were defined as IOP < 18 mmHg, without and with the use of antihypertensive medications, respectively.Results: The analysis included 51 eyes of 45 glaucoma patients (29 eyes in the non-AMT group and 22 in the AMT group). Mean IOP decreased from 24.72± 5.11 mmHg and 26.86± 10.62 mmHg preoperatively in non-AMT and AMT groups to 12.86± 4.22 mmHg and 12.60± 4.43 mmHg, respectively, at 12 months (p = 0.84). Postoperative number of medications decreased significantly in both groups. Absolute success was seen in 71% of non-AMT eyes and 55% of AMT eyes (p = 0.46), whereas relative success was obtained in 14% and 30%, respectively (p = 0.55). Reinterventions were needed in 28% of the eyes (11 bleb injection/needling and 4 Ahmed tube implantation) in the non-AMT group and in 27% of the AMT group (10 bleb injection/needling and 1 Ahmed tube implantation) (p = 0.89).Conclusion: Trabeculectomy combined with MMC and AMT did not show better results than trabeculectomy with MMC alone.Keywords: trabeculectomy, glaucoma, mitomycin C, amniotic membrane, antifibrotics, scarring

Background Locoregional stem cell delivery is very important for increasing the efficiency of cell therapy. Amnisite BA (Amnisite) is a freeze-dried amniotic membrane harvested from bovine placenta. The objective of this study was to investigate the retention of cells of the stromal vascular fraction (SVF) on Amnisite and to determine the effects of cell-loaded Amnisite in a porcine radiation-induced chronic wound model.Methods Initially, experiments were conducted to find the most suitable hydration and incubation conditions for the attachment of SVF cells extracted from pig fat to Amnisite. Before seeding, SVFs were labeled with PKH67. The SVF cell-loaded Amnisite (group S), Amnisite only (group A), and polyurethane foam (group C) were applied to treat radiation-induced chronic wounds in a porcine model. Biopsy was performed at 10, 14, and 21 days post-operation for histological analysis.Results Retaining the SVF on Amnisite required 30 minutes for hydration and 1 hour for incubation. A PKH67 fluorescence study showed that Amnisite successfully delivered the SVF to the wounds. In histological analysis, group S showed increased re-epithelialization and revascularization with decreased inflammation at 10 days post-operation.Conclusions SVFs had acceptable adherence on hydrated Amnisite, with successful cell delivery to a radiation-induced chronic wound model.

Recent reviews suggest that amniotic membrane products may accelerate healing of diabetic foot ulcers. A new dried human amniotic membrane (dHAM) has been used for ocular ulcers but not for diabetic foot ulcers. This was a multi‐centre, prospective, patient and observer blind, randomised controlled pilot trial, to investigate whether 2 weekly addition of the dHAM to standard care versus standard care alone increased the proportion of healed participants' index foot ulcers within 12 weeks. Thirty‐one people (mean age 59.8 years, 81% male, 87% type 2 diabetes) were randomised (15 dHAM, 16 usual care). Within 12 weeks, healing occurred in 4 (27%) ulcers in the dHAM group versus 1 (6.3%) usual care group (P = .1). Percentage wound area reduction was higher in the dHAM versus control group. (P = .0057). There was no difference in AEs between the two groups. Six participants allocated to dHAM correctly identified their treatment group, although 5 in usual care incorrectly thought they were in the intervention arm. This pilot trial result is encouraging showing that this dHAM preparation is safe and promising treatment. These results will be used to design a statistically powered, definitive double blind randomised controlled trial.

Objectives. Human amniotic membrane extract (AME) is known to contain numerous bioactive factors and anti-inflammatory substances. However, the anti-inflammatory effects of AME on the middle ear (ME) mucosa are unclear. This study assessed the effects of AME on the growth of the ME mucosa in response to bacterially-induced otitis media (OM).Methods. OM was induced by inoculating nontypeable Haemophilus influenzae (NTHi) into the ME cavity of rats. ME mucosal explants were cultured in AME concentrations of 0, 5, 10, or 50 μg/mL. The area of explant outgrowth was measured in culture and analyzed at 1, 3, 5, and 7 days after explantation. The expression of Ki-67, mucin 5AC (MUC5AC), tumor necrosis factor-α (TNF-α), and interleukin-10 (IL-10) in the explants was also evaluated using quantitative polymerase chain reaction (PCR) and immunocytochemistry (ICC).Results. The NTHi-induced ME mucosa growth increased gradually over the 7-day culture period in all explants at different AME concentrations. There was a trend for mucosal growth inhibition at higher concentrations of AME, although the growth was not significantly different among the groups until day 5. The ME mucosal explants treated with the 50 μg/mL concentration of AME showed significantly suppressed growth on postexplantation day 7 compared with other explants on the same day. PCR and ICC staining revealed that the expression of Ki-67, MUC5AC, TNF-α, and IL-10 further decreased in the explants with higher concentrations of AME than in those with lower concentrations of AME.Conclusion. Our results showed that higher concentrations of AME reduced the mucosal proliferative response in bacterial OM in rats. These findings provide evidence that AME has an influence on the inflammatory and proliferative responses to NTHi infection in ME mucosa.

Peripheral ulcerative keratitis (PUK) is a group of corneal disorders that cause peripheral corneal thinning, threatening globe integrity in advance stages. It is usually associated with systemic autoimmune diseases and management is based on local and systemic approaches. We present the case of a 47-year-old man with a previous history of bacterial keratitis in his left eye presenting with 1 month of bilateral ocular pain and redness. At examination, diffuse bilateral globe inflammation with paracentral corneal thinning in his left eye was observed. He was diagnosed with bilateral scleritis and PUK in his left eye. Workup for associated systemic autoimmune disease yielded negative results. The patient was started on pulses of intravenous methylprednisolone followed by oral prednisone failing to achieve sufficient control of the inflammatory syndrome. Subsequently, periodic intravenous cyclophosphamide was administered with a favorable response. A multilayer amniotic membrane graft was applied, but there was rapid melting with reabsorption of the tissue, resulting in extreme corneal thinning at the inferior paracentral cornea. A decentered 8.5-mm superficial anterior lamellar keratoplasty (SALK) was then performed obtaining the donor graft with a femtosecond laser but performing manual trepanation in the recipient. At 12 months, visual and biomicroscopic measures do not show deterioration and inflammation remains under control with oral azathioprine as maintenance regime. The management of PUK includes both systemic immunosuppression and tectonic procedures to preserve the globe integrity. Diverse surgical techniques have been attempted, but no definitive guidelines are available. Decentered large SALK is a simple technique that can yield acceptable visual results.

Background: Corneal neovascularization can result from many pathological processes affecting the ocular surface leading to disturbances and opacifications that reduce corneal clarity and may impact vision. In veterinary medicine, the use of topical corticosteroid is contraindicated in the presence of ulcerative keratitis, and there is sparse research regarding safe medical alternatives to inhibit corneal neovascularization in dogs to improve visual outcome.Aim: To investigate the pigment epithelium-derived factor (PEDF) concentration in equine amniotic membrane homogenate (EAMH) and its in-vitro vascular endothelial growth factor (VEGF) inhibition in tears of dogs with vascularized ulcerative keratitis.Methods: Homogenates from 10 equine amniotic membranes (AM) were analyzed by sandwich enzyme-linked immunosorbent assay (ELISA) for quantification of equine PEDF and VEGF. Forty tear samples were collected from both eyes of dogs diagnosed with vascularized ulcerative keratitis, and 50 samples from healthy dogs. Samples from affected eyes were allocated to G1 – affected undiluted tears; G2 – affected tears diluted with phosphate-buffer solution; G3 – affected tears treated with low-concentrated EAMH; and G4 – affected tears treated with high-concentrated EAMH. Tears from the unaffected contralateral eyes were composed in G5, while G6 was composed by tears from healthy dogs (control). The presence and levels of VEGF were evaluated in all groups by Western blot and ELISA.Results: The PEDF:VEGF ratio in EAMH was 110:1. An increase in VEGF levels was observed in tears from eyes with vascularized corneal ulcers (G1) as well as in contralateral tears (G5), compared to normal dogs (G6). Highconcentrated EAMH provided a greater decrease in VEGF levels in-vitro compared to low-concentrated EAMH.Conclusion: EAMHs exhibited high concentrations of PEDF in comparison to VEGF and were able to partially decrease VEGF levels in tears of dogs with vascularized ulcers, in-vitro. Our results suggest that VEGF concentration is elevated in tears of dogs with active vascularized ulcerative keratitis in both affected and contralateral eyes compared to that of healthy dogs. Keywords: Amniotic membrane, Corneal neovascularization, Dog, PEDF, VEGF.

We compared cellular viability between cryopreserved and lyopreserved amniotic membranes and clinical outcomes of the lyopreserved construct in a prospective cohort study of 40 patients with neuropathic foot ulcers. Patients received weekly application of lyopreserved membrane for 12 weeks with standard weekly debridement and offloading. We evaluated the proportion of foot ulcers that closed, time to closure, closure trajectories, and infection during therapy. We used chi‐square tests for dichotomous variables and independent t‐tests for continuous variables with an alpha of α = .10. Cellular viability was equivalent between cryo‐ and lyopreserved amniotic tissues. Clinically, 48% of subjects' wounds closed in an average of 40.0 days. Those that did not close were older (63 vs 59 years, P = .011) and larger ulcers at baseline (7.8 vs 1.6 cm2, P = .012). Significantly more patients who achieved closure reached a 50% wound area reduction in 4 weeks compared with non‐closed wounds (73.7% vs 47.6%, P = .093). There was no difference in the slope of the wound closure trajectories between closed and non‐closed wounds (0.124 and 0.159, P = .85), indicating the rate of closure was similar. The rate of closure was 0.60 mm/day (SD = 0.47) for wounds that closed and 0.50 mm/day (SD = 0.58) for wounds that did not close (P = .89).

Vyshnevska H. O., Kakabadze Z. Sh., Schnayder S. A., Zayats L. M., Tiron O. I., Vastyanov R. S. A comparative study on the effectiveness of application of different types of biological matrixes on mandibular alveolar mucosa lesion healing. Journal of Education, Health and Sport. 2020;10(6):382-393. eISSN 2391-8306. DOI http://dx.doi.org/10.12775/JEHS.2020.10.06.040 https://apcz.umk.pl/czasopisma/index.php/JEHS/article/view/JEHS.2020.10.06.040 https://zenodo.org/record/4056927 The journal has had 5 points in Ministry of Science and Higher Education parametric evaluation. § 8. 2) and § 12. 1. 2) 22.02.2019. © The Authors 2020; This article is published with open access at Licensee Open Journal Systems of Nicolaus Copernicus University in Torun, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author (s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non commercial license Share alike. (http://creativecommons.org/licenses/by-nc-sa/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 29.05.2020. Revised: 02.06.2020. Accepted: 30.06.2020. UDC 616.314-092.9:577.152.3 A COMPARATIVE STUDY ON THE EFFECTIVENESS OF APPLICATION OF DIFFERENT TYPES OF BIOLOGICAL MATRIXES ON MANDIBULAR ALVEOLAR MUCOSA LESION HEALING H. O. Vyshnevska1, Z. Sh. Kakabadze2, S. A. Schnayder1, L. M. Zayats3, O. I. Tiron4, R. S. Vastyanov4 1SI “Institute of Dentistry and Maxillofacial Surgery of the National Academy of Medical Sciences of Ukraine”, Odessa, Ukraine; 2Tbilisi State Medical University, Tbilisi, Georgia; 3Ivano-Frankivsk National Medical University, Ivano-Frankivsk, Ukraine 4Odessa National Medical University, Odessa, Ukraine Abstract Periodontal diseases is one of the most relevant and important problem in modern dentistry due to their increasing prevalence in population, association with the occurrence of focus of chronic infection, progressive development and irreversibility which, in case of severe damage of periodontal tissues, eventually leads to a loss of the number of teeth. All of the named factors eventually lead to the development of irreversible inflammatory-dystrophic and destructive changes in periodontal tissues. During the recent years the method of guided tissue regeneration that uses membranes is being successfully used for the treatment of periodontal diseases. A comparative study on the mandibular alveolar mucosa lesion healing during application of different types of biological matrixes containing decellularized amniotic membrane, decellularized amniotic membrane in combination with platelet-rich plasma (PRP) growth factors, and decellularized amniotic membrane in combination with PRP growth factors and hyaluronic acid was performed. The study proved the acceleration of the processes of wound area reduction. Under experimental conditions, the wound healing lasted from day 3 to day 5 of the study; however, the rate of reduction of the wound area was slower, which indicates an alterative stage of the acute inflammatory process. During days 5 – 7 (stage of regeneration, the 2nd phase of wound process) the wound area reduced rapidly and by day 14 a complete epithelization of the surgical wound was observed in all groups using decellularized amniotic membrane. In the group using the combination of PRP growth factors and hyaluronic acid the epithelization of wound was already observed on the day 5. In the control group healing occurres with scar formation. Key words: amniotic membrane; PRP growth factors; hyaluronic acid; epithelization; wound healing.

Introducción: La esclera es un tejido avascular compuesto por tejido conectivo denso cuya función principal es proteger las estructuras intraoculares. Existen diversas patologías que ponen en riesgo la integridad de dicho tejido y, en consecuencia, pueden provocar terribles secuelas con un pronóstico nefasto. El tratamiento sustitutivo forma parte de la terapia de dicha patología, siendo el injerto de esclerótica cadáver una opción biocompatible y biomimética a valorar. El injerto de esclerótica de cadáver es un tejido de fácil acceso y con buenos resultados post-quirúrgicos por lo que no solo es usado en el ámbito de la patología escleral, sino que también es utilizado para cirugía de oculoplástica, glaucoma o de retina. No obstante, el peligro de transmisión de priones o el déficit de suministro en algunos países hacen que este tejido sea suplantado por otros tejidos o técnicas quirúrgicas. Métodos: Los objetivos de este estudio son identificar la patología esclerótica susceptible de terapia sustitutiva en los últimos 5 años en el complejo hospitalario universitario de Granada, identificar los métodos empleados como terapéutica sustitutiva en dicha patología y evaluar la práctica de dicha terapia en el contexto de la bibliografía existente al respecto. Para ello, se aplicó un diseño retrospectivo en el que se realizó una búsqueda en los archivos de la unidad de documentación clínica del Hospital Clínico San Cecilio y Hospital Virgen de las Nieves, de los cuales se obtuvieron 9 casos clínicos. Resultados: Entre los 9 casos clínicos, solo 4 fueron sometidos a tratamiento quirúrgico, y de estos, 3 de ellos a terapia sustitutiva por membrana amniótica o injerto escleral. En los 5 casos restantes no se llevó a cabo ninguna técnica quirúrgica, no obstante, podría ser de gran valor el uso del refuerzo con injerto de esclerótica en los casos con escleromalacia para evitar complicaciones futuras. La esclerótica procedente de cadáver se postula como una opción con unos resultados prometedores; sin embargo, las líneas de investigación actuales apuestan por el desarrollo de un sustituto basado en las ventajas de la fibrina agarosa que supla los puntos negativos del tejido donante., Introduction: The sclera is an avascular tissue composed of dense connective tissue whose main function is to protect intraocular structures. There are several pathologies that jeopardize the integrity of this tissue and, consequently, can cause terrible consequences with a dire prognosis. Substitute treatment is part of the therapy of this pathology, with the donor sclera graft being a biocompatible and biomimetic option to be evaluated. The donor sclera graft is an easily accessible tissue with good post-surgical results, so it is not only used in the field of scleral pathology, but also used for oculoplastic, glaucoma or retinal surgery. However, the danger of prion transmission or the supply deficit in some countries causes that, other tissues or surgical techniques supplant this tissue. Methods: The objectives of this study are to identify the sclerotic pathology susceptible of replacement therapy in the last 5 years in the university hospital of Granada, identify the methods used as a replacement therapy in this pathology and evaluate the practice of such therapy. A retrospective design was applied and a search was made in the archives of the clinical documentation unit of the San Cecilio Clinical Hospital and Virgen de las Nieves Hospital. There were 9 clinical cases found. Results: Among the nine clinical cases, only 4 were subjected to surgical treatment, and of these, three of them for replacement therapy of donor sclera or amniotic membrane. In the remaining 5 cases did not carry out any surgical technique, however, it could be of great value using reinforcement sclera graft in cases with scleromalacia to avoid further complications. Sclera from cadaver is postulated as an option with promising results; however, current research committed to the development of a substitute based on the advantages of fibrin agarose that mitigates the negative points of the donor tissue.

In the majority of obstetric and gynaecological wards, the fetal membranes along with the rest of the placenta are usually discarded as medical waste. However, donating placenta to the Tissue Bank may provide a highly valuable source of graft material. One of the most common tissues used in clinical transplantation is amniotic membrane. The unique properties of amnion make it highly beneficial not only in preventing bacterial infections, protein loss, fluids and electrolytes disturbances, but also in reducing burn wound pain and accelerating healing processes. Due to its abundance of nutrients and low immunogenicity, it is often used as a skin substitute. Human amnion may be prepared in tissue banks, as biostatic or biovital grafts. In order to increase surgical treatment efficiency and to prevent the risk of potential transmission of infectious diseases, a number of methods of amniotic graft preparation have been developed. This paper presents the current clinical possibilities of applying human amnion and other types of placenta tissue grafts in the treatment of wounds of various etiology. It also describes new application areas of fetal membranes for other diseases. Placental grafts are extensively used as an alternative therapy in situations when standard treatment does not produce desirable outcomes. The properties of placental grafts including i.a. low cost, easy manipulation, capability of promoting cell migration and proliferation and stimulation of stem cell activity, making them highly beneficial in comparison to many other bioactive therapies. The alternative source of graft preparation may also be harvested from animal placenta, which makes it an alternative source of supply to human tissues.

AIM: To evaluate the efficacy of recombinant human nerve growth factor-loaded amniotic membrane (rhNGF-AM) on corneal epithelial and nerve regeneration in rabbit model. METHODS: Freshly prepared human amniotic membrane (AM) were immersed into PBS buffer containing 100 or 500 μg/mL rhNGF for 15, 30, and 60min at 4℃. The in vitro release kinetics of rhNGF was measured with ELISA. For in vivo evaluation, the AM were immersed with 500 μg/mL rhNGF for 30min. Fifty-seven rabbits were selected to establish corneal epithelial defect model. In addition to the 19 rabbits in control group, 38 rabbits received AM transplantation with or without rhNGF after the removal of central epithelium. Corneal epithelial defect area, sub-epithelial nerve fiber density, corneal sensitivity, rhNGF contents in resident AM and corneas were measured after the surgery. RESULTS: rhNGF was sustained release from the AM within 14d in vitro, with the positive correlation with initial immersion concentration. The immersion of AM in 500 μg/mL rhNGF for 30min achieved the most stable release within 14d. After transplantation in rabbit cornea, a high concentration of rhNGF in resident rhNGF-AM and cornea was maintained within 8d. Corneal epithelial healing, nerve fiber regeneration and the recovery of corneal sensitivity were significantly accelerated after the rhNGF-AM transplantation when compared to simple AM transplantation (all P

Previously, we reported a collagenase-based, animal product-free protocol for cultivated oral mucosal epithelial cell sheets for transplantation (COMET). Here, we reported the long-term outcomes of first 2 clinical cases. A 27-year-old man suffered from thermal burn, which resulted in symblepharon of lower fornix OD. COMET was performed, and the cornea remained clear with few peripheral NV and no more symblepharon 34 months postoperatively. Another 42-year-old man suffered from severe alkaline burn OD. He underwent COMET, followed by corneal transplantation half a year later. A biopsy taken two years after COMET showed stratified epithelium positive for keratin 4, 13, and 3 in the suprabasal layer. Staining for p63 and p75NTR was both positive in the basal layer. The graft remained clear up to post-OP 4 years. Our study confirmed the long-term survival of the transplanted OMECs, suggesting that collagenase-based spheroidal suspension culture is a promising technique for COMET.Trial registration ClinicalTrials.gov, ClinicalTrials.gov ID: NCT03943797 Registered 9 May 2019-Retrospectively registered, https://clinicaltrials.gov/ct2/show/NCT03943797.

This report describes the clinical features of a granuloma within the layers of amniotic membranes (AM) along with its surgical management and outcome. An eight-year-old boy underwent an AM graft in the right eye for an acute chemical injury. As there was slow healing of the ocular surface, the eye was subjected to a simple limbal epithelial transplantation (SLET). Although the corneal surface was well epithelialized, a superior symblepharon was noted six weeks following SLET. Excision of the fibrotic tissue with an AM graft was carried out. Growth of a grayish-white fibrotic lesion in the visual axis of the cornea was noted eight months after the second AM graft. The optical coherence tomography line scan revealed the location of the lesion to be anterior to the retained SLET AM. This lesion progressively increased in size, so an excision biopsy was carried out. The lesion excised in toto with the AM, and an optically clear plane was noted. Histopathology of the tissue revealed the presence of myofibroblasts, which possibly originated from the AM fibroblasts. At the three-year follow-up period, there was no recurrence of the growth, and the final visual acuity was 20/40. This is a rare report of a granuloma case arising within the layers of an AM. The underlying etiopathogenesis could be due to the multiple AM grafts that the patient underwent. These repeated grafts can incite an immune response and lead to the formation of a granuloma. The special staining and the restoration of the corneal clarity with a stable ocular surface suggest the AM origin of the mass lesion. Removal of the layer of the AM with the granuloma has good outcomes, with no recurrence on long-term follow-up.

Amniotic membrane grafts have some therapeutic potential for wounds healing. Early application of amniotic membrane turned out as beneficial in healing ulcers, burns, and dermal injuries. Since the second half of the 20th century, the autotransplants of amniotic/chorion tissue have been also used for the treatment of chronic neuropathic wounds, cornea surface injuries, pterygium and conjunctivochalasis, and dental and neurosurgical applications. The aim of this publication is to prepare a coherent overview of amniotic membrane derivatives use in the field of wound healing and also its efficacy. In total 60 publications and 39 posters from 2000-2020 were examined. In these examined publications of case studies with known study results was an assemblage of 1141 patients, and from this assemblage 977 were successfully cured. In case of posters, the assemblage is 570 patients and 513 successfully cured. From the investigated data it is clear that the treatment efficacy is very high-86% and 90%, respectively. Based on this information the use of the amniotic membrane for chronic wounds can be considered highly effective. info:eu-repo/semantics/openAccess

Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN) is a spectrum of rare, severe immunological blistering skin reactions which are triggered by medication intake or infections. The acute phase is characterized by necrolysis of the skin and desquamation of mucosa, primarily oral and ocular, with significant mortality rates. The chronic phase is characterized by multi-organ sequelae with increased rates of morbidity and reduced quality of life for patients who have survived the acute phase. Since the primary goal in the acute phase is saving the life of the patient, ocular involvement is often missed and a significant proportion of patients present to an ophthalmologist with the chronic ocular sequelae. In India, chronic ocular sequelae and low vision are observed in two-thirds of patients who present in the chronic phase of SJS/TEN. In the chronic phase of ocular involvement, there are definite windows of opportunity which if targeted with specific interventions such as scleral lenses and mucous membrane grafts can help reduce the incidence of corneal blindness and improve the quality of life for patients with SJS/TEN. Over the last decade, several studies from India have advanced the understanding of the natural course of ocular involvement in SJS/TEN and the outcomes of timely interventions in the chronic phase of the disease. We present an overview of the epidemiology of ocular complications of SJS/TEN in India, the specific challenges faced in the management of ocular complications in the acute stage and recent advances in management of the chronic ocular complications of the disease.

Ocular graft-versus-host disease (GVHD) is a severe complication of allogenic hematopoietic stem cell transplantation (HSCT). It is a term used to describe a spectrum of signs and symptoms including ocular surface inflammation, dry eye syndrome, lacrimal and meibomian gland dysfunction. We present a case of a 73-year-old man with chronic myeloblastic leukaemia and chronic GVHD. On examination, severe corneal thinning was detected in his left eye. We performed multi-layer amniotic membrane patching of the affected area, in an ambulatory setting, without using sutures or glue, but only a bandage contact lens to keep amniotic membranes attached. Three months post-amniotic-membrane-patching symptoms improved, corneal integrity was maintained, and corneal thickness increased significantly. Multi-layer amniotic membrane patching without glue and sutures may be sufficient enough to prevent further deterioration of corneal thinning and can be safely performed as an outpatient procedure, reducing the need for tectonic corneal transplantation.

Intrauterine adhesion (IUA) is a disease characterized by endometrial fibrosis caused by injury to the endometrium. In the present study, decellularized and lyophilized human amniotic membrane (DL-AM) material was transplanted in a rat model to explore the preventive effect against IUA. A total of 24 Sprague Dawley rats were randomly divided into an IUA (n=12) group and an IUA + DL-AM (n=12) group. To establish the model, the endometrium of the left uterus was scraped, while that of the right uterus was used as a control. In the IUA group, scraped uteri were sutured without any other treatment, whereas DL-AM was transplanted onto the scraped uteri in the IUA + DL-AM group. Uteri were resected for histological and immunohistochemical evaluation at 3, 7, 14 and 28 days after surgery. The results confirmed the development of IUA, which was accompanied by an increase in the rate of fibrotic area. Integral optical density (IOD) values of connective tissue growth factor (CTGF) were elevated in the IUA group, while matrix metalloproteinase-2 (MMP-2) decreased relative to the control group (P

Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) can cause significant damage to the ocular surface and eyelids. The sloughing and inflammation of the ocular mucosal epithelium during the acute phase may lead to scarring sequelae of the eyelids and ocular surface, resulting in pain and vision loss. Amniotic membrane transplantation (AMT) to the eyes and eyelids during the initial 1–2 weeks of the disease can decrease the chronic sequelae. The main development in the ophthalmologic treatment of SJS/TEN in the USA over the last 15 years has been the use of AMT on the ocular surface and eyelids during the acute phase. The evolution of AMT techniques, refinement of the evaluation of the eyes in acute SJS, and the efforts to increase the use of AMT in the USA are discussed.

The application of amniotic membrane (AM) on chronic wounds has proven very effective at resetting wound healing, particularly in re-epithelialization. Historically, several aspects of AM effect on wound healing have been evaluated using cell models. In keratinocytes, the presence of AM induces the activation of mitogen-activated protein (MAP) kinase and c-Jun N-terminal kinase (JNK) pathways, together with the high expression of c-Jun, an important transcription factor for the progression of the re-epithelialization tongue. In general, the levels of transforming growth factor (TGF)-β present in a wound are critical for the process of wound healing; they are elevated during the inflammation phase and remain high in some chronic wounds. Interestingly, the presence of AM, through epidermal growth factor (EGF) signaling, produces a fine-tuning of the TGF-β signaling pathway that re-conducts the stalled process of wound healing. However, the complete suppression of TGF-β signaling has proven negative for the AM stimulation of migration, suggesting that a minimal amount of TGF-β signaling is required for proper wound healing. Regarding migration machinery, AM contributes to the dynamics of focal adhesions, producing a high turnover and thus speeding up remodeling. This is clear because proteins, such as Paxillin, are activated upon treatment with AM. On top of this, AM also produces changes in the expression of Paxillin. Although we have made great progress in understanding the effects of AM on chronic wound healing, a long way is still ahead of us to fully comprehend its effects.

Purpose: To describe a novel technique of amniotic membrane (AM) patch graft in the management of double chamber treatment after big-bubble deep anterior lamellar keratoplasty (DALK). Case Report: A 35-year-old male patient with advanced keratoconus underwent big-bubble DALK. Manual lamellar dissection was done due to failed big-bubble. First-day postoperative double chamber was detected. Air bubbling and SF6 injection were tried without any success. Double chamber resolved by fixation of AM transplantation patch graft (1 × 1 mm) over the Descemet's membrane perforation with fibrin glue. Conclusion: Amniotic membrane patch graft can be used in the management of double chamber after DALK not responsive to intracameral gas injection.

Introduction Incontinence (up to 20%) and erectile dysfunction (up to 70%) occur frequently after radical prostatectomy (RP) in patients with localized prostate cancer. Human amniotic membrane (HAM) can improve tissue regeneration and functional outcome after RP owing to the growth factors and unique immune tolerance. Preliminary studies showed the potential value of HAM in the reconstruction of the urinary tract and nerve protection during RP. Methods A protocol is developed for a prospective, randomized, single-blind, single-surgeon, placebo-controlled exploration study of the efficacy and safety of dehydrated human amnion membrane placed around the neurovascular bundle (NVB) and vesicourethral anastomosis (VUA) during RP for the treatment of localized prostate cancer. Eligible for inclusion are patients with localized prostate cancer, requiring a surgical procedure and exclusion of preoperative incontinence and erectile dysfunction. The patients are randomized 1:1 to HAM vs. placebo and blinded during the study period. According to the T test with an alpha of 0.05 and a power of 80% and expecting a dropout of 20% of the patients, an adjusted sample size per arm of 164 patients is required. Planned Outcomes The primary outcome is a postoperative continence measured as 24-h pad test up to 12 months postoperatively. Secondary outcomes are potency, time of postoperative catheter removal, postoperative complications, and biochemical recurrence. The protocol for this randomized exploration study defines the conditions to assess the efficacy and safety of HAM application during RP in order to improve the postoperative functional outcome. This trial should pave the way for future studies of tissue engineering in an effort to reduce the morbidity of RP. Trial Registration Clinicaltrials.gov, identifier NCT03864939.

Objective: The aim of this study was to investigate the effects of human amniotic membrane (HAM) on fracture healing in an animal model. Methods: Standard tibial diaphysial fractures were created in twenty-eight Wistar-Albino rats and treated with intramedullary Kirschner wire (K-wire) and HAM (HAM (+) group) or K-wire only (HAM (–) group). Fracture healing was evaluated by histological analysis, radiologic X-ray views and callus diameter measurements at 3rd and 6th weeks postoperatively. Results: Fracture healing was histologically better in the HAM (+) group and the difference was statistically significant at both 3rd and 6th weeks postoperatively (p

Objective To evaluate the short-term safety and effectiveness of amniotic membrane/umbilical cord particulate (AMUC) in managing pain in patients with various severities of knee osteoarthritis (OA). Design Single-center, prospective, investigator-initiated pilot study. Setting Private practice. Subjects A total of 20 knee OA patients aged ≥18 years were enrolled with pain >40 mm, as determined by the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC)–A. Methods Patients received an ultrasound-guided, intra-articular injection of 50 mg of AMUC particulate reconstituted in 2 mL of preservative-free saline. All patients were then monitored at six weeks, 12 weeks, and 24 weeks postinjection. Patients who did not show >30% reduction in pain received a second injection of AMUC at six weeks. WOMAC, Patient Global Assessment, medication usage, and magnetic resonance imaging (MRI) were assessed. Results Knee OA pain significantly decreased from 74.3 ± 17.2 at baseline to 45.0 ± 25.4 at six weeks (P 30 kg/m2 (P = 0.025). MRI evaluation of the overall population revealed an improvement in the severity of bone marrow lesions in seven patients. No adverse events were observed. Conclusions AMUC particulate injection relieved pain and improved physical function in patients with symptomatic knee OA.

Corneal ulcerations are a medical emergency, and in recalcitrant cases, leading to perforation, a surgical ophthalmological emergency. The urgency of the treatment is dictated by the necessity of preventing complications that can lead to serious ocular morbidities. Medical treatment represents the first therapeutic approach and is a defining step in the further management of a patient with corneal ulceration. Multiple surgical strategies are available, but the option depends on the etiology and parameters of the ulceration: size, depth, and location.

Background Cartilage grafts are generally accepted for the restoration and reconstruction of nasal contours. The main concern that plastic surgeons may need to address after surgery pertains to the resorption and disfigurement of the grafted cartilage, especially in allogenic and heterogenic grafts. Methods A total of 12 white rabbits were divided into three groups according to the types of graft including autograft, allograft, and heterograft. We used three shapes of grafts, including block, crushed, and diced cartilage in the upper, middle, and lower rows. However, in each rabbit, these grafts were divided into two columns of wrapped and unwrapped grafts, with human amniotic membrane (HAM) grafted on each side of the rabbit's back. Results In total, 60 specimens underwent histopathological examination. No inflammation was observed in about 50% of the block-shaped conchal cartilages with HAM, and in 50%, less than 25 inflammatory cells per unit were seen. The prognosis and absorption of autograft specimens in block-shaped cartilages with HAM were significantly better compared with other shapes of cartilages with HAM and without HAM. The proliferation rate of fibroblasts in autograft and allograft specimens was more than that in heterograft specimens with HAM. Conclusion Our findings have demonstrated the new role of HAM in clinical applications, indicating that HAM may be used as a low-cost, easily accessible alternative for wrapping in cartilage grafts instead of fascia or surgicel in early future. It is useful for improving the long-term outcomes and decreasing the resorption rate.

Background: Premature rupture of membrane (PROM) is linked to significant adverse events in the prenatal, peripartum and neonatal period. Both, PROM and PPROM are associated with maternal and neonatal morbidities and mortalities. Severe oligohydramnios after PROM is associated with foetal abnormalities, pulmonary hypoplasia, respiratory distress syndromes, intrauterine growth restriction, intrauterine foetal death, foetal/neonatal sepsis, presence of meconium and an Apgar score lower than seven at five minutes. Objective: To determine the factors that influenced maternal and neonatal outcomes following a premature rupture of membranes. Subjects and Methods: This retrospective cross-sectional study was conducted over five years between 1st January 2012 and 31st December 2016 on mother neonate pair delivered at the University of Maiduguri Teaching Hospital Maiduguri. Results: During the period of the study there were 7200 deliveries at the labour ward of (UMTH), out of which 91 (1.3%) had PROM. Seventy three ( 80.2%) of the case files were retrieved and analysed. Thirty six women (49.3%) were delivered by emergency caesarean section (EMCS), while 37 (50.7%) were delivered vaginally (SVD). In the bivariate (crude) analysis, Apgar score < 7 at 5 minutes (P = 0.008, OR 95 % CL: 0.092 (0.011, 0.742), birth weight ≥ 2500g (P = 0.006, 8.944 (1.892, 42.284), and absence of APH (P = 0.007, 4.83 (1.440, 16.196) were factors modifying neonatal outcome. When adjusted regression with only factors with P value < 0.2 in crude analysis were done, only birth weight ≥ 2500 g (P = 0.024, 6.677 [1.286, 34.664]) and absence of APH (P = 0.038, 4.406 [1.085, 17.883]) were independent predictors of neonatal outcome. Thus, neonates with birth weight ≥ 2500 g were about 6.7 times more like to show favourable outcome than those with birth weight < 2500g while those without APH showed 4.4 times more likelihood for a favourable neonatal outcome. Antenatal corticosteroids use also was associated with favourable outcome as it had significantly improved preterm neonatal survival. Conclusion: Intervention with steroids, antibiotics in labour and delivery within 24 hours of PROM will greatly reduce maternal complications and enhances favourable neonatal outcome. Key words: Amniotic membrane; maternal corticosteroid; premature; preterm outcome.

The fetal membranes provide a supportive environment for the growing embryo and later fetus. Due to their versatile properties, the use of fetal membranes in tissue engineering and regenerative medicine is increasing in recent years. Moreover, as microbial infections present a crucial complication in various treatments, their antimicrobial properties are gaining more attention. The antimicrobial peptides (AMPs) are secreted by cells from various perinatal derivatives, including human amnio-chorionic membrane (hACM), human amniotic membrane (hAM), and human chorionic membrane (hCM). By exhibiting antibacterial, antifungal, antiviral, and antiprotozoal activities and immunomodulatory activities, they contribute to ensuring a healthy pregnancy and preventing complications. Several research groups investigated the antimicrobial properties of hACM, hAM, and hCM and their derivatives. These studies advanced basic knowledge of antimicrobial properties of perinatal derivatives and also provided an important insight into the potential of utilizing their antimicrobial properties in a clinical setting. After surveying the studies presenting assays on antimicrobial activity of hACM, hAM, and hCM, we identified several considerations to be taken into account when planning future studies and eventual translation of fetal membranes and their derivatives as antimicrobial agents from bench to bedside. Namely, (1) the standardization of hACM, hAM, and hCM preparation to guarantee rigorous antimicrobial activity, (2) standardization of the antimicrobial susceptibility testing methods to enable comparison of results between various studies, (3) investigation of the antimicrobial properties of fetal membranes and their derivatives in the in vivo setting, and (4) designation of donor criteria that enable the optimal donor selection. By taking these considerations into account, future studies will provide crucial information that will enable reaching the optimal treatment outcomes using the fetal membranes and their derivatives as antimicrobial agents.

Background: Toxic epidermal necrolysis (TEN) is a rare life-threatening disease that mainly affects the skin and mucous membranes, resulting from a toxic delayed-type hypersensitivity (DTH) reaction (type IV reaction) to the presence of foreign antigens such as drugs. The clinical symptoms are caused by pathophysiological processes leading to massive apoptosis of keratinocytes in the dermo-epidermal junction. This results in the formation of a bulla and subsequent separation of the entire epidermis with the exposure of the dermis. The current approach in the local therapy of TEN prefers the use of biological dressings, which helps provide several critical requirements for defect healing; in particular, it helps in the acceleration of the spontaneous wound closure (re-epithelialization) of the skin defect and the reduction of the risk of development of various complications and infections, such as the risk of pathological scar maturation. This paper is a case report of the use of a lyophilized amniotic membrane (AM) for accelerating wound healing in a patient with TEN.Case Presentation: We report a case of an 8-year-old girl transferred to our center with a histologically confirmed diagnosis of TEN. Despite the application of immunosuppressive therapy consisting of corticosteroids and intravenous immunoglobulins, we have observed disease progression and exfoliation of up to 60% of the total body surface area (TBSA). In the facial area, which is cosmetically privileged, we decided to use the lyophilized amniotic membrane (Amnioderm®) to cover up approximately 2% of the TBSA. Within 2 days after the application, we observed accelerated reepithelialisation, with rapid wound closure. We have not observed any side effects nor infections during the subsequent phases of wound healing. Skin defects in non-facial areas of the body were treated with synthetic dressings. When compared to the areas covered with the lyophilized AM, the healing process was prolonged.Conclusions: To our knowledge, this is the first case study using a lyophilized amniotic membrane in the treatment of a patient with TEN. The AM application in the cosmetically-privileged area (face), proved to be very efficient in the treatment of TEN patients. The use of this allogeneic material demonstrated excellent biocompatibility and caused a unique acceleration of epithelialization and wound healing, yielding also excellent long-term results. The current study opens broad possibilities for clinical application of the used material, the improvement of current therapy of patients with TEN and better outcomes and recovery of patients.

Background and objectives: the aim of this study was to analyze the efficacy of a modified “amnion-assisted conjunctival epithelial redirection (ACER)” technique for the treatment of partial limbal stem cell deficiency (LSCD). Materials and methods: the medical records of three patients with partial LSCD who underwent corneal surface reconstruction with modified ACER following superficial keratectomy were retrospectively studied. Briefly, in this technique, an inner amniotic membrane (AM) layer was applied on the corneal surface to promote corneal re-epithelialization. The outer AM layer was applied as a barrier to prevent the invasion of conjunctival epithelial cells into the cornea before the corneal surface was completely covered by corneal epithelial cells derived from the remaining intact limbal stem cells. Results: in all three cases, the outer AM layer successfully kept the conjunctival epithelium away from the corneal surface and prevented an admixture of conjunctival epithelial cells with corneal epithelial cells. In all three patients, the cornea was completely re-epithelized with epithelial cells derived from the remaining healthy limbal stem cells, and a clear visual axis was maintained without recurrence for a mean follow-up period of 37.3 ± 8.6 months. Conclusions: the preliminary results suggest that modified ACER appears to be a viable option for patients with partial LSCD.

International audience; Due to its intrinsic properties, there has been growing interest in human amniotic membrane (hAM) in recent years particularly for the treatment of ocular surface disorders and for wound healing. Herein, we investigate the potential use of hAM and amnion-chorion membrane (ACM) in oral surgery. Based on our analysis of the literature, it appears that their applications are very poorly defined. There are two options: implantation or use as a cover material graft. The oral cavity is submitted to various mechanical and biological stimulations that impair membrane stability and maintenance. Thus, some devices have been combined with the graft to secure its positioning and protect it in this location. This current opinion paper addresses in detail suitable procedures for hAM and ACM utilization in soft and hard tissue reconstruction in the oral cavity. We address their implantation and/or use as a covering, storage format, application side, size and number, multilayer use or folding, suture or use of additional protective covers, re-application and resorption/fate. We gathered evidence on pre- and post-surgical care and evaluation tools. Finally, we integrated ophthalmological and wound healing practices into the collected information. This review aims to help practitioners and researchers better understand the application of hAM and ACM in the oral cavity, a place less easily accessible than ocular or cutaneous surfaces. Additionally, it could be a useful reference in the generation of new ideas for the development of innovative protective covering, suturing or handling devices in this specific indication. Finally, this overview could be considered as a position paper to guide investigators to fulfill all the identified criteria in the future.

Purpose To report the outcomes of two cases of infectious scleritis with severe scleral thinning managed using combined mechanical debridement, cryopreserved umbilical amnion (UA) and amniotic membrane (AM) grafts, and antimicrobial therapy. Observations Two patients presented with severe infectious scleritis with progressive scleral melts secondary to Pseudomonas aeruginosa subsequent to pterygium surgery and intravitreal injection, respectively. Surgical debridement was followed by UA grafting, AM transplantation, and antimicrobial therapy. Epithelialization and vascularization were seen within 1–4 weeks post-operatively. At last follow up of 5–6 months, the structural integrity of the sclera remained preserved with no infection recurrence, graft necrosis, or wound dehiscence. Conclusions and importance This case series highlights the utilization of UA grafting along with AM transplantation and topical and systemic antibiotics to preserve the globe in cases of severe infectious scleritis.

International audience; Thanks to their biological properties, amniotic membrane (AM), and its derivatives are considered as an attractive reservoir of stem cells and biological scaffolds for bone regenerative medicine. The objective of this systematic review was to assess the benefit of using AM and amniotic membrane-derived products for bone regeneration. An electronic search of the MEDLINE-Pubmed database and the Scopus database was carried out and the selection of articles was performed following PRISMA guidelines. This systematic review included 42 articles taking into consideration the studies in which AM, amniotic-derived epithelial cells (AECs), and amniotic mesenchymal stromal cells (AMSCs) show promising results for bone regeneration in animal models. Moreover, this review also presents some commercialized products derived from AM and discusses their application modalities. Finally, AM therapeutic benefit is highlighted in the reported clinical studies. This study is the first one to systematically review the therapeutic benefits of AM and amniotic membrane-derived products for bone defect healing. The AM is a promising alternative to the commercially available membranes used for guided bone regeneration. Additionally, AECs and AMSCs associated with an appropriate scaffold may also be ideal candidates for tissue engineering strategies applied to bone healing. Here, we summarized these findings and highlighted the relevance of these different products for bone regeneration.

Background/aim Iatrogenic vas deferens injury is one of the most serious complications of operations in the inguinal region. Vasovasostomy is performed as treatment. However, stenosis is common after vasovasostomy. Oligospermia or azoospermia may develop and result in infertility. This study aimed to investigate the effect of amniotic membrane on healing in vas deferens injuries. Materials and methods Four groups consisting of 10 rats each were formed. No procedure was performed in Group-I. In Group- II, the left vas deferens was transected and left to spontaneous healing. In Group-III, the left vas deferens was transected, and end- to-end anastomosis was performed. In Group-IV, the left vas deferens was transected, end-to-end anastomosis was performed, and it was closed with a wrapping of amniotic membrane on the anastomosis line. Rats were sacrificed after 60 days, and each left vas deferens was evaluated. Lumen patency was checked by passing methylene blue through the vas deferens. Subsequently, the vas deferens was evaluated both macroscopically and histopathologically. Data were evaluated using SPSS version 21.0. p < 0.05 was considered statistically significant for all variables. Results The anastomosis lines in Group-IV healed better than those in Group-III, and less stenosis was observed. There were differences between the groups in terms of luminal patency (p = 0.009), adhesions to surrounding tissues (p = 0.02) and separation of the ends of the vas deferens (p = 0.03). Conclusion We observed improvement on luminal patency and histology of rat vas deferens injury after surrounding human amniotic membrane on the transected and repaired surface. Further studies are needed to apply this promising result on human beings.

Background The purpose is to perform a cost effectiveness analysis amniotic membrane vs. topical medications in the use of treating dry eye disease. A cost effectiveness analysis comparing amniotic membrane + other topical medications to topical cyclosporine A + other topical medications was evaluated using accepted decision tree modeling software. Methods TreeAge Pro 2019 software was used to evaluate the base case costs over a one year timeframe. Sensitivity analysis was performed on those variables which had the greatest effect on choosing one therapy versus the other based on cost. Monte Carlo simulation was run 1,000 times to determine the most effective, least costly alternative. Costs were evaluated from a societal level (direct + indirect). Quality of life utility scores were evaluated using known time tradeoffs from prior studies (scale 0–1; with 1 being perfect vision). Results Over a one year timeframe, the base case demonstrated that amniotic membrane + topical medications was the less expensive alternative and provided for incremental utilities versus topical cyclosporine + other medications (Cost/utility: $18,275/0.78 vs. $20,740/0.74). If examining direct costs only, topical cyclosporine was the least expensive option over a one year timeframe: $4,112 vs. $10,300. Sensitivity analysis demonstrated that in order for topical cyclosporine to be the less expensive alternative the following variables would need to be: 79% of amniotic membrane implants would need to be re-implanted at month 4 (for whatever reason); > $2677 per amniotic membrane implant procedure (Medicare reimbursement rate); > 96% positive response to topical cyclosporine A at month 4; > 58% positive response to topical cyclosporine A at month 6 and; Conclusion Based on improved outcomes using amniotic membrane, patient productivity was improved resulting in lower societal costs (less days lost from work). When considering the untoward effects of dry eye disease on societal costs, an improvement of the dry eye disease condition was accomplished most often with amniotic membrane.

BACKGROUND Minimizing inflammation on the ocular surface after pterygium surgery is essential to prevent recurrence. Currently, limited data are available for the comparison between freeze-dried amniotic membrane (FD-AM) and conjunctival–limbal graft (CLG). Therefore, this study aimed to assess conjunctival inflammation and the interleukin (IL)-6 levels in tears after pterygium excision between both techniques. METHODS This prospective, single-blinded, randomized clinical trial included 26 eyes of 26 patients with primary or recurrent pterygium and who were indicated for pterygium surgery at Cipto Mangunkusumo Hospital, Indonesia from October 2017 until February 2018. The patients were scheduled for pterygium excision with FD-AM or CLG based on block randomization. Conjunctival inflammation and the level of IL-6 in tears were measured on day-7 and -28 after surgery. RESULTS The 26 eyes (13 in each group) were subjected to a complete 1-month follow-up period. Day-7 after the surgery, severe inflammation occurred in 4/13 (31%) eyes having FD-AM and 8/13 eyes (62%) having CLG (p = 0.59). The IL-6 tear level increased similarly in FD-AM (22.85 pg/ml) and CLG (23.51 pg/ml) in (p = 0.78). No severe adverse events occurred. CONCLUSIONS Pterygium excision using FD-AM yield a milder inflammation than CLG on day-7 and -28 after surgery. The tear IL-6 levels similarly increased on day-7.

Amniotic membrane grafts (AMGs) are commonly used to treat a variety of ophthalmologic conditions. Complications exist with permanent tarsorrhaphies, including the risk of re-fusion following tarsorrhaphy separation. We report a novel application of amniotic graft in lieu of skin grafts to protect the exposed marginal surface during the initial re-epithelialization period following release of a permanent tarsorrhaphy. We present a 24-year-old man who sustained an 80% total body surface area burn from a motor vehicle accident 16 months prior to presentation at our Oculoplastic service for evaluation of residual lagophthalmos. His original permanent tarsorrhaphies were removed; however, re-fusion occurred temporally in both sides. During a second attempt, AMGs were secured over the eyelid margins, leading to a successful tarsorrhaphy takedown without re-fusion. Periocular burn injuries present particular challenges, as cicatricial changes continue to evolve and viable skin graft areas diminish with each successive graft. In the setting of recurrent auto-tarsorrhaphy, the AMG has shown to be a viable alternative to standard skin grafting. This case demonstrates excellent results in a skin graft sparing procedure that is effective and efficient. Amniotic membrane grafting reduces morbidity by foregoing skin graft donor sites and can achieve similar functional and cosmetic results to standard skin grafting with reduced overall surgical time. As such, AMGs have the potential to supplant standard skin grafting in cases of recurrent auto-tarsorrhaphy, particularly in the setting of diminished available healthy skin tissue.

Ocular chemical burns are potentially blinding ocular injuries and require urgent management. Amniotic membrane (AM) transplantation is an effective surgical treatment, one of the reasons is because AM is a rich source of growth factors that can promote epithelialization and wound healing. However, growth factors will be gradually lost and insufficient after preparation process and long-time storage, leading to unsatisfactory therapeutic effects. Herein, we present a modified AM (AM-HEP) for the supplement and sustained release of growth factor by surface grafting heparin for treatment of ocular chemical burns. Heparin grafting rate and stability, microstructure, physical property, and sustained release of epithelial growth factor (EGF) of AM-HEP were characterized. Biocompatibility and ability to promote corneal epithelial cell growth and migration were evaluated and compared with a biological amnion, which is available on the market in vitro. The therapeutic effects of AM-HEP combined with EGF (AM-HEP@EGF) in vivo had been evaluated in a model of mouse corneal alkali burn. The results indicated that heparin was introduced into AM and maintain stability over 3 weeks at 37°C. The modification process of AM-HEP did not affect microstructure and physical property after comparing with non-modified AM. EGF could be combined quickly and effectively with AM-HEP; the sustained release could last for more than 14 days. AM-HEP@EGF could significantly promote corneal epithelial cell growth and migration, compared with non-modified AM and control group. Faster corneal epithelialization was observed with the transplantation of AM-HEP@EGF in vivo, compared with the untreated control group. The corneas in the AM-HEP@EGF group have less inflammation and were more transparent than those in the control group. The results from in vitro and in vivo experiments demonstrated that AM-HEP@EGF could significantly enhance the therapeutic effects. Taken together, AM-HEP@EGF is exhibited to be a potent clinical application in corneal alkali burns through accelerating corneal epithelial wound healing.

This study aims to evaluate the changes of fragility and ultrastructure of amniotic membrane after cross-linking by UVA/riboflavin. Forty-nine fresh amniotic membranes were randomly divided into 3 groups. Eighteen were in group A (CX group) and immersed in 0.1% riboflavin solution for 10 min for UVA/riboflavin cross-linking. Sixteen were in group B (B2 group), soaked for 10 min with 0.1% riboflavin. After soaking, membranes in group A and B were transferred into corneal preservation solution. Fifteen pieces were in group C, directly into corneal preservation solution. The biomechanical and ultrastructural changes of the amniotic tissue before and after cross-linking were examined (CX group = 13, B2 group = 11, C group = 15). The amniotic membrane tissue of group A (n = 5) and B (n = 5) was transplanted into 16 eyes of the rabbits, respectively, and the dissolution time of the amniotic membrane tissue was investigated. After cross-linking, compared with the control group, the elastic modulus of the low-stress area of the amniotic membrane (Elow) was higher, while the elastic modulus of the high-stress area of the amniotic membrane (Ehigh) was lower, with no significant difference in the tensile strength. Also, the collagen fibers showed coarse and bamboo-like changes. In group A, amniotic membranes began to dissolve 4 weeks after conjunctiva transplantation, and all amniotic membranes were dissolved and absorbed 6 weeks after conjunctiva transplantation. In group B, some amniotic membrane tissues were still visible 6 weeks after conjunctiva transplantation. This study suggested that after amniotic membrane cross-linking, the brittleness was increased, the hardness was enhanced, and the morphology of the collagen fiber was changed. The cross-linked amniotic membrane showed resistance to tissue dissolution.

Background Diabetic foot ulcer is a complication with multiple aetiological factors which has a significant impact to patients’ lives and costs to the healthcare system. The potential of human amniotic membrane to act as an allograft has been studied in relation to this condition. Aim of this study is to evaluate the current scientific evidence on its effectiveness in healing diabetic foot ulcers. Methods Pubmed, Cochrane library, and Google scholar were searched using the search terms, “Amnion” OR “Placenta” AND “Diabetic foot”. (MeSH terms) in the title or the abstract field from 1st of January 2000 to 30th March 2020. The quality of published reports was assessed using standard methods. We searched for experimental and observational studies in terms of randomized control trials, prospective cohort, retrospective cohort studies and case series. Results When searched with Mesh terms, 12 citations in PubMed, 22 citations in Cochrane library and 30 in other data bases were found. After screening the studies and their reference lists, 12 studies met the inclusion criteria and the others were excluded. There were 8 randomized control trials (RCTs), 2 prospective studies and 2 retrospective studies employing different preparation methods of the amniotic membranes. A wide variation in study end points were noted. Majority of the RCTs (n = 7) were concluded with significantly higher wound closure rate compared to the conventional treatment groups. In prospective and retrospective studies, it was shown that large chronic ulcers which were resistant to closure with standard therapy achieved wound closure with amniotic membrane allografts. A meta-analysis could not be performed due to study heterogeneity, and publication bias was not assessed due to the small number of available studies which was not sufficient for accurate comparison. Conclusion Even though, the studies had some inherent heterogeneity due to different preparation methods, different study end points and outcome measurements. According to our review the current studies using amniotic membrane allografts give reliable evidence of reduction in healing time over conventional methods.

Purpose To report successful management of acute ocular graft-versus-host disease (oGVHD) by self-retained cryopreserved amniotic membrane (AM). Observations A 69-year-old male developed acute oGVHD following hematopoietic stem cell transplantation (HSCT) with severe dryness, pain, photophobia, and blurred vision, the right eye worse than the left eye. Despite topical artificial tears, corticosteroids, and bandage contact lens (BCL) for two weeks, his right eye deteriorated with increasing redness and blurred vision and presented with diffuse conjunctival inflammation, severe superficial punctate keratitis, and corneal and limbal epithelial defect. Hence, self-retained cryopreserved AM was applied in the right eye while conventional therapy was continued in the left eye. Placement of self-retained AM for 3-days resulted in resolution of symptoms, reduction in inflammation, and complete re-epithelialization of the corneal and limbal defect with the visual acuity improving from 20/500 to 20/70 in the right eye. In contrast, the visual acuity improved from 20/300 to 20/150 and remained inflamed with conventional therapy in the left eye. One-month post-AM placement, the right eye remained asymptomatic with the visual acuity improved to 20/30 without any additional therapy, whereas the left eye improved to 20/70 with conventional treatment of BCL, loteprednol, and artificial tears. Conclusions and Importance This case suggests application of self-retained AM can be an effective treatment to accelerate the restoration of vision and corneal surface health for acute oGVHD.

Background Tumours in mammary glands represent the most common neoplasia in bitches, as in humans. This high incidence results in part from the stimulation of sex hormones on these glands. Among mammary tumours, inflammatory carcinoma is the most aggressive, presenting a poor prognosis to surgical treatment and chemotherapy. One of the most widely used chemotherapy drugs for breast cancer treatment is doxorubicin (DOXO). Alternative therapies have been introduced in order to assist in these treatments; studies on treatments using stem cells have emerged, since they have anti-inflammatory and immunomodulatory properties. The aim of this study was to evaluate the effects of DOXO and canine amniotic membrane stem cells (AMCs) on the triple-negative canine inflammatory mammary carcinoma cell line IPC-366. Methods Four experimental groups were analysed: a control group without treatment; Group I with DOXO, Group II with AMC and Group III with an association of DOXO and AMCs. We performed the MTT assay with DOXO in order to select the best concentration for the experiments. The growth curve was performed with all groups (I-III) in order to verify the potential of treatments to reduce the growth of IPC-366. For the cell cycle, all groups (I-III) were tested using propidium iodide. While in the flow cytometry, antibodies to progesterone receptor (PR), estrogen receptor (ER), PCNA, VEGF, IL-10 and TGF-β1 were used. For steroidogenic pathway hormones, an ELISA assay was performed. Results The results showed that cells treated with 10 µg/mL DOXO showed a 71.64% reduction in cellular growth after 72 h of treatment. Reductions in the expression of VEGF and PCNA-3 were observed by flow cytometry in all treatments when compared to the control. The intracellular levels of ERs were also significantly increased in Group III (4.67% vs. 27.1%). Regarding to the levels of steroid hormones, significant increases in the levels of estradiol (E2) and estrone sulphate (S04E1) were observed in Groups I and III. On the other hand, Group II did not show differences in steroid hormone levels in relation to the control. We conclude that the association of DOXO with AMCs (Group III) promoted a reduction in cell growth and in the expression of proteins related to proliferation and angiogenesis in IPC-366 triple-negative cells. Conclusions This treatment promoted ER positive expression, suggesting that the accumulated oestrogen conducted these cells to a synergistic state, rendering these tumour cells responsive to ERs and susceptible to new hormonal cancer therapies.

Introduction: Early repair in patients affected by myelomeningocele (MMC) is of paramount importance in order to prevent infection, minimize neural tissue damage, and reduce mortality. Treatment must include duraplasty and possibly an adequate soft tissue coverage. Delayed surgery in MMC patients can be more tedious due to the less clear borders between the placode and the skin. Moreover, the risks of wound infection and breakdown increase significantly. Case Presentation: We present the unusual case of a large MMC in a 3-year-old patient treated by combining the recently described cryopreserved amniotic membrane (AM) as homograft for dural reconstruction and a bilateral Keystone flap for soft tissue reconstruction. Discussion: Thanks to its anti-inflammatory and elastic proprieties, the AM can play an important role in preventing adhesion between the reconstructed layers, thus reducing the risk of spinal cord tethering. The Keystone flap, at the same time, allows the wound tension to be distributed widely over the flap margins and not only along the midline, which overlies the duraplasty, enhancing the scar quality and lowering the risk of cerebrospinal fluid recurrence and wound dehiscence, with no donor site morbidity.

Acute acquired comitant esotropia (AACE) is an uncommon form of strabismus in older children and adults. It is characterized by acute onset concomitant esotropia and diplopia. It can occur as a result of monocular occlusion, vision loss, physical stress, or high myopia. We describe a child with chronic vernal keratoconjunctivitis with nonhealing shield ulcer who presented with AACE due to disruption of fusion following amniotic membrane transplantation. To the best of our knowledge, AACE following amniotic membrane transplantation has not been reported till date.

A randomised, controlled multicentre clinical trial was conducted at 14 wound care centres in the United States to confirm the efficacy of dehydrated human amnion/chorion membrane allograft (dHACM) for the treatment of chronic lower extremity ulcers in persons with diabetes. Patients with a lower extremity ulcer of at least 4 weeks duration were entered into a 2-week study run-in phase and treated with alginate wound dressings and appropriate offloading. Those with less than or equal to 25% wound closure after run-in were randomly assigned to receive weekly dHACM application in addition to offloading or standard of care with alginate wound dressings, for 12 weeks. A total of 110 patients were included in the intent-to-treat (ITT) analysis, with n = 54 in the dHACM group and n = 56 in the no-dHACM group. Of the participants, 98 completed the study per protocol, with 47 receiving dHACM and 51 not receiving dHACM. The primary study outcome was percentage of study ulcers completely healed in 12 weeks, with both ITT and per-protocol participants receiving weekly dHACM significantly more likely to completely heal than those not receiving dHACM (ITT-70% versus 50%, P = 0.0338, per-protocol-81% versus 55%, P = 0.0093). A Kaplan-Meier analysis was performed to compare the time-to-healing performance with/without dHACM, showing a significantly improved time to healing with the use of allograft, log-rank P < 0.0187. Cox regression analysis showed that dHACM-treated subjects were more than twice as likely to heal completely within 12 weeks than no-dHACM subjects (HR: 2.15, 95% confidence interval 1.30-3.57, P = 0.003). At the final follow up at 16 weeks, 95% of dHACM-healed ulcers and 86% of healed ulcers in the no-dHACM group remained closed. These results confirm that dHACM is an efficacious treatment for lower extremity ulcers in a heterogeneous patient population.

The amniotic membrane (AM) has a long history of use in the treatment of various diseases of the ocular surface. It contains pluripotent cells, highly organized collagen, anti-fibrotic and anti-inflammatory cytokines, immune-modulators, growth factors, and matrix proteins. It is used to promote corneal healing in severely damaged eyes. Recently, AM extract and AM extract eye drops have been successfully used in clinical applications, including dry eye and chemical burns. We provide an overview on the recent progress in the preparation, mechanisms of action, and use of AM extract/AM extract eye drops for corneal and external eye diseases.

This paper aims to discuss various therapeutical strategies (corneal cross-linking, amniotic membrane transplantation with or without autologous serum application, medical regenerative therapy) for treating ocular surface diseases according to medical indications, etiology, local and general status of the patient. Besides the evolution and treatment of the lesions induced by corneal foreign body, ocular burns, neurotrophic keratitis, pterygium removal, Mooren’s ulcer, this paper also follows and evaluates the migration, stratification and development of corneal epithelium. Of course, the success of the treatment depended on the therapeutic approach, the cause of the disease, the status of the eye and the patients’ compliance. All cases presented had good results, proving once again, that a well-chosen therapeutic approach ensures the improvement or cure for many ocular surface diseases nowadays. Abbreviations: BVCA = best corrected visual acuity, OCT = optical coherence tomography, CLX = corneal cross-linking

Conventional glaucoma surgeries, such as trabeculectomy and glaucoma drainage device (GDD) surgery, have been enhanced by surgeons to improve outcome and decrease complications. Over the last two decades, adjuncts, such as collagen matrix implants, fibrin adhesives, and amniotic membrane transplantation (AMT) have been found to be effective in modulating fibrosis and scarring during the wound-healing process, reducing postoperative inflammation, and repairing bleb leakage or conjunctival erosion. The use of these tools provides several advantages when used in trabeculectomy, GDD surgery, and surface reconstruction associated with glaucoma surgery complications. Their use will be discussed in this review. How to cite this article: Lu LJ, Hall L, Liu J. Improving Glaucoma Surgical Outcomes with Adjunct Tools. J Curr Glaucoma Pract 2018;12(1):19-28.

WOS: 000429421300004, PubMed ID: 29576892, Objectives: To investigate the long-term efficacy and results of surgical management of conjunctival melanoma reconstructed with amniotic membrane transplantation. Materials and Methods: Conjunctival melanoma in 10 patients (5 female, 5 male) was totally excised with adjunctive cryotherapy to the surgical margins, corneal epitheliectomy with absolute alcohol in cases of corneal involvement, lamellar sclerectomy in cases with episcleral involvement, and ocular surface grafting with cryopreserved amniotic membrane. Complications and tumor control rates were evaluated. Results: The mean age of the patients was 57.4 +/- 15.2 (range, 37-84) years. The mean diameter of the tumors was 15.5 +/- 4.9 (range, 10-25) mm and histopathologically confirmed complete excision was performed in all cases. Mild limbal stem cell deficiency (2 eyes) and subclinical symblepharon (3 eyes) were observed as long-term complications. In a mean follow-up of 56.7 +/- 40.4 (range, 30-132) months, only one local tumor recurrence was detected. Despite retreatment, exenteration was performed in this patient due to re-recurrence. One patient died due to disseminated metastasis despite the absence of local recurrence. Conclusion: In large conjunctival melanomas, reconstruction of the ocular surface is usually very challenging. The use of cryopreserved amniotic membrane for conjunctival defect repair is safe and effective with mild complications, and allows the excision of wider margins around the tumor.

Study Design: Narrative literature review. Objectives: Placental tissue, amniotic/chorionic membrane, and umbilical cord have seen a recent expansion in their clinical application in various fields of surgery. It is important for practicing surgeons to know the underlying science, especially as it relates to spine surgery, to understand the rationale and clinical indication, if any, for their usage. Methods: A literature search was performed using PubMed and MEDLINE databases to identify studies reporting the application of placental tissues as it relates to the practicing spine surgeon. Four areas of interest were identified and a comprehensive review was performed of available literature. Results: Clinical application of placental tissue holds promise with regard to treatment of intervertebral disc pathology, preventing epidural fibrosis, spinal dysraphism closure, and spinal cord injury; however, there is an overall paucity of high-quality evidence. As such, evidence-based guidelines for its clinical application are currently unavailable. Conclusions: There is no high-level clinical evidence to support the application of placental tissue for spinal surgery, although it does hold promise for several areas of interest for the practicing spine surgeon. High-quality research is needed to define the clinical effectiveness and indications of placental tissue as it relates to spine surgery.

Simple limbal epithelial transplantation (SLET) is an emerging technique for treating unilateral limbal stem cell deficiency. We report the high-resolution, anterior segment optical coherence tomography (OCT) features of the first 2 weeks of a patient undergoing SLET for an old acid injury of the right eye, repeatedly from postoperative day 1 through day 14. Three out of 11 explants with the subjacent human amniotic membrane (hAM) and the overlaid bandage contact lens were imaged. The hAM was intact and of the same thickness throughout the study period; the sub-hAM space increased from day 3 to 9 and disappeared by day 10; the explants started thinning from day 3 with the fibrin around them starting to decrease from day 2 and completely disappeared by day 4. Epithelialization occurred between day 8 and 14 and proceeded more rapidly towards the limbus than centrally. There was no change of the corneal stromal thickness or reflectivity. This case report uses high-definition, spectral-domain OCT to document the events on the ocular surface after a successful SLET surgery and opens up an avenue to study epithelialization in a convenient and noninvasive manner.