Effects of a human amniotic membrane extract on ARPE-19 cells.

BACKGROUND: Human Amniotic Membrane (hAM) is endowed with several biological activities and might be considered an optimal tool in surgical treatment for different ophthalmic pathologies. We pioneered the surgical use of hAM to treat retinal pathologies such as macular holes, tears, and retinal detachments, and to overcome photoreceptor damage in age-related macular degeneration. Although hAM contributed to improved outcomes, the mechanisms of its effects are not yet fully understood. The characterization and explanation of the effects of hAM would allow the adoption of this new natural product in different retinal pathologies, operative contexts, and hAM formulations. At this end, we studied the properties of a hAM extract (hAME) on the ARPE-19 cells. METHODS AND RESULTS: A non-denaturing sonication-based technique was developed to obtain a suitable hAME. Viability, proliferation, apoptosis, oxidative stress, and epithelial-mesenchymal transition (EMT) were studied in hAME-treated ARPE-19 cells. The hAME was able to increase ARPE-19 cell viability even in the presence of oxidative stress (H2O2, TBHP). Moreover, hAME prevented the expression of EMT features, such as EMT-related proteins, fibrotic foci formation, and migration induced by different cytokines. CONCLUSIONS: Our results demonstrate that the hAME retains most of the properties observed in the whole tissue by others. The hAME, other than providing a manageable research tool, could represent a cost-effective and abundant drug to treat retinal pathologies in the future.

Refractive error after combined phaco-vitrectomy: A multicentric study.

PURPOSE: To study the post-operative refractive error (RE) of patients undergoing combined phaco-vitrectomy and to find out which intraocular lens (IOL)-power formula had the best refractive outcomes. METHODS: In this retrospective multicentric study we compared the preoperative expected target with the postoperative RE of patients undergoing combined phaco-vitrectomy due to vitreomacular traction, macular pucker, full thickness macular hole or lamellar macular hole. A multinomial logistic regression was performed to compare the postoperative REs and the differences between expected and postoperative REs among the SRK-T, Olsen's and Holladay-2 formulas. The correlation between the difference in REs and IOL-power was also studied. RESULTS: Sixty-seven eyes with a mean axial length of 23.73 ± 1.21 mm were included. Forty-two (63%), 14 (21%) and 11 (16%) eyes were implanted with an IOL that was calculated respectively with SRK-T, Olsen's and the Holladay-2 formula. The mean preoperative expected- and post-operative REs were -0.16 ± 0.12D and -0.48 ± 0.17, respectively (p = 0.045). SRK-T and Holladay-2 formulas led to a significant myopic shift whereas Olsen's caused a significant hyperopic error, independently from the IOL power. CONCLUSION: Independently from the IOL power, none of the analyzed formulas is precise at calculating the post-operative RE.

Optic Disk Pit Maculopathy Treatment Using a Human Amniotic Membrane Patch: One-Year Results.

PURPOSE: To report the 1-year results of human amniotic membrane patch implantation for optic disc pit maculopathy. DESIGN: A prospective, consecutive, interventional study. METHODS: Eleven eyes of 11 patients affected by optic disc pit maculopathy associated with subretinal/intraretinal fluid were included in this single-institution study. A 25-gauge pars plana vitrectomy was performed in all cases, with an implant of a human amniotic membrane patch into the optic disc pit and air was injected as endotamponade. The primary study outcome was the subretinal and intraretinal fluid reabsorption. Secondary outcomes were visual acuity improvement and postoperative complications. RESULTS: Mean central retinal thickness gradually diminished from 512 ± 137 µm to 243 ± 19 µm, at the 12-month follow-up. The mean visual acuity improved from 20/80 at baseline to 20/32 at the 12-month follow-up. Complete fluid resorption occurred in 9 of 11 (81.8%) eyes and there was partial resorption in 2 eyes (18%). No subretinal fluid recurrence was observed during the 12-month follow-up. No intraoperative or postoperative complications were reported during the follow-ups. The amniotic membrane patch remained detectable inside the pit for the entire follow-up time. CONCLUSION: An amniotic membrane plug may be effective for improving optic disc pit maculopathy. All cases had an anatomical improvement and encouraging visual acuity recovery.

Management of submacular massive haemorrhage in age-related macular degeneration: comparison between subretinal transplant of human amniotic membrane and subretinal injection of tissue plasminogen activator.

PURPOSE: Macular neovascularization (MNV) can complicate age-related macular degeneration (AMD) and lead to severe visual acuity reduction. Massive submacular haemorrhage (SMH) is a sight-threatening complication of MNV and a challenge in the management of complications related to MNV in AMD since the effects of anti-vascular endothelial growth factor treatment alone are insufficient. Here, we evaluate the different postoperative outcomes of patients affected by MNV complicated by SMH that underwent subretinal implant of human amniotic membrane (hAM) or subretinal injection of tissue plasminogen activator (tPA). METHODS: This is a retrospective, consecutive, comparative, non-randomized interventional study. We included 44 eyes of 44 patients affected by AMD complicated by MNV and SMH. Twenty-two eyes underwent a pars plana vitrectomy (PPV), SMH and neovascular membrane removal, with a subretinal implant of hAM and silicone oil, and 22 eyes underwent PPV, subretinal injection of tPA, and 20% sulphur hexafluoride. The primary study outcome was visual acuity improvement. Secondary outcomes were postoperative complications, and MNV recurrence and optical coherence tomography (OCT)-Angiography parameters correlated with best-corrected visual acuity (BCVA). RESULTS: Mean preoperative BCVA was 1.9 logarithm of the minimal angle of resolution (logMAR) in the amniotic membrane-group and 2 logMAR in the tPA-group. The mean final BCVA values were 1.25 and 1.4 logMAR, respectively, with a statistically significant difference. Optical coherence tomography (OCT)-Angiography scan was be used to evaluate the retinal vascularization in the treated eye. CONCLUSION: Both techniques report similar VA improvements and postoperative complications. However, transplantation of hAM seems to have a significant benefit in inhibiting MNV recurrence.

Via pars plana anterior iris enclavation IOL fixation.

AIM: To describe a via pars plana anterior iris enclavation intraocular lens (IOL) fixation technique. METHODS: A total of 35 consecutive aphakic vitrectomised patients (average age 71.12±10.12y) underwent pars plana vitrectomy (PPV) and via pars plana anterior iris enclavation IOL fixation. RESULTS: The mean preoperative best corrected visual acuity (BCVA) was 0.11±0.14 logMAR, the mean postoperative BCVA was 0.07±0.11 logMAR. The preoperative mean spherical equivalent was 7.22±4.21 D. The final mean spherical equivalent was -0.25±0.15 D. No eyes had hypotony, retinal or choroidal detachment or endophthalmitis. CONCLUSION: This technique may be a safe and useful in the case of aphakia, and a prospective study would be useful to confirm this findings.

Combining cataract surgery with 25-gauge high-speed pars plana vitrectomy: A prospective study.

PURPOSE: To compare visual outcome and postoperative complications of combined 25-gauge pars plana vitrectomy and phacoemulsification with vitrectomy alone surgery in patients with epiretinal membrane and macular hole. METHODS: A total of 110 eyes (73 eyes with epiretinal membrane and 37 eyes with macular hole) were enrolled in this prospective study. The eyes were divided into two groups: Group A included 55 eyes which underwent phacovitrectomy at the same time and Group B included 55 eyes which underwent vitrectomy alone. Follow-up visits were at 1, 3, and 6 months. RESULTS: The best-corrected visual acuity correlation by analysis of variance measurement showed statistically non-significant differences between the two groups (p = 0.32). The post hoc analysis from baseline, 1°, 2°, and 3° follow-ups was not statistically significant (p > 0.05). The most common postoperative complication was cystoid macular edema that has been detected in 11 patients (10%) (seven eyes in Group A and four eyes in Group B): 2 patients (1.8%) developed a chronic macular edema. Intraocular hypertension occurred in five eyes (4.5%) (three in Group A and two in Group B). Four eyes (3.6%) underwent another surgical procedure for a persistent macular hole (two in Group A and two in Group B). The intraocular lens repositioning was performed one day after surgery in three eyes (2.7%) (Group A). The mean preoperative visual acuity was not different between the two groups (p = 0.80). CONCLUSION: No significative differences between combined surgery and vitrectomy alone have been detected, in terms of postoperative complications and visual outcome.

HUMAN AMNIOTIC MEMBRANE TO TREAT MACULAR HOLES THAT FAILED TO CLOSE, SULFUR HEXAFLUORIDE ENDOTAMPONADE VERSUS AIR ENDOTAMPONADE: A Prospective Comparative Study.

BACKGROUND/PURPOSE: To determinate the efficacy of the human amniotic membrane plugs with sulfur hexafluoride versus human amniotic membrane plug with air as endotamponade to treat macular holes that failed to close after vitrectomy plus internal limiting membrane peeling. Multimodal imaging was focused to evaluate preoperative features and postoperative changes. METHODS: Prospective interventional comparative study. Twenty eyes of 20 patients affected with macular hole that failed to close were divided into 2 groups: 10 eyes received an amniotic membrane plug with 20% sulfur hexafluoride tamponade and 10 eyes received an amniotic membrane plug with air tamponade. All eyes were studied using multimodal advanced diagnostic tools, such as spectral-domain optical coherence tomography, optical coherence tomography angiography, microperimetry, and adaptive optics to investigate the postoperative results. RESULTS: In both groups, all macular holes were found successfully closed after 12 months. Mean preoperative best-corrected visual acuity was 20/400 in the SF6 group and 20/250 in air group. Final mean best-corrected visual acuity was 20/63 in both groups. The superficial capillary plexus, studied using optical coherence tomography angiography, showed a statistically significant difference between the treated and the fellow eyes. Adaptive optics images revealed the presence of a photoreceptor cell mosaic in the area of the amniotic membrane plug. CONCLUSION: The human amniotic membrane combined with air endotamponade demonstrated its effectiveness to seal macular holes that failed to close after vitrectomy plus internal limiting membrane peeling. Advanced multimodal diagnostic imaging helped us to better understand the modifications associated with the use of the amniotic membrane in these cases.

OCT Angiography Findings in Macula-ON and Macula-OFF Rhegmatogenous Retinal Detachment: A Prospective Study.

Background: The aim of the study was to evaluate pre-operative and post-operative retinal vasculature using optical coherence tomography angiography (OCTA) in patients who underwent rhegmatogenous retinal detachment (RRD) surgery repair. Materials and Methods: A total of 33 eyes were included in this prospective consecutive observational study: 15 affected by macula-ON and 18 by macula-OFF RRD. Superficial (SCP), deep capillary plexus (DCP), and foveal avascular zone (FAZ) area variations were evaluated by OCTA and correlated with visual acuity (VA) during a six-month follow-up. Results: In the macula-ON group, the preoperative vascular density (VD) of the whole SCP (wSCP) on affected eyes was lower than that of the fellow eyes (p < 0.05); this difference disappeared at 6 months after surgery (p = 0.88). The wSCP VD and the parafoveal SCP (pfSCP) VD increased during follow-up (p < 0.05); moreover, the higher the preoperative wSCP and pfSCP VD, the better the baseline VA (p < 0.05). In the macula-OFF group, at the first and sixth months after surgery, the larger the FAZ, the lower the VA (p < 0.05). Conclusions: Macula-ON SCP VD affected preoperative VA, and it was lower than the fellow eye, but recovered over time. In the macula-OFF group, a larger FAZ area was related to a worse VA, as is the case in diabetes and in retinal vein occlusion (RVO).

Applications of the Amniotic Membrane in Vitreoretinal Surgery.

Recently, the use of the human amniotic membrane (hAM) has been extended to treat retinal disorders, such as macular holes that failed to close and retinal tears. The hAM has demonstrated the induction of a recovery process of the external retinal layers involving the external limiting membrane (ELM) and the ellipsoid zone (EZ). After that, the application of the hAM for retinal pathologies was extended to large macular tears, high myopic retinal detachment associated with MH, paravascular tears, serous macular detachment associated with optic pit, complicated retinal detachment and advanced age-related macular degeneration (AMD). The hAM has shown a potential in repairing retinal tissue through a regeneration process. This review aims to highlight the use of the hAM in various vitreo-retinal surgical fields, and to confront it with other cutting-edge surgical techniques used to treat challenging vitreo-retinal pathologies.

Human Amniotic Membrane Plug to Restore Age-Related Macular Degeneration Photoreceptor Damage.

PURPOSE: Age-related macular degeneration (AMD) is the leading cause of legal blindness in adults 65 years of age and older. Choroidal neovascularization (CNV) can complicate AMD and lead to severe visual acuity reduction. Despite the several treatments available, if the retinal pigment epithelium is damaged, we have to cope with the impossibility of restoring acceptable visual acuity using only medical treatments. DESIGN: Prospective, consecutive, interventional study. PARTICIPANTS: Eleven patients affected by AMD, 6 patients affected by CNV, and 5 patients affected by geographic atrophy. METHODS: All patients underwent a pars plana vitrectomy with subretinal implantation of human amniotic membrane (hAM) to induce photoreceptor regeneration and partial visual acuity restoration. MAIN OUTCOME MEASURES: Primary study outcome was visual acuity improvement. Secondary outcomes were multimodal imaging results. RESULTS: Mean preoperative best-corrected visual acuity (BCVA) was 20/2000 (2 logarithm of the minimum angle of resolution [logMAR]), and all the patients showed a BCVA of counting fingers or less. Mean final BCVA was 20/400 (1.31 logMAR), ranging from 20/2000 to 20/100 (2-0.7 logMAR). OCT angiography was used to measure retinal vascularization in the treated eye compared with the fellow eye. A high correlation between BCVA and deep vascular density was evidenced. Adaptive optics findings, obtained over the retinal area where the highest functionality was observed, were evaluated using microperimetry. The images showed possible photoreceptor presence over the hAM membrane. CONCLUSIONS: This work supports the feasibility and safety of the hAM to promote partial retinal function restoration 6 months after surgery with visual acuity improvement. The advanced diagnostics help to understand the interaction between the hAM and photoreceptors and suggest that photoreceptor regeneration may occur.

Morpho-Functional Evaluation of Full-Thickness Macular Holes by the Integration of Optical Coherence Tomography Angiography and Microperimetry.

(1) Objective: To use optical coherence tomography angiography (OCTA) and microperimetry (MP) to evaluate the correlation between retinal structure and function in patients with idiopathic, full-thickness macular holes (FTMHs) (2) Methods: This prospective, observational study included 11 eyes of 10 patients with FTMHs evaluated before surgery using OCTA and MP. MP sensitivity maps were superimposed and registered on slabs corresponding to superficial capillary plexus (SCP) and deep capillary plexus (DCP) on OCTA, and on the outer plexiform layer (OPL) and the Henle fiber layer (HFL) complex in en face OCT. On these maps, mean retinal sensitivity was calculated at 2° and 4°, all centered on the FTMH. Cystic cavity extension was assessed on the slab corresponding to the OPL + HFL complex in en face OCT and DCP in OCTA using the Image J software (Version 1.49v; National Institutes of Health, Bethesda, MD, USA); (3) Results: Absolute scotomas were observed corresponding to the FTMH. Additionally, rings of relative scotoma in the perilesional area were detected and correlated to the cystic spaces on en face OCT and OCTA. There was a significant correlation between reduced retinal sensitivity at 2° and 4° diameters around the FTMH and the extension of cystic areas (p < 0.01). There was a significant correlation between the extension of cystic cavities and BCVA (p < 0.01). (4) Conclusions: Morpho-functional analysis of FTMH using OCTA and MP, and the correlation between vascular abnormalities and impaired retinal sensitivity, may provide new, useful information. This integrated evaluation of FTMH may be useful to determine the function-structure correlation before and after vitreoretinal surgery, in order to gain a better understanding of the functional consequences induced by the morphological alterations, assessing outcomes in a more objective way, and potentially adding new surgical prognostic factors.

A human Amniotic Membrane plug to manage high myopic macular hole associated with retinal detachment.

OBJECTIVE: To assess the efficacy of the human amniotic membrane (hAM) to treat (HMMH) associated with retinal detachment (RD). MATERIAL AND METHODS: Ten eyes of 10 patients with recurrent HMMH and RD, who had already undergone one or more pars plana vitrectomy (PPV), underwent a PPV with an hAM plug implanted in the macular hole. The initial five patients enrolled were tamponaded with (SO) while the subsequent five patients with 10% octafluoropropane (C3 F8 ). Silicon oil was removed in all five patients 2 months later. No statistical differences were reported between the two groups. RESULTS: Final retinal reattachment was achieved in all the patients. BCVA improved from 1.73 logMAR to 0.94 logMAR after 6 months. No adverse events were registered during follow-up. CONCLUSION: An hAM plug is an efficient substrate to manage HMMH associated with RD resulting in encouraging visual acuity recovery.

IOL repositioning using iris sutures: a safe and effective technique.

This retrospective non-comparative consecutive case series study was conducted at Azienda Ospedaliera Universitaria Careggi, Florence, Italy and describes a useful intraocular lens (IOL) repositioning technique using iris sutures. In our study, 41 consecutive cases of posteriorly dislocated IOLs were surgically treated between January 2015 and May 2017. Six of the cases were post-traumatic luxations, and 20 patients had pseudoexfoliation syndrome. All the patients underwent pars plana vitrectomy and same IOL repositioning using iris sutures. The mean follow-up was 12.2mo. The mean preoperative best corrected visual acuity (BCVA) was 0.10±0.15 logMAR, whereas the mean postoperative BCVA was 0.08±0.14 logMAR. The mean postoperative BCVA did not change significantly from the preoperative BCVA. The final mean spherical equivalent was -0.44±0.49 SD. Three lenses (7.31%) were found tilted during post-operative follow-up. Two eyes (4.87%) had postoperative cystoid macular edema. No eyes had endophthalmitis, hypotony, retinal or choroidal detachment. The iris fixation technique seems to be a safe and valid option for the management of dislocated IOLs.

Perfluorodecalin Versus Densiron 68 Heavy Silicone Oil in the Management of Inferior Retinal Detachment Recurrence.

BACKGROUND AND OBJECTIVE: To assess the efficacy of perfluorodecalin (PFD) or Densiron 68 heavy silicone oil (HSO) in the management of inferior complex retinal detachment recurrence. PATIENTS AND METHODS: A retrospective, comparative consecutive case series study. Twenty-four eyes of 24 patients affected by inferior complex retinal detachment recurrence underwent pars plana vitrectomy with PFD or HSO as endotamponade. All patients recruited were affected by complicated inferior retinal detachments and had already undergone at least one vitreoretinal procedure. The primary endpoint was anatomical success with primary and secondary surgery. The secondary endpoints were functional outcome and inflammatory complications. RESULTS: Out of 24 cases of inferior retinal detachment recurrence, 12 were tamponed with PFD (PFD group) and 12 with HSO (D68 group). Retinal reattachment rate at first surgery was 50% for the D68 group and 66.6% for the PFD group. Final reattachment rate after two or more surgical operations was 91.6% for the PFD group and 83.3% for the D68 group. Best-corrected visual acuity improved in both groups from a mean of 1.00 logMAR (20/200; standard deviation [SD]: 1) to 0.60 logMAR (20/80; SD: 0.3), and from a mean of 1.81 logMAR (20/2000; SD: 1.1) to a mean of 2.00 logMAR (20/2,000; SD:1) for the PFD and D68 groups, respectively. CONCLUSION: Both the endotamponades used showed good results in solving inferior retinal detachment recurrence with a slightly better rate in the PFD group, but it was not statistically significant (P > .05). [Ophthalmic Surg Lasers Imaging Retina. 2019;50:274-280.].

An Internal Limiting Membrane Plug and Gas Endotamponade for Recurrent or Persistent Macular Hole.

INTRODUCTION: Recurrent or persistent macular holes (MHs) are rare today due to the tendency to carefully peel the internal limiting membrane. Conversely, their treatment is still a challenge for a vitreoretinal surgeon. MATERIALS AND METHODS: This is a retrospective, consecutive, and nonrandomized study of patients affected by recurrent or persistent MHs treated using small-gauge pars plana vitrectomy (25- or 23-gauge) and an autologous ILM plug, at the Eye Clinic of Azienda Ospedaliera Universitaria Careggi (Florence, Italy) between January 2016 and May 2018. We included 8 eyes of 8 patients in the study. Five patients had a recurrent MH while 3 had a persistent MH. The case series includes patients with myopic eyes and with large macular holes (>400 µ). Patients were followed up with ophthalmoscopic examinations and swept-source optical coherence tomography (SS-OCT). RESULTS: The mean age of the patients was 74 years (±4.81 standard deviation (SD)), 3 patients were men and 5 women. The average axial length was 26.28 mm (±2.84 SD). Four patients had an AL ≧ 26 mm. The mean MH diameter was 436.5 (±49.82 SD). Average preoperative best-corrected visual acuity (BCVA) was 0.81 logMAR (±0.16 SD) and 20/125 Snellen. The ILM plug has been found integrated in the MH in all the follow-ups. CONCLUSION: In our study, an ILM autologous macular transplant was used successfully in 5 cases of macular hole recurrence and 3 cases of macular hole persistence. The anatomical success was achieved in all the cases; 4 patients improved their BCVA, and 4 patients maintained it. No macular alterations such as RPE or retinal atrophy/dystrophy were observed after 6 months.

A human amniotic membrane plug to repair retinal detachment associated with large macular tear.

PURPOSE: To describe a surgical technique, using a human amniotic membrane (hAM) plug, in two cases of retinal detachment with a large macular tear. METHODS: Surgical technique description with surgical video. A 23-gauge pars plana vitrectomy with peripheral retinectomy was performed. An hAM plug was implanted under the neuroretina to seal the posterior retinal break. Standard silicone oil tamponade was performed at the end of the surgery. The patients were positioned face down after the operation for the first 2 weeks. Optical coherence tomography (OCT) scans were performed in the follow-ups. RESULTS: The 1 week postoperative OCT showed a neuroretina ingrowth over the hAM plug. The 3-month OCT showed a regenerated neurosensory retina entirely covering the hAM plug. Visual acuity improved from light perception to 20/400 (LogMAR 1,3) 3 months after the operation in both patients. The silicone oil was extracted 4 months after surgery, and no recurrences were observed. The patients' visual acuity remained stable at 20/400 after the silicone oil extraction. CONCLUSION: In these complex cases, hAM transplantation can be a valid option not only to help the retinal reattachment but also for a partial regenerative effect which, in these cases, was accompanied by a visual acuity recovery.