Muscle Grafts with Doxorubicin Pretreatment Produce "Empty Tubes" in the Basal Laminae, Promote Contentious Maturation of the Regenerated Axons, and Bridge 20-mm Sciatic Nerve Defects in Rats.

BACKGROUND: We newly developed a muscle graft that employs a doxorubicin pretreatment technique. The aims of this study were to reveal the biological and morphological features of the muscle tissue in the second week (Study I), to reveal the regeneration outcomes of functional and kinematic assessments of longer-term follow-up (16 weeks, Study II), and to make assessments of the muscle graft with doxorubicin pretreatment in the critical-sized nerve defect model (20 mm, Study III). METHODS: A total of 26 adult rats were used in this study. Doxorubicin treatment was accomplished by immersion in a doxorubicin solution for 10 minutes followed by a rinsing procedure. The rats were divided into three groups: the muscle graft with and without doxorubicin pretreatment (M-graft-w-Dox and M-graft-w/o-Dox) groups and the autologous nerve graft (N-graft) group. Assays of apoptosis, immunofluorescent histochemistry including CD68 (macrophage marker), scanning electron microscopy (SEM), morphometrical studies of the regenerated axons, nerve conduction studies, and kinematic studies were performed. RESULTS: The M-graft-w-Dox group contained significantly larger numbers of apoptotic cells and CD68-positive cells. SEM revealed the existence of the basal lamina, so called "empty tubes," in the M-graft-w-Dox group. Study II showed contentious maturation of the regenerated axons, especially in the compound muscle action potentials. Study III showed that even at 20 mm, the M-graft-w-Dox group promoted axonal regeneration and functional regeneration. CONCLUSION: The M-graft-w-Dox group showed superior regeneration results, and this easy and short-term procedure can expand the muscle graft clinical indication for the treatment of peripheral nerve defects.

Autologous internal limiting membrane transplant for recurrent idiopathic macular holes.

PURPOSE: To report a series of recurrent idiopathic macular holes treated by means of a free autologous internal limiting membrane flap and compare visual and anatomic results to a control group undergoing further internal limiting membrane peeling and novel gas tamponade. METHODS: Retrospective surgical series of 15 consecutive patients receiving autologous internal limiting membrane flap compared to 14 patients operated on for internal limiting membrane peeling enlargement. Autologous internal limiting membrane flap was created after brilliant blue G staining, internal limiting membrane lifting, perfluorocarbon bubble injection and creation of a wide internal limiting membrane free flap translocated underneath perfluorocarbon liquid, to the macular hole bed. Both groups were tamponated with 20% SF6 and positioned face down for 4 h a day for 3 days. RESULTS: Macular hole closed in 14/15 (93.3%) patients of the autologous internal limiting membrane group and 9/14 (64.2%) controls (p < 0.05). Visual acuity increased from 0.05 ± 0.03 to 0.23 ± 0.13 Snellen in the autologous internal limiting membrane group and from 0.05 ± 0.03 to 0.14 ± 0.10 Snellen of controls (p < 0.05 for both). Vision of the autologous internal limiting membrane group improved more than controls at 1 month (p = 0.043) and 3 months (p = 0.045). Inner segment/outer segment interruption at 3 months was smaller in the autologous internal limiting membrane group than controls, reducing from 1230 ± 288 µm at baseline to 611 ± 245 and 547 ± 204 µm at 3 months versus 1196 ± 362, 745 ± 222 and 705 ± 223 µm, respectively (p < 0.05). CONCLUSION: Autologous internal limiting membrane flap can effectively close recurrent idiopathic macular holes with a higher closure rate, smaller residual inner segment/outer segment line interruption and higher visual acuity at 3 months than previous standard of care.

ANATOMICAL AND VISUAL OUTCOMES OF INVERTED INTERNAL LIMITING MEMBRANE FLAP TECHNIQUE VERSUS INTERNAL LIMITING MEMBRANE PEELING IN MYOPIC MACULAR HOLE WITHOUT RETINAL DETACHMENT: A Preliminary Retrospective Study.

PURPOSE: To compare the results of vitrectomy with internal limiting membrane (ILM) peeling and inverted ILM flap for treating myopic macular hole without retinal detachment. METHODS: Twenty-eight eyes of 28 patients undergoing vitrectomy with either ILM peeling (n = 16) or inverted ILM flap technique (n = 12) were included. Outcomes were myopic macular hole closure by optical coherence tomography and visual acuity at 6 months and at the end of follow-up. RESULTS: Closure of myopic macular hole was achieved in 13 eyes (81.2%) of the ILM peeling group and in 11 eyes (91.7%) of the inverted ILM flap group. The median length of follow-up was 18 months in the peeling group and 10.3 in the inverted group. There were not statistically significant differences between restoration of the external limiting membrane, external limiting membrane and ellipsoid zone, and none of both layers between the two groups. The median best-corrected visual acuity (logarithm of minimal angle of resolution) at the end of follow-up was 0.25 (20/35 Snellen) in the peeling group and 0.4 (20/50) in the inverted group (P = 0.027). CONCLUSION: Both techniques were associated with high closure rates of myopic macular hole but the small sample size and the retrospective design prevents any claims of superiority of one technique over the other.

Folding the internal limiting membrane flap under perfluorocarbon liquid in large, chronic and myopic macular holes.

PURPOSE: The purpose of the present paper is to describe a surgical technique aimed at creating multiple layers of Internal Limiting Membrane (ILM) using Perfluorocarbon Liquid (PFCL) in order to favour the closure of large, chronic and myopic Macular Holes (MH). METHODS: Thirty patients belonging to 3 subgroups: large (> 500 µm), chronic (> 12 months) and myopic (> - 9 diopters), MHs, underwent surgery and completed 6 months follow-up. The ILM was engaged and peeled 360° around the MH, hinged to the rim and folded over the hole. A PFCL bubble spanning the vascular arcades was then injected and the ILM is grasped repeatedly to fold the distal edge towards the MH centre, creating multiple ILM layers over the MH. RESULTS: MH closed in 26/30 cases (86.6%) with no significant difference among subgroups. Vision improved 2.57 ± 1.56 Snellen lines from LogMAR 1.50 ± 1.19 to 1.19 ± 1.32 (p < 0.01). Ellipsoid Zone (EZ) interruption width reduced from 1129 ± 439 µm to 258 ± 507 µm (p < 0.001) and correlated to pattern of MH closure, post-BCVA and line improvement (p < 0.001 in all cases). DISCUSSION: The use PFCL allows multiple ILM layers and resulted in a high closure rate. Pattern of MH closure differs from those previously described leaving a plug of ILM tissue that interrupts retinal architecture often only in the inner layers. PFCL gravity and hydrophobicity displace aqueous while the intensely polar opposite faces of the ILM attract each other. The folded ILM plugs MH and bridges the gap and may help glial cell proliferation and migration.

Internal Limiting Membrane Transposition for Persistent Macular Holes Using Double Layers of Viscoelastic.

Internal limiting membrane (ILM) grafting provides a useful option for repair of large and refractory macular holes that fail to close following prior ILM removal. However, current ILM graft techniques are associated with several challenges that may result in failure, most notably the difficulty in maintaining the graft in situ. In this video, the authors describe their modified technique for ILM grafting using a double layer of viscoelastic for stabilization in situ during the procedure. Four of five eyes managed with this technique demonstrated type 1 closure, and all eyes demonstrated improvement in visual acuity.

Dynamic intraoperative optical coherence tomography for inverted internal limiting membrane flap technique in large macular hole surgery.

BACKGROUND/OBJECTIVES: To assess the efficacy of dynamic intraoperative spectral-domain optical coherence tomography (iSD-OCT) imaging for inverted internal limiting membrane (ILM) flap technique (IILMFT) in large macular hole (MH) surgery. SUBJECTS/METHODS: Prospective, non-randomized, observational study was conducted on 8 eyes of 7 patients with large, chronic and recurrent MHs, which were treated by pars plana vitrectomy (PPV) with IILMFT. All patients underwent standard pre- and postoperative examination. The iSD-OCT imaging was performed using microscope integrated systems before, during, and after ILM peeling. The iSD-OCT data were post-processed using graphic software and reviewed for tissue behavior and instruments position. RESULTS: The real-time iSD-OCT-assisted IILMFT allowed for real-time imaging of the entire surgery with visualization of the MH, vitreoretinal instruments, and all steps of inverted ILM flap formation. In spite of shadowing created by the steel instruments, it was possible to follow and control the distance between the instrument tips and retinal layers. Dynamic imaging of the surgical maneuvers including ILM peeling and mechanical apposition of MH edges revealed the iatrogenic impact on the retina (depression and appearance of hyporeflective zones). iSD-OCT imaging could confirm the proper position of the inverted ILM flap at the very end of the surgery after fluid-air exchange. CONCLUSIONS: iSD-OCT imaging is an effective tool for learning and performing a well-controlled and safe inverted ILM flap technique in patients with large MH. Clinical significance of the structural iSD-OCT findings has to be further studied.

Urea-De-Epithelialized Human Amniotic Membrane for Ocular Surface Reconstruction.

The conjunctiva is a clear tissue covering the white part of the eye and lines the back of the eyelids. Conjunctival diseases, such as symblepharon, cause inflammation, discharges, and photophobia. The treatment often requires excision of large parts of conjunctiva. Tissue engineering of conjunctival cells using human amniotic membrane (HAM) denuded of its epithelium as a basement membrane scaffold has been shown to be effective for covering conjunctival defects. However, most epithelial denudation protocols are time-consuming and expensive or compromise HAM's basement membrane structure and matrix components. We have previously described a method to de-epithelialize HAM using ice-cold urea (uHAM). In this report, we used this method to provide tissue-engineered constructs with cultivated conjunctival epithelial cells on uHAM in two patients, one with a giant conjunctival nevus and the other with a large symblepharon. Autologous conjunctival epithelial cells harvested from incisional biopsies of these two patients were cultured on the uHAM scaffold. The transplantation of tissue-engineered constructs to patients' ocular surface immediately after the removal of lesions showed successful reconstruction of the ocular surface. Postoperatively, there were neither recurrence of lesions nor epithelial defects throughout the follow-up (up to 7 and 19 months, respectively). This report highlights the translational potential of an efficient and inexpensive method to prepare de-epithelialized HAM as a basement membrane scaffold for cell-based tissue-engineered treatments of ocular surface disorders. Stem Cells Translational Medicine 2019;8:620&626.

One-Year Results after Internal Limiting Membrane Flap Transposition for Surgical Repair of Macular Holes with Respect to Microperimetry.

PURPOSE: Internal limiting membrane (ILM) flap transposition in surgical repair of macular holes represents a new technique offering good anatomical success rates with large macular holes. The aim of the study was to examine microperimetric outcomes 1 year after ILM flap transposition for surgical repair of macular holes. METHODS: Patients with idi-opathic macular holes scheduled for 23-G pars plana vitrectomy with ILM peeling, ILM flap transposition, and SF6 tamponade were examined in a prospective case series. Distance corrected visual acuity (DCVA), optical coherence tomography, and microperimetry were measured before and 1 year after surgery. RESULTS: In all eyes, closure of the macular hole could be achieved. While foveal sensitivity improved in 71%, perifoveal sensitivity improved in 86% of the cases. DCVA improved in 83% and was unchanged in 17% of the cases. CONCLUSIONS: Beside visual acuity, foveal and perifoveal macular sensitivity improved, underlining the functional success of surgery after ILM peeling with ILM flap transposition.

Autologous internal limiting membrane flap for retinal detachment due to posterior retinal tears over choroidal atrophy in highly myopic eyes.

PURPOSE: To review a series of highly myopic eyes with retinal detachment undergoing pars plana vitrectomy with autologous internal limiting membrane (ILM) flap placed over posterior retinal breaks located in areas of choroidal atrophy. METHODS: Retrospective review of 13 consecutive patients receiving pars plana vitrectomy with ILM flap over causative breaks, compared with 19 controls receiving the same surgery with ILM peeling but no ILM flap. Main outcome measures included anatomical success rate, visual acuity, number of surgeries and the rate of silicone oil removal. RESULTS: Patients in the ILM group required 2.08±0.37 interventions versus 2.58±0.75 in the control group (p=0.037). One (1/13; 7.6%) patient in the ILM group required additional unplanned surgery versus 8/19 (42.10%) in the control group (p=0.038). Final anatomical success rate defined as attached retina after silicone oil (SiO) removal was 13/13 in the I-ILM group and 14/19 (73.6%) in the control group (p=0.052). No patients (0/13) in the I-ILM group retained SiO at the end of follow-up versus 4/19 (21.1%) patients in the control group (p=0.061). Best-corrected visual acuity at the end of follow-up was logMAR 0.65±0.36 (20/91 Snellen) in the ILM group and logMAR 0.89±0.44 (20/158 Snellen) in the control group (p=0.20). CONCLUSION: Autologous ILM may help seal posterior retinal breaks and improve the surgical prognosis of retinal detachment due to breaks located over areas of choroidal atrophy within the myopic staphyloma.

Microstructural retinal regeneration after internal limiting membrane flap surgery for repair of large macular holes: a 1-year follow-up study.

PURPOSE: To evaluate long-term change in functional and structural outcomes after successful repair of large macular holes (MH) with internal limiting membrane (ILM) flap techniques. METHODS: Eleven consecutive patients were reviewed over a 1-year time period after the successful repair of large MH with ILM flap techniques. SD-optical coherence tomography (SD-OCT) images were taken to assess the anatomical outcome after surgery, while the best-corrected visual acuity (BCVA) was tested using Snellen charts to evaluate the functional outcome. Each patient was evaluated at 1, 6 and 12 months after surgery, respectively. RESULTS: All cases achieved complete anatomical closure. All patients showed a microstructural regeneration of the retina with a decrease in ellipsoid zone defects over the 1-year follow-up. Functionally, as compared to baseline, all of the patients showed improvements in best-corrected visual acuity of 1-4 lines at the final examination after 12 months post-operatively. CONCLUSIONS: Long-term results show further improvement in the best-corrected visual acuity as well as further microstructural regeneration of the retina and decrease in ellipsoid zone defects over time. The exact mechanism, which promotes closure of the macular hole and reconstruction of the ellipsoid zone after internal Limiting Membrane autograft surgery, still remains unknown.

Spare some internal limiting membrane for later: free ILM patch and neurosensory retina graft.

PURPOSE: Editorial to De Giacinto et al case report on free autologous neurosensory retina patch. METHODS: Literature review and experts' opinion RESULTS: In the present issue, De Giacinto et al describe a free autologous neurosensory retina patch to close a chronic macular hole. This new technique was made necessary by an extended internal limiting membrane peeling during the first surgery, that prevented grafting a patch of internal limiting membrane when the hole did not close. We hereby review pros and cons of patching a chronic macular hole with an internal limiting membrane patch, as well as the importance of not over-enlarging a peeling. DISCUSSION: Internal limiting membrane patch can be considered in chronic macular holes. It may not be an option in cases of over-enlargement of a previous peel; free autologous neurosensory retina patch may be a valid alternative in such cases.

Comparison of Inverted ILM-Stuffing Technique and ILM Peeling Alone for Optic Disc Pit-Associated Maculopathy: Long-Term Results.

BACKGROUND AND OBJECTIVE: To evaluate long-term outcomes of inverted internal limiting membrane (ILM) stuffing into the optic disc pit for optic disc pit maculopathy (ODP-M) compared with vitrectomy with ILM peeling alone. PATIENTS AND METHODS: Twelve eyes of 12 patients who underwent vitrectomy for ODP-M and followed up more than 12 months were included. We retrospectively analyzed outcomes of inverted ILM flap stuffing into ODP (group 1, n = 6) with ILM peeling alone (group 2, n = 6). RESULTS: At 12 months, both groups showed significant improvement in best-corrected visual acuity and central macular thickness. On comparison, significantly faster resolution of maculoschisis was found in group 1 compared with group 2 (P = .012). CONCLUSIONS: Pars plana vitrectomy (PPV) with ILM peeling and PPV with inverted ILM stuffing are effective treatment modalities for ODP-M, with inverted ILM stuffing showing faster resolution of maculoschisis. [Ophthalmic Surg Lasers Imaging Retina. 2018;49:e226-e232.].

27-Gauge Via Pars Plana Vitrectomy With Autologous ILM Transplantation for Optic Pit Disc Maculopathy.

To report a case of a serous macular detachment associated with optic pit that did not improve after a previous vitrectomy with internal limiting membrane (ILM) peeling, peripapillary laser, and gas tamponade. The authors performed a lens-sparing 27-gauge pars plana vitrectomy with autologous transplantation of ILM inside the optic nerve pit and gas tamponade. The subretinal fluid gradually resolved and visual acuity improved to 20/20 at the tenth month after surgery. The authors did not observe a recurrence of subretinal fluid during the 14 months of follow-up. Autologous transplantation of the ILM may be effective to repair recurring optic disc pit maculopathy. [Ophthalmic Surg Lasers Imaging Retina. 2018;49:712-714.].

Contralateral Autologous Internal Limiting Membrane Transplantation for Closure of a Refractory Macular Hole: Surgical Technique.

BACKGROUND AND OBJECTIVE: To determine the efficacy of a new surgical technique involving use of a contralateral autologous internal limiting membrane (ILM) free flap for closure of a refractory macular hole (MH). PATIENTS AND METHODS: Closure could not be achieved after several MH surgeries in a 72-year-old woman with an epiretinal membrane (ERM) in the right eye and a chronic MH in the left eye. Therefore, the authors performed surgery on the left eye using a contralateral autologous ILM free flap obtained during ERM surgery performed on the right eye. RESULTS: The refractory MH had disappeared, and the patient's corrected visual acuity was improved in both eyes by 4 months after surgery. CONCLUSION: When a suitable free flap cannot be obtained from the operated eye, contralateral autologous ILM transplantation has the potential to improve functional and anatomical outcomes in an eye with a refractory MH. [Ophthalmic Surg Lasers Imaging Retina. 2018;49:e75-e77.].

Non-inverted pedicle internal limiting membrane transposition for large macular holes.

PURPOSE: The purpose of this study is to investigate the effectiveness of a new surgical technique of non-inverted pedicle internal limiting membrane (ILM) transposition for the treatment of eyes with large macular hole. METHODS: This is a retrospective, consecutive, interventional case series. Twelve eyes of 12 consecutive patients who underwent vitrectomy for the treatment of a large macular hole (MH size > 400 µm) were treated. ILM was peeled and left with a pedicle attached to the superior temporal retina. The macular hole was covered by transposition of the pedicle ILM in a non-inverted way. Preoperative and postoperative best-corrected visual acuity (BCVA), SD-OCT image, macular sensitivity by microperimetry, and multifocal electroretinogram (mERG) response were evaluated. All of the patients were followed for more than 3 months. RESULTS: Postoperative OCT examination confirmed 11 of 12 macular hole closed (91.7%). Six macular hole filled with silicone oil closed as early as the next day. The postoperative BCVA significantly increased compared with preoperative BCVA (P = 0.002). The improvement of macular sensitivity within 2° and 8° circle was also statistical significant (P = 0.018 and P = 0.017, respectively). Fixation stability, shown as the percentage of fixation point within the 2° circle and 4° circle, was remarkably improved (P = 0.017 and P = 0.018, respectively). The R1/R2 and R1/R4 ring ratios also increased significantly as compared with that of baseline. CONCLUSION: These findings indicate that the non-inverted pedicle ILM transposition results in a high incidence of anatomic closure with good visual outcome for the treatment of large macular hole.

Active Cone Regeneration Following Autologous Internal Limiting Membrane 'Chunk' Transplantation in Optic Disc Pit-Associated Maculopathy.

This case report aims to study active cone regeneration and its functional correlation in optic disc pit maculopathy (ODPM) with a large outer macular defect treated with autologous internal limiting membrane (ILM) "chunk" transplantation. Outcome analysis was done using spectral-domain optical coherence tomography (SD-OCT), adaptive optics, and microperimetry. Visual acuity improved from counting fingers at 2 meters to 6/18 Snellen equivalent postoperatively, whereas SD-OCT demonstrated closure of outer subfoveal defect. Adaptive optics imaging revealed maximum active cone concentration of 14,232/mm2 at the fovea, and fixation stability improved to 61% and 90% (P1 and P2, respectively) on microperimetry. To conclude, autologous ILM "chunk" transplantation provides an additional therapeutic option in treatment of ODPM with large outer macular defect. [Ophthalmic Surg Lasers Imaging Retina. 2018;49:152-155.].

Inverted Internal Limiting Membrane Technique Maintains Macular Hole Closure in Retinal Detachment Following Macular Hole Repair.

Recent advances in macular hole surgery have significantly improved the rates of anatomical success. One important advance is the dye-assisted peeling of the internal limiting membrane (ILM) around the hole. In large holes (<400 µ), sometimes, ILM peeling alone is not enough to achieve closure, and this may be improved by inverting part of the peeled membrane and using it to cover the macular hole or inverting and tucking the ILM flap into the hole. In this case report, a patient with large full-thickness macular hole underwent a hole repair with an inverted ILM flap technique. The eye developed a subsequent retinal detachment with large collection of subretinal fluid beneath the repaired hole, but the hole remained intact, suggesting that the adhesive forces binding an inverted flap to the edges of a repaired macular hole appear to be strong enough to maintain structural integrity of the closed hole even in the presence of a macula-involving retinal detachment with large subretinal fluid collection.