Descemet's membrane substrate from human donor lens anterior capsule

Kopsachilis, Nikolaos; Tsinopoulos, Ioannis; Tourtas, Theofilos; Kruse, Friedrich E.; Luessen, U. Welge

doi:10.1111/j.1442-9071.2011.02678.x

Cited by 22 publications

(25 citation statements)

References 30 publications

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“…Transplanting the LC with the epithelial side down resulted in a more rapid recovery and a transparent cornea at 6 months when compared with control eyes. LC has also been used successfully as a scaffold for the expansion of both limbal epithelial stem cells and HCEnC in vitro (Kopsachilis et al, ; Lachaud et al, ; Navaratnam et al, ; Yoeruek et al, ). These data suggested that LC could eventually be used for human transplantation, but prior to in vivo use, further characterization is required.…”

Section: Discussionmentioning

confidence: 99%

“…Anterior capsular material could potentially be obtained as a by‐product of cataract surgery, one of the most frequently performed surgeries in the world. Preliminary studies have investigated the growth of corneal derived cells on the LC, namely, corneal epithelial and endothelial cells (Albert et al, ; Kopsachilis, Tsinopoulos, Tourtas, Kruse, & Luessen, ; Yoeruek et al, ). These studies however did not examine the additional physical properties of the LC that could make it suitable as a corneal endothelial scaffold.…”

Section: Introductionmentioning

confidence: 99%

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Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering

Bogerd

Dhubhghaill

Zakaria

2018

J Tissue Eng Regen Med

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The idea of transplanting a sheet of laboratory‐grown corneal endothelium dates back to 1978; however, the ideal scaffold is still lacking. We hypothesized that human crystalline lens capsules (LCs) could qualify as a scaffold and aimed to characterize the properties of this material for endothelial tissue engineering. LCs were isolated from donor eyes, stored at −80 °C, and decellularized with water and trypsin‐EDTA. The decellularization was investigated by nuclear staining and counting and the capsule thickness was determined by optical coherence tomography and compared with Descemet's membrane (DM). Transparency was examined by spectrometry, and collagenase degradation was performed to evaluate its resistance to degradation. Cell‐scaffold interaction was assessed by measuring focal adhesions surface area on LC and plastic. Finally, primary corneal endothelial cells were grown on LCs to validate the phenotype. Trypsin‐EDTA decellularized most effectively, removing 99% of cells. The mean LC thickness was 35.76 ± 0.43 μm, whereas DM measured 25.93 ± 0.26 μm (p < .0001). Light transmission was 90% for both LC and DM. On a collagenase challenge, LC and amniotic membrane were digested after 13 hr, whereas DM was digested after 17 hr. The surface area of focal adhesions for cells grown on coated LCs was at least double that compared with other conditions, whereas tight junctions, ion pumps, and hexagonal morphology were well maintained when endothelial cells were cultured on LCs. In conclusion, LCs demonstrate excellent scaffolding properties for tissue engineering and sustain the cell phenotype and can be considered a suitable substrate for ocular tissue engineering or as a template for future scaffolds.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering

Bogerd

Dhubhghaill

Zakaria

2018

J Tissue Eng Regen Med

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“…Recently, the human lens capsule of donor globes was used as a scaffold for cultivation for human endothelial cells (Kopsachilis et al. ). In the present series, we therefore used the porcine lens capsule, which notably formed a roll when submerged in fluid (similarly to the stripped‐off human DM); in addition, it was resistant to tearing and easy to prepare.…”

Section: Discussionmentioning

confidence: 99%

A simple ex vivo model for teaching Descemet membrane endothelial keratoplasty

Droutsas

Petrak

Melles

et al. 2014

Acta Ophthalmologica

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ABSTRACT.Purpose: To describe a simple ex vivo porcine model for teaching Descemet Membrane Endothelial Keratoplasty (DMEK). Methods: Twenty freshly enucleated porcine eyes were used to create a pseudograft from the porcine lens capsule and implant it into an intact porcine globe using the standardized 'no-touch' technique of the Netherlands Institute for Innovative Ocular Surgery (NIIOS). Results: The DMEK pseudo-graft could be inserted, centred, unfolded and appositioned in all cases. Conclusions: Practicing DMEK surgery under wetlab conditions was feasible by the described simple and low-cost model. This new tool may facilitate the adaptation of DMEK by cornea surgeons.

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“…8 Previous in vitro studies have reported the use of denuded DM and human anterior lens capsules (HALCs), as well as bioengineered matrices consisting of collagen or gelatin, or a combination of biopolymers, as potential carriers for cultured CECs. [9][10][11][12][13][14][15][16][17] These cell-carrier constructs should preferably be similar to an "original" DMEK-graft, since it has been reported for Descemet stripping (automated) endothelial keratoplasty (DSEK/DSAEK) that thicker grafts may interfere with visual outcome 18 and the biomechanical properties of cell-carrier constructs should allow implantation.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of the Suitability of Biocompatible Carriers as Artificial Transplants Using Cultured Porcine Corneal Endothelial Cells

Spinozzi

Miron

Bruinsma

et al. 2018

Current Eye Research

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Purpose/Aim: Evaluating the suitability of bioengineered collagen sheets and human anterior lens capsules (HALCs) as carriers for cultivated porcine corneal endothelial cells (pCECs) and in vitro assessment of the cell-carrier sheets as tissue-engineered grafts for Descemet membrane endothelial keratoplasty (DMEK). Materials and Methods: pCECs were isolated, cultured up to P2 and seeded onto LinkCell™ bioengineered matrices of 20 µm (LK20) or 100 µm (LK100) thickness, and on HALC. During expansion, pCEC viability and morphology were assessed by light microscopy. ZO-1 and Na + /K +-ATPase expression was investigated by immunohistochemistry. Biomechanical properties of pCEC-carrier constructs were evaluated by simulating DMEK surgery in vitro using an artificial anterior chamber (AC) and a human donor cornea without Descemet membrane (DM). Results: During in vitro expansion, cultured pCECs retained their proliferative capacity, as shown by the positive staining for proliferative marker Ki67, and a high cell viability rate (96 ± 5%). pCECs seeded on all carriers formed a monolayer of hexagonal, tightly packed cells that expressed ZO-1 and Na + /K +-ATPase. During in vitro surgery, pCEC-LK20 and pCEC-LK100 constructs were handled like Descemet stripping endothelial keratoplasty (DSEK) grafts, i.e. folded like a "taco" for insertion because of challenges related to rolling and sticking of the grafts in the injector. pCEC-HALC constructs behaved similar to the DMEK reference model during implantation and unfolding in the artificial AC, showing good adhesion to the bare stroma. Conclusions: In vitro DMEK surgery showed HALC as the most suitable carrier for cultivated pCECs with good intraoperative graft handling. LK20 carrier showed good biocompatibility, but required a DSEKadapted surgical protocol. Both carriers might be notional candidates for potential future clinical applications.

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Descemet's membrane substrate from human donor lens anterior capsule

Abstract: Human donor anterior lens capsule might therefore be a potential scaffold for the ex vivo expansion of human corneal endothelial cells.

Cited by 22 publications

References 30 publications

Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering

Characterizing human decellularized crystalline lens capsules as a scaffold for corneal endothelial tissue engineering

A simple ex vivo model for teaching Descemet membrane endothelial keratoplasty

Evaluation of the Suitability of Biocompatible Carriers as Artificial Transplants Using Cultured Porcine Corneal Endothelial Cells

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