Engineering a Corneal Stromal Equivalent Using a Novel Multilayered Fabrication Assembly Technique

Fernández‐Pérez, Julia; Madden, Peter; Ahearne, Mark

doi:10.1089/ten.tea.2020.0019

Cited by 17 publications

(19 citation statements)

References 50 publications

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“…These include cell-based approaches, biomimetic hydrogels, and molecular approaches. Various studies have shown the promise of different hydrogels (with and without cells), as a viable alternative to stromal replacement with donor tissue (33)(34)(35)(36). Others have shown the importance of various secretory molecules like exosomes (37), anti-TGF-β (7,(38)(39), anti-PDFG (7,(40)(41), and HGF (42)(43) in preventing or reverting the corneal scars.…”

Section: Discussionmentioning

confidence: 99%

Safety and toxicity of human limbus-derived stromal/mesenchymal stem cells with and without alginate encapsulation for clinical application

Damala

Pakalapati

Singh

et al. 2021

Preprint

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Background The purpose of this study was to assess the ocular and systemic toxicity of topically applied human limbus-derived stromal/mesenchymal stem cells (hLMSCs) with and without alginate encapsulation as per Indian regulatory guidelines for stem cell therapy. Methods The hLMSCs were obtained from cadaveric corneoscleral rims and expanded in a current good manufacturing practice compliant laboratory. The hLMSCs were checked for viability, chromosomal stability, growth kinetics, contamination, and endotoxin levels. Cells with (En+ hLMSCs) or without (En− hLMSCs) alginate encapsulation were used for the animal experiments. The study involved 3 groups of 6 New Zealand white rabbits each, which underwent corneal wounding followed by treatment with sham (G1), En− hLMSCs (G2), and En+ hLMSCs cells (G3). Ophthalmic assessment including intraocular pressure (IOP), blood investigations and inflammatory marker (IL-6, TNF-α, IgE) expression in serum and tears were assessed on days 1, 7, 14, 21, and day 28. At the end of 28 days, the animals were sacrificed, and the organs were subjected to histopathological examination. Results The hLMSCs had 88.33 ± 2.37% viability at the end of 6 hours and 78.21 ± 1.47% at the end of 24 hours. The cells showed positive expression for the stem-cell biomarkers (p63α, Pax6, and ABCG2), extracellular matrix marker (Col-III) and mesenchymal biomarkers (VIM, CD73, CD90 and CD105). No contamination by the Mycoplasma species was found in either of the En-/En + hLMSCs and the levels of bacterial endotoxins in the En- hLMSCs and En + hLMSCs cell suspension was found be within the permissible levels (≤ 0.12 EU/mL). Ophthalmic examination showed no significant difference in IOP, corneal clarity and conjunctival congestion between the three groups at every time point. Haematological parameters were comparable between the three groups. The inflammatory markers in tear and serum (TNF-α and IL-6) were not significantly elevated in the groups receiving En+/En− hLMSCs. Histological examination did not show any abnormality in the ocular or corneal tissue, and the viscera. Conclusions The results of the study show that hLMSCs do not cause any local or systemic toxicity in recipients, implying that these cells are safe for clinical use and their efficacy can be assessed in human clinical trials.

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Section: Discussionmentioning

confidence: 99%

Safety and toxicity of human limbus-derived stromal/mesenchymal stem cells with and without alginate encapsulation for clinical application

Damala

Pakalapati

Singh

et al. 2021

Preprint

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“…Solutions should be used at room temperature. 5. Wash the eye globes 3 times for 2 min each in PBS and keep moist during further stages.…”

Section: Methodsmentioning

confidence: 99%

Multilayered Fabrication Assembly Technique to Engineer a Corneal Stromal Equivalent

Fernández‐Pérez¹,

Madden²,

Ahearne³

2021

BIO-PROTOCOL

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Tissue engineering has emerged as a strategy to combat the donor shortage of human corneas for transplantation. Synthetic corneal substitutes are currently unable to support the normal phenotype of human cells and so decellularized animal corneas have been deployed to more closely provide the topographical and biochemical cues to promote cell attachment and function. Although full thickness decellularized corneas can support corneal cells, the cells are slow to populate the scaffold and density declines from the surface. To avoid these problems, this protocol describes the stacking of alternate layers of decellularized porcine corneal sheets and cell-laden collagen hydrogel to produce a corneal construct. The sheets are obtained by cryosectioning porcine corneas, decellularizing them with detergents and nucleases and finally air drying for storage and ease of manufacture. Corneal stromal cells are then encapsulated in a collagen type I solution and cast between these sheets. This protocol presents a rapid method to ensure high cellularity throughout the construct using tissue-derived materials alone.

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“…18,[27][28][29][30][31] In recent years, there have also been an increased effort in developing bioengineered scaffolds for replacing the need for native corneas. [32][33][34] These scaffolds are composed of either natural biomaterials (e.g., type 1 collagen, silk, and gelatin) or synthetic materials (e.g., polyvinyl alcohol and polyethylene glycol) or a mix of both, and have the ability to help deliver cell therapy, including HCEnC therapy. 32,33 However, these biomimetic scaffolds are subject to considerable translational and regulatory challenges, which will not be discussed in this review.…”

Section: Overview Of Corneal Endothelial Diseases and Potential Impact Of Cell Therapymentioning

confidence: 99%

Translational and Regulatory Challenges of Corneal Endothelial Cell Therapy: A Global Perspective

Ting

Peh

Adnan

2022

Tissue Engineering Part B: Reviews

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Cell therapies are emerging as a unique class of clinical therapeutics in medicine. In 2015, Holoclar (ex vivo expanded autologous human corneal epithelial cells containing stem cells) gained the regulatory approval for treating limbal stem cell deficiency after chemical eye burn. This has set a precedent in ophthalmology and in medicine, reinforcing the therapeutic promise of cell therapy. However, to generalize and commercialize cell therapies on a global scale, stringent translational and regulatory requirements need to be fulfilled at both local and international levels. Over the past decade, the Singapore group has taken significant steps in developing human corneal endothelial cell (HCEnC) therapy for treating corneal endothelial diseases, which are currently the leading indication for corneal transplantation in many countries. Successful development of HCEnC therapy may serve as a novel solution to the current global shortage of donor corneas. Based on the experience in Singapore, this review aims to provide a global perspective on the translational and regulatory challenges for bench-to-bedside translation of cell therapy. Specifically, we discussed about the characterization of the critical quality attributes (CQA), the challenges that can affect the CQA, and the variations in the regulatory framework embedded within different regions, including Singapore, Europe, and the United States.

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Engineering a Corneal Stromal Equivalent Using a Novel Multilayered Fabrication Assembly Technique

Cited by 17 publications

References 50 publications

Safety and toxicity of human limbus-derived stromal/mesenchymal stem cells with and without alginate encapsulation for clinical application

Safety and toxicity of human limbus-derived stromal/mesenchymal stem cells with and without alginate encapsulation for clinical application

Multilayered Fabrication Assembly Technique to Engineer a Corneal Stromal Equivalent

Translational and Regulatory Challenges of Corneal Endothelial Cell Therapy: A Global Perspective

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