In-situ porcine corneal matrix hydrogel as ocular surface bandage (2023)

Table of Contents

The Ocular Surface Abstract Purpose Methods Results Conclusions Graphical abstract Introduction Section snippets Fabrication of COMatrix ocular bandage hydrogel COMatrix hydrogel enhances attachment and proliferation of HCECs and attenuates TNF-α expression Discussion Conclusion Funding/support Commercial relationships disclosures Acknowledgement References (54) Regulation of corneal collagen fibrillogenesis in vitro by corneal proteoglycan (lumican and decorin) core proteins Exp Eye Res Management strategies for persistent epithelial defects of the cornea Saudi J Ophthalmol Current status of corneal xenotransplantation Int J Surg Simple physical model of collagen fibrillogenesis based on diffusion limited aggregation J Mol Biol Prior inhibition of AKT phosphorylation by BX795 can define a safer strategy to prevent herpes simplex virus-1 infection of the eye Ocul Surf Decellularized porcine conjunctiva as an alternative substrate for tissue-engineered epithelialized conjunctiva Ocul Surf Extracellular matrix hydrogels from decellularized tissues: structure and function Acta Biomater Decellularized porcine cornea-derived hydrogels for the regeneration of epithelium and stroma in focal corneal defects Ocul Surf Bio-orthogonally crosslinked hyaluronate-collagen hydrogel for suture-free corneal defect repair Biomaterials The application of human amniotic membrane in the surgical management of limbal stem cell deficiency Ocul Surf Corneal stromal wound healing: major regulators and therapeutic targets Ocul Surf Phase II randomized, double-masked, vehicle-controlled trial of recombinant human nerve growth factor for neurotrophic keratitis Ophthalmology Growth factors and corneal epithelial wound healing Brain Res Bull Progress in corneal wound healing Prog Retin Eye Res The impact of hyperglycemia on the corneal epithelium: molecular mechanisms and insight Ocul Surf Strategies for reconstructing the limbal stem cell niche Ocul Surf Limbal and corneal epithelial homeostasis Curr Opin Ophthalmol Corneal epithelial wound healing Exp Biol Med Corneal wound healing effects of mesenchymal stem cell secretome delivered within a viscoelastic gel carrier Stem Cells Transl Med Simultaneous interpenetrating polymer network of collagen and hyaluronic acid as an in situ-forming corneal defect filler Chem Mater In situ-forming collagen hydrogel crosslinked via multi-functional PEG as a matrix therapy for corneal defects Sci Rep Shear-induced alignment of collagen fibrils using 3D cell printing for corneal stroma tissue engineering Biofabrication Characterization of cornea-specific bioink: high transparency, improved in vivo safety J Tissue Eng Early observation of extracellular matrix-derived hydrogels for corneal stroma regeneration Tissue Eng C Methods Solubilized extracellular matrix bioscaffolds derived from diverse source tissues differentially influence macrophage phenotype J Biomed Mater Res Therapeutic effects of lyophilized conditioned-medium derived from corneal mesenchymal stromal cells on corneal epithelial wound healing Curr Eye Res Down-regulation of Notch signaling during corneal epithelial proliferation Mol Vis Cited by (13) Dual-crosslinked regenerative hydrogel for sutureless long-term repair of corneal defect Functionalized hydrogels in ophthalmic applications: Ocular inflammation, corneal injuries, vitreous substitutes and intravitreal injection Decellularized extracellular matrix: New promising and challenging biomaterials for regenerative medicine Critical media attributes in E-beam sterilization of corneal tissue Supramolecular host-guest hyaluronic acid hydrogels enhance corneal wound healing through dynamic spatiotemporal effects Preparation and biological evaluation of decellularized dermal matrix hydrogel Recommended articles (6) Targeted steroid ointment application to the lid margins in ocular graft-versus-host disease associated blepharitis treatment Long-term outcomes of the MICOF keratoprosthesis surgery Unapproved drugs – A fix to an unintended consequence of an FDA policy Human meibum and tear film derived (O-acyl)-omega-hydroxy fatty acids in meibomian gland dysfunction Ocular redness – II: Progress in development of therapeutics for the management of conjunctival hyperemia Chronic ocular complications in lamotrigine vs. trimethoprim-sulfamethoxazole induced Stevens-Johnson syndrome/toxic epidermal necrolysis

The Ocular Surface

Volume 21,

July 2021

, Pages 27-36

Author links open overlay panel, , , , , , , , ,

https://doi.org/10.1016/j.jtos.2021.04.004 Get rights and content

Abstract

Purpose

Bioactive substrates can be used therapeutically to enhance wound healing. Here, we evaluated the effect of an in-situ thermoresponsive hydrogel from decellularized porcine cornea ECM, COMatrix (COrnea Matrix), for application as an ocular surface bandage for corneal epithelial defects.

Methods

COMatrix hydrogel was fabricated from decellularized porcine corneas. The effects of COMatrix hydrogel on attachment and proliferation of human corneal epithelial cells (HCECs) were evaluated in vitro. The effect of COMatrix on the expressions of the inflammatory genes, IL-1β, TNF-α, and IL-6 was assessed by RT-PCR. The in-situ application and also repairing effects of COMatrix hydrogel as an ocular bandage was studied in a murine model of corneal epithelial wound. The eyes were examined by optical coherence tomography (OCT) and slit-lamp microscopy in vivo and by histology and immunofluorescence post-mortem.

Results

In vitro, COMatrix hydrogel significantly enhanced the attachment and proliferation of HCECs relative to control. HCECs exposed to COMatrix had less induced expression of TNF-α (P<0.05). In vivo, COMatrix formed a uniform hydrogel that adhered to the murine ocular surface after in-situ curing. Corneal epithelial wound closure was significantly accelerated by COMatrix hydrogel compared to control (P<0.01). There was significant increase in the expression of proliferation marker Ki-67 in wounded corneal epithelium by COMatrix hydrogel compared to control (P<0.05).

Conclusions

COMatrix hydrogel is a naturally derived bioactive material with potential application as an ocular surface bandage to enhance epithelial wound healing.

Graphical abstract

In-situ porcine corneal matrix hydrogel as ocular surface bandage (3)

Introduction

The stratified non-keratinized squamous epithelium of the cornea plays a critical role in preserving corneal function by providing a barrier against environmental damage and inhibiting angiogenesis and modulating immune response [1]. The corneal epithelium is dynamically rejuvenated following normal shedding of superficial cells or epithelial wounds due to trauma or infections. Corneal epithelial regeneration is orchestrated and regulated by many factors including cross-talk between corneal cells, immune cells, extracellular matrix (ECM) and secreted factors (e.g. growth factors) [2,3].

Many studies have shown the corneal epithelial restorative effects of growth factors such as epidermal growth factor (EGF), nerve growth factor (NGF) and hepatocyte growth factors (HGF), as well as secretomes from mesenchymal stem cells and extracellular-matrix (ECM)-rich scaffolds such as amnion [[4], [5], [6], [7], [8], [9], [10]]. Other groups have pursued the application of in-situ chemically cross-linked natural biomaterials such as collagen and hyaluronic acid for regeneration of corneal stroma and epithelium [[11], [12], [13]]. While there are several reports on the use of processed decellularized ECMs for focal corneal stromal regeneration [[14], [15], [16], [17]], the potential of corneal ECM hydrogel for enhancing the corneal epithelial wound repair as an ocular bandage, has not been studied. Processed (partially-digested) decellularized ECMs have been fabricated from several tissues including urinary bladder, heart, intestine, brain, testis, bone, cartilage, tendon, conjunctiva, and adipose tissue [[18], [19], [20]]. Processed ECMs not only possess regenerative effects, but also readily form in-situ thermoresponsive hydrogels with no further manipulations. We recently characterized the rheological behavior and the protein composition of a thermoresponsive hydrogel fabricated from decellularized porcine cornea ECM [54].

Cornea-derived ECM hydrogel can potentially enhance wound healing as an ocular bandage by providing a protective barrier that simultaneously provides factors to promote epithelial regeneration [54]. In this study, we evaluated the healing potential of fabricated in-situ thermoresponsive hydrogel, COMatrix (COrneal Matrix) from decellularized porcine cornea, as an ocular bandage to promote healing of corneal epithelial wounds. We investigated the effects of COMatrix hydrogel on attachment, proliferation, and inflammatory responses of human corneal epithelial cells (HCECs) in vitro while providing proof of concept application of COMatrix hydrogel as an in-situ ocular surface bandage in a murine model of corneal epithelial wound healing.

Section snippets

Fabrication of COMatrix ocular bandage hydrogel

Fresh intact porcine eyeballs were obtained from a certified abattoir (Park Packing Co. Inc., Chicago, IL). Under sterile conditions, the porcine corneas (PCs) were dissected and washed with Phosphate Buffered Saline (PBS, 1x) containing 1% gentamicin, 1% penicillin and 1% streptomycin.

The decellularization process for PCs were performed as described before with some modifications [15]. The PCs were cut into pieces with an average size of 2×2 mm². The tissue pieces were first stirred in 20mM

COMatrix hydrogel enhances attachment and proliferation of HCECs and attenuates TNF-α expression

The fabricated COMatrix hydrogel before and after thermal (heat) gelation and its transparency are shown in Fig. 1B. The bioactivity of COMatrix hydrogel was evaluated using in-vitro attachment, proliferation, and inflammatory cytokine induction assays on human corneal epithelial cells. To assess the interaction of human corneal epithelial cells with COMatrix, the attachment of cells to plates coated with hydrogel was compared to uncoated control plates (Fig. 2A). Microscopic observation of

Discussion

In this study, an in-situ thermoresponsive and transparent ocular bandage hydrogel, COMatrix, was fabricated for application to the ocular surface to promote regeneration of the corneal epithelium. The induction of cell proliferation by COMatrix hydrogel was shown in-vitro, followed by a proof of concept application as an ocular surface bandage in a murine corneal epithelial wound healing model.

Corneal epithelial wound healing is a dynamic process involving proliferation, migration, adhesion,

Conclusion

In this study, an in-situ thermoresponsive and transparent ocular bandage hydrogel, COMatrix, was fabricated from decellularized porcine cornea ECM as a regenerative biomaterial for corneal epithelial repair. The in-vitro and in-vivo experiments have shown the corneal epithelial regenerative effects of COMatrix hydrogel which provide proof of concept and potential for clinical translation. COMatrix hydrogel is a promising biomaterial which can potentially be used as an easy-to-apply ocular

Funding/support

This work was supported by R01 EY024349 (ARD), Core Grant for Vision Research EY01792 (MIR) from NEI/NIH; Unrestricted Grant to the Department and Physician-Scientist Award both from Research to Prevent Blindness; Eversight.

The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Commercial relationships disclosures

The authors have no commercial disclosure related to this work.

Acknowledgement

The authors would like to appreciate Lauren Kalinoski for the illustrations.

This study is extracted from PhD dissertation of G.Y.

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