Decellularized Human Cornea for Reconstructing the Corneal Epithelium and Anterior Stroma

Shafiq, M. Zubair; Gemeinhart, Richard A.; Yue, Beatrice Y.J.T.; Djalilian, Ali R.

doi:10.1089/ten.tec.2011.0072

Cited by 109 publications

(130 citation statements)

References 39 publications

(36 reference statements)

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“…These agents were selected due to their decellularization power and in agreement with previous reports describing the use of detergents for decellularization of animal corneas or other organs. SDS and Triton X-100 have been widely applied to decellularize corneas, 12,[17][18][19][20][21] and Igepal and BAK were selected for the first time to determine their usefulness for cornea decellularization. Igepal has been previously used for decellularizing heart valves, 22 pericardium, 23 has never been used as a decellularization agent; nonetheless, it is a potent detergent employed as preservative in many drugs, like glaucoma eye drops, which associates high levels of cytotoxicity.…”

Section: Discussionmentioning

confidence: 99%

“…Corneas were washed thoroughly with 10% antibiotic-antimycotic solution (Invitrogen-Gibco, Carlsbad, CA) in phosphate-buffered saline (PBS) for 10 minutes and then washed in PBS. Several independent decellularization protocols based on benzalkonium chloride (BAK), Igepal, SDS, or Triton X-100 (all of them from Sigma-Aldrich, Steinheim, Germany) were applied to the NPCs at different concentrations (0.01%, 0.05%, and 0.1%) and times (12,24, and 48 hours) for each of the agents used, except for control NPCs that remained as native nondecellularized corneas. Afterwards, porcine corneas were washed with PBS for 24 hours.…”

Section: Decellularization Of Porcine Corneasmentioning

confidence: 99%

“…Decellularized porcine corneas (DPC) were then placed between two pieces of absorbent paper and incubated in a dry chamber at 608C for 1 hour to eliminate the excess liquid. In total, 33 different study groups were established, and three samples were analyzed per condition: native nondecellularized corneas; corneas decellularized with BAK at three concentrations (0.01%, 0.05%, and 0.1%) for three different times (12,24, and 48 hours); corneas decellularized with Igepal at three concentrations (0.01%, 0.05%, and 0.1%) for three different times (12,24, and 48 hours); corneas decellularized with SDS at three concentrations (0.01%, 0.05%, and 0.1%) for three different times (12,24, and 48 hours); corneas decellularized with Triton X-100 at three concentrations (0.01%, 0.05%, and 0.1%) for three different times (12,24, and 48 hours).…”

Section: Decellularization Of Porcine Corneasmentioning

confidence: 99%

“…11 In a previous work, we demonstrated the efficacy of sodium dodecyl sulfate (SDS) and sodium chloride (NaCl) to decellularize porcine corneal stromas after using dispase II. 10 Afterwards, Shafiq et al 12 modified our method based on NaCl by adding nucleases to the protocol, and they demonstrated that this protocol could be associated to undetectable levels of cellular material in human corneas, supporting both fibroblast and epithelial cell growth in vitro. On the other hand, we recently demonstrated that the use of decellularized extracellular matrix (ECM) obtained from extraocular tissues, such as the small intestine could permit generating bioengineered cornea substitutes with potential clinical usefulness and translational perspectives.…”

Section: Introductionmentioning

confidence: 99%

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Effects of Detergent-Based Protocols on Decellularization of Corneas With Sclerocorneal Limbus. Evaluation of Regional Differences

González‐Andrades

Carriel

Rivera-Izquierdo

et al. 2015

Trans. Vis. Sci. Tech.

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Citation: González-Andrades M, Carriel V, Rivera-Izquierdo M, et al. Effects of detergent-based protocols on decelluarization of corneas with sclerocorneal limbus. Evaluation of regional differences. Tran Vis Sci Tech. 2014;4(2):13, http://tvstjournal. org/doi/full/10.1167/tvst. 4.2.13, doi: 10.1167/tvst.4.2.13 Purpose: In this work, we decellularized whole porcine corneas including the sclerocorneal limbus (SCL) and we evaluated regional differences in order to identify an efficient method to decellularize whole corneas for future clinical use. Methods:We analyzed the efficiency of four decellularization protocols based on benzalkonium chloride (BAK), Igepal, sodium dodecyl sulfate (SDS), and Triton X-100 detergents on whole porcine corneas.Results: Results showed that the decellularization efficiency of most protocols was low, with the specific protocol resulting in more efficient levels of decellularization being 0.1% SDS for 48 hours, especially in the medium and posterior cornea regions. A significant correlation was found between the decellularization efficiency and the concentration of agent used (P ¼ 0.0174; r ¼ 0.1540), but not for the incubation time (P . 0.05). The analysis of cornea components preservation demonstrated that all protocols were able to preserve the integrity of the Bowman's layer and Descemet's membrane. Although the collagen structure was partially altered, the global decellularization groups showing highest preservation of the ECM collagen contents and orientation were Igepal and SDS, with differences among the three regions of the cornea. All global groups showed high levels of proteoglycan and glycoprotein preservation after decellularization, with the best results were found in the SDS group followed by the Igepal group.Conclusions: These results suggest that very powerful protocols are necessary for whole-cornea decellularization. For the generation of lamelar corneas for clinical use, decellularization regional differences should be taken into account.Translational Relevance: Decellularized whole corneas may be potential therapeutic agents for lamelar keratoplasty.

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Section: Discussionmentioning

confidence: 99%

Section: Decellularization Of Porcine Corneasmentioning

confidence: 99%

Section: Decellularization Of Porcine Corneasmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effects of Detergent-Based Protocols on Decellularization of Corneas With Sclerocorneal Limbus. Evaluation of Regional Differences

González‐Andrades

Carriel

Rivera-Izquierdo

et al. 2015

Trans. Vis. Sci. Tech.

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“…The native goat-lung tissue (control) containing cells showed positive nuclear staining by DAPI stain. These methods have been used repeatedly by several researchers for assessment of cell removal from the decellularized tissues [61][62][63]. DNA quantification and DAPI staining thus verified the efficient removal of cells from the goat-lung tissue demonstrating the applicability of decellularized goatlung matrix as a scaffold.…”

Section: Decellularization Of Goat-lung Matrix and Its Verificationmentioning

confidence: 96%

Antibacterial activity and composition of decellularized goat lung extracellular matrix for its tissue engineering applications

Gupta¹,

Dinda²,

Mishra³

2017

Biol Eng Med

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In this study, scaffold was fabricated from goat-lung of cadaver goat by decellularization method for its potential application in tissue engineering. The cellular components of goat-lung were removed by employing trypsin-SDS based method of decellularization. DNA quantification and DAPI staining verified the effective removal of native cells from the goat-lung. Verhoeff-Van Gieson (VVG) staining, H&E staining and Periodic-Acid Schiff (PAS) staining histologically assessed the composition of native and decellularized goat-lung matrix-demonstrating the presence of collagen, elastin and glycoproteins in the decellularized goat-lung matrix. Antibacterial activity of decellularized goat-lung matrix was evaluated against gram-negative (Escherichiacoli) and gram-positive (Staphylococcusaureus) bacteria, which shows the presence of naturally occurring bioactive peptides-exhibiting the antibacterial activity against E. coli for up to 9h and against S. aureus for up to 5h. The decellularized goat-lung matrix, as demonstrated by MTT assay, was found to be biocompatibile to L929 mouse fibroblasts, HepG2, human skin derived mesenchymal stem cells (hS-MSCs), osteoblasts and BMMSCs. All the above results confer the applicability of the decellularized goat-lung matrix as a potential scaffold for tissue engineering applications.

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Designing Scaffolds for Corneal Regeneration

Ahearne

Fernández‐Pérez

Masterton

et al. 2020

Adv Funct Materials

121

109

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Corneal blindness is one of the most common causes of vision loss worldwide, affecting millions of people. To treat these patients, researchers have been examining different approaches to engineer corneal scaffolds suitable for transplantation. Scaffolds have been developed to replace part or all of the cornea depending on the patient requirements. Both acellular and cell-seeded scaffolds have been tested in animal models. Materials that have been under investigation for manufacturing scaffolds include collagen, silk fibroin, amniotic membrane, decellularized cornea, fibrin, chitosan, gelatin, agarose, alginate, and hyaluronic acid in addition to several synthetic polymers. Different combinations of materials, fiber crosslinking techniques, and incorporation of bioactive molecules have also been examined. Factors such as the physical properties, cytocompatibility, degradation behavior, and optical characteristics have to be considered when selecting a suitable scaffold material. Recent advancements in materials fabrication techniques such as bioprinting, electrospinning, and different collagen alignment techniques, allow scaffolds to be generated that more accurately mimic the structure of the corneal stroma. A number of scaffolds have commenced clinical trials to determine their suitability for corneal regeneration.

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Decellularized Human Cornea for Reconstructing the Corneal Epithelium and Anterior Stroma

Cited by 109 publications

References 39 publications

Effects of Detergent-Based Protocols on Decellularization of Corneas With Sclerocorneal Limbus. Evaluation of Regional Differences

Effects of Detergent-Based Protocols on Decellularization of Corneas With Sclerocorneal Limbus. Evaluation of Regional Differences

Antibacterial activity and composition of decellularized goat lung extracellular matrix for its tissue engineering applications

Designing Scaffolds for Corneal Regeneration

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