Reconstruction of corneal stroma with decellularized porcine xenografts in a rabbit model

Abstract

ABSTRACT.Purpose: To evaluate the potential use of decellularized porcine stromal matrix (PSM) for reconstruction of corneal stroma in a rabbit model. Methods: Ten chinchilla bastard rabbit corneas were exposed to a circular half-thickness keratotomy with a 3.0 mm diameter at the central cornea. Porcine corneas were decellularized using hypotonic tris buffer, ethylene diamine tetraacetic acid (EDTA, 0.1%), aprotinin (10 K IU ⁄ ml) and 0.3% sodium dodecyl sulphate (SDS). The 3.0 mm in diameter decellularized corneal stromal xenograft was inserted into the pocket, and the incision was closed with four 10.0 nylon sutures. Clinical photographs were taken at day 1, day 7, day 30 and on a monthly basis for up to 6 months after transplantation. Six months after surgery, the rabbits were killed and eyes were enucleated. Haematoxylin-eosin (HE) and 4,6-diamidino-2-phenylindole (DAPI) staining were performed to confirm the complete removal of the corneal cells after decellularization of porcine corneas and repopulation with rabbit cells. Alcian blue staining was performed for analysing the structure of the extracellular matrix (ECM). Results: Efficient elimination of porcine cells was achieved by our decellularization protocol and confirmed via HE and DAPI stainings. Moreover, the major histoarchitectural ECM structure had been maintained as visualized by the alcian blue stain. Finally, the PSM was biocompatible with the host's epithelium evidenced as a regrowth covering the exposed xenograft. Conclusions: This novel technique of tissue engineering may provide one of many solutions to addressing anterior corneal pathological conditions in the face of a shortage of human corneal material.

A Fish Scale–Derived Collagen Matrix as Artificial Cornea in Rats: Properties and Potential

“…These corneal substitutes range from completely synthetic prostheses, which primarily aim to restore the cornea's refractive function, 7,8 and tissue-engineered cell-based constructs, 9 to hydrogels and scaffolds that facilitate the regeneration of the host tissue. [10][11][12] Although much progress has been realized, keratoprostheses have not reached widespread use. 8 At the moment, they are too expensive and complex for routine use in developing countries, where the need for implants is highest.…”

“…A decellularized porcine corneal matrix as a xenographic scaffold for corneal regeneration has been studied for several years by several research groups, as it closely resembles the human corneal stromal organization. 12,20,21 Indeed, such porcine matrices may offer a relative inexpensive and widely available alternative to human donor corneas. Results from clinical trials are not yet available, but a first phase I clinical trial is currently running.…”

Dextran Preserves Native Corneal Structure During Decellularization

“…The uniform diameter and high level of lateral order of these fibrils gives the cornea its transparency (26). A simple method to evaluate the maintenance of this architecture, that is often employed, is to soak corneas in glycerol after decellularization (20,56,64) and evaluate changes macroscopically and by measuring light transmittance. Transparency was enhanced in all corneas after soaking in glycerol, however the dextran treated corneas had a similar level of light transmittance to the native cornea that was also treated in glycerol suggesting that the histoarchitecture of the stroma was maintained for these groups, similar results were obtained in the HHP studies using dextran.…”

In-body tissue-engineered collagenous connective tissue membranes (BIOSHEETs) for potential corneal stromal substitution

“…The extracellular matrix of the corneal stroma consists primarily of collagen type I with a lower amount of collagen type IV and proteoglycans (Knupp et al, 2009;Hassell and Birk, 2010). Therefore, collagenous biomaterials such as collagen gels, and acellular porcine corneas are widely used as potential corneal substitutes (Liu et al, 2007;Fagerholm et al, 2009Fagerholm et al, , 2010Merrett et al, 2009;Duncan et al, 2010;Tanaka et al, 2011;Xiao et al, 2011;Yoeruek et al, 2011). As alternative approach, autogenic transplantation of collagen-rich tissues such as ligaments, tendons, and skin have been widely applied for the replacement of damaged tissue (Jackson and Simon, 2002;Antonogiannakis et al, 2005;de Vries Reilingh et al, 2007;Carey et al, 2009).…”

“…However, biological integration with the surrounding recipient tissue is still a major problem, and restoration of sensory or physiological corneal function has not been achieved. Alternatively, many groups have attempted to develop tissue-engineered corneal equivalents (Liu et al, 2007;Fagerholm et al, 2009Fagerholm et al, , 2010Merrett et al, 2009;Duncan et al, 2010;Tanaka et al, 2011;Xiao et al, 2011;Yoeruek et al, 2011). An ideal scaffold for a tissue-engineered corneal equivalent should be biocompatible, non-immunogenic, non-mutagenic, sufficiently strong to withstand surgical procedures and optically clear (Feinberg, 2012).…”