Evaluation of Healing Potential of Tissue Engineered Construct in Rabbit Corneal Ulcer Model
Loading...
Date
2023
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
SKUAST Kashmir
Abstract
The present study was aimed to explore the feasibility of development of the natural Acellular Corneal Stromal Disc (ACSD) and its biocompatibility and utility with the bone marrow derived mesenchymal stem cells (BM-MSCs) in repairing induced corneal stromal defect (ulcer) in rabbit model. The study was conducted in two phases viz., A) In vitro development and characterization of ovine sheep decellularized corneal matrix, and evaluation of its biocompatibility with the rabbit cryopreserved BM-MSCs through histopathology, MTT assay and scanning electron microscopy (SEM). B) In vivo evaluation of healing potential of the BM-MSCs seeded ACSD in the rabbit corneal ulcer defect (5 mm diameter) model. The animals (n=48 rabbits) were divided into four (4) groups (n=12 per group). Group I (Control, received no treatment), group II (standard clinical treatment), group III (transplanted with ACSD) and group IV (transplanted with ACSD seeded with cryopreserved allogeneic BM-MSCs). The animals were observed regularly and evaluated at days 14 and 28 using fluorescein dye test, ultrasonography, slit lamp biomicroscopy, histopathology, SEM, and relevant gene (keratocan and ALDH1A1) expression analysis. In vitro study revealed that ACSD were almost completely decellularized and supported the growth and infiltration of BM-MSCs. In vivo experimental corneal ulcer study showed transplantation of ACSD seeded BM-MSCs reduced corneal ulcer size together with improvement in corneal opacity and corneal neovascularization as compared to ACSD (Group III), standard clinical treatment (group II) and control. Ulcer healing evaluated with fluorescein dye test and histopathology revealed better corneal healing in group IV at days 14 and 28 as compared to other three groups. The gene expression revealed that keratocan and ALDH1A1 were highly expressed in group IV as compared to other groups. Corneal ultrasonography showed near normal corneal thickness in groups II and IV at the end of the experimental trial (day 28). In compliance to ultrasonography, SEM also revealed lower cellular infiltration in groups II and IV and regenerated corneal tissue maintained continuity and had least crevices in group IV at the end of study. It is worth mentioning that the complete normalization of corneal ulcer at day 28 was not seen in any group including the group IV. It was concluded that development of sheep corneal tissue engineered construct (ACSD) is a feasible option that supports growth of BM-MSCs. The cell-scaffold assembly potentially heals corneal ulcer although the time required for complete corneal ulcer healing remains to be seen.
Description
Keywords
Gene expression, Mesenchymal stem cells, Corneal ulcer, Fluorescein Dye Test, Ultrasonography, Biomicroscopy, Decellularized, Healing, Tissue engineered, Crevices, Veterinary Surgery and Radiology