Abstract
Limbal Stem Cell Deficiency (LSCD) is a very serious and painful disease that often results in impaired vision. Cultivation of limbal stem cells for clinical application is usually performed on carriers such as amniotic membrane or surgical fibrin gel. Transplantation of these grafts is associated with the risk of local postoperative infection that can destroy the graft and devoid therapeutic benefit. For this reason, electrospun scaffolds are good alternatives, as proven to mimic the natural cells surroundings, while their fabrication technique is versatile with regard to polymer functionalization and scaffolds architecture. This study considers the development of poly(ε-caprolactone) (PCL) immune-compatible and biodegradable electrospun scaffolds, comprising cefuroxime (CF) or titanium dioxide (TiO2) active components, that provide both bactericidal activity against eye infections and support of limbal stem cells growth in vitro. The PCL/CF scaffolds were prepared by blend electrospinning, while functionalization with the TiO2 particles was performed by ultrasonic post-processing treatment. The fabricated scaffolds were evaluated in regard to their physical structure, wetting ability, static and dynamic mechanical behaviour, antimicrobial efficiency and drug release, through scanning electron microscopy, water contact angle measurement, tensile testing and dynamic mechanical analysis, antimicrobial tests and UV-Vis spectroscopy, respectively. Human limbal stem cells, isolated from surgical remains of human cadaveric cornea, were cultured on the PCL/CF and PCL/TiO2 scaffolds and further identified through immunocytochemistry in terms of cell type thus were stained against p63 marker for limbal stem cells, a nuclear transcription factor and cytokeratin 3 (CK3), a corneal epithelial differentiation marker. The electrospun PCL/CF and PCL/TiO2 successfully supported the adhesion, proliferation and differentiation of the cultivated limbal cells and provided the antimicrobial effect against Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans.
Highlights
Tissue damaged in pathological process or after some trauma is usually treated with autografts, allografts or implantable devices
The results obtained in this study study are comparable to other similar studies concerning tissue engineering application
The study focuses on the development of electrospun PCL/CF and PCL/TiO2 scaffolds with an embedded antimicrobials against common pathogens causing eye infections, to support the adhesion, proliferation and growth of cultivated limbal stem cells for corneal tissue regeneration
Summary
Tissue damaged in pathological process or after some trauma is usually treated with autografts, allografts or implantable devices. Tissue engineering is an interdisciplinary field that emerged almost three decades ago with the goal to offer solutions to the issues mentioned above [1]. It concerns the replacement of injured tissues via living cells, temporarily grown onto a scaffolding material. Engineered tissue should acquire architectural complexity of targeted anatomical site. It must be porous enough for the host cells to infiltrate and deposit their own components of extracellular matrix (ECM). A scaffold should provide a tissue specific niche for stem cells to grow and exchange activating and inhibitory signals via autocrine, paracrine and endocrine modes [7]
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