Poly(ε-caprolactone) Titanium Dioxide and Cefuroxime Antimicrobial Scaffolds for Cultivation of Human Limbal Stem Cells

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.

Bioactivity Comparison of Electrospun PCL Mats and Liver Extracellular Matrix as Scaffolds for HepG2 Cells

“…Electrospinning techniques offer the possibility to tailor scaffolds of various chemical and physical properties as well as topography [ 25 ]. The PCL mats used in this work were electrospun with the same parameters applied in our previous work with primary cells [ 6 ]. To obtain ECM scaffolds, decellularization of pork liver tissue was carried out using slightly adapted protocol described in [ 8 , 14 ].…”

“…PCL is aliphatic polyester composed of repeated units of hexanoate. Although its biocompatibility is rather low, its physical properties and adjustable biodegradability, including ease of blending and copolymerisation, make it a desirable cell scaffold material as well as drug delivery particles [ 4 , 5 , 6 ]. The major limitation of PCL matrices is hydrophobicity, which might affect cell adhesion efficiency and duration of biodegradability.…”

Trends of Corneal Transplantation in Adults from 2010 to 2019 in East China: A 10-Year Experience

“…Nevertheless, the long-term outcome of KLA is not satisfactory [25]. With the rapid development of stem cell therapy, KLA has been gradually replaced by ex vivo cultivated limbal epithelial transplantation [26] and simple limbal epithelial transplantation [27, 28].…”

Profile of biological characterizations and clinical application of corneal stem/progenitor cells

“…However, due to the disadvantages of HAM (such as high thickness, variable transparency, biodegradation and immunosuppression), it is urgent for us to find alternative cell carriers/scaffolds to substitute for HAM. At present, alternative cell carriers/scaffolds contain modified HAM, collagen[ 159 , 160 ], fibrin[ 161 ], siloxane hydrogel contact lenses, human lens capsules, poly (ε-caprolactone)[ 162 ], onion epithelial membrane scaffolds[ 163 ], carboxymethyl cellulose (CMC)[ 164 ], electrospun polycaprolactone/gelatine nanocomposites[ 165 ] and other emerging materials. The modified HAM will be described in detail below.…”