• Combination of Microstereolithography and Electrospinning to Produce Membranes Equipped with Niches for Corneal Regeneration

      Ortega, Í.; Sefat, Farshid; Deshpande, P.; Paterson, T.; Ramachandran, C.; Ryan, A.J.; MacNeil, S.; Claeyssens, F. (2014)
      We report a technique for the fabrication of micropockets within electrospun membranes in which to study cell behavior. Specifically, we describe a combination of microstereolithography and electrospinning for the production of PLGA (Poly(lactide-co-glycolide)) corneal biomaterial devices equipped with microfeatures.
    • An “off-the shelf” Synthetic Membrane to Simplify Regeneration of Damaged Corneas

      Sefat, Farshid; Ortega, Í.; McKean, R.; Deshpande, P.; Ramachandran, C.; Hill, C.J.; Tzokov, S.B.; Claeyssens, F.; Sangwan, V.S.; Ryan, A.J.; et al. (2014)
      Our overall aim is to develop a synthetic off-the-shelf alternative to human amniotic membrane which is currently used for delivering cultured limbal stem cells to the cornea in patients who suffer scarring of the cornea because of the loss of limbal stem cells. We have recently reported that both cultured cells and limbal explants grow well on electrospun Poly(D,L-lactide-co-glycolide) (PLGA) (44 kg/mol) with a 50:50 ratio of lactide and glycolide and sterilized with γ-irradiation. Prior to undertaking a clinical study our immediate aim now is to achieve long term storage of the membranes in convenient to use packaging. Membranes were electrospun from Poly(D,L-lactide-co-glycolide) (44 kg/mol) with a 50:50 ratio of lactide and glycolide and sterilized with γ-irradiation and then stored dry (with desiccant) for several months at -80°C and -20°C , Room temperature (UK and India), 37°C and 50°C. We explored the contribution of vacuum sealing and the use of a medical grade bag (PET/Foil/LDPE) to achieve a longer shelf life. Confirmation of membranes being suitable for clinical use was obtained by culturing tissue explants on membranes post storage. When scaffolds were stored dry the rate of breakdown was both temperature and time dependent. At -20°C and -80°C there was no change in fiber diameter over 18 months of storage, and membranes were stable for 12 months at 4°C while at 50°C (above the transition temperature for PLGA) scaffolds lost integrity after several weeks. The use of vacuum packaging and a medical grade bag both improved the storage shelf-life of the scaffolds. The impact of temperature on storage is summarized beneath. We report that this synthetic membrane can be used as an off-the-shelf or-out-of-the freezer alternative to the amniotic membrane for corneal regeneration.