Treatment of Ocular Surface Injuries by Limbal and Mesenchymal Stem Cells Growing on Nanofiber Scaffolds

Abstract

Stem cell (SC) therapy represents a promising approach to treat a wide variety of injuries, inherited diseases, or acquired SC deficiencies. One of the major problems associated with SC therapy remains the absence of a suitable matrix for SC growth and transfer. We describe here the growth and metabolic characteristics of mouse limbal stem cells (LSCs) and mesenchymal stem cells (MSCs) growing on 3D nanofiber scaffolds fabricated from polyamide 6/12 (PA6/12). The nanofibers were prepared by the original needleless electrospun Nanospider technology, which enables to create nanofibers of defined diameter, porosity, and a basis weight. Copolymer PA6/12 was selected on the basis of the stability of its nanofibers in aqueous solutions, its biocompatibility, and its superior properties as a matrix for the growth of LSCs, MSCs, and corneal epithelial and endothelial cell lines. The morphology, growth properties, and viability of cells grown on PA6/12 nanofibers were comparable with those grown on plastic. LSCs labeled with the fluorescent dye PKH26 and grown on PA6/12 nanofibers were transferred onto the damaged ocular surface, where their seeding and survival were monitored. Cotransfer of LSCs with MSCs, which have immunosuppressive properties, significantly inhibited local inflammatory reactions and supported the healing process. The results thus show that nanofibers prepared from copolymer PA6/12 represent a convenient scaffold for growth of LSCs and MSCs and transfer to treat SC deficiencies and various ocular surface injuries.

The Role of Mouse Mesenchymal Stem Cells in Differentiation of Naive T-Cells into Anti-Inflammatory Regulatory T-Cell or Proinflammatory Helper T-Cell 17 Population

“…The residual population of MSCs was characterized by a uniform spindle-shaped morphology, by the absence of CD11b + and CD45 + cells and by expression of phenotypic markers associated with murine MSCs. These purified MSCs had the ability to specifically differentiate into adipocytes and osteoblasts and effectively inhibit T-cell proliferation, as it has been described for MSCs [6,29].…”

“…Quantitative real-time polymerase chain reaction (PCR) was performed on an iCycler (BioRad, Hercules, CA) as we have previously described [28,29]. The primers used for amplification were as follows:…”

Controlled gentamicin release from multi-layered electrospun nanofibrous structures of various thicknesses

“…In our previous studies, 35,36 we demonstrated that nanofibers produced by needleless technology represent a convenient scaffold for cell cultivation and subsequent transfer to damaged tissue. The multilayered nanofibers studied in this work may offer a promising combination of such inherent properties of electrospun nanofibers with controlled drug release for developing suitable scaffolds for wound dressing applications.…”

Stem Cells in Ophthalmology

“…in the eye have shown that electrospun nanofibers can serve as a scaffold to grow and transfer limbal stem cells in the treatment of limbal stem cell deficiency (Zajicova et al, 2010). As biological discoveries identify key factors to stem cell survival and technological advances provide new biomaterials, the potential for improved treatment is dramatic.…”

“…It is possible this frill of conjunctiva possesses factors which enhance clinical outcomes of transplantation. Co-transfer of limbal stem cells with other stem cells, such as mesenchymal cells, may also lead to improved outcomes by inhibiting local inflammation and improving healing through immunosuppressive properties of these cells (Zajicova et al, 2010). Furthermore, human amniotic membrane may serve as a niche for limbal stem cells, possibly through the suppression of apoptosis and expression of certain growth factors (Tseng et al, 1998).…”