Topical administration of orbital fat-derived stem cells promotes corneal tissue regeneration

Abstract

IntroductionTopical administration of eye drops is the major route for drug delivery to the cornea. Orbital fat-derived stem cells (OFSCs) possess an in vitro corneal epithelial differentiation capacity. Both the safety and immunomodulatory ability of systemic OFSC transplantation were demonstrated in our previous work. In this study, we investigated the safety, therapeutic effect, and mechanism(s) of topical OFSC administration in an extensive alkali-induced corneal wound.MethodsCorneal injury was created by contact of a piece of 0.5 N NaOH-containing filter paper on the corneal surface of a male Balb/c mouse for 30 s. The area of the filter paper covered the central 70% or 100% of the corneal surface. OFSCs (2 × 105) in 5 μl phosphate-buffered saline (PBS) were given by topical administration (T) twice a day or by two intralimbal (IL) injections in the right cornea, while 5 μl of PBS in the left cornea served as the control.ResultsTopical OFSCs promoted corneal re-epithelialization of both the limbal-sparing and limbal-involved corneal wounds. In the first three days, topical OFSCs significantly reduced alkali-induced corneal edema and stromal infiltration according to a histopathological examination. Immunohistochemistry and immunofluorescence staining revealed that transplanted cells were easily detectable in the corneal epithelium, limbal epithelium and stroma, but only some of transplanted cells at the limbal epithelium had differentiated into cytokeratin 3-expressing cells. OFSCs did not alter neutrophil (Ly6G) levels in the cornea, but significantly reduced macrophage (CD68) infiltration and inducible nitrous oxide synthetase (iNOS) production during acute corneal injury as quantified by a Western blot analysis. Continuous topical administration of OFSCs for seven days improved corneal transparency, and this was accompanied by diffuse stromal engraftment of transplanted cells and differentiation into p63-expressing cells at the limbal area. The therapeutic effect of the topical administration of OFSCs was superior to that of the IL injection. OFSCs from the IL injection clustered in the limbal area and central corneal epithelium, which was associated with a persistent corneal haze.ConclusionsTopical OFSC administration is a simple, non-surgical route for stem cell delivery to promote corneal tissue regeneration through ameliorating acute inflammation and corneal epithelial differentiation. The limbal area serves as a niche for OFSCs differentiating into corneal epithelial cells in the first week, while the stroma is a potential site for anti-inflammation of OFSCs. Inhibition of corneal inflammation is related to corneal transparency.

Age-Related Changes in the Regenerative Potential of Adipose-Derived Stem Cells Isolated from the Prominent Fat Pads in Human Lower Eyelids

“…The multi-lineage differentiation capacity of OASCs has shown great therapeutic potential in the field of ophthalmology and regenerative medicine [6–9]. Because such promising applications involve the treatment of diseases and conditions found in an aging population, hindered by stem cell senescence and dysfunction in the natural aging process, the donor or patient age must be accounted for before the autologous stem-cell-based therapeutics entering the clinic.…”

“…The existence of human orbital adipose-derived stem cells (OASCs) also has been proven. In addition to differentiating into adipocytes, chondrocytes and osteoblasts, these cells also can differentiate towards the corneal epithelial, smooth muscle and neuronal lineages under specific inductive conditions, showing great therapeutic potential in treating the orbital and ocular diseases [6–9]. …”

Progress in corneal wound healing

“…were tried as alternative sources of autologous stem cell-based therapy for corneal diseases and wound healing (Ono et al 2007; Yang et al, 2008; Kumagai et al, 2010; Meyer-Blazejewska, et al 2011; Yan et al, 2014). Adipose-derived, orbital fat-derived, and immature dental pulp stem cells have been used for successful re-epithelialization of chemically burned cornea (Monteiro et al, 2009; Lin et al, 2013b; Zeppieri et al, 2013). Additionally, mesenchymal stem cells (MSCs) were also considered as a source of corneal cells to treat burns and LSCD (Ma et al, 2006; Reinshagen et al, 2011; Pinarli et al, 2014).…”

Local Injection of Mesenchymal Stem Cells Protects Testicular Torsion-induced Germ Cell Injury

“…Orbital fat-derived stem cells (OFSCs) are MSCs isolated from human orbital fat tissue [14] and their therapeutic effects on acute tissue injury have been demonstrated via paracrine tissue support, immunomodulation, and differentiation ability in our previous experimental studies [15][16][17]. In this study, rats received surgery with 720°of unilateral testicular torsion for 3 hours, and local injection of OFSCs 30 minutes before surgical detorsion was performed.…”