Abstract
Corneal opacities are a leading cause of global blindness. They are conventionally treated by the transplantation of donor corneal tissue, which is, restricted by a worldwide donor material shortage and allograft rejection. Autologous adult stem cells with a potential to differentiate into corneal stromal keratocytes (CSKs) could offer a suitable choice of cells for regenerative cell therapy. Postnatal periodontal ligament (PDL) contains a population of adult stem cells, which has a similar embryological origin as CSK, that is cranial neural crest. We harvested PDL cells from young adult teeth extracted because of non‐functional or orthodontic reason and differentiated them towards CSK phenotype using a two‐step protocol with spheroid formation followed by growth factor and cytokine induction in a stromal environment (human amnion stroma and porcine corneal stroma). Our results showed that the PDL‐differentiated CSK‐like cells expressed CSK markers (CD34, ALDH3A1, keratocan, lumican, CHST6, B3GNT7 and Col8A2) and had minimal expression of genes related to fibrosis and other lineages (vasculogenesis, adipogenesis, myogenesis, epitheliogenesis, neurogenesis and hematogenesis). Introduction of PDL spheroids into the stroma of porcine corneas resulted in extensive migration of cells inside the host stroma after 14‐day organ culture. Their quiescent nature and uniform cell distribution resembled to that of mature CSKs inside the native stroma. Our results demonstrated the potential translation of PDL cells for regenerative corneal cell therapy for corneal opacities.
Highlights
The cornea provides a physical and biological barrier to protect inner eye tissues and its transparency allows efficient light transmission and refraction for normal vision and visual acuity
This study showed that human postnatal periodontal ligament (PDL) containing neural crest (NC)-derived stem cells were able to differentiate and assume corneal stromal keratocytes (CSKs) phenotype under a two-step protocol involving spheroid formation followed by growth factor and cytokine induction on a stromal niche
Our results demonstrated the potential translation of PDL cells for regenerative corneal cell therapy
Summary
The cornea provides a physical and biological barrier to protect inner eye tissues and its transparency allows efficient light transmission and refraction for normal vision and visual acuity. The corneal stroma contains collagen fibrils (mainly type I) aligned in the form of lamellae, which run in orthogonal orientations to provide the corneal strength and transparency.[1] Corneal stromal keratocytes (CSKs) located between collagen lamellae synthesize and deposit collagens and keratan sulphate proteoglycans Our results have provided useful information to demonstrate PDL as a novel and autologous cell source for regenerative corneal cell therapy
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