The aim of this study was to investigate the transdifferentiation potential of murine vibrissa hair follicle (HF) stem cells into corneal epithelial-like cells through modulation by corneal- or limbus-specific microenvironmental factors. Adult epithelial stem cells were isolated from the HF bulge region by mechanical dissection or fluorescence-activated cell sorting using antibodies to α6 integrin, enriched by clonal expansion, and subcultivated on various extracellular matrices (type IV collagen, laminin-1, laminin-5, fibronectin) and in different conditioned media derived from central and peripheral corneal fibroblasts, limbal stromal fibroblasts, and 3T3 fibroblasts. Cellular phenotype and differentiation were evaluated by light and electron microscopy, real-time reverse transcription-polymerase chain reaction, immunocytochemistry, and Western blotting, using antibodies against putative stem cell markers (K15, α6 integrin) and differentiation markers characteristic for corneal epithelium (K12, Pax6) or epidermis (K10). Using laminin-5, a major component of the corneo-limbal basement membrane zone, and conditioned medium from limbal stromal fibroblasts, clonally enriched HF stem and progenitor cells adhered rapidly and formed regularly arranged stratified cell sheets. Conditioned medium derived from limbal fibroblasts markedly upregulated expression of cornea-specific K12 and Pax6 on the mRNA and protein level, whereas expression of the epidermal keratinocyte marker K10 was strongly downregulated. These findings suggest that adult HF epithelial stem cells are capable of differentiating into corneal epithelial-like cells in vitro when exposed to a limbus-specific microenvironment. Therefore, the HF may be an easily accessible alternative therapeutic source of autologous adult stem cells for replacement of the corneal epithelium and restoration of visual function in patients with ocular surface disorders.

No abstract

P>Ocular manifestations are very common in all types of mucopolysaccharidoses (MPS) and often lead to visual impairment. They arise as a result of the accumulation of glycosaminoglycans deposits in ocular tissues or secondary to increased intracranial pressure. Typical ocular features in MPS include corneal clouding, retinopathy, glaucoma, optic disc swelling, optic atrophy, and ocular motility and refractive error problems. This paper reviews the ocular features in patients with MPS, discusses the diagnosis of these ocular features and the diagnostic problems that may arise in patients (children) with MPS, and highlights the central role ophthalmologists may play in the diagnosis and follow-up of these patients

The findings provide evidence for a specific upregulation, production, and deposition of collagen types III and XVI, agrin, TGFBI, and clusterin in late-onset FCD and thus point to the importance of matrix alterations in the pathophysiology of FCD.

Venous bupivacaine plasma concentrations were measured in six neonates and infants aged 4 days to 3.9 mo (mean, 2.1 mo) and 10 infants and children aged 9 mo to 6 yr (mean, 3.1 yr) after administration of an initial bolus of 0.5 mL/kg bupivacaine 0.25%, followed by a continuous infusion of local anesthetic (0.25 mL.kg-1.h-1) over a period of 4 h (first hour: bupivacaine 0.25%, then reduced to 0.125%). Plasma concentrations of local anesthetic measured at 180 min and 300 min after beginning of bupivacaine administration were significantly higher in younger infants when compared to older infants and children (180 min: 0.67 +/- 0.24 micrograms/mL [0.25-0.97] vs 0.27 +/- 0.11 micrograms/mL [0.19-0.55], P < 0.01; 300 min: 0.86 +/- 0.36 micrograms/mL [0.35-1.25] vs 0.34 +/- 0.12 micrograms/mL [0.18-0.57], P < 0.01). The results of our study show that despite applying the same dosage of epidural bupivacaine significantly higher plasma concentrations were seen after short periods of continuous infusion in infants up to 4 mo than in children older than 9 mo.