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almudena del hierro, Ana Boto, Ignacio García Gomez, Mariano García-Arranz, Alfredo Insausti García, Félix Armada; Reconstruction of the ocular surface with adipose-derived stem cells (ASCs). Invest. Ophthalmol. Vis. Sci. 2013;54(15):1404.
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To evaluate the ability and safety of the adipose-derived stem cells (ASCs) as treatment to reconstruction of the corneal surface of rabbits with experimentally induced limbal stem cell deficiency (LSCD).We compare two different procedures: perilimbar subconjunctival injection versus application under an amniotic membrane patch.
Rabbit allogenic ASCs were isolated, cultured on plastic plates and identified by inmunostaining and flow cytometry. Limbal stem-cell deficiency (LSCD) was induced in 16 rabbits by alkali burns and mechanical debridement. One week after the injury, eyes were divided as follow: Group 1: 1A (n=8, left eyes) for subconjuctival-limbal injection of 1.6 x 106 ASCs/400 μl distributed in four limbal quadrants and 1B (n=8, right eyes) for injection of 100 μl of saline serum per quadrant; Group 2: 2A (n=8, Left eyes) for application of 1x106 ASCs under the stromal layer of amniotic membrane and over the injured corneas and 2B (n=8, right eyes) for transplantation of amniotic membrane. Each eye was examined under microscope every week before sacrifice. Rabbits were euthanized after 4 weeks and studied for inflammation (H&E staining), fibrosis (α-smooth muscle actin immunostaining), corneal inmunophenotype (Citokeratine 3) and inmunodetection of ASCs(GFP).
Incidence of perforations and calcifications was lower in eyes which had received ASCs by two procedures (p<0,05). Eyes treated by injection of ASCs had an epithelial defect closure faster than the other groups (p<0,05).The lowest levels of inflamatory cells at limbus were found at Group 1A and lowest inflamation in cornea corresponded at Group 2A. No differences were observed in arresting corneal neovessel ingrowth between groups. Corneal stromal fibrosis was higher in groups where ASCs were applied. CK3 was observed in no eyes. ASCs immunolocalization was observed at 24 hs after limbar injection and at two and four weeks in group which used amniotic membrane. No side effects were found in eyes which had received ASCs.
Complications were lower in eyes that received ASCs.Inflammation was lower where ASCs were located. No adverse reactions were observed in eyes that received ASCs.In conclusion, the ASCs therapy by limbal injection or using amniotic membrane was a feasible and safe procedure for the keratopathy secondary to LDSC by chemically burned .
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