Rafael Martínez-Carrasco1, Luis Ignacio Sánchez-Abarca2, Cristina Nieto-Gómez3, Elisabet Martín García3, Fermín Sánchez-Guijo4, Pablo Argüeso5, José Aijón6, Emiliano Hernández-Galilea7, Almudena Velasco6. 1. Institute for Biomedical Research of Salamanca (IBSAL), Salamanca, 37007, Spain; Department Cell Biology & Pathology, University of Salamanca, Salamanca, 37007, Spain; INCyL, University of Salamanca, Salamanca, 37007, Spain. Electronic address: rafawallace@usal.es. 2. Institute for Biomedical Research of Salamanca (IBSAL), Salamanca, 37007, Spain; Department of Hematology, IBSAL-University Hospital of Salamanca, Salamanca, 37007, Spain; Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, Spain. 3. Department of Surgery, Ophthalmology Service, University Hospital of Salamanca, University of Salamanca, Salamanca, 37007, Spain. 4. Institute for Biomedical Research of Salamanca (IBSAL), Salamanca, 37007, Spain; Department of Hematology, IBSAL-University Hospital of Salamanca, Salamanca, 37007, Spain; Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León, Spain; RETIC TerCel, y CIBERONC, Instituto de Salud Carlos III (ISCIII), Spain. 5. Schepens Eye Research Institute of Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, United States. 6. Institute for Biomedical Research of Salamanca (IBSAL), Salamanca, 37007, Spain; Department Cell Biology & Pathology, University of Salamanca, Salamanca, 37007, Spain; INCyL, University of Salamanca, Salamanca, 37007, Spain. 7. Institute for Biomedical Research of Salamanca (IBSAL), Salamanca, 37007, Spain; Department of Surgery, Ophthalmology Service, University Hospital of Salamanca, University of Salamanca, Salamanca, 37007, Spain.
Abstract
PURPOSE: To evaluate the therapeutic effect of subconjunctival injection of human mesenchymal stromal cells (hMSCs) in the cornea of mice with graft versus host disease (GVHD). METHODS: GVHD was induced in mice after hematopoietic stem cell transplantation (HSCT) between MHC-mismatched mouse strains. Subconjunctival injection of hMSCs was applied at day 10 post-HSCT. Infiltration of CD3+ cells in the cornea and epithelial alterations were analyzed by immunofluorescence. Tear was assessed using the PRT test and TearLab Osmolarity System. qPCR was used to evaluate changes in cytokines, Pax6 and Sprr1b expression. To evaluate the effect of irradiation, we analyzed the expression of these genes in TBI mice. RESULTS: Immune cell invasion occurs in mice with GVHD, as shown by the presence of CD3+ cells in the cornea. Interestingly, eyes treated with hMSC did not present CD3+ cells. Tear osmolarity was increased in GVHD eyes, but not in treated eyes. TNFa expression was highly increased in all corneas except in Control and treated eyes. Pax6 in corneal epithelium showed a similar pattern in GVHD and Control mice, and its gene expression was enhanced in GVHD corneas. In contrast, Pax6 was reduced in GVHD + MSC corneas. We also found an increase in SPRR1B staining in GVHD eyes that was lower in GVHD + MSC mice, demonstrating that corneal keratinization is less frequent after treatment with hMSC. CONCLUSIONS: The treatment with hMSCs by subconjunctival injection is effective in reducing corneal inflammation and squamous metaplasia in ocular GVHD (oGVHD). Local treatment with hMSCs is a promising strategy for oGVHD.
PURPOSE: To evaluate the therapeutic effect of subconjunctival injection of human mesenchymal stromal cells (hMSCs) in the cornea of mice with graft versus host disease (GVHD). METHODS:GVHD was induced in mice after hematopoietic stem cell transplantation (HSCT) between MHC-mismatched mouse strains. Subconjunctival injection of hMSCs was applied at day 10 post-HSCT. Infiltration of CD3+ cells in the cornea and epithelial alterations were analyzed by immunofluorescence. Tear was assessed using the PRT test and TearLab Osmolarity System. qPCR was used to evaluate changes in cytokines, Pax6 and Sprr1b expression. To evaluate the effect of irradiation, we analyzed the expression of these genes in TBI mice. RESULTS: Immune cell invasion occurs in mice with GVHD, as shown by the presence of CD3+ cells in the cornea. Interestingly, eyes treated with hMSC did not present CD3+ cells. Tear osmolarity was increased in GVHD eyes, but not in treated eyes. TNFa expression was highly increased in all corneas except in Control and treated eyes. Pax6 in corneal epithelium showed a similar pattern in GVHD and Control mice, and its gene expression was enhanced in GVHD corneas. In contrast, Pax6 was reduced in GVHD + MSC corneas. We also found an increase in SPRR1B staining in GVHD eyes that was lower in GVHD + MSC mice, demonstrating that corneal keratinization is less frequent after treatment with hMSC. CONCLUSIONS: The treatment with hMSCs by subconjunctival injection is effective in reducing corneal inflammation and squamous metaplasia in ocular GVHD (oGVHD). Local treatment with hMSCs is a promising strategy for oGVHD.
Authors: Sara Galindo; Ana de la Mata; Marina López-Paniagua; Jose M Herreras; Inmaculada Pérez; Margarita Calonge; Teresa Nieto-Miguel Journal: Stem Cell Res Ther Date: 2021-01-13 Impact factor: 6.832
Authors: Ilham Putra; Xiang Shen; Khandaker N Anwar; Behnam Rabiee; Ravand Samaeekia; Enmar Almazyad; Pushpanjali Giri; Sayena Jabbehdari; Mohammed R Hayat; Abdelrahman M Elhusseiny; Mahmood Ghassemi; Nadim Mahmud; Deepak P Edward; Charlotte E Joslin; Mark I Rosenblatt; Reza Dana; Medi Eslani; Peiman Hematti; Ali R Djalilian Journal: Transl Vis Sci Technol Date: 2021-08-12 Impact factor: 3.283