In vivo confocal microscopy evaluation of infiltrated immune cells in corneal stroma treated with cell therapy in advanced keratoconus

Mona El Zarif; Karim Abdul Jawad; Jorge L Alió; Nehman Makdissy; María P De Miguel

doi:10.1186/s12348-024-00385-2

In vivo confocal microscopy evaluation of infiltrated immune cells in corneal stroma treated with cell therapy in advanced keratoconus

J Ophthalmic Inflamm Infect. 2024 Jan 26;14(1):5. doi: 10.1186/s12348-024-00385-2.

Authors

Mona El Zarif^{1

2

3}, Karim Abdul Jawad¹, Jorge L Alió^{2

4}, Nehman Makdissy⁵, María P De Miguel⁶

Affiliations

¹ Optica General, Saida, Lebanon.
² Division of Ophthalmology, Universidad Miguel Hernández, Alicante, Spain.
³ Doctoral School of Sciences and Technology, Lebanese University, Hadath, Lebanon.
⁴ Cornea, Cataract and Refractive Surgery Unit, Vissum (Miranza Group), Alicante, Spain.
⁵ Genomic Surveillance and Biotherapy GSBT, Faculty of Sciences, Lebanese University, RasMaska, Lebanon. nehman.makdissy@ul.edu.lb.
⁶ Cell Engineering Laboratory, IdiPAZ, La Paz Hospital Health Research Institute, Madrid, Spain. mariapdemiguel@gmail.com.

Abstract

Purpose: This study investigates immune cell (ICs) infiltration in advanced keratoconus patients undergoing autologous adipose-derived adult stem cell (ADASC) therapy with recellularized human donor corneal laminas (CL).

Methods: A prospective clinical trial included fourteen patients divided into three groups: G-1, ADASCs; G-2, decellularized CL (dCL); and G-3, dCL recellularized with ADASCs (ADASCs-rCL). Infiltrated ICs were assessed using in vivo confocal microscopy (IVCM) at 1,3,6, and12 months post-transplant.

Results: Infiltrated ICs, encompassing granulocytes and agranulocytes, were observed across all groups, categorized by luminosity, structure, and area. Stromal ICs infiltration ranged from 1.19% to 6.62%, with a consistent increase in group-related cell density (F = 10.68, P < .0001), independent of post-op time (F = 0.77, P = 0.511); the most substantial variations were observed in G-3 at 6 and 12 months (2.0 and 1.87-fold, respectively). Similarly, significant size increases were more group-dependent (F = 5.76, P < .005) rather than time-dependent (F = 2.84, P < .05); G-3 exhibited significant increases at 6 and 12 months (3.70-fold and 2.52-fold, respectively). A lamina-induced shift in IC size occurred (F = 110.23, P < .0001), primarily with 50-100 μm² sizes and up to larger cells > 300μm², presumably macrophages, notably in G-3, indicating a potential role in tissue repair and remodeling, explaining reductions in cells remnants < 50μm².

Conclusions: ADASCs-rCL therapy may lead to increased IC infiltration compared to ADASCs alone, impacting cell distribution and size due to the presence of the lamina. The findings reveal intricate immune patterns shaped by the corneal microenvironment and highlight the importance of understanding immune responses for the development of future therapeutic strategies.

Keywords: Adipose-derived adult stem cells; Cell infiltration; Cell therapy; Clinical trial; Corneal laminas; Immune cells; Immune response; In vivo confocal microscopy; Keratoconus; Tissue repair.