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Presentation type	Oral presentation
Title	In Vitro Comparative Evaluation of Autologous Serum Enhanced with Amniotic Membrane Extract Eye Drops (ASRAM) for Persistent Corneal Defects

Purpose	To compare the anti-inflammatory and regenerative properties of a novel formulation, Autologous Serum enhanced with Amniotic Membrane extract Eye Drops (ASRAM), with Autologous Serum Eye Drops (ASED) and Amniotic Membrane Extract Eye Drops (AMEED)
Methods	Human cornea-derived mesenchymal stromal cells (cMSCs) were exposed to a diluted inflammatory cocktail. Immunomodulatory effects of ASED, AMEED and ASRAM were assessed by flow cytometry (CD40, CD54, CD274, HLA-ABC). Regenerative capacity was evaluated using scratch assays, including Mitomycin-C-treated cMSCs to distinguish migration from proliferation. Growth factors and anti-inflammatory mediators (EGF, bFGF, TIMP-1, IL-1RA) were quantified by ELISA in AMEED prepared after varying amniotic membrane soaking durations.
Results	ASRAM significantly reduced CD54 and HLA-ABC expression compared with ASED, with effects comparable to AMEED. Scratch assays showed faster wound closure with ASRAM and AMEED versus ASED at 48h with no statistically significant difference. Mitomycin-C pretreatment completely abolished wound closure under all conditions, confirming a proliferation-dependent repair mechanism. EGF and bFGF levels increased significantly with longer AM soaking (days 2 and 4, p<0.01) without enhancing anti-inflammatory activity, while IL-1RA and TIMP-1 remained stable across soaking durations.
Conclusion	ASRAM showed anti-inflammatory activity superior to ASED and comparable to AMEED in inflamed cMSCs, with a similar trend toward faster wound closure. These in vitro findings support ASRAM's potential as an enhanced biologic therapy for corneal epithelial repair, warranting further investigation.
Conflict of interest	No

Authors 1

Last name	DAOUDI
Initials	I
Department	Hôpital Erasme
City	Brussels

Authors 2

Last name	AFIFI
Initials	Y
Department	Hôpital Erasme
City	Brussels

Authors 3

Last name	HAKAMI
Initials	S
Department	Hôpital Erasme
City	Brussels

Authors 4

Last name	MOUJANE
Initials	F
Department	Hôpital Erasme
City	Brussels

Authors 5

Last name	MOTULSKY
Initials	E
Department	Hôpital Erasme
City	Brussels