Retrospective series compares single- and triple-layer decellularized amniotic membrane in DED

A retrospective clinical series published in Clinical Optometry evaluated long-term outcomes of a dehydrated, decellularized basement membrane (DBM) allograft in patients with moderate to severe dry eye disease (DED) and persistent corneal epithelial defects.

The study’s first author is Melanie Denton, OD, of the University of Alabama School of Optometry; Department of Optometry and Vision Science in Birmingham, Alabama.

DED is characterized by tear film instability, ocular surface inflammation, and epithelial compromise, which can predispose patients to complications such as acute keratitis and persistent corneal epithelial defects. Amniotic membrane therapy (AMT) has been used in refractory cases to protect the ocular surface, modulate inflammation and fibrosis, and promote epithelial healing.

“AMT has been increasingly adopted as a therapy since it acts as a protective covering preventing mechanical disruption, modulates inflammation and fibrosis, supports corneal epithelial migration, and helps restore surface homeostasis in treatment-refractory DED and related epithelial disease,” Denton stated in the study.

The study compared single-layer (Biovance SL) and 3-layer (Biovance Ocular 3L) DBM constructs applied with pressure patching. Seventeen eyes from 9 patients met inclusion criteria. Nine eyes received the single-layer graft and eight received the 3-layer graft. Corneal fluorescein staining, graded using the Oxford scale (0–5), was assessed at baseline and at 1, 3, and 6 months.

Both groups demonstrated statistically significant reductions in corneal staining at 3 and 6 months compared with baseline. The single-layer DBM showed sustained improvement of 42–48% at 3 and 6 months, respectively. The 3-layer DBM demonstrated sustained improvement of 57–89% at the same time points. At 6 months, the reduction in corneal staining in the three-layer group was statistically significantly greater than that observed in the single-layer group (p=0.0036). No significant difference between groups was observed at 3 months.

Complete resolution of corneal staining at 6 months was reported in 22% of eyes treated with the single-layer graft and 75% of eyes treated with the three-layer graft. All single-layer grafts were fully incorporated within 24 hours, and all three-layer grafts within 72 hours. No adverse events, including ocular infection, pain, or allergic response, were observed.

Denton noted that the basement membrane is the key structural component of amniotic tissue involved in re-epithelialization, containing extracellular matrix proteins and signaling molecules that support epithelial adhesion and migration. The 3-layer construct consists of laminated decellularized basement membrane without heat or freeze-drying. Prior in vitro research cited in the study suggests the three-layer graft may promote a less pro-inflammatory cytokine profile and slower biodegradation compared with other amniotic membrane platforms.

Denton concluded that both single- and 3-layer DBM constructs were associated with sustained improvements in corneal epithelial integrity over six months, with the 3-layer graft demonstrating greater magnitude and durability of effect. She suggested multilayer DBM may offer enhanced long-term efficacy in DED-related epithelial disease.

“DBMs that preserve the native amniotic basement membrane architecture offer both practical and biologic advantages, including reduced immunogenicity, retention of key structural and signaling components of the extracellular matrix, and facilitation of epithelial cell adhesion, migration, and differentiation,” Denton stated in the study.

The study’s limitations include its retrospective design, small sample size, potential selection bias, variability in visit timing, and baseline differences between groups.

Reference:

1. Denton M. Comparison of effectiveness of Biovance single and triple layer decellularized basement membranes for treatment of ocular surface disease: A retrospective study. Clinical Optometry. 2026;18. https://www.dovepress.com/article/download/111693