Research Article
Open Access
Human Amniotic Membrane Scaffold Use in Wound Care: Macrophage Activation and Healing Outcomes
Stephen Brigido1*, Nicole Protzman2, Chila Ferdinand3, Nancy Lisa2,3, Biadge Kefale3
1Research and Technology, Lonza, Walkersville, MD, USA
2Process Development, Lonza, Walkersville, MD, USA
3Research and Technology, Lonza, Walkersville, MD, USA
2Process Development, Lonza, Walkersville, MD, USA
3Research and Technology, Lonza, Walkersville, MD, USA
Brigido S, Protzman N, Ferdinand C, Lisa N, Kefale B. Human Amniotic Membrane Scaffold Use in Wound Care: Macrophage Activation and Healing Outcomes, Accounts of Biotechnology Research. 2023, Vol. 11 No. 1: 103
Abstract
Effective wound healing using biomaterials depends on the presence of extracellular
matrix (ECM) components that support and guide the body’s natural regenerative
processes. Macrophages, key immune cells derived from monocytes, play a pivotal
role in both the initiation and resolution phases of tissue repair. During early
wound healing, macrophages drive the inflammatory response, whereas in later
stages, they contribute to tissue remodeling and resolution. Chronic wounds often
result from dysregulated macrophage activation, which impairs normal healing.
This study aimed to evaluate the influence of a decellularized, dehydrated human
amniotic membrane (DDHAM) on monocyte-to-macrophage differentiation
and activation in vitro. Human monocytes were isolated from peripheral blood
of healthy donors and cultured on standard tissue culture plates (CB), collagen
type I-coated plates (COL), and DDHAM-coated plates. Proinflammatory (M1)
macrophage differentiation was induced using granulocyte-macrophage colonystimulating
factor (GM-CSF) followed by activation with a proinflammatory
stimulus comprising lipopolysaccharide (LPS) and interferon-gamma (IFN-ɣ).
The results demonstrated that DDHAM significantly enhanced monocyte
differentiation compared to CB and COL controls, as evidenced by increased cell
size, viability, expression of macrophage-specific genes, and secretion of soluble
factors. Notably, macrophages differentiated on DDHAM and activated with
inflammatory signals exhibited a marked reduction in expression of several LPSinducible
NF-κB target genes, with IL12β, encoding the IL12p40 subunit, showing
the most pronounced downregulation (p < 0.001). Mechanistic studies revealed
that DDHAM-mediated effects on differentiation were dependent on β2 integrins.
Collectively, these findings provide the first evidence that DDHAM can modulate
macrophage behavior by promoting polarization toward an M2 phenotype, which
is associated with tissue regeneration, vascular remodeling, and resolution of
inflammation, supporting its potential as a scaffold to enhance wound repair.
matrix (ECM) components that support and guide the body’s natural regenerative
processes. Macrophages, key immune cells derived from monocytes, play a pivotal
role in both the initiation and resolution phases of tissue repair. During early
wound healing, macrophages drive the inflammatory response, whereas in later
stages, they contribute to tissue remodeling and resolution. Chronic wounds often
result from dysregulated macrophage activation, which impairs normal healing.
This study aimed to evaluate the influence of a decellularized, dehydrated human
amniotic membrane (DDHAM) on monocyte-to-macrophage differentiation
and activation in vitro. Human monocytes were isolated from peripheral blood
of healthy donors and cultured on standard tissue culture plates (CB), collagen
type I-coated plates (COL), and DDHAM-coated plates. Proinflammatory (M1)
macrophage differentiation was induced using granulocyte-macrophage colonystimulating
factor (GM-CSF) followed by activation with a proinflammatory
stimulus comprising lipopolysaccharide (LPS) and interferon-gamma (IFN-ɣ).
The results demonstrated that DDHAM significantly enhanced monocyte
differentiation compared to CB and COL controls, as evidenced by increased cell
size, viability, expression of macrophage-specific genes, and secretion of soluble
factors. Notably, macrophages differentiated on DDHAM and activated with
inflammatory signals exhibited a marked reduction in expression of several LPSinducible
NF-κB target genes, with IL12β, encoding the IL12p40 subunit, showing
the most pronounced downregulation (p < 0.001). Mechanistic studies revealed
that DDHAM-mediated effects on differentiation were dependent on β2 integrins.
Collectively, these findings provide the first evidence that DDHAM can modulate
macrophage behavior by promoting polarization toward an M2 phenotype, which
is associated with tissue regeneration, vascular remodeling, and resolution of
inflammation, supporting its potential as a scaffold to enhance wound repair.
Keywords
Biomaterials; Decellularization; Human amniotic membrane; DDHAM; Extracellular matrix; Macrophages; Monocytes; Differentiation; Polarization; Scaffold; Wound healing.