Developing a pro-angiogenic placenta derived amniochorionic scaffold with two exposed basement membranes as substrates for cultivating endothelial cells

Abstract

Decellularized and de-epithelialized placenta membranes have widely been used as scaffolds and grafts in tissue engineering and regenerative medicine. Exceptional pro-angiogenic and biomechanical properties and low immunogenicity have made the amniochorionic membrane a unique substrate which provides an enriched niche for cellular growth. Herein, an optimized combination of enzymatic solutions (based on streptokinase) with mechanical scrapping is used to remove the amniotic epithelium and chorion trophoblastic layer, which resulted in exposing the basement membranes of both sides without their separation and subsequent damages to the in-between spongy layer. Biomechanical and biodegradability properties, endothelial proliferation capacity, and in vivo pro-angiogenic capabilities of the substrate were also evaluated. Histological staining, immunohistochemistry (IHC) staining for collagen IV, and scanning electron microscope demonstrated that the underlying amniotic and chorionic basement membranes remained intact while the epithelial and trophoblastic layers were entirely removed without considerable damage to basement membranes. The biomechanical evaluation showed that the scaffold is suturable. Proliferation assay, real-time polymerase chain reaction for endothelial adhesion molecules, and IHC demonstrated that both side basement membranes could support the growth of endothelial cells without altering endothelial characteristics. The dorsal skinfold chamber animal model indicated that both side basement membranes could promote angiogenesis. This bi-sided substrate with two exposed surfaces for cultivating various cells would have potential applications in the skin, cardiac, vascularized composite allografts, and microvascular tissue engineering.

Decellularized placental sponge as an excellent carrier for human mesenchymal stem cells and management of full-thickness skin wounds: an in vitro and in vivo study

“…They also demonstrated that 3D print placental scaffold could upregulate the expression of CD31+/CD34 + in HUVEC cells and support their culture and growth in vitro. The angiogenesis potential of placental-derived scaffold was also con rmed by Shariatzadeh and coworkers [52]. According to their results, the placental-derived scaffold supported endothelial cell growth in vitro and promoted neovascularization in vivo.…”

Decellularized Placental Sponge Seeded with Human Mesenchymal Stem Cells Improves Deep Skin Wound Healing in the Animal Model

“…The angiogenesis potential of the placental-derived scaffold was also confirmed by Shariatzadeh and co-workers. 46 According to their results, the placental-derived scaffold supported endothelial cell growth in vitro and promoted neovascularization in vivo.…”

Immunogenicity of decellularized extracellular matrix scaffolds: a bottleneck in tissue engineering and regenerative medicine

“…Decellularization inevitably depletes GAGs and other ECM compositions and drives scaffolds with inferior mechanical strength. To address these challenges, crosslinking has proved its efficacy in promoting mechanical stability and inhibiting the rapid degradation of decellularized tissues, providing sufficient time for angiogenesis by endothelial cells and new ECM synthesis [195,196]. Taken together, crosslinking is a double-edged sword, and accurate control of the degradation rate is required to achieve an equilibrium between graft stability and functional remodeling [190,195].…”