The Generation of Hybrid Electrospun Nanofiber Layer with Extracellular Matrix Derived from Human Pluripotent Stem Cells, for Regenerative Medicine Applications

Abstract

Extracellular matrix (ECM) has been utilized as a biological scaffold for tissue engineering applications in a variety of body systems, due to its bioactivity and biocompatibility. In the current study we developed a modified protocol for the efficient and reproducible derivation of mesenchymal progenitor cells (MPCs) from human embryonic stem cells as well as human induced pluripotent stem cells (hiPSCs) originating from hair follicle keratinocytes (HFKTs). ECM was produced from these MPCs and characterized in comparison to adipose mesenchymal stem cell ECM, demonstrating robust ECM generation by the excised HFKT-iPSC-MPCs. Exploiting the advantages of electrospinning we generated two types of electrospun biodegradable nanofiber layers (NFLs), fabricated from polycaprolactone (PCL) and poly(lactic-co-glycolic acid) (PLGA), which provide mechanical support for cell seeding and ECM generation. Elucidating the optimized decellularization treatment we were able to generate an available "off-the-shelf" implantable product (NFL-ECM). Using rat subcutaneous transplantation model we demonstrate that this stem-cell-derived construct is biocompatible and biodegradable and holds great potential for tissue regeneration applications.

Extracellular Matrix‐Based Biohybrid Materials for Engineering Compliant, Matrix‐Dense Tissues

“…By decellularizing the composite after a fixed time, the result is self‐termed an available “off‐the‐shelf” implantable product. This composite demonstrated biodegradability and biocompatibility in a rat subcutaneous model, and supported advanced cellular infiltration and habitation compared to uncoated PCL/PLGA scaffolds . Along similar lines, another study utilized a Vicryl knitted mesh made of polyglactin 910 (a 90:10 copolymer of glycolic acid and lactic acid (PLGA)), and cultured either mesenchymal stem cells, normal human articular chondrocytes, or normal human dermal fibroblasts onto both sides of the PLGA mesh.…”

“…Without intervention, ECM‐based materials are most often degraded in vivo, and are associated with constructive tissue remodeling and minimal fibrosis . As of yet, the specific cell signaling events by which ECM biomaterials modulate the host macrophage population toward a more constructive remodeling phenotype are not fully understood . In several examples, the presence of ECM components was shown to influence the initial response of the immune system to prohealing, despite the polymer components that would be expected to individually illicit an inflammatory response .…”

Nanofibrous Scaffolds for Skin Tissue Engineering and Wound Healing Based on Synthetic Polymers

“…The PLGA nanofibers were also modified with ECM components. In a study by Shtrichman et al [93], the PLGA nanofibrous scaffolds were modified with ECM deposited on these scaffolds by mesenchymal progenitor cells, derived from human embryonic stem cells, and human induced pluripotent stem cells, originating from hair follicle keratinocytes, which were cultured on the scaffolds and removed by subsequent decellularization. Subcutaneous implantation of the ECM-modified scaffolds in rats then showed that this stem cell-derived construct is biocompatible, biodegradable, and holds great potential for tissue regeneration applications.…”

Engineering Cell‐Derived Matrices: From 3D Models to Advanced Personalized Therapies

“…This process can be applied as a template for the growth and remodeling of the ECM. [50,51] Interestingly, Zhou et al reported that CDMs promoted peripheral nerve repair when L929 mouse fibroblast cells from adipose tissue were seeded onto ≈10 S cm −1 conductive mats combining electrospun poly(L-lactic acid) (PLLA) fibers and electrochemically deposited polypyrrole (PPy) nanoparticles. When rat PC12 cells were seeded onto the combined PLLA/PPy/ECM decellularized scaffolds (containing laminin, fibronectin, and collagen), their neurites were seen to differentiate and protrude.…”