Tissue Engineered Neural Constructs Composed of Neural Precursor Cells, Recombinant Spidroin and PRP for Neural...

Area of Science:

• Biomaterials Science
• Neuroscience
• Tissue Engineering

Background:

• Developing effective strategies for neural precursor cell (NPC) delivery and survival is crucial for regenerative medicine.
• Existing methods often face challenges with cell integration, differentiation, and long-term viability in vivo.
• Novel biomaterials are needed to support neural tissue regeneration and functional recovery.

Purpose of the Study:

• To design and evaluate a novel two-component matrix (SPRPix) for encapsulating directly reprogrammed human neural precursor cells (drNPCs).
• To assess the biocompatibility and efficacy of the SPRPix matrix in promoting drNPC survival, differentiation, and neural tissue formation in vivo.
• To explore the potential of SPRPix as a therapeutic scaffold for spinal cord injury (SCI) treatment.

Main Methods:

• Fabrication of a composite scaffold (rSS-PCL) using electrospun recombinant spidroins (rS1/9, rS2/12) and polycaprolactone (PCL).
• Incorporation of platelet-rich plasma (PRP) as a liquid matrix component with the rSS-PCL scaffold.
• In vitro assessment of drNPC proliferation and differentiation within the SPRPix matrix.
• In vivo implantation of SPRPix-loaded drNPCs into the brain and spinal cord of Rhesus macaque monkeys for biocompatibility and survival studies.

Main Results:

• The SPRPix matrix, combining spidroin and PRP, significantly promoted drNPC proliferation and neurogenesis, forming neural tissue organoids.
• Differentiated drNPCs expressed neuronal markers (βIII-tubulin, MAP2) and astrocyte markers (GFAP), indicating successful neuronal development.
• Microfibrils within the SPRPix matrix provided guidance cues, aligning neuronal processes.
• In vivo studies demonstrated excellent biocompatibility with no astroglial or microglial reactions.
• Implanted human drNPCs survived for 3 months and differentiated into MAP2-positive neurons in non-human primates.

Conclusions:

• The SPRPix matrix is a promising biomaterial for encapsulating and supporting the survival and differentiation of drNPCs.
• The combination of spidroin and PRP enhances neurogenesis and promotes the formation of functional neural tissue.
• The observed biocompatibility and neural differentiation in vivo suggest significant potential for SPRPix in treating spinal cord injuries.