A Proteomic Approach to Determine Stem Cell Skeletal Differentiation Signature on Additive Manufactured Scaffolds

Area of Science:

• Biomaterials Science
• Tissue Engineering
• Proteomics

Background:

• Understanding cell-material interactions on porous biomaterials is crucial for tissue engineering.
• Additive manufacturing offers precise control over scaffold composition and design.
• Traditional biochemical assays have limitations in evaluating scaffold performance for specific tissue applications.

Purpose of the Study:

• To compare the chondrogenic and osteogenic potential of three additive manufactured polymers: polycaprolactone (PCL), poly(ethylene oxide)-poly(butylene terephthalate) (PEOT/PBT), and polylactic acid (PLA).
• To utilize proteomics to decipher cell-material interactions and cell differentiation on these scaffolds using human bone marrow-derived mesenchymal stem cells (hBMSCs).
• To identify suitable materials for specific tissue engineering applications based on cellular response.

Main Methods:

• Fabrication of 3D scaffolds using PCL, PEOT/PBT, and PLA via additive manufacturing.
• Characterization of scaffold properties including hydrophilicity and Young's modulus.
• In vitro evaluation of chondrogenic and osteogenic potential using hBMSCs, including biochemical assays and comprehensive proteomics analysis.

Main Results:

• PEOT/PBT and PLA scaffolds demonstrated higher chondrogenic potential (glycosaminoglycan and collagen deposition) compared to PCL.
• PLA and PEOT/PBT scaffolds showed enhanced osteogenic potential (calcium deposition and alkaline phosphatase activity), particularly PLA.
• Proteomics pathway analysis revealed that PCL did not induce differentiation-related pathways, while PEOT/PBT and PLA modulated key pathways for chondrogenesis and osteogenesis. PEOT/PBT supported chondrogenesis, and PLA promoted both chondrogenesis and osteogenesis, including angiogenesis.

Conclusions:

• PEOT/PBT is a promising candidate for cartilage and osteochondral applications, capable of driving hBMSC differentiation without growth factors.
• PLA is suitable for both cartilage and bone applications, promoting chondrogenic and osteogenic protein expression, and its angiogenic protein upregulation makes it ideal for bone regeneration requiring vascularization.
• Proteomics is a powerful tool for understanding cell-material interactions and guiding the selection of optimal biomaterials for specific tissue engineering applications.