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Acoustic Patterning of Growth Factor for Three-Dimensional Tissue Engineering.

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  • 1Department of Orthopedic Surgery, School of Medicine, Stanford University, Stanford, California.

Tissue Engineering. Part A
|January 18, 2020
PubMed
Summary

This study introduces a novel method combining additive manufacturing and acoustic droplet ejection to precisely control growth factor distribution on 3D scaffolds. This technique enhances tissue engineering constructs for improved cell differentiation and tissue formation.

Keywords:
3D tissue engineeringacoustic patterningadditive manufacturingbiofabricationgrowth factorpolycaprolactone (PCL)

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Area of Science:

  • Biomaterials Engineering
  • Tissue Engineering
  • Additive Manufacturing

Background:

  • Precise control over biological cue presentation is crucial for effective tissue engineering.
  • Current methods for incorporating growth factors (GFs) onto 3D scaffolds have limitations in spatial and temporal control.
  • There is a significant need for advanced techniques to improve GF distribution in tissue engineering constructs.

Purpose of the Study:

  • To develop and evaluate a combined additive manufacturing (AM) and acoustic droplet ejection (ADE) process for controlled GF patterning on 3D scaffolds.
  • To investigate the distribution and retention of recombinant human bone morphogenetic protein-2 (rhBMP-2) on polycaprolactone (PCL)-based tissue engineering constructs (TECs).
  • To assess the impact of controlled GF patterning on osteogenic differentiation of C2C12 cells.

Main Methods:

  • Utilized AM and ADE technologies to precisely deposit rhBMP-2 onto PCL-based TECs.
  • Employed three substrate types: PCL-N-hydroxysuccinimide (NHS), alkali-treated PCL (PCL-NaOH), and fibrin-coated PCL (PCL-Fibrin).
  • Analyzed GF spot size, spacing, retention after washing, and induced cellular differentiation.

Main Results:

  • Achieved patterned deposition of BMP-2 spots with approximately 250 μm diameter and 700 μm spacing.
  • An initial BMP-2 concentration >300 μg/L was required for detectable GF retention post-washing.
  • Osteogenic differentiation of C2C12 cells was observed at initial BMP-2 concentrations >750 μg/L, with spatial correlation to GF patterns.

Conclusions:

  • The developed AM-ADE process enables highly controlled GF patterning on 3D scaffolds.
  • This technology holds significant promise for creating advanced TECs with tailored biological cue presentation.
  • Controlled GF patterning can enhance cell behavior, leading to improved tissue formation in vitro and in vivo.