An Innovative Collagen-Based Cell-Printing Method for Obtaining Human Adipose Stem Cell-Laden Structures Consisting of Core-Sheath Structures for Tissue Engineering

Biomacromolecules. 2016 Apr 11;17(4):1365-75. doi: 10.1021/acs.biomac.5b01764. Epub 2016 Mar 25.

Abstract

Three-dimensional (3D) cell printing processes have been used widely in various tissue engineering applications due to the efficient embedding of living cells in appropriately designed micro- or macro-structures. However, there are several issues to overcome, such as the limited choice of bioinks and tailor-made fabricating strategies. Here, we suggest a new, innovative cell-printing process, supplemented with a core-sheath nozzle and an aerosol cross-linking method, to obtain multilayered cell-laden mesh structure and a newly considered collagen-based cell-laden bioink. To obtain a mechanically and biologically enhanced cell-laden structure, we used collagen-bioink in the core region, and also used pure alginate in the sheath region to protect the cells in the collagen during the printing and cross-linking process and support the 3D cell-laden mesh structure. To achieve the most appropriate conditions for fabricating cell-embedded cylindrical core-sheath struts, various processing conditions, including weight fractions of the cross-linking agent and pneumatic pressure in the core region, were tested. The fabricated 3D MG63-laden mesh structure showed significantly higher cell viability (92 ± 3%) compared with that (83 ± 4%) of the control, obtained using a general alginate-based cell-printing process. To expand the feasibility to stem cell-embedded structures, we fabricated a cell-laden mesh structure consisting of core (cell-laden collagen)/sheath (pure alginate) using human adipose stem cells (hASCs). Using the selected processing conditions, we could achieve a stable 3D hASC-laden mesh structure. The fabricated cell-laden 3D core-sheath structure exhibited outstanding cell viability (91%) compared to that (83%) of an alginate-based hASC-laden mesh structure (control), and more efficient hepatogenic differentiations (albumin: ∼ 1.7-fold, TDO-2: ∼ 7.6-fold) were observed versus the control. The selection of collagen-bioink and the new printing strategy could lead to an efficient way to achieve 3D cell-laden mesh structures that mimic the anatomical architecture of a patient's defective region.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipose Tissue / cytology*
  • Alginates / chemistry
  • Cell Survival
  • Collagen / chemistry
  • Glucuronic Acid / chemistry
  • Hexuronic Acids / chemistry
  • Humans
  • Printing, Three-Dimensional / instrumentation*
  • Stem Cells / cytology*
  • Tissue Engineering / methods*
  • Tissue Scaffolds / chemistry*

Substances

  • Alginates
  • Hexuronic Acids
  • Glucuronic Acid
  • Collagen