Histological evaluation of tendon formation using a scaffold-free three-dimensional-bioprinted construct of human dermal fibroblasts under in vitro static tensile culture.
Regen Ther
; 11: 47-55, 2019 Dec.
Article
in En
| MEDLINE
| ID: mdl-31193148
ABSTRACT
INTRODUCTION:
Tendon tissue engineering requires scaffold-free techniques for safe and long-term clinical applications and to explore alternative cell sources to tenocytes. Therefore, we histologically assessed tendon formation in a scaffold-free Bio-three-dimensional (3D) construct developed from normal human dermal fibroblasts (NHDFs) using our Bio-3D printer system under tensile culture in vitro.METHODS:
Scaffold-free ring-like tissues were constructed from 120 multicellular spheroids comprising NHDFs using a bio-3D printer. Ring-like tissues were cultured in vitro under static tensile-loading with or without in-house tensile devices (tension-loaded and tension-free groups), with increases in tensile strength applied weekly to the tensile-loaded group. After a 4 or 8-week culture on the device, we evaluated histological findings according to tendon-maturing score and immunohistological findings of the middle portion of the tissues for both groups (n = 4, respectively).RESULTS:
Histology of the tension-loaded group revealed longitudinally aligned collagen fibers with increased collagen deposition and spindle-shaped cells with prolonged culture. By contrast, the tension-free group showed no organized cell arrangement or collagen fiber structure. Additionally, the tension-loaded group showed a significantly improved tendon-maturing score as compared with that for the tension-free group at week 8. Moreover, immunohistochemistry revealed tenascin C distribution with a parallel arrangement in the tensile-loading direction at week 8 in the tension-loaded group, which exhibited stronger scleraxis-staining intensity than that observed in the tension-free group at weeks 4 and 8.CONCLUSIONS:
The NHDF-generated scaffold-free Bio-3D construct underwent remodeling and formed tendon-like structures under tensile culture in vitro.
3D, three-dimensional; ECM, extracellular matrix; ESCs, embryonic stem cells; H&E, hematoxylin and eosin; HDF, human dermal fibroblast; Human dermal fibroblast; In vitro study; MCSs, multicellular spheroids; Multicellular spheroid; NHDFs, normal HDFs; Scaffold-free; TDSCs, tendon-derived stem cells; TGF, transforming growth factor; Tendon formation; Tensile culture
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Collection:
01-internacional
Database:
MEDLINE
Language:
En
Journal:
Regen Ther
Year:
2019
Document type:
Article
Affiliation country: