New Bioink Derived from Neonatal Chicken Bone Marrow Cells and Its 3D-Bioprinted Niche for Osteogenic Stimulators

Abstract

This study examined whether neonatal chicken bone marrow cells (cBMCs) could support the osteogenesis of human stromal cells in a three-dimensional (3D) extracellular bioprinting niche. The majority (>95%) of 4-day-old cBMCs subcultured 5 times were positive for osteochondrogenesis-related genes (Col I, Col II, Col X, aggrecan, Sox9, osterix, Bmp2, osteocalcin, Runx2, and osteopontin) and their related proteins (Sox9, collagen type I, and collagen type II). LC–MS/MS analysis demonstrated that cBMC-conditioned medium (c-medium) contained proteins related to bone regeneration, such as periostin and members of the TGF-β family. Next, a significant increase in osteogenesis was detected in three human adipose tissue-derived stromal cell (hASC) lines, after exposure to c-medium concentrates in 2D culture (p < 0.05). To evaluate biological function in a 3D environment, we employed the cBMC-derived bioactive components as a cell-supporting biomaterial in collagen bioink, which was printed to construct a 3D hASC-laden scaffold for observing osteogenesis. Complete osteogenesis was detected in vitro. Moreover, after transplantation of the hASC-laden structure into rats, prominent bone formation was observed compared with that in control rats receiving scaffold-free hASC transplantation. These results demonstrated that substance(s) secreted by chick bone marrow cells clearly activated the osteogenesis of hASCs in 2D- or 3D-niches.

Bioengineered Living Bone Grafts—A Concise Review on Bioreactors and Production Techniques In Vitro

“…In another study, Kosik-Kozioł et al [57] showed that scaffolds composed of gelatin methacrylamide, alginate, β-TCP, and human BMDSCs increased the expression of ALP and bone gamma-carboxyglutamate protein (BGLAP) genes. In turn, Yang et al [58] developed a novel bioink composed of collagen, human ADSCs, and neonatal chicken BMDSCs-conditioned medium that contained bioactive components supporting bone restoration, such as TGF-β and periostin. In vitro experiments showed that 3D bioprinted hADSC-based constructs increased ALP activity, ECM mineralization, and the expression of osteogenesis-related genes (runt-related transcription factor 2 (RUNX2), COL 1, ALP, BMP-2, OCN, and OPN).…”

Three-Dimensional Avian Hematopoietic Stem Cell Cultures as a Model for Studying Disease Pathogenesis

“…Very little attention has been given to the use of 3D cell cultures in avian models. It has been studied predominantly in embryonal cells, such as primordial germ cells ( Chen et al, 2018 ), intestinal epithelial cells ( Pierzchalska et al, 2019 ), or chicken bone marrow cells as support for osteogenesis of human stromal cells ( Yang et al, 2020 ). Therefore, our current knowledge about 3D stem cell cultures comes from studies in mammals.…”