Effect of Calcium-Infiltrated Hydroxyapatite Scaffolds on the Hematopoietic Fate of Human Umbilical Vein Endothelial Cells

Abstract

Foamed hydroxyapatite offers a three-dimensional scaffold for the development of bone constructs, mimicking perfectly the in vivo bone structure. In vivo, calcium release at the surface is assumed to provide a locally increased gradient supporting the maintenance of the hematopoietic stem cells niche. We fabricated hydroxyapatite scaffolds with high surface calcium concentration by infiltration, and used human umbilical vein endothelial cells (HUVECs) as a model to study the effects on hematopoietic lineage direction. HUVECs are umbilical vein-derived and thus possess progenitor characteristics, with a prospective potential to give rise to hematopoietic lineages. HUVECs were cultured for long term on three-dimensional porous hydroxyapatite scaffolds, which were either infiltrated biphasic foams or untreated. Controls were cultured in two-dimensional dishes. The release of calcium into culture medium was determined, and cells were analyzed for typical hematopoietic and endothelial gene expressions, surface markers by flow cytometry, and hematopoietic potential using colony-forming unit assays. Our results indicate that the biphasic foams promoted a hematopoietic lineage direction of HUVECs, suggesting an improved in vivo-like scaffold for hematopoietic bone tissue engineering.

Modulating electrospun polycaprolactone scaffold morphology and composition to alter endothelial cell proliferation and angiogenic gene response

“…et al . noted a doubling in the relative gene expression of VEGF from HUVECs between 7 and 14 days when cultured on hydroxyapatite scaffolds [ 53 ]. The same applies for MMP1, MMP2 and TIMP2, whereby all four morphologies showed a relative decrease in gene expression for these genes after 12 days compared to 6 days.…”

“…This trend has been noted before when studying HUVECs. Zhang, G. et al noted a doubling in the relative gene expression of VEGF from HUVECs between 7 and 14 days when cultured on hydroxyapatite scaffolds [53]. The same applies for MMP1, MMP2 https://doi.org/10.1371/journal.pone.0240332.g007…”

Comprehensive phenotyping of endothelial cells using flow cytometry 2: Human

“…Pan‐endothelial markers are similar between species, except for human‐specific Uea‐1 [2‐4]. Human‐specific dump channels include CD90 to exclude mesenchymal cells and CD235a to exclude erythrocytes [2,5‐9]. The listed dump channel markers can be replaced with other established markers for the dump subset(s).…”

Journey into Bone Models: A Review

“…The following are available online at , Table S1: Overview on scaffold based bone models published between 2007 and 2017, Table S2: Summary of bioreactors suitable for bone models, Table S3: Summary of microfluidic bone models. References [ 221 , 222 , 223 , 224 , 225 , 226 , 227 , 228 , 229 , 230 , 231 , 232 , 233 , 234 , 235 , 236 , 237 , 238 , 239 , 240 , 241 , 242 , 243 , 244 , 245 , 246 , 247 , 248 , 249 , 250 , 251 , 252 , 253 , 254 , 255 , 256 ] are cited in the supplementary materials.…”