Page 211 - IJB-10-5

International Journal of Bioprinting Biomimetic osteochondral scaffold

(Boster Biological Technology, China) diluted at 1:500 and ± standard deviation. For statistical comparisons, the
incubated at 4°C overnight. The subsequent steps were Student’s t-test was used for two groups, and one-way
performed as described in Section 2.11. To investigate analysis of variance (ANOVA) was used for multiple
the production of glycosaminoglycan (GAG), after 21 groups. Additionally, p < 0.05 was considered statistically
days of cultivation, toluidine blue and alcian blue were significant. Three replicates were performed for
used to stain the fixed cell-laden osteochondral scaffolds. each analysis.
Briefly, after washing with DPBS, samples were fixed with
4% PFA for 30 min. The samples were then treated with 3. Results
1% toluidine blue and 1% alcian blue working solutions, 3.1. Design of osteochondral scaffolds with
respectively. Furthermore, RT-qPCR was performed spatiotemporally-controlled growth factor release
as described in Section 2.11. The primer sequences are In this study, an engineered osteochondral tissue was
displayed in Table 1. designed to facilitate osteochondral tissue regeneration
(Figure 1). The tissue consists of a subchondral layer, an
2.13. Statistical analysis interface layer, and a cartilage layer, and the tissue was
All experiments included at least three replicates for further seeded with MSCs with varied organizational
each sample, and numerical data are expressed as mean morphologies. Cryogenic 3D-printed BMP-2/TCP/PLGA

Figure 1. Schematic illustration of the development of engineered osteochondral tissue with the spatiotemporally controlled delivery strategy for
osteochondral regeneration. (A) The schematic biofabrication process of the tri-phasic osteochondral scaffold. (B) The cultivation of engineered
osteochondral tissue. (C) The spatiotemporally controlled delivery strategy for osteochondral regeneration. Abbreviations: BMP-2, bone morphogenetic
protein-2; FGF-18, fibroblast growth factor-18; GelMA, gelatin methacrylate; P(DLLA-TMC), poly(d,l-lactic acid-co-trimethylene carbonate); PLGA/β-
TCP, poly(lactic-co-glycolic acid)/β-tricalcium phosphate; rBMSC, rat bone marrow mesenchymal stem cell.

Volume 10 Issue 5 (2024) 203 doi: 10.36922/ijb.3229