RESUMO
Polycaprolactone (PCL) scaffolds have recently been developed via efficient and green supercritical carbon dioxide (scCO2) melt-state foaming. However, previously reported gas-foamed scaffolds sometimes showed insufficient interconnectivity or pore size for tissue engineering. In this study, we have correlated the thermal and rheological properties of PCL scaffolds with their porous morphology by studying four foamed samples with varied molecular weight (MW), and particularly aimed to clarify the required properties for the fabrication of scaffolds with favorable interconnected macropores. DSC and rheological tests indicate that samples show a delayed crystallization and enhanced complex viscosity with the increasing of MW. After foaming, scaffolds (27 kDa in weight-average molecular weight) show a favorable morphology (pore size = 70-180 µm, porosity = 90% and interconnectivity = 96%), where the lowest melt strength favors the generation of interconnected macropore, and the most rapid crystallization provides proper foamability. The scaffolds (27 kDa) also possess the highest Young's modulus. More importantly, owing to the sufficient room and favorable material transportation provided by highly interconnected macropores, cells onto the optimized scaffolds (27 kDa) perform vigorous proliferation and superior adhesion and ingrowth, indicating its potential for regeneration applications. Furthermore, our findings provide new insights into the morphological control of porous scaffolds fabricated by scCO2 foaming, and are highly relevant to a broader community that is focusing on polymer foaming.
RESUMO
Bone tissue engineering has substantial potential for the treatment of massive bone defects; however, efficient vascularization coupled with bone regeneration still remains a challenge in this field. In the current study, supercritical carbon dioxide (scCO2) foaming technique was adopted to fabricate mesoporous bioactive glasses (MBGs) particle-poly (lactic-co-glycolic acid) (PLGA) composite scaffolds with appropriate mechanical and degradation properties as well as in vitro bioactivity. The MBG-PLGA scaffolds incorporating the bioactive lipid FTY720 (designated as FTY/MBG-PLGA) exhibited simultaneously sustained release of the bioactive lipid and ions. In addition to providing a favorable microenvironment for cellular adhesion and proliferation, FTY/MBG-PLGA scaffolds significantly facilitated the in vitro osteogenic differentiation of rBMSCs and also markedly stimulated the upregulation of Hif-1α expression via the activation of the Erk1/2 pathway, which mediated the osteogenic and pro-angiogenic effects on rBMSCs. Furthermore, FTY/MBG-PLGA extracts induced superior in vitro angiogenic performance of HUVECs. In vivo evaluation of critical-sized rat calvarial bone defects indicated that FTY/MBG-PLGA scaffolds potently promoted vascularized bone regeneration. Notably, the significantly enhanced formation of type H vessels (CD31hiEmcnhi neo-vessels) was observed in newly formed bone tissue in FTY/MBG-PLGA group, strongly suggesting that FTY720 and therapeutic ions released from the scaffolds synergistically induced more type H vessel formation, which indicated the coupling of angiogenesis and osteogenesis to achieve efficiently vascularized bone regeneration. Overall, the results indicated that the foamed porous MBG-PLGA scaffolds incorporating bioactive lipids achieved desirable vascularization-coupled bone formation and could be a promising strategy for bone regenerative medicine. STATEMENT OF SIGNIFICANCE: Efficacious coupling of vascularizationandbone formation is critical for the restoration of large bone defects. Anoveltechnique was used to fabricate composite scaffolds incorporating bioactive lipids which possessedsynergistic cues of bioactive lipids and therapeutic ions to potently promotebone regenerationas well as vascularization. The underlying molecular mechanism for the osteogenic and pro-angiogenic effects of the compositescaffolds was unveiled. Interestingly, the scaffolds were furtherfoundto enhance the formation oftype H capillarieswithin the bone healing microenvironment to couple angiogenesis to osteogenesis to achieve satisfyingvascularizedbone regeneration.These findings provide a novel strategy to develop efficiently vascularized engineering constructs to treat massive bone defects.
