Today, various reconstruction strategies are applied after breast cancer surgery, including implants, microsurgical free tissue transfer, and lipofilling. Nevertheless, these procedures are associated with various risks and limitations. A technique which is gaining more attention is lipofilling. However, it is typically not suggested for large defects due to volume retention uncertainty related to fat absorption, that require multiple injection procedures. Therefore, in the present manuscript the use of 3D (bio)printing technology in the field of adipose tissue engineering was investigated. More specifically, polyester-hydrogel hybrid scaffolds were developed. To this end, as polyester material, a photocurable low molar mass (i.e. around 2500 g mol-1)-1 ) acrylate-endcapped urethane-based polyesters (AUP's), with a poly (D,L lactic acid)/poly (epsilon-caprolactone) (PDLLA/PCL) copolymer backbone was synthesized, providing structural integrity to the final construct. These polyesters were processed into porous 3D scaffolds using an indirect 3D printing method taking advantage of a sacrificial poly (vinyl alcohol) (PVA) as produced by fused filament fabrication (FFF). Cubic porous scaffolds were obtained with pore size around 1000 mu m and strut diameters around 300 mu m and a compression modulus close to 500 kPa. Next, the obtained scaffolds were filled with a shear-thinning gelatin-based hydrogel containing primary human bone marrow-derived stem cells (hBMSCs). Next, the homogenously hBMSCs-laden constructs were cultured up to three weeks, to investigate adipogenic differentiation based on expression of lineage-specific biomarkers, both on gene and on protein level. qRT-PCR and Western blotting showed an initial up-regulation of PPAR-gamma gamma followed by a decrease towards week 3, while expression of ADP and LP gradually increased throughout the entire experiment. Finally, a progressive augmentation in intracellular lipid accumulation was observed over the entire incubation period, indicating the formation of mature adipocytes. In conclusion, the cell-laden scaffolds enabled differentiation of hBMSCs into the adipogenic lineage. This hybrid approach proved well-suited for tissue engineering applications, presenting an innovative pathway towards adipose tissue reconstruction.
Combining photocrosslinkable polyester-based scaffolds with a cell-encapsulated shear-thinning gelatin hydrogel as a hybrid strategy for adipose tissue reconstruction
Alessio, N.;La Gatta, A.;Schiraldi, C.;
2024
Abstract
Today, various reconstruction strategies are applied after breast cancer surgery, including implants, microsurgical free tissue transfer, and lipofilling. Nevertheless, these procedures are associated with various risks and limitations. A technique which is gaining more attention is lipofilling. However, it is typically not suggested for large defects due to volume retention uncertainty related to fat absorption, that require multiple injection procedures. Therefore, in the present manuscript the use of 3D (bio)printing technology in the field of adipose tissue engineering was investigated. More specifically, polyester-hydrogel hybrid scaffolds were developed. To this end, as polyester material, a photocurable low molar mass (i.e. around 2500 g mol-1)-1 ) acrylate-endcapped urethane-based polyesters (AUP's), with a poly (D,L lactic acid)/poly (epsilon-caprolactone) (PDLLA/PCL) copolymer backbone was synthesized, providing structural integrity to the final construct. These polyesters were processed into porous 3D scaffolds using an indirect 3D printing method taking advantage of a sacrificial poly (vinyl alcohol) (PVA) as produced by fused filament fabrication (FFF). Cubic porous scaffolds were obtained with pore size around 1000 mu m and strut diameters around 300 mu m and a compression modulus close to 500 kPa. Next, the obtained scaffolds were filled with a shear-thinning gelatin-based hydrogel containing primary human bone marrow-derived stem cells (hBMSCs). Next, the homogenously hBMSCs-laden constructs were cultured up to three weeks, to investigate adipogenic differentiation based on expression of lineage-specific biomarkers, both on gene and on protein level. qRT-PCR and Western blotting showed an initial up-regulation of PPAR-gamma gamma followed by a decrease towards week 3, while expression of ADP and LP gradually increased throughout the entire experiment. Finally, a progressive augmentation in intracellular lipid accumulation was observed over the entire incubation period, indicating the formation of mature adipocytes. In conclusion, the cell-laden scaffolds enabled differentiation of hBMSCs into the adipogenic lineage. This hybrid approach proved well-suited for tissue engineering applications, presenting an innovative pathway towards adipose tissue reconstruction.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
