Toward mimicking the bone structure: design of novel hierarchical scaffolds with a tailored radial porosity gradient

Abstract

Guiding bone regeneration poses still unmet challenges due to several drawbacks of current standard treatments in the clinics. A possible solution may rely on the use of three-dimensional scaffolds with optimized structural properties in combination with human mesenchymal stem cells (hMSCs). Bone presents a radial gradient structure from the outside, where the cortical bone is more compact (porosity ranging from 5% to 10%), toward the inner part, where the cancellous bone is more porous (porosity ranging from 50% to 90%). Here, we present a new scaffold design with a built-in gradient in porosity, which approximate the radial bone structure. The pores of the outer ring were 500 μm, the ones in the middle zone were 750 μm and the inner part presented pores of 1000 μm. The porosity of each scaffold region resembled the gradient present in bone, with the outer ring having a porosity of 29.6% ± 5%, the middle and inner regions a porosity of 50.8% ± 8.1% and 77.6% ± 3.2% respectively. hMSCs behavior was analyzed in terms of growth, extracellular matrix deposition and differentiation toward the osteogenic lineage. A trend was displayed by the hMSCs residing in different zones of the gradient scaffolds after 7, 14 and 28 days of culture in mineralization medium. Osteogenic differentiation was influenced by pore size and location in scaffolds displaying a radial porosity gradient. Cell differentiation was confirmed by gene expression with upregulation of Runx2 and bone sialoprotein markers. Mineralization staining further confirmed the maturation of cell differentiation, as indicated by the presence of calcium and phosphate mineral deposits.

Improving cell distribution on 3D additive manufactured scaffolds through engineered seeding media density and viscosity

“…Interestingly, ALP activity increased from day 14 to day 28 in cells seeded with CS method. Concordantly with our results, a peak of ALP activity after 28 days of culture on scaffolds seeded with hMSCs via the CS method has previously been reported [66,67]. In contrast, ALP decreased from day 14 to day 28 in cells seeded with MS methods despite the macroM type.…”

Bioreactor culture duration of engineered constructs influences bone formation by mesenchymal stem cells

“…This phenomenon merits further investigation using composite scaffolds with consistent pore size and geometry. Others who modulate pore size, distribution, and interconnectivity using additive manufacturing to regulate geometry reported the favorable effect of a porosity gradient within scaffolds on MSC osteogenic differentiation [50–53]. …”

Porous Scaffold Design for Additive Manufacturing in Orthopedics: A Review

“…Shi et al (2017) made a three-layer gradient scaffold through changing struct diameter, in which the porosity was from 68.5% to 88.2% with the Young modulus of 12-18 GPa. Another method was to connect the completely dense layer and porous layer to form a gradient scaffold (Di Luca et al, 2016;Fousova et al, 2017). By changing the diameter of the rhombus cell struts, the scaffold was made into three different porosity and pore (Zhang et al, 2019).…”

“…The results showed that alkaline phosphatase (ALP) activity was higher in the high porosity of the same scaffold. Their study suggested that porosity can influence the differentiation of hMSCs into osteoblasts (Di Luca et al, 2016). Another study investigated the behavior of hMSCs in different porosity regions of gradient scaffolds.…”