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Recent advances in bone tissue engineering have shown promise for bone repair post osteosarcoma excision. However, conflicting research on mesenchymal stem cells (MSCs) has raised concerns about their potential to either promote or inhibit tumor cell proliferation. It is necessary to thoroughly understand the interactions between MSCs and tumor cells. Most previous studies only focused on the interactions between cells within the tumor tissues. It has been challenging to develop an in vitro model of osteosarcoma excision sites replicating the complexity of the bone microenvironment and cell distribution. In this work, we designed and fabricated modular bioceramic scaffolds to assemble into a co-culture model. Because of the bone-like composition and mechanical property, tricalcium phosphate bioceramic could mimic the bone microenvironment and recapitulate the cell-extracellular matrix interaction. Moreover, the properties for easy assembly enabled the modular units to mimic the spatial distribution of cells in the osteosarcoma excision site. Under this co-culture model, MSCs showed a noticeable tumor-stimulating effect with a potential risk of tumor recurrence. In addition, tumor cells also could inhibit the osteogenic ability of MSCs. To undermine the stimulating effects of MSCs on tumor cells, we present the methods of pre-differentiated MSCs, which had lower expression of IL-8 and higher expression of osteogenic proteins. Both in vitro and in vivo studies confirm that pre-differentiated MSCs could maintain high osteogenic capacity without promoting tumor growth, offering a promising approach for MSCs' application in bone regeneration. Overall, 3D modular scaffolds provide a valuable tool for constructing hard tissue in vitro models. Bone tissue engineering using mesenchymal stem cells (MSCs) and biomaterials has shown promise for bone repair post osteosarcoma excision. However, conflicting researches on MSCs have raised concerns about their potential to either promote or inhibit tumor cell proliferation. It remains challenges to develop in vitro models to investigate cell interactions, especially of osteosarcoma with high hardness and special composition of bone tissue. In this work, modular bioceramic scaffolds were fabricated and assembled to co-culture models. The interactions between MSCs and MG-63 were manifested as tumor-stimulating and osteogenesis-inhibiting, which means potential risk of tumor recurrence. To undermine the stimulating effect, pre-differentiation method was proposed to maintain high osteogenic capacity without tumor-stimulating, offering a promising approach for MSCs' application in bone regeneration. [Display omitted] [ABSTRACT FROM AUTHOR]

Developmental engineering (DE) involves culturing various cells on modular scaffolds (MSs), yielding modular tissues (MTs) assembled into three-dimensional (3D) tissues, mimicking developmental biology. This study employs an integrated approach, merging experimental and mathematical methods to investigate the biological processes in MT cultivation and assembly. Human dermal fibroblasts (HDFs) were cultured on tissue culture plastics, poly(lactic acid) (PLA) discs with regular open structures, or spherical poly(methyl methacrylate) (PMMA) MSs, respectively. Notably, HDFs exhibited flattened spindle shapes when adhered to solid surfaces, and complex 3D structures when migrating into the structured voids of PLA discs or interstitial spaces between aggregated PMMA MSs, showcasing coordinated colonization of porous scaffolds. Empirical investigations led to power law models simulating density-dependent cell growth on solid surfaces or voids. Concurrently, a modified diffusion model was applied to simulate oxygen diffusion within tissues cultured on solid surfaces or porous structures. These mathematical models were subsequently combined to explore the influences of initial cell seeding density, culture duration, and oxygen diffusion on MT cultivation and assembly. The findings underscored the intricate interplay of factors influencing MT design for tissue assembly. The integrated approach provides insights into mechanistic aspects, informing bioprocess design for manufacturing MTs and 3D tissues in DE. [ABSTRACT FROM AUTHOR]

Developmental engineering (DE) aims to culture mammalian cells on corresponding modular scaffolds (scale: micron to millimeter), then assemble these into functional tissues imitating natural developmental biology processes. This research intended to investigate the influences of polymeric particles on modular tissue cultures. When poly(methyl methacrylate) (PMMA), poly(lactic acid) (PLA) and polystyrene (PS) particles (diameter: 5–100 µm) were fabricated and submerged in culture medium in tissue culture plastics (TCPs) for modular tissue cultures, the majority of adjacent PMMA, some PLA but no PS particles aggregated. Human dermal fibroblasts (HDFs) could be directly seeded onto large (diameter: 30–100 µm) PMMA particles, but not small (diameter: 5–20 µm) PMMA, nor all the PLA and PS particles. During tissue cultures, HDFs migrated from the TCPs surfaces onto all the particles, while the clustered PMMA or PLA particles were colonized by HDFs into modular tissues with varying sizes. Further comparisons revealed that HDFs utilized the same cell bridging and stacking strategies to colonize single or clustered polymeric particles, and the finely controlled open pores, corners and gaps on 3D-printed PLA discs. These observed cell–scaffold interactions, which were then used to evaluate the adaptation of microcarrier-based cell expansion technologies for modular tissue manufacturing in DE. [ABSTRACT FROM AUTHOR]

[Display omitted] • The optimized modular reinforced scaffold with enhanced mechanical properties is designed. • The optimization significantly improves the mechanical properties of the scaffold. • The optimization of the module is based on the defect shape. • The finite element model of customized alveolar bone defect is developed. • This kind of scaffold can be useful for treatment of critical-sized defects. A modular reinforced bone scaffold with enhanced mechanical properties has recently been developed by our group. It includes: 1) A load-bearing module: a skeleton which is made of a slowly degradable material, undertaking mechanical necessities of the scaffold, and 2) A bio-reactive module: a porous and biodegradable component undertaking biological necessities of the scaffold. The load-bearing module is placed into the bio-reactive module to reinforce it. This paper is dedicated to optimizing the load-bearing module for a certain customized alveolar bone defect. More specifically, a 3D-printed skeleton, made of polycaprolactone (PCL), is optimized based on the boundary conditions of the defect shape using the finite element method (FEM) to minimize the weight (to minimize the amount of PCL) and maximize the mechanical properties and porosity of the skeleton. Gelatin foam has been incorporated into the optimized skeleton through the aminolysis process to form the bio-reactive module. The mechanical characterization confirmed that the optimized load-bearing module has a bridge-like shape and can significantly improve the mechanical properties of the scaffold. Also, in vitro studies showed that the fabricated scaffold can improve cell proliferation and osteogenesis. This kind of scaffold can be useful for the treatment of critical-sized defects. [ABSTRACT FROM AUTHOR]

In the past decade, modular scaffolds prepared by assembling biocompatible and biodegradable building blocks (e.g. microspheres) have found promising applications in tissue engineering (TE) towards the repair/regeneration of damaged and impaired tissues. Nevertheless, to date this approach has failed to be transferred to the clinic due to technological limitations regarding microspheres patterning, a crucial issue for the control of scaffold strength, vascularization and integration in vivo . In this work, we propose a robust and reliable approach to address this issue through the fabrication of polycaprolactone (PCL) microsphere-based scaffolds with in-silico designed microarchitectures and high compression moduli. The scaffold fabrication technique consists of four main steps, starting with the manufacture of uniform PCL microspheres by fluidic emulsion technique. In the second step, patterned polydimethylsiloxane (PDMS) moulds were prepared by soft lithography. Then, layers of 500 µ m PCL microspheres with geometrically inspired patterns were obtained by casting the microspheres onto PDMS moulds followed by their thermal sintering. Finally, three-dimensional porous scaffolds were built by the alignment, stacking and sintering of multiple (up to six) layers. The so prepared scaffolds showed excellent morphological and microstructural fidelity with respect to the in-silico models, and mechanical compression properties suitable for load bearing TE applications. Designed porosity and pore size features enabled in vitro human endothelial cells adhesion and growth as well as tissue integration and blood vessels invasion in vivo . Our results highlighted the strong impact of spatial patterning of microspheres on modular scaffolds response, and pay the way about the possibility to fabricate in silico-designed structures featuring biomimetic composition and architectures for specific TE purposes., (© 2022 IOP Publishing Ltd.)