The scaffold-free isogenic artificial tracheas produced by a bio-3D printer could be utilized as tracheal grafts in rats.

Current scaffold‐based tissue engineering approaches are subject to several limitations, such as design inflexibility, poor cytocompatibility, toxicity, and post‐transplant degradation. Thus, scaffold‐free tissue‐engineered structures can be a promising solution to overcome the issues associated with classical scaffold‐based materials in clinical transplantation. The present study seeks to optimize the culture conditions and cell combinations used to generate scaffold‐free structures using a Bio‐3D printing system. Human cartilage cells, human fibroblasts, human umbilical vein endothelial cells, and human mesenchymal stem cells from bone marrow are aggregated into spheroids and placed into a Bio‐3D printing system with dedicated needles positioned according to 3D configuration data, to develop scaffold‐free trachea‐like tubes. Culturing the Bio‐3D‐printed structures with proper flow of specific medium in a bioreactor facilitates the rearrangement and self‐organization of cells, improving physical strength and tissue function. The Bio‐3D‐printed tissue forms small‐diameter trachea‐like tubes that are implanted into rats with the support of catheters. It is confirmed that the tubes are viable in vivo and that the tracheal epithelium and capillaries proliferate. This tissue‐engineered, scaffold‐free, tubular structure can represent a significant step toward clinical application of bioengineered organs.

Lung transplantation is the last option for the treatment of end stage chronic lung disorders. Because the shortage of donor lung organs represents the main hurdle, lung regeneration has been considered to overcome this hurdle. Recellularization of decellularized organ scaffold is a promising option for organ regeneration. Although detergents are ordinarily used for decellularization, other approaches are possible. Here we used high alkaline (pH12) sodium hydroxide (NaOH)-PBS solution without detergents for lung decellularization and compared the efficacy on DNA elimination and ECM preservation with detergent based decellularization solutions CHAPS and SDS. Immunohistochemical image analysis showed that cell components were removed by NaOH solution as well as other detergents. A Collagen and GAG assay showed that the collagen reduction of the NaOH group was comparable to that of the CHAPS and SDS groups. However, DNA reduction was more significant in the NaOH group than in other groups (p < 0.0001). The recellularization of HUVEC revealed cell attachment was not inferior to that of the SDS group. Ex vivo functional analysis showed 100% oxygen ventilation increased oxygen partial pressure as artificial hemoglobin vesicle-PBS solution passed through regenerated lungs in the SDS or NaOH group. It was concluded that the NaOH-PBS based decellularization solution was comparable to ordinal decellularizaton solutions and competitive in cost effectiveness and residues in the decellularized scaffold negligible, thus providing another potential option to detergent for future clinical usage.

Recrystallization of polycrystalline silicon (poly-Si) film by excimer laser annealing (ELA) is discussed by considering the experimental results that the three stages of nucleation, textured grain growth and secondary grain growth were observed. Although the phenomenon of nucleation in the amorphous silicon (a-Si) is understood by considering crystallization from the super cooled liquid, the growth mechanisms of the textured grain and secondary grain are not understood by this, because the melting point of poly-Si which has already been formed on the entire surface during these growth stages is higher than that of a-Si. The recrystallization mechanism considering the dislocation movement is introduced to investigate the present phenomenon. It also clarifies the reason why secondary grain growth occurs under the critical conditions of laser irradiation energy and shot number. The feasibility of nucleation through the super cooled liquid is also discussed.

Various strategies have been attempted to replace esophageal defects with natural or artificial substitutes using tissue engineering. However, these methods have not yet reached clinical application because of the high risks related to their immunogenicity or insufficient biocompatibility. In this study, we developed a scaffold-free structure with a mixture of cell types using bio-three-dimensional (3D) printing technology and assessed its characteristics in vitro and in vivo after transplantation into rats. Normal human dermal fibroblasts, human esophageal smooth muscle cells, human bone marrow-derived mesenchymal stem cells, and human umbilical vein endothelial cells were purchased and used as a cell source. After the preparation of multicellular spheroids, esophageal-like tube structures were prepared by bio-3D printing. The structures were matured in a bioreactor and transplanted into 10-12-week-old F344 male rats as esophageal grafts under general anesthesia. Mechanical and histochemical assessment of the structures were performed. Among 4 types of structures evaluated, those with the larger proportion of mesenchymal stem cells tended to show greater strength and expansion on mechanical testing and highly expressed α-smooth muscle actin and vascular endothelial growth factor on immunohistochemistry. Therefore, the structure with the larger proportion of mesenchymal stem cells was selected for transplantation. The scaffold-free structures had sufficient strength for transplantation between the esophagus and stomach using silicon stents. The structures were maintained in vivo for 30 days after transplantation. Smooth muscle cells were maintained, and flat epithelium extended and covered the inner surface of the lumen. Food had also passed through the structure. These results suggested that the esophagus-like scaffold-free tubular structures created using bio-3D printing could hold promise as a substitute for the repair of esophageal defects.