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3D printed hydrogels with oxidized cellulose nanofibers and silk fibroin for the proliferation of lung epithelial stem cells.

ABSTRACT: A novel biomaterial ink consisting of regenerated silk fibroin (SF) and 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-oxidized bacterial cellulose (OBC) nanofibrils was developed for 3D printing lung tissue scaffold. Silk fibroin backbones were cross-linked using horseradish peroxide/H₂O₂ to form printed hydrogel scaffolds. OBC with a concentration of 7wt% increased the viscosity of inks during the printing process and further improved the shape fidelity of the scaffolds. Rheological measurements and image analyses were performed to evaluate inks printability and print shape fidelity. Three-dimensional construct with ten layers could be printed with ink of 1SF-2OBC (SF/OBC?=?1/2, w/w). The composite hydrogel of 1SF-1OBC (SF/OBC?=?1/1, w/w) printed at 25 °C exhibited a significantly improved compressive strength of 267?±?13 kPa and a compressive stiffness of 325?±?14 kPa at 30% strain, respectively. The optimized printing parameters for 1SF-1OBC were 0.3 bar of printing pressure, 45 mm/s of printing speed and 410 ?m of nozzle diameter. Furthermore, OBC nanofibrils could be induced to align along the print lines over 60% degree of orientation, which were analyzed by SEM and X-ray diffraction. The orientation of OBC nanofibrils along print lines provided physical cues for guiding the orientation of lung epithelial stem cells, which maintained the ability to proliferate and kept epithelial phenotype after 7 days' culture. The 3D printed SF-OBC scaffolds are promising for applications in lung tissue engineering.

SUBMITTER: Huang L

PROVIDER: S-EPMC7590576 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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3D printed hydrogels with oxidized cellulose nanofibers and silk fibroin for the proliferation of lung epithelial stem cells.

Huang Li L Yuan Wei W Hong Yue Y Fan Suna S Yao Xiang X Ren Tao T Song Lujie L Yang Gesheng G Zhang Yaopeng Y

Cellulose (London, England) 20201026 1

A novel biomaterial ink consisting of regenerated silk fibroin (SF) and 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-oxidized bacterial cellulose (OBC) nanofibrils was developed for 3D printing lung tissue scaffold. Silk fibroin backbones were cross-linked using horseradish peroxide/H<sub>2</sub>O<sub>2</sub> to form printed hydrogel scaffolds. OBC with a concentration of 7wt% increased the viscosity of inks during the printing process and further improved the shape fidelity of the scaffolds. Rh ...[more]

PMID: 33132545

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Project description:The repair of bone defects remains a major challenge in the clinic, and treatment requires bone grafts or bone replacement materials. Existing biomaterials have many limitations and cannot meet the various needs of clinical applications. To treat bone defects, we constructed a nanohydroxyapatite (nHA)/methylacrylylated silk fibroin (MASF) composite biological scaffold using photocurable 3D printing technology. In this study, scanning electron microscopy (SEM) was used to detect the changes in the morphological structure of the composite scaffold with different contents of nanohydroxyapatite, and FTIR was used to detect the functional groups and chemical bonds in the composite scaffold to determine the specific components of the scaffold. In in vitro experiments, bone marrow mesenchymal stem cells from SD rats were cocultured with scaffolds soaking solution, and the cytotoxicity, cell proliferation, Western blot analysis, Quantitative real-time PCR analysis, bone alkaline phosphatase activity and alizarin red staining of scaffolds were detected to determine the biocompatibility of scaffolds and the effect of promoting proliferation and osteogenesis of bone marrow mesenchymal stem cells in vitro. In the in vivo experiment, the skull defect was constructed by adult SD rats, and the scaffold was implanted into the skull defect site. After 4 weeks and 8 weeks of culture, the specific osteogenic effect of the scaffold in the skull defect site was detected by animal micro-CT, hematoxylin and eosin (HE) staining and Masson's staining. Through the analysis of the morphological structure of the scaffold, we found that the frame supported good retention of the lamellar structure of silk fibroin, when mixed with nHA, the surface of the stent was rougher, the cell contact area increased, and cell adhesion and lamellar microstructure for cell migration and proliferation of the microenvironment provided a better space. FTIR results showed that the scaffold completely retained the β -folded structure of silk fibroin, and the scaffold composite was present without obvious impurities. The staining results of live/dead cells showed that the constructed scaffolds had no significant cytotoxicity, and thw CCK-8 assay also showed that the constructed scaffolds had good biocompatibility. The results of osteogenic induction showed that the scaffold had good osteogenic induction ability. Moreover, the results also showed that the scaffold with a MASF: nHA ratio of 1: 0.5 (SFH) showed better osteogenic ability. The micro-CT and bone histometric results were consistent with the in vitro results after stent implantation, and there was more bone formation at the bone defect site in the SFH group.This research used photocurable 3D printing technology to successfully build an osteogenesis bracket. The results show that the constructed nHA/MASF biological composite material, has good biocompatibility and good osteogenesis function. At the same time, in the microenvironment, the material can also promote bone defect repair and can potentially be used as a bone defect filling material for bone regeneration applications.

Dataset Information

3D printed hydrogels with oxidized cellulose nanofibers and silk fibroin for the proliferation of lung epithelial stem cells.

Publications

3D printed hydrogels with oxidized cellulose nanofibers and silk fibroin for the proliferation of lung epithelial stem cells.

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