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Polydopamine coating with static magnetic field promotes the osteogenic differentiation of human bone-derived mesenchymal stem cells on three-dimensional printed porous titanium scaffolds by upregulation of the BMP-Smads signaling pathway

ABSTRACT: Bone regeneration has always been a hot topic for orthopedic surgeons. The role of polydopamine coating in promoting bone regeneration has attracted much attention. Static magnetic field (SMF) is considered an effective and noninvasive treatment for enhancing bone regeneration. However, the effect of polydopamine combined with SMF on bone regeneration on scaffolds is not clear. The aim of this study was to investigate the effects and potential mechanism of polydopamine coating combined with SMF on bone regeneration in three-dimensional printed scaffolds. The polydopamine coating (pTi group) was applied onto porous Ti6Al4V scaffolds (Ti group). Surface characterization was performed by scanning electron microscopy. The 100 mT SMF environment (pTi-SMF group) was established to enhance osteogenic differentiation of human bone-derived mesenchymal stem cells (hBMSCs) on polydopamine coating scaffolds. The cell viability and proliferation were significantly enhanced in the SMF environment (pTi-SMF vs. Ti: P=0.005). Improved morphology (pTi-SMF vs. pTi: P=0.024, pTi-SMF vs. Ti: P=0.001) and adhesion (Ti: x?±s=1.585±0.324; pTi: x?±s=2.164±0.314; pTi-SMF: x?±s=4.634±0.247, P<0.001) of hBMSCs were observed in the pTi-SMF group. The high expression of osteogenesis-related RNA and protein (ALP: Ti, x?±s=1.249±0.218; pTi, x?±s=2.503±0.209; pTi-SMF, x?±s=2.810±0.246. OCN: Ti, x?±s=1.483±0.304; pTi, x?±s=3.636±0.322; pTi-SMF, x?±s=4.641±0.278. Runx2: Ti, x?±s=1.372±0.227; pTi, x?±s=3.054±0.229; pTi-SMF, x?±s=3.914±0.253) was found in the pTi-SMF group (pTi-SMF vs. Ti: P<0.001). Proteomics was applied to explore the osteogenic mechanism of polydopamine coating combined with SMF. A total of 147 different proteins were identified between the pTi-SMF and Ti group. The osteogenic effect might be associated with the BMP-Smads signaling pathway (pTi-SMF vs. Ti: BMPR1A, P=0.001; BMPR2, P<0.001; Smad4, P=0.001; Smad1/5/8, P=0.008). In conclusion, the osteogenic differentiation of hBMSCs on polydopamine coating scaffolds could be enhanced by SMF stimulation by upregulation of the BMP-Smads signaling pathway.

SUBMITTER: Kong L

PROVIDER: S-EPMC7791488 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Project description:Biocatalysis is increasingly becoming an alternative method for the synthesis of industrially relevant complex molecules. This can be realized by using enzyme immobilized polysaccharide-based 3D scaffolds as compatible carriers, with defined properties. Especially, immobilization of either single or multiple enzymes on a 3D printed polysaccharide scaffold, exhibiting well-organized interconnected porous structure and morphology, is a versatile approach to access the performance of industrially important enzymes. Here, we demonstrated the use of nanocellulose-based 3D porous scaffolds for the immobilization of glycosyltransferases, responsible for glycosylation in natural biosynthesis. The scaffolds were produced using an ink containing nanofibrillated cellulose (NFC), carboxymethyl cellulose (CMC), and citric acid. Direct-ink-writing 3D printing followed by freeze-drying and dehydrothermal treatment at elevated temperature resulted in chemically cross-linked scaffolds, featuring tunable negative charges (2.2-5.0 mmol/g), pore sizes (10-800 μm), fluid uptake capacity, and exceptional dimensional and mechanical stability in the wet state. The negatively charged scaffolds were applied to immobilize two sugar nucleotide-dependent glycosyltransferases (C-glycosyltransferase, Zbasic2-CGT; sucrose synthase, Zbasic2-SuSy), each harboring a cationic binding module (Zbasic2) to promote charge-based enzyme adsorption. Both enzymes were immobilized at ∼30 mg of protein/g of dry carrier (∼20% yield), independent of the scaffold used. Their specific activities were 0.50 U/mg (Zbasic2-CGT) and 0.19 U/mg (Zbasic2-SuSy), corresponding to an efficacy of 37 and 18%, respectively, compared to the soluble enzymes. The glycosyltransferases were coimmobilized and shown to be active in a cascade reaction to give the natural C-glycoside nothofagin from phloretin (1.0 mM; ∼95% conversion). All enzyme bound scaffolds showed reusability of a maximum of 5 consecutive reactions. These results suggest that the 3D printed and cross-linked NFC/CMC-based scaffolds could present a class of solid carriers for enzyme (co)-immobilization, with promising applications in glycosyltransferase-catalyzed synthesis and other fields of biocatalysis.

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Polydopamine coating with static magnetic field promotes the osteogenic differentiation of human bone-derived mesenchymal stem cells on three-dimensional printed porous titanium scaffolds by upregulation of the BMP-Smads signaling pathway

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