Thymosin beta 10 loaded ZIF-8/sericin hydrogel promoting angiogenesis and osteogenesis for bone regeneration.

Angiogenesis is pivotal for osteogenesis during bone regeneration. A hydrogel that promotes both angiogenesis and osteogenesis is essential in bone tissue engineering. However, creating scaffolds with the ideal balance of biodegradability, osteogenic, and angiogenic properties poses a challenge. Thymosin beta 10 (TMSB10), known for its dual role in angiogenesis and osteogenesis differentiation, faces limitations due to protein activity preservation. To tackle this issue, ZIF-8 was engineered as a carrier for TMSB10 (TMSB10@ZIF-8), and subsequently integrated into the self-assembled sericin hydrogel. The efficacy of the composite hydrogel in bone repair was assessed using a rat cranial defect model. Characterization of the nanocomposites confirmed the successful synthesis of TMSB10@ZIF-8, with a TMSB10 encapsulation efficiency of 88.21 %. The sustained release of TMSB10 from TMSB10@ZIF-8 has significantly enhanced tube formation in human umbilical vein endothelial cells (HUVECs) in vitro and promoted angiogenesis in the chicken chorioallantoic membrane (CAM) model in vivo. It has markedly improved the osteogenic differentiation ability of MC 3 T3-E1 cells in vitro. 8 weeks post-implantation, the TMSB10@ZIF-8/ Sericin hydrogel group exhibited significant bone healing (86.77 ± 8.91 %), outperforming controls. Thus, the TMSB10@ZIF-8/Sericin hydrogel, leveraging ZIF-8 for TMSB10 delivery, emerges as a promising bone regeneration scaffold with substantial clinical application potential.

Degradation Kinetic Studies of BSA@ZIF-8 Nanoparticles with Various Zinc Precursors, Metal-to-Ligand Ratios, and pH Conditions.

Nanosized zeolitic imidazolate framework particles (ZIF-8 nanoparticles [NPs]) have strong potential as effective carriers for both in vivo and in vitro protein drug delivery. Synthesis of ZIF-8 and stability of protein encapsulation within ZIF-8 are affected by several factors, notably the metal ion source and molar ratio. To systematically investigate these factors, we investigated such effects using BSA as a model test protein to synthesize BSA@ZIF-8 NPs at various metal-to-ligand (M:L) ratios. SEM, FTIR, XRD, and DLS were applied to characterize the morphology and structure of BSA@ZIF-8 NPs and their effects on protein loading capacity. Degradation kinetics and protein release behavior of BSA@ZIF-8 NPs were evaluated at pH 5.0 (to simulate the tumor environment) and pH 7.4 (to mimic the blood environment). Our objective was to define optimal combinations of the high protein loading rate and rapid release under varying pH conditions, and we found that (i) the yield of BSA@ZIF-8 NPs decreased as the M:L ratio increased, but the protein content increased. This highlights the need to strike a balance between cost-effectiveness and practicality when selecting ZIF-8 NPs with different molar ratios for protein-based drug formulation. (ii) BSA@ZIF-8 NPs exhibited cruciate flower-like shapes when synthesized using Zn(NO3)2 as the zinc precursor at M:L ratios of 1:16 or 1:20. In all other cases, the NPs displayed a regular rhombic dodecahedral structure. Notably, the release behavior of the NPs did not differ significantly between these morphologies. (iii) When Zn(OAc)2 was used as the zinc precursor, the synthesized ZIF-8 NPs exhibited a smaller size compared to the Zn(NO3)2-derived ZIF-8 NPs. (iv) The release rate and amount of BSA protein were higher at pH 5.0 compared to pH 7.4. (v) Among the different formulations, BSA@ZIF-8 with an M:L ratio of 1:16 at pH 5.0 was observed to have a shorter time to reach a plateau (0.5 h) and higher protein release, making it suitable for locally rapid administration in a tumor environment. BSA@ZIF-8 prepared from Zn(OAc)2 at an M:L ratio of 1:140 showed the slower release of BSA protein over a 24-h period, indicating its suitability for sustained release delivery. In conclusion, our findings provide a useful basis for the practical application of ZIF-8 NPs in protein-based drug delivery systems.

Lelliottia steviae sp. nov. isolated from Stevia rebaudiana Bertoni.

A Gram-negative, aerobic, chemoheterotrophic, rod-shaped, and motile bacterium, designated as LST-1T, was isolated from wild Stevia rebaudiana Bertoni and subjected to a polyphasic taxonomic analysis. The LST-1 strain grew optimally at 37 °C and pH 6.0-7.0 in the presence of 0.5% (w/v) NaCl. Phylogenetic analysis based on the 16S rDNA sequence indicated that LST-1 is closely related to Lelliottia jeotgali PFL01T (99.85%), Lelliottia nimipressuralis LMG10245T (98.82%), and Lelliottia amnigena LMG2784T (98.54%). Multi-locus sequence typing of concatenated partial atpD, infB, gyrB, and rpoB genes was performed to improve the resolution, and clear distinctions between the closest related type strains were observed. The results of average nucleotide identify analyses and DNA-DNA hybridization with four species (16S rDNA similarity > 98.65%) were less than 90 and 40%, respectively, verifying the distinct characteristics from other species of Lelliottia. The cellular fatty acid profile of the strain consisted of C16:0, Summed Feature3, and Summed Feature8 (possibly 16:1 w6c/16:1 w7c and 18:1 w6c) as major components. The major polar lipids included phosphatidylethanolamine, phosphatidylglycerol, an aminophospholipid, three non-characteristic phospholipids, and a non-characteristic lipid. The genome of LST-1T was 4,611,055 bp in size, with a G + C content of 55.02%. The unique combination of several phenotypic, chemotaxonomic, and genomic characteristics proved that strain LST-1T belongs to a novel species, for which the name Lelliottia steviae sp. nov. is proposed. The type strain is LST-1T (= CGMCC 1.19175T = JCM 34938T).Repositories: The genbank accession numbers for the 16S rRNA gene and genome sequences of strain LST-1T are MZ497264 and CP063663, respectively.