2-N, 6-O sulfated chitosan evokes periosteal stem cells for bone regeneration.

Musculoskeletal injuries and bone defects represent a significant clinical challenge, necessitating innovative approaches for effective bone tissue regeneration. In this study, we investigated the potential of harnessing periosteal stem cells (PSCs) and glycosaminoglycan (GAG)-mimicking materials for in situ bone regeneration. Our findings demonstrated that the introduction of 2-N, 6-O sulfated chitosan (26SCS), a GAG-like polysaccharide, enriched PSCs and promoted robust osteogenesis at the defect area. Mechanistically, 26SCS amplifies the biological effect of endogenous platelet-derived growth factor-BB (PDGF-BB) through enhancing the interaction between PDGF-BB and its receptor PDGFRß abundantly expressed on PSCs, resulting in strengthened PSC proliferation and osteogenic differentiation. As a result, 26SCS effectively improved bone defect repair, even in an osteoporotic mouse model with lowered PDGF-BB level and diminished regenerative potential. Our findings suggested the significant potential of GAG-like biomaterials in regulating PSC behavior, which holds great promise for addressing osteoporotic bone defect repair in future applications.

A BMP-2-triggered in vivo osteo-organoid for cell therapy.

Cell therapies and regenerative medicine interventions require an adequate source of therapeutic cells. Here, we demonstrate that constructing in vivo osteo-organoids by implanting bone morphogenetic protein-2-loaded scaffolds into the internal muscle pocket near the femur of mice supports the growth and subsequent harvest of therapeutically useful cells including hematopoietic stem/progenitor cells (HSPCs), mesenchymal stem cells (MSCs), lymphocytes, and myeloid cells. Profiling of the in vivo osteo-organoid maturation process delineated three stages-fibroproliferation, osteochondral differentiation, and marrow generation-each of which entailed obvious changes in the organoid structure and cell type distribution. The MSCs harvested from the osteochondral differentiation stage mitigated carbon tetrachloride (CCl4)-induced chronic liver fibrosis in mice, while HSPCs and immune cells harvested during the marrow generation stage rapidly and effectively reconstituted the impaired peripheral and solid immune organs of irradiated mice. These findings demonstrate the therapeutic potentials of in vivo osteo-organoid-derived cells in cell therapies.

Hybrid bismuth oxide-graphine oxide nanomaterials improve the signal-to-noise response of small molecules analyzed by matrix assisted laser desorption ionization-time-of-flight mass spectrometry.

Matrix-assisted laser desorption ionization (MALDI) has received increasing attention for the analysis small molecules. Nanomaterials are frequently used as the matrix in LDI, and various inorganic materials have been developed, particularly those based on thermally-driven positive DI mechanisms. However, the unwanted detection of alkali metal ion adducts in the positive ion mode can compromise small molecule identification. Here, we report the synthesis and application of a novel hybrid bismuth oxide-graphene oxide (Bi2O3@GO) semiconductor matrix for the analysis of small molecules by LDI-time-of-flight mass spectrometry (TOF-MS) operating in the negative ion mode. The structure of the semiconductor nanomaterial was characterized using conventional methods and its performance for the detection of small molecules (e.g., amino acids, fatty acids, sugars and other small molecules) was compared with traditional DI matrices (e.g., cyano-4-hydroxycinnamic acid, 2,5-dihydroxybenzoic acid, 9-aminoacridine and GO). The results showed that the negative ion LDI-TOF MS of small molecules on Bi2O3@GO were free of matrix-related interferences, and possessed good signal intensity and repeatability. Application of Bi2O3@GO to the quantitative determination of glucose in human serum and soft drinks confirmed that the hybrid matrix could also be applied to complex samples. Conclusions drawn from the experimental results, computational chemistry calculations, and previous studies, suggesting that interfacial photogenerated thermal electron transfer and capture are key processes in the LDI mechanism.

Harvest of functional mesenchymal stem cells derived from in vivo osteo-organoids.

Bone marrow-derived mesenchymal stem cells (BM-MSCs) play a crucial role in stem cell therapy and are extensively used in regenerative medicine research. However, current methods for harvesting BM-MSCs present challenges, including a low yield of primary cells, long time of in vitro expansion, and diminished differentiation capability after passaging. Meanwhile mesenchymal stem cells (MSCs) recovered from cell banks also face issues like toxic effects of cryopreservation media. In this study, we provide a detailed protocol for the isolation and evaluation of MSCs derived from in vivo osteo-organoids, presenting an alternative to autologous MSCs. We used recombinant human bone morphogenetic protein 2-loaded gelatin sponge scaffolds to construct in vivo osteo-organoids, which were stable sources of MSCs with large quantity, high purity, and strong stemness. Compared with protocols using bone marrow, our protocol can obtain large numbers of high-purity MSCs in a shorter time (6 days vs. 12 days for obtaining passage 1 MSCs) while maintaining higher stemness. Notably, we found that the in vivo osteo-organoid-derived MSCs exhibited stronger anti-replicative senescence capacity during passage and amplification, compared to BM-MSCs. The use of osteo-organoid-derived MSCs addresses the conflict between the limitations of autologous cells and the risks associated with allogeneic sources in stem cell transplantation. Consequently, our protocol emerges as a superior alternative for both stem cell research and tissue engineering.

Study on Absorption, Distribution, Metabolism, and Excretion Properties of Novel Insecticidal GABA Receptor Antagonist, Pyraquinil, in Diamondback Moth Combining MALDI Mass Spectrometry Imaging and High-Resolution Mass Spectrometry.

A thorough understanding of absorption, distribution, metabolism, and excretion (ADME) of insecticide candidates is essential in insecticide development and structural optimization. Here, ADME of pyraquinil, a novel insecticidal GABA receptor antagonist, in Plutella xylostella larvae during the accumulation phase and depuration phase was investigated separately using a combination of UHPLC-Q-Orbitrap, HPLC-MS/MS, and MALDI-MSI. Five new metabolites of pyraquinil were identified, and a metabolic pathway was proposed. The oxidative metabolite (pyraquinil-sulfone) was identified as the main metabolite and confirmed by its standard. Quantitative results showed that pyraquinil was taken up by the larvae rapidly and then undergone a cytochrome P450s-mediated oxidative transformation into pyraquinil-sulfone. Both fecal excretion and oxidative metabolism were demonstrated to be predominant ways to eliminate pyraquinil in P. xylostella larvae during accumulation, while oxidative metabolism followed by fecal excretion was probably the major pathway during depuration. MALDI-MSI revealed that pyraquinil was homogeneously distributed in the larvae, while pyraquinil-sulfone presented a continuous enrichment in the midgut during accumulation. Conversely, pyraquinil-sulfone located in hemolymph can be preferentially eliminated during depuration, suggesting its tissue tropism. It improves the understanding of the fate of pyraquinil in P. xylostella and provides useful information for insecticidal mechanism elucidation and structural optimization of pyraquinil.

Construction of developmentally inspired periosteum-like tissue for bone regeneration.

The periosteum, a highly vascularized thin tissue, has excellent osteogenic and bone regenerative abilities. The generation of periosteum-mimicking tissue has become a novel strategy for bone defect repair and regeneration, especially in critical-sized bone defects caused by trauma and bone tumor resection. Here, we utilized a bone morphogenetic protein-2 (BMP-2)-loaded scaffold to create periosteum-like tissue (PT) in vivo, mimicking the mesenchymal condensation during native long bone development. We found that BMP-2-induced endochondral ossification plays an indispensable role in the construction of PTs. Moreover, we confirmed that BMP-2-induced PTs exhibit a similar architecture to the periosteum and harbor abundant functional periosteum-like tissue-derived cells (PTDCs), blood vessels, and osteochondral progenitor cells. Interestingly, we found that the addition of chondroitin sulfate (CS), an essential component of the extracellular matrix (ECM), could further increase the abundance and enhance the function of recruited PTDCs from the PTs and finally increase the regenerative capacity of the PTs in autologous transplantation assays, even in old mice. This novel biomimetic strategy for generating PT through in vivo endochondral ossification deserves further clinical translation.