Current diabetes therapies often fail to address the loss of insulin-producing β-cells. Bone marrow mesenchymal stem cells (BMSCs) can differentiate into insulin-producing cells (IPCs), but functional maturity remains limited. Toll-like receptor 9 (TLR9) agonists may enhance BMSC maturation. To investigate the ability of TLR9 agonists to promote the maturation of BMSCs in diabetic rats. BMSCs were isolated from rat bone marrow and identified by trilineage differentiation tests and flow cytometry. BMSCs were then differentiated into IPCs and treated with different concentrations of TLR9 agonists. PCR detected transcriptional levels of pancreatic endocrine-related genes. Diabetic model in rats was established using streptozotocin. BMSCs were encapsulated in TheraCyte capsules and subcutaneously transplanted into diabetic rats. Intraperitoneal injection of TLR9 agonists was performed, with serum insulin, C-peptide and fasting blood glucose (FBG) levels monitored in the transplanted rats. The recovered BMSCs were further characterized by immunofluorescence staining after the rats were killed. Differentiated BMSCs displayed characteristics of IPCs, such as the ability to secrete insulin and C-peptide in vitro. Further, when exposed to gradient concentrations of TLR9 agonists, differentiated MSCs exhibited enhanced expression of pancreatic-associated endocrine genes and transcription factor levels. In vivo, differentiated BMSC transplantation combined with TLR9 agonist administration notably improved diabetic conditions, as evidenced by suppressed FBG levels and elevated serum insulin and C-peptide levels, comparable to those in the normal group. However, withdrawal of TLR9 agonists attenuated the therapeutic effects. Finally, immunofluorescence staining confirmed that a TLR9 agonist promoted IPC maturation, as evidenced by co-expression of insulin and C-peptide in vivo. TLR9 agonists promote the differentiation and functional maturation of BMSCs into IPCs, thereby improving metabolic parameters in diabetic rats. This "stem cell-immunomodulator" strategy provides a foundation for developing functional β-cell replacement therapies.

It was found that pressure can promote the regeneration and repair of cartilage defects based on bone marrow mesenchymal stem cells (BMSCs). Since the compressive microenvironment of the cartilage in vivo may change with different movement, the mechanobiological effects of different compressive condition on BMSCs, especially the impact for its chondrogenic differentiation and influence on the cartilage microenvironment, is what we concerned about. Rat BMSCs were cultured and subjected to various types of pressure stimulation for 1h. The Cell Counting Kit-8 (CCK-8) assay was used to analyze cell proliferation, flow cytometry was employed to assess the cell cycle and apoptosis, confocal microscopy was used to observe the cytoskeleton, and transmission electron microscopy was performed to examine the cellular ultrastructure. RT-PCR was used to identify chondrogenic differentiation markers. Apoptotic vesicles derived from BMSCs were isolated by ultracentrifugation, and differentially expressed microRNAs in these vesicles under - 40kPa compression were identified by transcriptome sequencing. Specific pressure conditions promoted the proliferation of BMSCs, with dynamic pressure showing a stronger proliferative effect than static pressure. Higher static negative pressure (- 40kPa) significantly increased the spreading area of BMSCs. Dynamic pressure is stronger than static pressure in promoting cytoskeletal rearrangement, stress fiber formation, and cartilage marker expression in BMSCs. Flow cytometry and transmission electron microscopy results show that both - 40kPa static and 90kPa dynamic pressures promote BMSCs apoptosis to some extent. Under - 40kPa static negative pressure, the differentially expressed microRNAs in BMSCs-derived apoptotic vesicles are involved in stem cell maintenance and chondrogenic proliferation processes. Static negative pressure (- 40kPa) induces apoptosis in bone marrow mesenchymal stem cells (BMSCs). Notably, compared to chemical induction with staurosporine (STS), BMSCs subjected to - 40kPa mechanical stimulation display distinct microRNA expression profiles within apoptotic vesicles, specifically enriched in microRNAs implicated in stem cell fate determination and cartilage regeneration. These findings offer valuable insights into biomechanical strategies for optimizing tissue-engineered cartilage repair.

Jiawei Qiwei Baizhu Powder (JQBP) has been widely applied in clinical practice for the treatment of pediatric short stature associated with spleen-kidney deficiency and dampness accumulation, showing favorable therapeutic outcomes. Bone marrow mesenchymal stem cells (BMSCs) play a pivotal role in skeletal growth and development through their proliferative and osteogenic differentiation capacities. This study aimed to explore the potential cellular mechanisms underlying the therapeutic effects of JQBP on pediatric short stature by investigating the effects of JQBP-containing serum on the proliferation and early osteogenic differentiation of rat BMSCs in-vitro. JQBP-containing serum and normal control serum were prepared using a serum pharmacology approach. BMSCs were isolated from Sprague-Dawley rats using the whole bone marrow adherence method and identified by flow cytometry. Cell proliferation under different serum concentrations was dynamically assessed using the CCK-8 assay. Based on proliferation outcomes, the optimal serum concentration was selected for osteogenic induction experiments. After 7 days of osteogenic induction, early osteogenic differentiation was evaluated by alkaline phosphatase (ALP) staining and quantitative grayscale analysis. CCK-8 assays demonstrated that JQBP-containing serum significantly promoted BMSC proliferation, with the 1% concentration showing the most stable and sustained proliferative effect. Under osteogenic induction conditions, ALP staining revealed markedly enhanced ALP activity in the 1% JQBP-containing serum group compared with the induction-only group and the 1% normal serum group (P < 0.05). Quantitative grayscale analysis further confirmed significantly lower grayscale values in the 1% JQBP-containing serum group, indicating the strongest ALP activity among all groups. JQBP-containing serum effectively promotes BMSC proliferation and enhances early osteogenic differentiation in-vitro. These findings suggest that JQBP may facilitate bone growth by directly activating osteogenic precursor cells, providing experimental evidence for integrating traditional Chinese medicine into modern growth-related therapeutics.

Cholesterol-driven mitochondrial rejuvenation by quercetin nanotherapeutics restores implant osseointegration in diabetes.

The safety of an extracellular vesicle (EV) enriched secretome from bone marrow mesenchymal stem cells (BM-MSCs) was evaluated in a multi-site, prospective, expanded access trial as potential treatment for respiratory failure due to COVID-19. Subjects (103) received up to three doses of 15 mL IP every 72h. The primary outcome was all cause 60-day mortality. Secondary outcomes included serious adverse events. One TEAE (grade 1 hyperpigmentation at the infusion site) related to IP occurred. 60-day mortality was 29% for all patients, 22.4% in patients < 65 years and 41.7% in patients ≥ 65 years. Mean ventilation free days was 40.8 for all patients, 44.7 days in patients < 65 years and 33.4 days in patients ≥ 65 years. Median time to hospital (IQR) discharge by Kaplan-Meier was 11 (5.0-NR) days in all patients, 9 (5.0-NR) days in patients < 65 years and 19 (5.5-NR) days in patients ≥ 65 years. The IP (15 mL dose) is safe in patients with severe or critical COVID-19 respiratory failure. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT04657458 (2020-12-08).

Osteoarthritis (OA) is a common degenerative joint disease closely associated with obesity. Adipokines secreted by adipose tissue play critical roles in disease progression. This study focuses on the adipokine leptin and investigates its relationship with knee osteoarthritis. Analysis of subchondral bone tissue from OA patients revealed decreased leptin receptor expression, enhanced osteogenic activity, and a significant increase in H-type vessel number. Concurrently, serum leptin levels were markedly elevated in OA patients. Animal experiments further demonstrated that leptin-knockout obese mice exhibited significantly milder OA, reduced H-type vessel formation, and decreased osteogenic activity. In contrast, mice with high serum leptin levels showed more severe joint destruction and vascular hyperplasia. Invitro cell experiments indicated that leptin activates the PI3K/AKT signalling pathway in bone marrow mesenchymal stem cells (BMSCs), increasing the secretion of angiogenic factors such as VEGF and HIF-1α. Conditioned medium from leptin-treated BMSCs promoted tube formation and expression of H-type vessel markers (EMCN/CD31) in mouse umbilical vein endothelial cells. Inhibition of the PI3K pathway using Ly294002 in BMSCs or ZSTK474 in mice abolished leptin's effects. These results suggest that obesity-induced hyperleptinemia drives aberrant H-type vessel angiogenesis and active bone remodelling in subchondral bone via the PI3K/AKT pathway, thereby accelerating OA progression.

Kidney tissue injury is clinically very common, and how to effectively repair renal damage is a critical scientific issue that urgently requires attention. This study focuses on exploring alternative therapeutic strategies for renal repair. Specifically, it aims to compare the differentiation capabilities of metanephric mesenchymal cells (MMC), bone marrow mesenchymal stem cells (BMSC), and adipose-derived mesenchymal stem cells (AMSC) into epithelial cells, and to investigate the role and mechanism of catalpol, an active component of the traditional Chinese medicine Rehmannia glutinosa, in enhancing the epithelial differentiation of MMC. The epithelial differentiation potential of AMSC, BMSC, and MMC was compared under epithelial induction medium with or without catalpol supplementation. MMC were cultured in epithelial induction medium, and the expression of epithelial marker E-Cadherin, mesenchymal markers α-SMA and renal epithelial-specific marker AQP2 were assessed using Western Blot and immunofluorescence staining to evaluate the epithelial differentiation capacity of MMC and the enhancing effect of catalpol. RNA-seq was performed to identify signaling pathways activated by catalpol in MMC. The canonical Wnt pathway inhibitor MSAB was added to determine whether the pro-epithelial differentiation effect of catalpol was inhibited. AMSC and BMSC exhibited limited epithelial differentiation potential, which was not enhanced by catalpol. Under epithelial induction medium, MMC differentiated into epithelial cells, with increased expression of E-Cadherin and decreased expression of α-SMA. Catalpol further enhanced this effect and promoted the expression of the renal epithelial marker AQP2. RNA-seq revealed that the canonical Wnt pathway was activated in MMC. The pro-epithelial differentiation effect of catalpol was inhibited by MSAB, accompanied by decreased expression of E-Cadherin and AQP2 and increased expression of α-SMA. Catalpol promotes the differentiation of MMC into renal epithelial cells by activating the canonical Wnt pathway.

Activating the SDF-1/CXCR4 axis: Notoginsenoside R1-Functionalized zinc scaffolds accelerate fracture healing and angiogenesis in diabetic osteoporosis.

Exosomal miRNA let-7i-5p alleviates asthma triggered by RSV-induced exosomes by regulating dendritic cell autophagy via the MITF/DAP1/P70S6K pathway.

Adipocyte hypertrophy is an obesity-related metabolic dysfunction, which is frequently associated with decreased mitochondrial activity during adipocyte development. The current study aimed to assess the potential of epoxy clerodane diterpene (ECD) (IUPAC: 5R, 10R)-4R, 8R-dihydroxy-2S, 3R:15, 16-diepoxycleroda-13(16), 17, 12S:18,1S-dilactone) extracted from Cassia tora on adipocyte differentiation, lipid accumulation, mitochondrial function, and inflammation. Human bone marrow mesenchymal stem cells (hMSCs) were stimulated into adipocytes using the standard differentiation medium. The methodological design included the evaluation of ECD cytotoxicity, lipid accumulation, mitochondrial membrane potential, qRT-PCR, and ELISA. ECD didn't significantly reduce the cellular viability; however, it decreased lipid accumulation by 65%, 87%, and 87.5% at doses of 2, 4, and 8 μM, respectively. Also, at 4 µM of ECD, it decreased adipocyte hypertrophy, increased mitochondrial membrane potential, raised the expression of thermogenesis-related genes (UCP-1, PPARγC1α, SREBP1c), decreased the expression of adipogenic proteins (C/EBPα, PPARγ), increased adiponectin levels, and reduced inflammatory markers (IL-4, TNF-α) compared to untreated controls. Thus, ECD may hold tremendous promise as a natural agent for controlling adipogenesis, and its impacts on lipid metabolism, mitochondrial function, and inflammatory responses demonstrate its potential for therapeutic use in the treatment of obesity and associated metabolic diseases.

3D-printed PLA/β-TCP scaffold functionalized with PDA and neobavaisoflavone-loaded PEG hydrogel for enhanced bone repair.

The soft tissue healing diamond (ST-Diamond) concept: A translational framework for tendon and ligament regeneration.

Risperidone induces osteoporosis and neuropsychiatric treatment resistance via SMPD1-lysosome-mediated ferroptosis: dual rescue by active vitamin D analog ED-71.

Elevated levels of reactive oxygen species (ROS) in the bone defect area impede bone regeneration and repair. Although titanium and its alloys are widely used for bone defect repair, they lack bioactivity and the ability to scavenge ROS at the defect site, leading to poor osseointegration. To address this, surface modification of Ti alloys with coatings is a common strategy. This study aimed to investigate the osteogenic and antioxidant effects of a novel glutathione (GSH)-coated Ti screw (TiGSH) on bone regeneration. TiGSH was fabricated using a PDA intermediary layer, followed by chelation with GSH. In vitro studies demonstrated sustained GSH release from TiGSH over 15 days. In vivo studies revealed that TiGSH did not alter the levels of GSH in the plasma of rats, but it significantly increased the GSH content at the bone defect site. TiGSH effectively scavenged DPPH• radicals and reduced intracellular ROS levels in bone marrow mesenchymal stem cells (BMSCs), promoting their proliferation, osteogenic differentiation, and mineralization. In vivo experiments in a rat bone defect model showed that TiGSH significantly reduced the level of oxidative stress at the bone defect site, thereby enhancing the bone regeneration and osseointegration capabilities of the titanium screw. Meanwhile, TiGSH did not cause any adverse effects in the rats. Overall, TiGSH demonstrates promising potential as a biocompatible and osteoinductive material for bone defect repair, with sustained local GSH release mitigating oxidative stress and promoting bone regeneration.

Biodegradable Mg2+-releasing piezoelectric scaffold for segmental bone defect repair.

Iron oxide nanoparticles-driven mitochondrial renewal rejuvenates the aged bone marrow niche.

PTEN-knockdown modified T2DM ADSCs-derived extracellular vesicles promote T2DM implant osseointegration by enhancing the osteogenic differentiation of bone marrow mesenchymal stem cells.

DNA tetrahedron-mediated vascular targeted delivery of astragaloside IV enhances distraction osteogenesis via PI3K/AKT/FOXO pathway.

The core pathological mechanism of steroid-induced osteonecrosis of the femoral head (SONFH) is an "immune freeze" microenvironment-where corticosteroids continuously drive macrophages toward a pro-inflammatory M1 phenotype, inhibiting the conversion to reparative M2 macrophages, thereby impairing bone regeneration. To address this bottleneck, this study was inspired by the hierarchical pore structure of coral. Using low-temperature deposition 3D printing technology, we constructed an immunoreprogramming biomimetic scaffold that integrates multi-walled carbon nanotubes (MWCNT) and nano-hydroxyapatite (nHA) (MWCNT bionic scaffold). The scaffold leverages the active immune regulatory function of MWCNT to activate the PI3K-AKT signaling pathway, driving macrophages to transition from the M1 to M2 phenotype, effectively breaking the "immunological freeze" state and reshaping the pro-regenerative bone immune microenvironment. Meanwhile, the nHA component provides a biomimetic mineralization matrix and sustained release of calcium and phosphate ions, synergizing with the nanofiber structure of MWCNT to promote the migration and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and angiogenesis. In vivo experiments confirm that scaffold implantation reverses the local "immune freeze" state, drives macrophage polarization toward the M2 phenotype, reduces inflammatory responses, and enhances the maturation and functional vascular network formation of new bone matrix in bone defect areas, ultimately achieving bone structural reconstruction. The coral-inspired immunoreprogramming strategy proposed in this study provides a new strategy for targeting the regulation of the pathological microenvironment in SONFH.

Colorectal cancer (CRC) is the third most common cancer in the world. Bone metastasis, which occurs in advanced cancer, causes severe damage and accelerates patient death. TROP2, encoded by the TACSTD2 gene, is highly expressed in multiple types of cancer and is associated with tumor proliferation, invasion and metastasis. However, the role of TROP2 in CRC is unclear. TROP2 expression was detected in tumor tissues of CRC patients with or without bone metastases, as well as CRC cell lines. The functional role of TROP2 on CRC cells was validated by in vitro cell culture models and in vivo mouse experiments. Comparative proteomics was used to discover the interactome of TROP2, and later confirmed by immunoprecipitation and immunofluorescent studies. The therapeutic role of TROP2-targeted agent in CRC was verified by intratibial injection mouse models. TROP2 was highly expressed in the tumor tissues of CRC patients, with largely negative expression in the normal counterparts. CRC patients with bone metastasis had even higher TROP2 expression in the tumors than those with primary carcinoma only. We demonstrated that TROP2 promoted the proliferation and migration of CRC cells by in vitro and in vivo experiments, and the pro-invasive and pro-adhesive effects of TROP2 in CRC cells could be enhanced by co-culturing with bone marrow mesenchymal stem cells (BMSCs). Mechanistically, we validated that TROP2 was able to interact with fibronectin (FN) expressed by BMSCs, which increased the expressions of α5, αV, and β3 integrin subunits and facilitated the expression of integrin-linked kinase (ILK) on CRC cells. This interaction was required for the downstream activation of PI3K-Akt signaling and the CRC cell adhesion to BMSCs. Furthermore, intratibial injection mouse experiments verified that TROP2 promoted the colonization of CRC cells in bone marrow, and the combination of the TROP2-targeted antibody-drug conjugate, Sacituzumab Govitecan, and anti-integrin α5β1 monoclonal antibody, Volociximab, was effective in inhibiting bone colonization of CRC cells. TROP2 promotes CRC bone metastasis by interacting with bone marrow-derived FN-integrin axis. Through its adhesive function, TROP2 may coordinate with the specialized bone niche to facilitate metastatic colonization and establish a permissive environment for stromal-tumor interactions. Targeting this axis offers a promising therapeutic strategy for managing bone metastasis in CRC and potentially other TROP2-overexpressing malignancies.