ABSTRACT
BACKGROUND: The treatment of bone loss has posed a challenge to clinicians for decades. Thus, it is of great significance to identify more effective methods for bone regeneration. However, the role and mechanisms of long non-coding RNA small nucleolar RNA host gene 5 (SNHG5) during osteogenic differentiation remain unclear. METHODS: We investigated the function of SNHG5, Yin Yang 1 (YY1), miR-212-3p and growth differentiation factor 5 (GDF5) in osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) in vitro and in vivo. Molecular mechanisms were clarified by chromatin immunoprecipitation assay and dual luciferase reporter assay. RESULTS: We found SNHG5 expression was upregulated during osteogenesis of hBMSCs. Knockdown of SNHG5 in hBMSCs inhibited osteogenic differentiation while overexpression of SNHG5 promoted osteogenesis. Moreover, YY1 transcription factor directly bound to the promoter region of SNHG5 and regulated SNHG5 expression to promote osteogenesis. Dual luciferase reporter assay confirmed that SNHG5 acted as a miR-212-3p sponge and miR-212-3p directly targeted GDF5 and further activated Smad1/5/8 phosphorylation. miR-212-3p inhibited osteogenic differentiation, while GDF5 promoted osteogenic differentiation of hBMSCs. In addition, calvarial defect experiments showed knockdown of SNHG5 and GDF5 inhibited new bone formation in vivo. CONCLUSION: Our results demonstrated that the novel pathway YY1/SNHG5/miR-212-3p/GDF5/Smad regulates osteogenic differentiation of hBMSCs and may serve as a potential target for the treatment of bone loss.
Subject(s)
Mesenchymal Stem Cells , MicroRNAs , Osteogenesis , RNA, Long Noncoding , Growth Differentiation Factor 5/genetics , Growth Differentiation Factor 5/metabolism , Humans , Mesenchymal Stem Cells/cytology , MicroRNAs/genetics , RNA, Long Noncoding/geneticsABSTRACT
Tumor vaccines based on synthetic human papillomavirus (HPV) oncoprotein E7 and/or E6 peptides have shown encouraging results in preclinical model studies and human clinical trials. However, the clinical efficacy may be limited by the disadvantages of vulnerability to enzymatic degradation and low immunogenicity of peptides. To further improve the potency of vaccine, we developed a poly(lactide-co-glycolide)-acid (PLGA) nanoparticle, which encapsulated the antigenic peptide HPV16 E744-62, and used adenosine triphosphate (ATP), one of the most important intracellular metabolites and an endogenous extracellular danger signal for the immune system, as a new adjuvant component. The results showed that PLGA nanoparticles increased the in vivo stability, lymph node accumulation, and dendritic cell (DC) uptake of the E7 peptide; in addition, ATP further increased the migration, nanoparticle uptake, and maturation of DCs. Preventive immunization with ATP-adjuvanted nanoparticles completely abolished the growth of TC-1 tumors in mice and produced long-lasting immunity against tumor rechallenge. When tumors were fully established, therapeutic immunization with ATP-adjuvanted nanoparticles still significantly inhibited tumor progression. Mechanistically, ATP-adjuvanted nanoparticles significantly improved the systemic generation of antitumor effector cells, boosted the local functional status of these cells in tumors, and suppressed the generation and tumor infiltration of immunosuppressive Treg cells and myeloid-derived suppressor cells. These findings indicate that ATP is an effective vaccine adjuvant and that nanoparticles adjuvanted with ATP were able to elicit robust antitumor cellular immunity, which may provide a promising therapeutic vaccine candidate for the treatment of clinical malignancies, such as cervical cancer.
Subject(s)
Adenosine Triphosphate/metabolism , Cancer Vaccines/immunology , Immunity, Cellular , Nanoparticles/chemistry , Polylactic Acid-Polyglycolic Acid Copolymer/chemistry , Adenosine Triphosphate/chemistry , Amino Acid Sequence , Animals , Antigen-Presenting Cells/immunology , Antigen-Presenting Cells/metabolism , Cancer Vaccines/therapeutic use , Cell Line, Tumor , Dendritic Cells/immunology , Dendritic Cells/metabolism , Female , Humans , Mice , Mice, Inbred C57BL , Neoplasms/pathology , Neoplasms/therapy , Papillomavirus E7 Proteins/chemistry , Papillomavirus E7 Proteins/immunology , Peptides/chemistry , Peptides/immunology , Peptides/metabolism , Transplantation, HeterologousABSTRACT
In our study, plant polyphenol-inspired chemistry is explored to nano-engineer the topological and chemical structures of commercial melamine sponge surface for preparing superhydrophobic sponges. Briefly, tannic acid (TA, a typical plant polyphenol) is applied to induce the co-assembly of silica nanoparticles (SiO2) and silver ions (Ag+) to form SiO2@TA@Ag nanostructures on a melamine sponge surface. After further chemical fluorination, the superhydrophobic sponge with a "lotus leaf-mimic" surface is formed. Surface topological/chemical structures, superhydrophobic property and anti-combustion characteristics of the sponge are examined by a series of characterization techniques, including scanning electron microscopy, X-ray photoelectron spectroscopy, water contact angle measurements, combustion/heating test, etc. The superhydrophobic sponge presents an adsorption capacity of 69-153 times of its own weight toward various oils/organic solvents, and exhibits excellent recycling ability evidenced by over 100-cycled uses. Continuous oil/water separation apparatus is also set up through equipping the superhydrophobic sponge on a peristaltic pump, realizing the clean-up of oils and organic solvents from water continuously. Together with the facile, easy-to-scale-up and substrate non-selective features of plant polyphenol-inspired chemistry, the superhydrophobic sponge and the surface nano-engineering method would hold great promise for the effective treatment of oil spillages and organic discharges, achieving high sustainability to energy and environment.