Platelet-derived biomaterial with hyaluronic acid alleviates temporal-mandibular joint osteoarthritis: clinical trial from dish to human.

BACKGROUND: Bioactive materials have now raised considerable attention for the treatment of osteoarthritis (OA), such as knee OA, rheumatoid OA, and temporomandibular joint (TMJ) OA. TMJ-OA is a common disease associated with an imbalance of cartilage regeneration, tissue inflammation, and disability in mouth movement. Recently, biological materials or molecules have been developed for TMJ-OA therapy; however, ideal treatment is still lacking. In this study, we used the combination of a human platelet rich plasma with hyaluronic acid (hPRP/HA) for TMJ-OA therapy to perform a clinical trial in dish to humans. METHOD: Herein, hPRP was prepared, and the hPRP/HA combined concentration was optimized by MTT assay. For the clinical trial in dish, pro-inflammatory-induced in-vitro and in-vivo mimic 3D TMJ-OA models were created, and proliferation, gene expression, alcian blue staining, and IHC were used to evaluate chondrocyte regeneration. For the animal studies, complete Freund's adjuvant (CFA) was used to induce the TMJ-OA rat model, and condyle and disc regeneration were investigated through MRI. For the clinical trial in humans, 12 patients with TMJ-OA who had disc displacement and pain were enrolled. The disc displacement and pain at baseline and six months were measured by MRI, and clinical assessment, respectively. RESULTS: Combined hPRP/HA treatment ameliorated the proinflammatory-induced TMJ-OA model and promoted chondrocyte proliferation by activating SOX9, collagen type I/II, and aggrecan. TMJ-OA pathology-related inflammatory factors were efficiently downregulated with hPRP/HA treatment. Moreover, condylar cartilage was regenerated by hPRP/HA treatment in a proinflammatory-induced 3D neocartilage TMJ-OA-like model. During the animal studies, hPRP/HA treatment strongly repaired the condyle and disc in a CFA-induced TMJ-OA rat model. Furthermore, we performed a clinical trial in humans, and the MRI data demonstrated that after 6 months of treatment, hPRP/HA regenerated the condylar cartilage, reduced disc displacement, alleviated pain, and increased the maximum mouth opening (MMO). Overall, clinical trials in dish to human results revealed that hPRP/HA promoted cartilage regeneration, inhibited inflammation, reduced pain, and increased joint function in TMJ-OA. CONCLUSION: Conclusively, this study highlighted the therapeutic potential of the hPRP and HA combination for TMJ-OA therapy, with detailed evidence from bench to bedside. Trial registration Taipei Medical University Hospital (TMU-JIRB No. N201711041). Registered 24 November 2017. https://tmujcrc.tmu.edu.tw/inquiry_general.php .

Correlation Between Cephalometric Measures and End-of-Treatment Facial Attractiveness.

Sixty-nine experienced Chinese orthodontists evaluated 108 Chinese patients' facial attractiveness from set of photographs (frontal, lateral, and frontal smiling photos) taken at the end of orthodontic treatment. These 108 patients, which contained an equal number of patients with Class I, II, and III malocclusion, were randomly selected from 6 orthodontic treatment centers throughout China. Spearman rank-order correlation coefficients (rs) analyses were performed to examine agreement in ranking between all judge pairs. Pearson correlation and multivariate regression were performed to examine the correlation between cephalometric measures and end-of-treatment Photo Attractiveness Rank.96.68% judge pairs showed moderate correlated (+0.4 ≤ rs < +0.7) subjective rankings. Cephalometric measures significantly correlated with end-of-treatment Photo Attractiveness Rank included interincisal angle (r = 0.330, P < 0.05), L1/MP° (r = 0.386, P < 0.05), L1-NBmm (r = 0.451, P < 0.01), L1/NB° (r = 0.374, P < 0.05), and profile angle (r = 0.353, P < 0.05) in Class I patients with an explained variance of 32.8%, and ANB angle (r = 0.432, P < 0.01), angle of convexity (r = 0.448, P < 0.01), profile angle (r = 0.488, P < 0.01), Li to E-line (r = 0.374, P < 0.05), Li to B-line (r = 0.543, P < 0.01), and Z angle (r = 0.543, P < 0.01) in Class II patient with an explained variance of 43.3%.There was less association than expected between objective measurements on the lateral cephalograms and clinicians' rankings of facial attractiveness on clinical photography in Chinese patients. Straight-stand lower incisor was desired for facial attractiveness of Class I malocclusion; and sagittal relationship and lip prominence influence the esthetics of Class II malocclusion in Chinese population.

Superior intratumoral penetration of paclitaxel nanodots strengthens tumor restriction and metastasis prevention.

Recently discovered intratumoral diffusion resistance, together with poor solubility and nontargeted distribution of chemotherapeutic drugs, has significantly impaired the performance of cancer treatments. By developing a well-designed droplet-confined/cryodesiccation-driven crystallization approach, we herein report the successful preparation of nanocrystallites of insoluble chemotherapeutic drug paclitaxel (PTX) in forms of nanodots (NDs, ≈10 nm) and nanoparticles (NPs, ≈70 nm) with considerably high drug loading capacity. Superficially coated Pluronic F127 is demonstrated to endow the both PTX nanocrystallites with excellent water solubility and prevent undesired phagocyte uptake. Further decoration with tumor-penetrating peptide iRGD, as expected, indiscriminatively facilitates tumor cell uptake in traditional monolayer cell culture model. On the contrary, distinctly enhanced performances in inward penetration and ensuing elimination of 3D multicellular tumor spheroids are achieved by iRGD-NDs rather than iRGD-NPs, revealing the significant influence of particle size variation in nanoscale. In vivo experiments verify that, although efficient tumor enrichment is achieved by all nanocrystallites, only the iRGD-grafted nanocrystallites of ultranano size realize thorough intratumoral delivery and reach cancer stem cells, which are concealed inside the tumor core. Consequently, much strengthened restriction on progress and metastasis of orthotopic 4T1 mammary adenocarcinoma is achieved in murine model, in sharp contrast to commercial PTX formulation Taxol.

Development of loop-mediated isothermal amplification assay for specific and rapid detection of differential goat pox virus and sheep pox virus.

BACKGROUND: Capripox viruses are economically important pathogens in goat and sheep producing areas of the world, with specific focus on goat pox virus (GTPV), sheep pox virus (SPPV) and the Lumpy Skin Disease virus (LSDV). Clinically, sheep pox and goat pox have the same symptoms and cannot be distinguished serologically. This presents a real need for a rapid, inexpensive, and easy to operate and maintain genotyping tool to facilitate accurate disease diagnosis and surveillance for better management of Capripox outbreaks. RESULTS: A LAMP method was developed for the specific differential detection of GTPV and SPPV using three sets of LAMP primers designed on the basis of ITR sequences. Reactions were performed at 62°C for either 45 or 60 min, and specificity confirmed by successful differential detection of several GTPV and SPPV isolates. No cross reactivity with Orf virus, foot-and-mouth disease virus (FMDV), A. marginale Lushi isolate, Mycoplasma mycoides subsp. capri, Chlamydophila psittaci, Theileria ovis, T. luwenshuni, T. uilenbergi or Babesia sp was noted. RFLP-PCR analysis of 135 preserved epidemic materials revealed 48 samples infected with goat pox and 87 infected with sheep pox, with LAMP test results showing a positive detection for all samples. When utilizing GTPV and SPPV genomic DNA, the universal LAMP primers (GSPV) and GTPV LAMP primers displayed a 100% detection rate; while the SPPV LAMP detection rate was 98.8%, consistent with the laboratory tested results. CONCLUSIONS: In summary, the three sets of LAMP primers when combined provide an analytically robust method able to fully distinguish between GTPV and SPPV. The presented LAMP method provides a specific, sensitive and rapid diagnostic tool for the distinction of GTPV and SPPV infections, with the potential to be standardized as a detection method for Capripox viruses in endemic areas.

[Correlation study on lignin contents of Schisandra chinensis and ecological factors].

The contents of schisandrin, schizandrin A, B and C were determined by HPLC, and the effects of the climate factors and altitude on lignin contents were analyzed in order to select the optimal cultivation area of S. chinensis. The lignin contents were analyzed by HPLC using a ZORBAX SB-C18 column (4.6 mm x 250 mm, 5 microm). The column temperature and detection wave length were set at 35 degrees C and 254 nm, respectively. Methanol-water was used as the mobile phase in gradient elution mode and the flow-rate was 1.0 mL min(-1). The method had a good repeatability, stability and accuracy. The correlation of climate factors and lignins contents was analyzed by SPSS software. The results showed that the schizandrin A content in S. chinensis fruits were higher than 0.4% in Ji'an, Liuhe, Antu and Fusong in Jilin province, which met the quality requirement. It had significant linear negative correlation relationship between schisandrin, schizandrin A, B and altitude, the contents decreased with the increase of altitude. The significant negative linear fitting coefficient was 0.844 1 between schisandrin and altitude; but it had not significant correlation between schizandrin C and altitude. A significant positive correlation of climate factors and the contents of S. chinensis lignins were mainly the temperature factors (the average annual temperature, the highest temperature in July, the average temperature in July, the highest temperature in January, the average temperature in January) and precipitation factor (average annual precipitation), which reveals that higher temperature and precipitation were helpful to the formation and accumulation of lignins of S. chinensis. So the cultivation area of S. chinensis should be in the low elevations region with warm and rainy climate.

Recent advances of smart materials for ocular drug delivery.

Owing to the variety and complexity of ocular diseases and the natural ocular barriers, drug therapy for ocular diseases has significant limitations, such as poor drug targeting to the site of the disease, poor drug penetration, and short drug retention time in the vitreous body. With the development of biotechnology, biomedical materials have reached the "smart" stage. To date, despite their inability to overcome all the aforementioned drawbacks, a variety of smart materials have been widely tested to treat various ocular diseases. This review analyses the most recent developments in multiple smart materials (inorganic particles, polymeric particles, lipid-based particles, hydrogels, and devices) to treat common ocular diseases and discusses the future directions and perspectives regarding clinical translation issues. This review can help researchers rationally design more smart materials for specific ocular applications.

The orchestration of cellular and humoral responses is facilitated by divergent intracellular antigen trafficking in nanoparticle-based therapeutic vaccine.

Therapeutic vaccination for the treatment of chronic hepatitis B is promising but has so far shown limited clinical efficacy. Herein, we employ polylactide nanoparticles (NPs) as the vaccine adjuvant and systematically explore their effect on activation of specific immunity and the underlying theoretical mechanisms. In vitro studies show that hepatitis B surface antigen (HBsAg) accumulates in antigen-presenting cells (APCs) to a larger content (270%) with the assistant of NP in comparison with the pure-antigen group. Besides the elevated costimulators (CD80/86) and increased major histocompatibility complex (MHC) II molecules, the MHC I molecules are also found upregulated. This result is mostly owing to the divergent antigen trafficking ways of NP-antigen in APCs, especially for the escape of exogenous HBsAg from the lysosomes to the cytosol. Interestingly, the MHC I level is downregulated in alum-antigen group, indicating a possible reason for its inefficiency in priming cellular response. Further in vivo experiments establish that NP-antigen group indeed enhances the CD8(+) CTL cytotoxicity and IFN-γ cytokine secretion. Meanwhile, specific antibody titer is also upregulated, and even surpasses that of the commercialized alum-antigen. All these results strongly support that NP-based antigen promotes an orchestration of cellular and humoral immune response, exhibiting favorable intrinsic properties to be applied in therapeutic vaccines.

Nickel-titanium shape-memory sawtooth-arm embracing fixator for periprosthetic femoral fractures.

PURPOSE: We reviewed data to determine outcomes for 21 consecutive Vancouver type B1 or type C periprosthetic fractures that we treated between 2001 and 2008 using a nickel-titanium shape-memory sawtooth-arm embracing fixator. METHODS: The study participants were 12 men and 9 women (mean age, 70.8 years; range, 42-85 years). The average duration of follow-up monitoring was 39.7 months (range, 1-78 months). In five cases, cables and screws were used for further stabilisation. No bone grafting was performed for any of the patients. RESULTS: Results were satisfactory, except for one patient who died one month after surgery from a cause unrelated to arthroplasty. Bone union was achieved in the remaining 20 cases within an average of 5.25 months. No implant failures or malunions occurred in any of the patients. The average Harris hip score at the final follow-up examination was 79.3 points. CONCLUSIONS: Our results show that the embracing fixator is a valid alternative treatment for Vancouver type B1 or type C periprosthetic femoral fractures.

[Application of Wechat public platform in teaching reform of prosthodontics].

PURPOSE: To explore the law of development and existing problems in prosthodontics teaching in the new era, through flipped classroom teaching based on WeChat public platform. METHODS: WeChat public number was applied and WeChat groups were set up among the students of grade 2013, and 93 students were divided into 20 WeChat groups, with 4 to 5 students in each group. The main platform on reform of prosthodontics teaching was WeChat app, supplemented by platforms of Lediaocha and Youkaoshi. Teachers published courseware and learning resources with WeChat public number and WeChat groups. Two-dimensional codes on questionnaires and tests which were generated by Lediaocha and Youkaoshi could be published with WeChat and the final learning effectiveness was compared using SPSS 19.0 software package for t test. RESULTS: The results showed that 59.1% students admitted WeChat teaching effective, only 5.4% students believed ineffective. Based upon partial chapters of prosthodontics, the score of classroom test (91.35±4.45) was significantly higher than that of pre-class test(90.14±5.03, P＜0.05). CONCLUSIONS: The reform of flipped classroom based on WeChat platform in prosthodontics have some advantages, such as more flexibility in teaching form and time arrangement, promoting students' learner autonomy, and increasing students' motivation and effects of learning prosthodontics.

Transforming growth factor-ß: An early predictor of a functional cure in chronic hepatitis B treated with interferon.

BACKGROUND: The relationship between the serum transforming growth factor (TGF)-ß level and HBsAg loss has not been clearly elaborated in patients with chronic hepatitis B (CHB). METHODS: Two cohorts of patients with CHB were studied. Cohort A: A total of 207 hepatitis B e antigen (HBeAg)-negative CHB patients who finished ≥1 year nucleos(t)ide analogue monotherapy and sequentially received PEGylated interferon treatment for less than 96 weeks were included. Cohort B: Forty HBeAg-positive patients who initially received entecavir therapy for at least 96 weeks were included. Their viral markers and serum TGF-ß levels were measured at different time points during therapy. RESULTS: The levels of serum TGF-ß and HBsAg (0-24 W) were significantly lower in the patients who had HBsAg< 0.05 IU/mL at 48 weeks than in patients who did not in cohort A. We got the same results when we further divided the patients into subgroups according to the initial HBsAg cut-off values (1000 IU/mL, 100 IU/mL, 50 IU/mL) in cohort A. However, HBeAg seroconversion did not lead to the downregulation of TGF-ß levels. The levels of serum TGF-ß were significantly correlated with HBsAg quantitation in cohort A (12-24 W) but not in cohort B (0-48 W). The levels of TGF-ß at week 12 could be used as an early index to predict a functional cure (AUC=0.818) as well as the levels of HBsAg itself (AUC=0.882) in HBeAg-negative chronic hepatitis B patients treated with PEGylated interferon. CONCLUSIONS: The levels of serum TGF-ß were significantly associated with HBsAg loss but not with HBeAg seroconversion and could be used as an early index to predict a functional cure in CHB patients treated with PEGylated interferon.

Porous quaternized chitosan nanoparticles containing paclitaxel nanocrystals improved therapeutic efficacy in non-small-cell lung cancer after oral administration.

Clinical application of paclitaxel (PTX) is limited because of its poor solubility in aqueous media. To overcome this hurdle, we devised an oral delivery system by encapsulating PTX into N-((2-hydroxy-3-trimethylammonium) propyl) chitosan chloride (HTCC) nanoparticles. These nanoparticles were small (~130 nm), had a narrow size distribution, and displayed high loading efficiency owing to the homogeneous distribution of PTX nanocrystals. The matrix hydrophilicity and porous structure of the obtained nanoparticles accelerated their degradation and improved drug release. In vitro and in vivo transport experiments had proved that the presence of positive charges enhanced the intestinal permeability of these nanoparticles. Further in vitro experiment of cytotoxicity showed that the PTX-loaded HTCC nanoparticle (HTCC-NP:PTX) was more effective than native PTX owing to enhanced cellular uptake. Drug distribution in tissues and in vivo imaging studies confirmed the preferred accumulation of HTCC-NP:PTX in subcutaneous tumor tissue. Subsequent tumor xenograft assays demonstrated the promising therapeutic effect of HTCC-NP:PTX on inhibition of tumor growth and induction of apoptosis in tumor cells. Additional investigation into side effects revealed that HTCC-NP:PTX caused lower Cremophor EL-associated toxicities compared with Taxol. These results strongly supported the notion that HTCC nanoparticle (HTCC-NP) is a promising candidate as an oral carrier of PTX for cancer therapy.

pH-sensitive and mucoadhesive microspheres for duodenum-specific drug delivery system.

Particulate systems that could deliver drug specifically to duodenum have not yet been reported. The aim of this study was to develop a novel duodenum-specific drug delivery system based on thiolated chitosan and hydroxypropyl methylcellulose acetate maleate (HPMCAM) for the duodenal ulcer application. Berberine hydrochloride was used as model drug. Thiolated chitosan was synthesized and further used for the preparation of mucoadhesive microspheres. HPMCAM, which is insoluble below pH 3.0 was synthesized and used for the coating of thiolated chitosan microspheres (TCM). The resulting thiolated chitosan immobilized on chitosan was 268.21 ± 18 µmol/g. In vitro mucoadhesion study showed that the mucoadhesion property of TCM was better than that of chitosan microspheres. Morphological observation showed that the HPMCAM coating would maintain its integrity in simulated gastric fluid (SGF) for 2 h and dissolved quickly in simulated pathological duodenal fluid (SPDF; pH 3.3). In vitro drug release studies showed that only 4.75% of the drug was released in SGF for 2 h, while nearly 90% of the drug was released within 6 h after transferring into SPDF.

Evaluation Preparation of Apatinib-Loaded Polymer Nanoparticles and Its Effect in the Treatment of Advanced Ovarian Cancer.

Ovarian cancer is a common gynecological malignant tumor, second only to cervical cancer and uterine body cancer. In China, ovarian cancer has the highest mortality rate in gynecological tumors. Due to the rapid spread of cancer cells, the prognosis is relatively poor. Because the ovarian epithelial cancer is relatively insidious, there are no obvious clinical manifestations in the early stage. At present, apatinib chemotherapy is widely used, which can reduce the disease of patients by inhibiting the migration and proliferation of vascular endothelial cells. Fluorescence imaging is widely used in biomedical imaging. Organic fluorescent dyes are stable in nature and can be linked to a variety of molecules. They are often used in targeted imaging and therapeutic research. A cyanine dye is an organic molecule formed by two nitrogen-containing aromatic heterocycles connected by a polymethine bridge. Because neither MR contrast agents nor fluorescent dyes are targeted, specific biomolecules and contrast agents are often used in the research to diagnose and treat tumors. In this paper, a polymer nanoparticle loaded with apatinib was prepared and its therapeutic effect in advanced ovarian cancer was explored. The results show that nanoparticles loaded with apatinib are more effective than ordinary therapeutic drugs.

Prophylactic vaccine delivery systems against epidemic infectious diseases.

Prophylactic vaccines have evolved from traditional whole-cell vaccines to safer subunit vaccines. However, subunit vaccines still face problems, such as poor immunogenicity and low efficiency, while traditional adjuvants are usually unable to meet specific response needs. Advanced delivery vectors are important to overcome these barriers; they have favorable safety and effectiveness, tunable properties, precise location, and immunomodulatory capabilities. Nevertheless, there has been no systematic summary of the delivery systems to cover a wide range of infectious pathogens. We herein summarized and compared the delivery systems for major or epidemic infectious diseases caused by bacteria, viruses, fungi, and parasites. We also included the newly licensed vaccines (e.g., COVID-19 vaccines) and those close to licensure. Furthermore, we highlighted advanced delivery systems with high efficiency, cross-protection, or long-term protection against epidemic pathogens, and we put forward prospects and thoughts on the development of future prophylactic vaccines.

[Orofacial myofunctional therapy improves facial morphology of children with obstructive sleep apnea after adenotonsillectomy].

PURPOSE: This study investigated the effectiveness of orofacial myofunctional therapy(OMT) in improving facial morphology of children with obstructive sleep apnea (OSA) after adenotonsillectomy (AT). METHODS: Ten children aged from 4-7 years with persistent oral breathing for more than 1 month after adenotonsillectomy were chosen to receive orofacial myofunctional therapy. The patients were required to take photos before and after orofacial myofunctional therapy. In order to compare the soft changes before and after OMT treatment, twelve representative mark points were selected and used for proportion and angle measurements. Graphpad Prism 8 statistical software was used for statistical analysis, to compare the differences in facial morphology of patients before and after treatment. RESULTS: Compared with before OMT, a significant difference was found in the proportion of Sn-Ls/Sn-Stms(P=0.0002), Sn-Stms/Sn-Me'(P<0.05), as well as in the angle of Gs-Sn-Pos (P<0.05), nasolabial angle(P=0.0005), mentolabial angle (P=0.0026) after OMT treatment. CONCLUSIONS: Orofacial myofunctional therapy can be considered as an effective complementary treatment for OSA patients with oral breathing after adenotonsillectomy.

[Study of the root position of palatally malposed maxillary lateral incisor based on cone-beam CT images].

PURPOSE: The goal of this investigation was to measure and analyze the root position of palatally malposed maxillary lateral incisor based on cone-beam CT(CBCT)images, in order to provide references for orthodontists to move this kind of teeth scientifically. METHODS: CBCT data from 200 patients meeting the selection criteria with palatally malposed maxillary lateral incisor were investigated in this study. The root was divided into eight equal parts by length, then T1 to T8 were orderly pointed from root apex to alveolar ridge crest. The labial and palatal bone thickness at each point was measured; meanwhile, the angle between the long axis of the tooth and that of the alveolar bone was measured.The data was analyzed using SPSS 19.0 software package. RESULTS: The mean labial bone thicknesses at all researched points were less than 1.00 mm, except for point T1,T7 and T8. The mean labial bone thicknesses at point T3, T4 and T5 were the thinnest, which were all less than 0.5 mm(P<0.05). The mean thicknesses of labial bone gradually increased from T4 to T8(P<0.05). The mean palatal bone thicknesses were all more than 1.00 mm at the eight points,the mean thicknesses of palatal bone gradually increased from T8 to T1(P<0.05). All the angulations between the long axes of teeth and those of the alveolar bone were negative, indicating the root was close to the labial alveolar wall. The average angulation was minus 31.06 degrees. CONCLUSIONS: Results suggest that the root of palatally malposed maxillary lateral incisor is close to the labial wall of the alveolar bone, its labial alveolar bone is frequently quite thin or even deficient, especially in the zone between 1/4 root length to the root apex and the mid-root. If we move the palatally malposed maxillary lateral incisor labially, it is better to choose tipping movement, instead of bodily movement, in order to avoid serious bone fenestration and dehiscence or root absorption.

HIF-1α activator DMOG inhibits alveolar bone resorption in murine periodontitis by regulating macrophage polarization.

Periodontitis is initiated by serious and sustained bacterial infection and ultimately results in chronic immune-mediated inflammation, tissue destruction, and bone loss. The pathogenesis of periodontitis remains unclear. Host immunological responses to periodontal bacteria ultimately determine the severity and mechanisms governing periodontitis progression. This study aimed to clarify the effect of the hypoxia-inducible factor-1α (HIF-1α) activator dimethyloxalylglycine (DMOG) on a mouse periodontitis model and its underlying role in macrophage polarization. qRT-PCR analysis showed that DMOG inhibited the M1-like polarization of both RAW264.7 macrophages and murine bone marrow macrophages (BMMs) and downregulated TNF-α, IL-6, CD86, and MCP-1 expression in vitro. Immunofluorescence staining and flow cytometry also confirmed the less percentage of F4/80 + CD86 + cells after DMOG treatment. The phosphorylation of NF-κB pathway was also inhibited by DMOG with higher level of HIF-1α expression. Furthermore, mice treated with DMOG showed decreased alveolar bone resorption in the experimental periodontitis model, with significant increases in alveolar bone volume/tissue volume (BV/TV) and bone mineral density (BMD). DMOG treatment of mice decreased the ratio of M1/M2 (CD86+/CD206+) macrophages in periodontal tissues, resulting in the downregulation of proinflammatory cytokines such as TNF-α and IL-6 and increased levels of anti-inflammatory factors such as IL-4 and IL-10. DMOG treatment promoted the number of HIF-1α-positive cells in periodontal tissues. This study demonstrated the cell-specific roles of DMOG in macrophage polarization in vitro and provided insight into the mechanism underlying the protective effect of DMOG in a model of periodontitis.

Platelet-derived biomaterials-mediated improvement of bone injury through migratory ability of embryonic fibroblasts: in vitro and in vivo evidence.

Bony injuries lead to compromised skeletal functional ability which further increase in aging population due to decreased bone mineral density. Therefore, we aimed to investigate the therapeutic potential of platelet-derived biomaterials (PDB) against bone injury. Specifically, we assessed the impact of PDB on osteo-inductive characteristics and migration of mouse embryonic fibroblasts (MEFs). Osteogenic lineage, matrix mineralization and cell migration were determined by gene markers (RUNX2, OPN and OCN), alizarin Red S staining, and migration markers (FAK, pFAK and Src) and EMT markers, respectively. The therapeutic impact of TGF-ß1, a key component of PDB, was confirmed by employing inhibitor of TGF-ß receptor I (Ti). Molecular imaging-based in vivo cellular migration in mice was determined by establishing bone injury at right femurs. Results showed that PDB markedly increased expression of osteogenic markers, matrix mineralization, migration and EMT markers, revealing higher osteogenic and migratory potential of PDB-treated MEFs. In vivo cell migration was manifested by expression of migratory factors, SDF-1 and CXCR4. Compared to control, PDB-treated mice exhibited higher bone density and volume. Ti treatment inhibited both migration and osteogenic potential of MEFs, affirming impact of TGF-ß1. Collectively, our study clearly indicated PDB-rescued bone injury through enhancing migratory potential of MEFs and osteogenesis.

Platelet-Derived Biomaterials Inhibit Nicotine-Induced Intervertebral Disc Degeneration Through Regulating IGF-1/AKT/IRS-1 Signaling Axis.

Apart from aging process, adult intervertebral disc (IVD) undergoes various degenerative processes. However, the nicotine has not been well identified as a contributing etiology. According to a few studies, nicotine ingestion through smoking, air or clothing may significantly accumulate in active as well as passive smokers. Since nicotine has been demonstrated to adversely impact various physiological processes, such as sympathetic nervous system, leading to impaired vasculature and cellular apoptosis, we aimed to investigate whether nicotine could induce IVD degeneration. In particular, we evaluated dose-dependent impact of nicotine in vitro to simulate its chronic accumulation, which was later treated by platelet-derived biomaterials (PDB). Further, during in vivo studies, mice were subcutaneously administered with nicotine to examine IVD-associated pathologic changes. The results revealed that nicotine could significantly reduce chondrocytes and chondrogenic indicators (Sox, Col II and aggrecan). Mice with nicotine treatment also exhibited malformed IVD structure with decreased Col II as well as proteoglycans, which was significantly increased after PDB administration for 4 weeks. Mechanistically, PDB significantly restored the levels of IGF-1 signaling proteins, particularly pIGF-1 R, pAKT, and IRS-1, modulating ECM synthesis by chondrocytes. Conclusively, the PDB impart reparative and tissue regenerative processes by inhibiting nicotine-initiated IVD degeneration, through regulating IGF-1/AKT/IRS-1 signaling axis.

Cytosolic delivery of HBsAg and enhanced cellular immunity by pH-responsive liposome.

It remains a major challenge to prime the cytotoxic T lymphocytes (CTLs) response for the treatment of Hepatitis B virus (HBV) infection. Inspired by an important natural biological behavior, membrane fusion, we constructed a pH-responsive nanocarrier (HBsAg&CpG@Lip) with a membrane fusion capacity for HBsAg intracellular delivery and subsequent process of CTLs. In the in vitro experiments, HBsAg&CpG@Lip greatly promoted the cellular uptake of HBsAg and the activation of BMDCs. It induced the cytosolic release of HBsAg in BMDCs, which was in conformity with the membrane fusion capacity of HBsAg&CpG@Lip. Such a capacity was improved by 4.4-fold in pH = 5.0 than that in pH = 7.4 at 60 min, which was calculated through fluorescence resonance energy transfer (FRET) efficiency. Furthermore, the cross-presentation activity induced by liposome at 1 µg/mL was equivalent to that by soluble antigen at 2000 µg/mL, which indicated an advantage for the liposome in cytosolic antigen delivery and potent CTLs activation. In respect of the transport of liposomes into draining lymph nodes (DLNs), the recruitment of liposome+migratory DCs at 24 h raised 11-fold than that at 3 h, suggesting that liposomes were mainly carried by migratory DCs. Meanwhile, HBsAg&CpG@Lip also elicited the activation of DCs and the generation of Tfh cells in DLNs. After immunization, HBsAg&CpG@Lip induced a higher anti-HBsAg IgG and ratio of IgG2c/IgG1 than that by HBsAg or Alum+HBsAg group. It also augmented a strong CTLs response as expected. Specifically, stimulation with HBsAg&CpG@Lip increased the number of IFN-γ-secreting splenocytes, the proportion of CD107α+CD8+and FasL+CD8+ T cells and the secretion of Granzyme B, which verified the design of membrane fusion in vivo. In summary, this design for HBsAg cytosolic delivery exhibits great benefits for triggering CTLs, opening an avenue for the development of a potent therapeutic HBV vaccine.