Immunomodulatory differences between mesenchymal stem cells from different oral tissues.

Mesenchymal stem cells (MSCs) have recently been identified as having potentially therapeutic immunomodulatory properties. MSCs isolated from different oral tissues have similar morphology and immunophenotypes, however, direct comparisons of their gene expression and immunomodulatory properties have not been conducted. We isolated alveolar bone-derived MSCs (aBMSCs), dental pulp stem cells (DPSCs) and gingiva-derived MSCs (GMSCs) from the same patients and compared their immunophenotypes and transcriptomes. Additionally, we compared their production of soluble immunomodulatory cytokines as well as their immunoregulatory properties in coculture with THP-1 human monocytic cells. RNA sequencing revealed distinct gene expression in DPSCs while aBMSCs and GMSCs had less differentially expressed genes. DPSCs also had significantly less secretion of osteopontin compared to aBMSCs and GMSCs. Finally, DPSCs did not exhibit an immunosuppresive effect on THP-1 cells to the same degree as aBMSCs and GMSCs. These findings demonstrate that MSCs from different oral tissues have distinct transcriptomes and immunoregulatory properties.

Dual-enzyme inhibiting nanomedicines for enhanced cancer chemodynamic therapy by inducing intratumoral acidosis.

Deficiency of endogenous hydrogen peroxide and insufficient intracellular acidity are usually two important factors limiting chemodynamic therapy (CDT). Here we report a glutathione-responsive nanomedicine that can provide a suitable environment for CDT by inhibiting dual-enzymes simultaneously. The nanomedicine is constructed by encapsulation of a novel hydrogen sulfide donor in nanomicelle assembled by glutathione-responsive amphiphilic polymer. In response to intracellular glutathione, the nanomedicine can efficiently release the active ingredients hydrogen sulfide, carbonic anhydrase inhibitor and ferrocene. The hydrogen sulfide can increase the concentrations of hydrogen peroxide and lactic acid by inhibiting catalase and enhancing glycolysis. The carbonic anhydrase inhibitor can further induce intratumoral acidosis by inhibiting the function of carbonic anhydrase IX. Therefore, the nanomedicine can provide more efficient reaction conditions for the ferrocene-mediated Fenton reaction to generate abundant toxic hydroxyl radicals. In vivo results show that the combination of enhanced CDT and acidosis can effectively inhibit tumor growth. This design of nanomedicine provides a promising dual-enzyme inhibiting strategy to enhance antitumor efficacy of CDT.

Imaging Structural and Electrical Changes of Aging Cells Using Scanning Ion Conductance Microscopy.

The local charge density and distribution of extracellular membranes play a crucial role in the various cellular processes, such as regulation and localization of membrane proteins, electrophysiological signal transduction, transcriptional control, cell growth, and cell death. In this study, a novel scanning ion conductance microscopy-based method is employed to extracellular membrane mapping. This method allows to not only visualize the dynamic topography and surface charge distribution around individual cells, but also distinguish the charge difference. To validate the accuracy and effectiveness of this method, the charge density on model sample surfaces are initially manipulated and the charge sensing mechanism using finite element modeling (FEM) is explored subsequently. By applying this method, both the extracellular charge distributions and topography structures of normal and senescent human dental pulp stem cells (hDPSCs) are able to monitor. Interestingly, it is observed that the surface charge became significantly more negative after cellular senescence. This innovative approach enables us to gain valuable insights into surface charge changes during cellular senescence, which can contribute to a better understanding of the underlying mechanisms and potential therapeutic strategies for age-related diseases.

Three-dimensional printing of clinical scale and personalized calcium phosphate scaffolds for alveolar bone reconstruction.

OBJECTIVE: Alveolar bone defects can be highly variable in their morphology and, as the defect size increases, they become more challenging to treat with currently available therapeutics and biomaterials. This investigation sought to devise a protocol for fabricating customized clinical scale and patient-specific, bioceramic scaffolds for reconstruction of large alveolar bone defects. METHODS: Two types of calcium phosphate (CaP)-based bioceramic scaffolds (alginate/ß-TCP and hydroxyapatite/α-TCP, hereafter referred to as hybrid CaP and Osteoink™, respectively) were designed, 3D printed, and their biocompatibility with alveolar bone marrow stem cells and mechanical properties were determined. Following scaffold optimization, a workflow was developed to use cone beam computed tomographic (CBCT) imaging to design and 3D print, defect-specific bioceramic scaffolds for clinical-scale bone defects. RESULTS: Osteoink™ scaffolds had the highest compressive strength when compared to hybrid CaP with different infill orientation. In cell culture medium, hybrid CaP degradation resulted in decreased pH (6.3) and toxicity to stem cells; however, OsteoInk™ scaffolds maintained a stable pH (7.2) in culture and passed the ISO standard for cytotoxicity. Finally, a clinically feasible laboratory workflow was developed and evaluated using CBCT imaging to engineer customized and defect-specific CaP scaffolds using OsteoInk™. It was determined that printed scaffolds had a high degree of accuracy to fit the respective clinical defects for which they were designed (0.27 mm morphological deviation of printed scaffolds from digital design). SIGNIFICANCE: From patient to patient, large alveolar bone defects are difficult to treat due to high variability in their complex morphologies and architecture. Our findings shows that Osteoink™ is a biocompatible material for 3D printing of clinically acceptable, patient-specific scaffolds with precision-fit for use in alveolar bone reconstructive procedures. Collectively, emerging digital technologies including CBCT imaging, 3D surgical planning, and (bio)printing can be integrated to address this unmet clinical challenge.

Experimental study on anchorage performance of rockbolts by adding steel aggregates into resin anchoring agents.

Pull-out testing was carried out to evaluate the effects of shape, size and concentration of steel aggregates on anchorage performance. Steel grit with particle sizes of 1.5, 2.0, and 2.8 mm and steel shot diameters of 1.4, 2.0, and 2.5 mm were used as steel aggregates and were added into the resin anchoring agent. For each kind of steel aggregate, either 30, 40 or 50 aggregates were used to evaluate the effects of different steel aggregate densities. Anchorage specimens were prepared using ϕ20mm rebar bolts and steel sleeves. Compressive and shear strengths of resin containing steel aggregates, the pullout curve, and the circumferential strain of the sleeves were measured, and the energy consumption was calculated. Results show that compressive and shear strengths of resin containing steel grit and steel shot are increased by 8.4%-17.0% compared to pure resin. For the aggregate numbers of 30, 40 and 50, the anchoring force is increased by 7.9%, 7.5% and 6.5%; energy consumption is increased by 19.2%, 15.0% and 18.6%; and the circumferential strain of the specimen is increased by 28.4%, 25.1% and 39.5%, respectively. The effect of aggregate size on anchoring performance is significant; that is, the aggregate sizes of 1.4~1.5, 2.0 and 2.5~2.8 mm increase the anchoring force, energy consumption and sleeve circumferential strain by 8.5%, 4.6% and 8.7%, 16.0%, 8.4% and 28.4%, and 17.9%, 23.3% and 51.9%, respectively. The relationships of the anchoring force, energy consumption, and circumferential strain with steel aggregate quantity and size are formulated. Results show that the addition of steel aggregates increases the compressive and shear strengths of the resin, and steel aggregate quantity and size have significant impact on anchoring performance. This paper provides the basis for optimization of resin anchoring agents used in the mining industry. The impact of anchoring agent shear strength and residual shear strength on the anchoring effect were also discussed based on the failure analysis of the anchoring section.

3-year follow-up after balloon sinuplasty in children with chronic rhinosinusitis.

OBJECTIVE: Balloon sinuplasty could restore nasal sinus drainage and airflow, limiting the damages and lowering the risks. This study aimed to assess the 3-year clinical efficacy, safety, and satisfactory degree of balloon sinuplasty through multiple aspects. Also the experience about treatment for postoperative adhesion was described. MATERIALS AND METHODS: A study was performed in 30 children who had failed medical therapy, and received Balloon sinuplasty of selected sinuses. Data in 3-year follow-up including questionnaire, VAS, CT, and nasal endoscopy findings were collected. Also postoperative nose-related medications or auxiliary therapies, revision surgery, and incidence of complications were collected. The satisfactory degree of children and parents were assessed. RESULTS: Compared to preoperative, the VAS scores, questionnaire (SN-5 or SNOT-22) scores and Lund-Mackay scores were significantly lower. In the 3-year follow-up, most of them did not require nose-related medications or auxiliary therapies, and were free of symptoms, or the symptoms did not affect their daily activities. None of the 30 children had complications of facial pain, teeth numbness, facial deformity, and dysosmia. However, nasal cavity adhesion still remained the most frequent. The satisfactory degree was relatively high. CONCLUSION: The 3-year follow-up demonstrated that balloon sinuplasty had high clinical efficacy and safety, the satisfactory degree of children and parents remained relatively high. Improved the upper respiratory infection and increased the immunity, which were considered effective methods to reduce the risk of recurrence. Nasal spraying of steroids and nasal irrigation in three months after surgery cold help prevent the nasal cavity adhesion.

Hyperbranched poly(ß-amino ester) based polyplex nanopaticles for delivery of CRISPR/Cas9 system and treatment of HPV infection associated cervical cancer.

Persistent high-risk HPV infection is the main factor for cervical cancer. HPV E7 oncogene plays an important role in HPV carcinogenesis. Down-regulation of E7 oncogene expression could induce growth inhibition in HPV-positive cells and thus treats HPV related cervical cancer. Here we developed a non-virus gene vector based on poly(amide-amine)-poly(ß-amino ester) hyperbranched copolymer (hPPC) for the delivery of CRISPR/Cas9 system to specifically cleave HPV E7 oncogene in HPV-positive cervical cancer cells. The diameter of polyplex nanoparticles (NPs) formed by hPPCs/linear poly(ß-amino ester) (PBAE) and plasmids were approximately 300 nm. These hPPCs/PBAE-green fluorescence protein plasmids polyplex NPs showed high transfection efficiency and low toxicity in cells and mouse organs. By cleaving HPV16 E7 oncogene, reducing the expression of HPV16 E7 protein and increasing intracellular retinoblastoma 1 (RB1) amount, hPPCs/PBAE-CRISPR/Cas9 therapeutic plasmids polyplex NPs, especially highly branched hPPC1-plasmids polyplex NPs, exhibited strong growth inhibition of cervical cancer cells in vitro and xenograft tumors in nude mice. Together, the hPPCs/PBAE polyplex NPs to deliver HPV16 E7 targeted CRISPR/Cas9 system in this study could potentially be applied to treat HPV-related cervical cancer.

An effective vaginal gel to deliver CRISPR/Cas9 system encapsulated in poly (ß-amino ester) nanoparticles for vaginal gene therapy.

BACKGROUND: Gene therapy has held promises for treating specific genetic diseases. However, the key to clinical application depends on effective gene delivery. METHODS: Using a large animal model, we developed two pharmaceutical formulations for gene delivery in the pigs' vagina, which were made up of poly (ß-amino ester) (PBAE)-plasmid polyplex nanoparticles (NPs) based two gel materials, modified montmorillonite (mMMT) and hectorite (HTT). FINDINGS: By conducting flow cytometry of the cervical cells, we found that PBAE-GFP-NPs-mMMT gel was more efficient than PBAE-GFP-NPs-HTT gel in delivering exogenous DNA intravaginally. Next, we designed specific CRISPR/SpCas9 sgRNAs targeting porcine endogenous retroviruses (PERVs) and evaluated the genome editing efficacy in vivo. We discovered that PERV copy number in vaginal epithelium could be significantly reduced by the local delivery of the PBAE-SpCas9/sgRNA NPs-mMMT gel. Comparable genome editing results were also obtained by high-fidelity version of SpCas9, SpCas9-HF1 and eSpCas9, in the mMMT gel. Further, we confirmed that the expression of topically delivered SpCas9 was limited to the vagina/cervix and did not diffuse to nearby organs, which was relatively safe with low toxicity. INTERPRETATION: Our data suggested that the PBAE-NPs mMMT vaginal gel is an effective preparation for local gene therapy, yielding insights into novel therapeutic approaches to sexually transmitted disease in the genital tract. FUNDING: This work was supported by the National Science and Technology Major Project of the Ministry of science and technology of China (No. 2018ZX10301402); the National Natural Science Foundation of China (81761148025, 81871473 and 81402158); Guangzhou Science and Technology Programme (No. 201704020093); National Ten Thousand Plan-Young Top Talents of China, Fundamental Research Funds for the Central Universities (17ykzd15 and 19ykyjs07); Three Big Constructions-Supercomputing Application Cultivation Projects sponsored by National Supercomputer Center In Guangzhou; the National Research FFoundation (NRF) South Africa under BRICS Multilateral Joint Call for Proposals; grant 17-54-80078 from the Russian Foundation for Basic Research.

FaceWarehouse: a 3D facial expression database for visual computing.

We present FaceWarehouse, a database of 3D facial expressions for visual computing applications. We use Kinect, an off-the-shelf RGBD camera, to capture 150 individuals aged 7-80 from various ethnic backgrounds. For each person, we captured the RGBD data of her different expressions, including the neutral expression and 19 other expressions such as mouth-opening, smile, kiss, etc. For every RGBD raw data record, a set of facial feature points on the color image such as eye corners, mouth contour, and the nose tip are automatically localized, and manually adjusted if better accuracy is required. We then deform a template facial mesh to fit the depth data as closely as possible while matching the feature points on the color image to their corresponding points on the mesh. Starting from these fitted face meshes, we construct a set of individual-specific expression blendshapes for each person. These meshes with consistent topology are assembled as a rank-3 tensor to build a bilinear face model with two attributes: identity and expression. Compared with previous 3D facial databases, for every person in our database, there is a much richer matching collection of expressions, enabling depiction of most human facial actions. We demonstrate the potential of FaceWarehouse for visual computing with four applications: facial image manipulation, face component transfer, real-time performance-based facial image animation, and facial animation retargeting from video to image.