Predictive Analysis in Oral Cancer Immunotherapy: Profiling Dual PD-L1-Positive Extracellular Vesicle Subtypes with Step-Wedge Microfluidic Chips.

PD-L1-positive extracellular vesicles (PD-L1+ EVs) play a pivotal role as predictive biomarkers in cancer immunotherapy. These vesicles, originating from immune cells (I-PD-L1+ EVs) and tumor cells (T-PD-L1+ EVs), hold distinct clinical predictive values, emphasizing the importance of deeply differentiating the PD-L1+ EV subtypes for effective liquid biopsy analyses. However, current methods such as ELISA lack the ability to differentiate their cellular sources. In this study, a novel step-wedge microfluidic chip that combines magnetic microsphere separation with single-layer fluorescence counting is developed. This chip integrates magnetic microspheres modified with anti-PD-L1 antibodies and fluorescent nanoparticles targeting EpCAM (tumor cell marker) or CD45 (immunocyte marker), enabling simultaneous quantification and sensitive analysis of PD-L1+ EV subpopulations in oral squamous cell carcinoma (OSCC) patients' saliva without background interference. Analysis results indicate reduced levels of I-PD-L1+ EVs in OSCC patients compared to those in healthy individuals, with varying levels of heterogeneous PD-L1+ EVs observed among different patient groups. During immunotherapy, responders exhibit decreased levels of total PD-L1+ EVs and T-PD-L1+ EVs, accompanied by reduced levels of I-PD-L1+ EVs. Conversely, nonresponders show increased levels of I-PD-L1+ EVs. Utilizing the step-wedge microfluidic chip allows for simultaneous detection of PD-L1+ EV subtypes, facilitating the precise prediction of oral cancer immunotherapy outcomes.

The Minhang Pediatric Biobank cohort study: protocol overview and baseline characteristics.

BACKGROUND: Little has been done to establish biobanks for studying the environment and lifestyle risk factors for diseases among the school-age children. The Minhang Pediatric Biobank (MPB) cohort study aims to identify factors associated with health and diseases of school-aged children living in the urban or suburban area of Shanghai. METHODS: This population-based cohort study was started in all sub-districts/towns of Minhang district of Shanghai in 2014. First-grade students in elementary school were enrolled during the time of their routine physical examinations, with self-administered questionnaires completed by their primary caregivers. Additional information was extracted from multiple health information systems. Urine and saliva samples were collected during the baseline survey and follow-up visits. RESULTS: At the end of 2014 academic year, a total number of 8412 children and their parents were recruited, including 4339 boys and 4073 girls. All the participants completed the baseline survey and physical examination, and 7128 urine and 2767 saliva samples were collected. The five most prevalent childhood diseases in this population were dental caries, bronchitis, pneumonia, asthma and overweight/obese. CONCLUSIONS: The MPB cohort has been successfully established, serving as a useful platform for future research relating to the genetic, environmental and lifestyle risk factors for childhood diseases.

Bacterial exclusion and wound healing potential of horizontal platelet-rich fibrin (H-PRF) membranes when compared to 2 commercially available collagen membranes.

OBJECTIVES: The aim of the present study was to compare the barrier function during bacterial invasion and wound healing properties of 3 commonly used membranes including horizontal platelet-rich fibrin (H-PRF) against two commercially available resorbable collagen membranes. MATERIALS AND METHODS: H-PRF membranes were prepared by collecting venous blood from 3 healthy volunteers using a 700 g for 8-min centrifugation protocol followed by compression into membranes. To evaluate their barrier function, 3 groups (H-PRF membrane, collagen membrane A (Bio-Gide, Geistlich), collagen membrane B (Megreen, Shanxi Ruisheng Biotechnology Co) were placed between an inner chamber and outer chamber and inoculated with S. aureus. At 2 h, 24 h, and 48 h post-inoculation, cultures from the inner and outer chambers were assessed for bacterial CFUs. Then, scanning electron microscope (SEM) was utilized to visualized the morphological destruction by bacteria of the inner and outer surfaces of the membranes. To assess the wound healing properties of each membrane, leachates from each group were applied to human gingival fibroblasts (HGF) and a scratch assay was performed at 24 h and 48 h. RESULTS: S. aureus showed a minimal bacterial attachment or invasion rate through either collagen membranes at 2 h post-inoculation, yet over time demonstrated rapid degradation, especially on the rougher surface. While PRF demonstrated higher number of CFUs after 2 h, no significant penetration/degradation of the H-PRF membranes was observed at 24 h and 48 h in the H-PRF group. Both collagen membranes demonstrated significant morphological changes 48 h post-bacterial innoculation, while minimal obvious morphological changes were observed in the H-PRF group. The wound healing assay also demonstrated significantly better wound closure rates in the H-PRF group. CONCLUSION: H-PRF membranes exhibited better barrier function towards S. aureus over 2 days of innoculation and better wound healing ability when compared to two commercially available collagen membranes. CLINICAL RELEVANCE: This study provides further evidence for the application of H-PRF membranes during guided bone regeneration by minimizing bacterial invasion. Furthermore, H-PRF membranes have significantly better ability to promote wound healing.

2-methoxyethylacrylate modified polysulfone membrane and its blood compatibility.

Hydrophilic material 2-methoxyethylacrylate (MEA) was grafted onto polysulfone (PSF) membrane via Michael addition reaction. The 1H nuclear magnetic resonance (1H NMR), Attenuated total reflectance-Fourier transform infrared spectroscope (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS) characterization of the modified membrane proved that MEA had been successfully grafted onto PSF membrane surface. The water contact angle of the membrane surface was tested. The results showed that the water contact angle changed from 76° to 59.5°, which means that the hydrophilicity of the modified membrane was improved. A series of blood compatibility tests including bovine serum protein adsorption, platelet adhesion, prothrombin time (PT), partial thromboplastin time (APTT) and thrombin time (TT) were carried out on PSF membrane and the modified PSF membrane with highest grafted density of MEA. All of the results indicate that MEA plays an important role in improving the blood compatibility of PSF membrane.

The accuracy of cone-beam computed tomography in assessing maxillary molar furcation involvement.

AIM: The aim of this study was to investigate the accuracy of cone-beam computed tomography (CBCT) in assessing maxillary molar furcation involvement (FI). MATERIALS AND METHODS: Fifteen patients with generalized chronic periodontitis after initial therapy were recruited. CBCT was performed in maxillary molars with probing pocket depths of ≥6 mm and advanced FI, and CBCT images were analysed. Furcation surgery was performed in 20 maxillary molars. Lastly, intra-surgical FI assessments were compared with CBCT-based data. RESULTS: Intra-surgical findings confirmed 82.4% of the CBCT data, with a weighted kappa of 0.917. The agreement between both assessments was the highest in buccal furcation entrances, followed by distopalatal and mesiopalatal furcation entrances. Of the four parameters tested of detailed root anatomy and furcation morphology, the mean length of the root trunk and the width of the furcation entrance revealed by CBCT were consistent with their respective intra-surgical values (p > 0.05). Horizontal bone loss and vertical bone loss were underestimated by CBCT relative to their respective intra-surgical classifications (p ≤ 0.05). CONCLUSIONS: Cone-beam computed tomography images demonstrate a high accuracy in assessing the loss of periodontal tissue of the FI and root morphologies in maxillary molars.

Novel approach to soft tissue regeneration: in vitro study of compound hyaluronic acid and horizontal platelet-rich fibrin combination.

OBJECTIVE: This study aims to develop a compound biomaterial to achieve effective soft tissue regeneration. METHODOLOGY: Compound hyaluronic acid (CHA) and liquid horizontal-platelet-rich fibrin (H-PRF) were mixed at a ratio of 1:1 to form a CHA-PRF gel. Human gingival fibroblasts (HGFs) were used in this study. The effect of CHA, H-PRF, and the CHA-PRF gel on cell viability was evaluated by CCK-8 assays. Then, the effect of CHA, H-PRF, and the CHA-PRF gel on collagen formation and deposition was evaluated by qRT‒PCR and immunofluorescence analysis. Finally, qRT‒PCR, immunofluorescence analysis, Transwell assays, and scratch wound-healing assays were performed to determine how CHA, H-PRF, and the CHA-PRF gel affect the migration of HGFs. RESULTS: The combination of CHA and H-PRF shortened the coagulation time of liquid H-PRF. Compared to the pure CHA and H-PRF group, the CHA-PRF group exhibited the highest cell proliferation at all time points, as shown by the CCK-8 assay. Col1a and FAK were expressed at the highest levels in the CHA-PRF group, as shown by qRT‒PCR. CHA and PRF could stimulate collagen formation and HGF migration, as observed by fluorescence microscopy analysis of COL1 and F-actin and Transwell and scratch healing assays. CONCLUSION: The CHA-PRF group exhibited greater potential to promote soft tissue regeneration by inducing cell proliferation, collagen synthesis, and migration in HGFs than the pure CHA or H-PRF group. CHA-PRF can serve as a great candidate for use alone or in combination with autografts in periodontal or peri-implant soft tissue regeneration.

Efficacy of bone grafts in jaw cystic lesions: A systematic review.

BACKGROUND: Bone grafts have been applied for many years in orthopedic surgery to assist with bone repair for defects or bone discontinuity caused by trauma and tumors as well as periodontal defects. Jaw cysts are another common benign disease of the maxillofacial region which may lead to pathological bone fracture, loss of teeth, and infection. However, whether bone grafts are beneficial for bone regeneration in jaw cystic lesions and when bone grafts should be used remains unclear. AIM: To study the efficacy of bone grafts compared to spontaneous healing in the treatment of jaw cystic lesions. METHODS: A literature search was performed in Medline, Cochrane Library and Embase to identify related articles published in English in the last ten years. The following key words and MeSH terms were used: "jaw cyst", "cystic lesion", "odontogenic cyst", "periapical cyst", "dentigerous cyst", "follicular cyst", "keratocyst", "treatment", "surgery", "bone graft", "enucleation", "cystectomy", and "bone regeneration". Case reports, clinical trials, clinical studies, observational studies and randomized controlled trials were included. Study quality was evaluated. RESULTS: Ten studies (n = 10) met the inclusion criteria. Five studies reported spontaneous bone healing after enucleation, three studies investigated the efficacy of various bone grafts, and two randomized comparative studies focused on the comparison between spontaneous healing and bone grafting. Over 90% of bone regeneration occurred within 6 mo after bone grafting. The bone regeneration rate after cystectomy showed great variation, ranging from 50% to 100% after 6 mo, but reaching over 90% after 12 mo. CONCLUSION: While the long-term superiority of bone grafting compared with spontaneous healing after cystectomy is unclear, bone grafts accelerate the process of healing and significantly increase bone quality.

The hemocompatibility of the modified polysulfone membrane with 4-(chloromethyl)benzoic acid and sulfonated hydroxypropyl chitosan.

Polysulfone (PSf) membrane is widely employed in blood purification fields, but the blood compatibility of PSf membrane is not adequate. To improve the hemocompatibility of PSf membrane, 4-(chloromethyl)benzoic acid (CMBA) and sulfonated hydroxypropyl chitosan (SHPCS) were grafted onto PSf membrane surface. In our strategy, CMBA was firstly grafted on the PSf membrane surface through the Friedel-Crafts alkylation reaction, and the product was named BAPSf membrane. Then, SHPCS was grafted onto the BAPSf membrane surface by esterification, and the product was named SHPCS-BAPSf membrane. The effects of temperature and reaction time on the productivity of BAPSf and the grafting density of carboxyl and the effects of reaction time on the grafting density of SHPCS grafted onto the BAPSf membrane surface were studied. The SHPCS-BAPSf membranes are investigated by ATR-FTIR, XPS, contact angle measurements and evaluated by blood compatibility in vitro. The results reveal that the hydrophilicity of SHPCS-BAPSf membranes were grealy improved and the evaluation of protein adsorption, hemolysis test, platelet adhesion plasma recalcification time(PRT), activated partial thromboplastin time(APTT), prothrombin time(PT) and thrombin time(TT) confirmed that the SHPCS-BAPSf membranes have remarkable blood compatibility.

Graphdiyne-modified TiO2 nanofibers with osteoinductive and enhanced photocatalytic antibacterial activities to prevent implant infection.

Titanium implants have been widely used in bone tissue engineering for decades. However, orthopedic implant-associated infections increase the risk of implant failure and even lead to amputation in severe cases. Although TiO2 has photocatalytic activity to produce reactive oxygen species (ROS), the recombination of generated electrons and holes limits its antibacterial ability. Here, we describe a graphdiyne (GDY) composite TiO2 nanofiber that combats implant infections through enhanced photocatalysis and prolonged antibacterial ability. In addition, GDY-modified TiO2 nanofibers exert superior biocompatibility and osteoinductive abilities for cell adhesion and differentiation, thus contributing to the bone tissue regeneration process in drug-resistant bacteria-induced implant infection.

2-methoxyethylacrylate modified polyurethane membrane and its blood compatibility.

Hydrophilic material 2-methoxyethylacrylate (MEA) was grafted onto polyurethane (PU) membrane via Michael addition reaction. Fourier transform infrared spectroscope (FTIR) and X-ray photoelectron spectroscopy (XPS) characterizations of the modified membrane proved that MEA was successfully grafted onto PU membrane surface. The water contact angle of the modified PU membrane decreased from 86° to 48° compared with unmodified PU membrane, which means that the hydrophilicity of the modified membrane was greatly improved. A series of blood compatibility tests including bovine serum protein adsorption, platelet adhesion, hemolysis assay, plasma recalacification time, prothrombin time (PT), partial thromboplastin time (APTT) and thrombin time (TT) were carried out on PU membrane and the modified PU membrane with highest grafted density of MEA. The combined results indicate that MEA plays an important role in improving the blood compatibility of PU membrane.

Novel approach to soft tissue regeneration: in vitro study of compound hyaluronic acid and horizontal platelet-rich fibrin combination

Abstract Objective This study aims to develop a compound biomaterial to achieve effective soft tissue regeneration. Methodology Compound hyaluronic acid (CHA) and liquid horizontal-platelet-rich fibrin (H-PRF) were mixed at a ratio of 1:1 to form a CHA-PRF gel. Human gingival fibroblasts (HGFs) were used in this study. The effect of CHA, H-PRF, and the CHA-PRF gel on cell viability was evaluated by CCK-8 assays. Then, the effect of CHA, H-PRF, and the CHA-PRF gel on collagen formation and deposition was evaluated by qRT‒PCR and immunofluorescence analysis. Finally, qRT‒PCR, immunofluorescence analysis, Transwell assays, and scratch wound-healing assays were performed to determine how CHA, H-PRF, and the CHA-PRF gel affect the migration of HGFs. Results The combination of CHA and H-PRF shortened the coagulation time of liquid H-PRF. Compared to the pure CHA and H-PRF group, the CHA-PRF group exhibited the highest cell proliferation at all time points, as shown by the CCK-8 assay. Col1a and FAK were expressed at the highest levels in the CHA-PRF group, as shown by qRT‒PCR. CHA and PRF could stimulate collagen formation and HGF migration, as observed by fluorescence microscopy analysis of COL1 and F-actin and Transwell and scratch healing assays. Conclusion The CHA-PRF group exhibited greater potential to promote soft tissue regeneration by inducing cell proliferation, collagen synthesis, and migration in HGFs than the pure CHA or H-PRF group. CHA-PRF can serve as a great candidate for use alone or in combination with autografts in periodontal or peri-implant soft tissue regeneration.

A novel kind of polysulfone material with excellent biocompatibility modified by the sulfonated hydroxypropyl chitosan.

In order to ameliorate the biocompatibility of polysulfone (PSf), sulfonated hydroxypropyl chitosan (SHPCS) was grafted from PSf membrane material by Schiff-Base reaction. The original and modified membranes were characterized by attenuated total reflectance-Fourier transform infrared spectroscope (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), water contact angle (WCA) measurement, tensile strength test and antibacterial test in vitro, and the results indicated that the PSf modified by SHPCS (PSf-SHPCS) was synthesized successfully, the hydrophilicity of PSf-SHPCS membrane was improved to a great extent, all the membranes possessed good stability in physiological condition and the PSf-SHPCS membrane had good antibacterial property. Protein adsorption, platelet adhesion, hemolysis assay, plasma recalcification time, activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT) and whole blood clotting time were executed to evaluate the hemocompatibility of membranes decorated by SHPCS, and the results demonstrated that the modified membrane had fine hemocompatibility.

Characterization, catalyzed water oxidation and anticancer activities of a NIR BODIPY-Mn polymer.

To obtain near-IR absorbing biomaterials as fluorescence cellular imaging and anticancer agents for hypoxic cancer cell, a nano NIR fluorescence Mn(III/IV) polymer (PMnD) was spectroscopically characterized. The PMnD shows strong emission at 661nm when excited with 643nm. Furthermore, PMnD can catalyze water oxidation to generate dioxygen when irradiated by red LED light (10W). In particular, the PMnD can enter into HepG-2 cells and mitochondria. Both anticancer activity and the inhibition of the expression of HIF-1α for PMnD were concentration dependent. Our results demonstrate that PMnD can be developed as mitochondria targeted imaging agents and new inhibitors for HIF-1 in hypoxic cancer cells.

Fractionation of rapeseed straw by hydrothermal/dilute acid pretreatment combined with alkali post-treatment for improving its enzymatic hydrolysis.

The aim of the research was to evaluate the effect of combined treatments on fermentable sugar production from rapeseed straw. An optimum condition was found to be the combination of hydrothermal pretreatment at 180°C for 45min and post-treatment by 2% NaOH at 100°C for 2h, which was based on the quantity of monosaccharides released during enzymatic hydrolysis. As compared with the raw material without treatment, the combination of hydrothermal pretreatment and alkali post-treatment resulted in a significant increase of the saccharification rate by 5.9times. This process potentially turned rapeseed straw into value added products in accordance with the biorefinery concept.

Enhanced biocompatibility and antibacterial property of polyurethane materials modified with citric acid and chitosan.

Citric acid (CA) and chitosan (CS) were covalently immobilized on polyurethane (PU) materials to improve the biocompatibility and antibacterial property. The polyurethane pre-polymer with isocyanate group was synthesized by one pot method, and then grafted with citric acid, followed by blending with polyethersulfone (PES) to prepare the blend membrane by phase-inversion method so that chitosan can be grafted from the membrane via esterification and acylation reactions eventually. The native and modified membranes were characterized by attenuated total reflectance-Fourier transform infrared spectroscope, X-ray photoelectron spectroscopy, scanning electron microscopy, water contact angle measurement, and tensile strength test. Protein adsorption, platelet adhesion, hemolysis assay, activated partial thromboplastin time, prothrombin time, thrombin time, and adsorption of Ca(2+) were executed to evaluate the blood compatibility of the membranes decorated by CA and CS. Particularly, the antibacterial activities on the modified membranes were evaluated based on a vitro antibacterial test. It could be concluded that the modified membrane had good anticoagulant property and antibacterial property.

Synthesis, characterization, cell imaging and anti-tumor activity of multifunctional nanoparticles.

Most anticancer complexes are unable to differentiate between diseased and healthy cells, systemic toxicity and undesired side effects can result. In the current study, a PEG and RGD peptides functionalized fluorescent dye Rhodamine B isothiocyanate (RBITC) doped magnetic silica nanoparticle (MnFe(3)O(4)@SiO(2)-PEG-RGD), carrying a anticancer superparamagnetic Mn(II) complex, was synthesized and characterized using spectroscopic methods. The multifunctional nanoparticles (MnFe(3)O(4)@SiO(2)-PEG-RGD) can image HepG-2 cells and differentiate between HepG-2 and WRL-68 cells based on T(1) MR imaging technology. The in vitro fluorescence image and inhibition assay on the proliferation of HeLa cells indicate that MnFe(3)O(4)@SiO(2)-PEG-RGD nanoparticles can effectively reach the tumor site, be internalized by endocytosis and then retain in cancer cells due to the retention effect of nanoparticles. This study demonstrated that a PEG and RGD peptides functionalized silica nanoparticle was a good carrier for the anticancer complexes, and the anticancer complexes loaded multifunctional nanoparticles could be developed as special agents in monitoring therapy of cancer.

Preparation of liposomal brucine and its pharmaceutical/pharmacodynamic characterization.

AIM: To prepare a novel transdermal preparation of liposomal brucine (LB) and investigate its pharmaceutical/pharmacodynamic characterization. METHODS: LB was prepared by a modified ethanol-dripping method. Its drug encapsulation efficiency (EE), particle size, in vitro release, and skin permeation were studied. Furthermore, a safety evaluation and pharmacodynamic analysis of LB, including acute dermal toxicity, skin irritation, and analgesic and anti-inflammatory effects were investigated. RESULTS: the EE of LB was 72% and the mean particle size of the liposomes was 55.4 nm. The in vitro release profile indicated that less than 68% of the encapsulated brucine was released in 10 h. A skin permeation study showed that compared with the free brucine, LB exhibited higher cumulative drug permeation through the skin and lower drug accumulation in skin tissue, indicative of an obvious promotion of skin permeation with liposomal encapsulation. The acute dermal LD50 of LB was greater than 100 mg/kg (brucine content) and skin irritation tests revealed that LB had no irritation to both integrity and broken skin. A pharmacodynamic evaluation of LB was performed by xylene-induced mouse ear edema test and acetic acid-induced writhing test at the dosage of 1.5, 3, and 6 mg/kg, respectively. The results showed that anti-inflammatory activities and analgesic effects of brucine encapsulated were significantly higher than that of the free brucine (P<0.01). Moreover, LB maintained a remarkably longer antiinflammatory and analgesic duration. CONCLUSION: It can be proposed that LB prepared here could represent a safe, effective and promising transdermal formulation for analgesic and anti-inflammatory effects.