Effect of Nanoparticle Curvature on Its Interaction with Serum Proteins.

The size or the curvature of nanoparticles (NPs) plays an important role in regulating the composition of the protein corona. However, the molecular mechanisms of how curvature affects the interaction of NPs with serum proteins still remain elusive. In this study, we employ all-atom molecular dynamics simulations to investigate the interactions between two typical serum proteins and PEGylated Au NPs with three different surface curvatures (0, 0.1, and 0.5 nm-1, respectively). The results show that for proteins with a regular shape, the binding strength between the serum protein and Au NPs decreases with increasing curvature. For irregularly shaped proteins with noticeable grooves, the binding strength between the protein and Au NPs does not change obviously with increasing curvature in the cases of smaller curvature. However, as the curvature continues to increase, Au NPs may act as ligands firmly adsorbed in the protein grooves, significantly enhancing the binding strength. Overall, our findings suggest that the impact of NP curvature on protein adsorption may be nonmonotonic, which may provide useful guidelines for better design of functionalized NPs in biomedical applications.

Fatigue behavior of endodontically treated maxillary premolars with MOD defects under different minimally invasive restorations.

OBJECTIVES: To analyze the stress distribution and subsequent fracture resistance of human maxillary premolars with mesial-occlusal-distal (MOD) defects restored with different minimally invasive restorations. MATERIALS AND METHODS: Seventy non-carious human maxillary premolars were selected and divided into seven groups (n = 10). Ten teeth without further preparation served as control. The remaining teeth were endodontically treated and received three restorative patterns: inlays without cusp coverage (I), onlays with palatal coverage (O), overlays with both buccal and palatal coverage (Ov). Lithium disilicate glass ceramics (EM) and machinable composite resin (LU) were used for restoration. Specimens were tested under cycling loading with tongue direction of 45° for 1.2 × 106 cycles at a 50-N load and 2.0-Hz frequency. The survival time and two fracture mode classifications were assessed. Three-dimensional models of each group were designed. The magnitude and pattern of stresses were analyzed under the same condition of the in vitro test using finite element stress analysis. RESULTS: Although the overlay model pattern produced more favorable stress distribution, three restorative patterns restored with the same material had no difference in survival curves (P > 0.05). Only the survival curve of the EM-Ov group had no statistical difference with that of the control group (P > 0.05). EM groups presented mainly interface adhesive failure, while LU groups were mainly material cohesive failure. CONCLUSION: For the endodontically treated maxillary premolars with MOD defect, the lithium disilicate glass ceramic overlay pattern can reach the best restorative effect. CLINICAL RELEVANCE: Comparing with restorative pattern, restorative material had a greater influence on the minimally invasive restoration of posterior teeth.

The role of a multidisciplinary team in the management of portal hypertension.

BACKGROUND: Gastroesophageal variceal hemorrhage is the most severe complication of portal hypertension, with a high mortality rate. The current recommendations for gastroesophageal varices include pharmacological treatment, endoscopic treatment, transjugular intrahepatic portosystemic shunt (TIPS) placement, and splenectomy with devascularization surgery. Multidisciplinary team (MDT) comprises of a group of medical experts and specialists across a range of disciplines, providing personalized and targeted patient care tailored to each individual's condition, circumstances, and expectations. METHODS: Patients referred to the MDT clinic since its establishment in September 2014 were prospectively enrolled and followed-up for at least 12 months. Patient baseline characteristics, treatment methods, outcome and survival were compared to non-MDT patients retrieved from a prospectively maintained database with propensity score matching. RESULTS: Propensity-score matching (PSM) was carried out to balance available covariates, resulting in 58 MDT patients vs. 111 non-MDT patients. Overall survival and variceal rebleed was compared between the two groups. The rate of variceal rebleed was significantly higher in the non-MDT group, while no difference in overall survival was observed. CONCLUSIONS: This study is the first to investigate the role of a multidisciplinary team in the management of gastroesophageal varices secondary to portal hypertension. Patients treated based on MDT clinic recommendations had a significantly lower risk for variceal rebleed.

Controlling the Interaction of Nanoparticles with Cell Membranes by the Polymeric Tether.

The well control over the cell-nanoparticle interaction can be of great importance and necessity for different biomedical applications. In this work, we propose a new and simple way (i.e., polymeric tether) to tuning the interaction between nanoparticles and cell membranes by dissipative particle dynamics simulations. It is found that the linked nanoparticles (via polymeric tether) can show some cooperation during the cellular uptake and thereby have a higher wrapping degree than the single nanoparticle. The effect of the property of the polymer on the wrapping is also investigated, and it is found that the length, rigidity, and hydrophobicity of the polymer play an important role. More interestingly, the uptake of linked nanoparticles could be adjusted to the firm adhesion via two rigid polymeric tethers. The present study may provide some useful guidelines for novel design of functional nanomaterials in the experiments.

Preparation of Polymer@AuNPs with Droplets Approach for Sensing Serum Copper Ions.

A microfluidic droplet synthesis approach for the preparation of poly N-isopropylacrylamide protected gold nanoparticles (PNIPAm@AuNPs) was presented here. Well-dispersed PNIPAm@AuNPs could be generated within 8 min. On the basis of the aggregation-induced UV-vis adsorption intensity increasing mechanism, the PNIPAm@AuNPs-based colorimetric probe displayed high sensitivity and good selectivity for sensing copper ions. A linear calibration of relative UV-vis adsorption intensity increasing versus copper ions concentration was obtained within 5.0-750.0 µM, and the limit of detection was 2.5 µM. Furthermore, after copper ions were injected in rat, a metabolic assay was developed with the proposed probe. The results indicated that the droplet microfluidic synthesis system could provide a new way for preparation of polymer@AuNPs with good polydispersity index and showed great potential of polymer@AuNPs-based sensing probe for application in biological and clinical analysis.

Enhanced blood-brain barrier transmigration using a novel transferrin embedded fluorescent magneto-liposome nanoformulation.

The blood-brain barrier (BBB) is considered as the primary impediment barrier for most drugs. Delivering therapeutic agents to the brain is still a big challenge to date. In our study, a dual mechanism, receptor mediation combined with external non-invasive magnetic force, was incorporated into ferrous magnet-based liposomes for BBB transmigration enhancement. The homogenous magnetic nanoparticles (MNPs), with a size of ∼10 nm, were synthesized and confirmed by TEM and XRD respectively. The classical magnetism assay showed the presence of the characteristic superparamagnetic property. These MNPs encapsulated in PEGylated fluorescent liposomes as magneto-liposomes (MLs) showed mono-dispersion, ∼130 ± 10 nm diameter, by dynamic laser scattering (DLS) using the lipid-extrusion technique. Remarkably, a magnetite encapsulation efficiency of nearly 60% was achieved. Moreover, the luminescence and hydrodynamic size of the MLs was stable for over two months at 4 ° C. Additionally, the integrity of the ML structure remained unaffected through 120 rounds of circulation mimicking human blood fluid. After biocompatibility confirmation by cytotoxicity evaluation, these fluorescent MLs were further embedded with transferrin and applied to an in vitro BBB transmigration study in the presence or absence of external magnetic force. Comparing with magnetic force- or transferrin receptor-mediated transportation alone, their synergy resulted in 50-100% increased transmigration without affecting the BBB integrity. Consequently, confocal microscopy and iron concentration in BBB-composed cells further confirmed the higher cellular uptake of ML particles due to the synergic effect. Thus, our multifunctional liposomal magnetic nanocarriers possess great potential in particle transmigration across the BBB and may have a bright future in drug delivery to the brain.

Hardness gradients of dual-polymerized flowable composite resins in simulated root canals.

STATEMENT OF PROBLEM: Information is lacking of the polymerization depth of dual-polymerized flowable composite resin foundation materials in simulated root canals. PURPOSE: The purpose of this study was to investigate the hardness gradients and the polymerization depth of dual-polymerized flowable composite resin foundation materials in simulated root canals. MATERIAL AND METHODS: Slots in steel split cylinders with 1 open end were filled with the following 6 materials: Luxa Core, Para Core, Clearfil DC Core, Multi Core Flow, Gradia Core, and Core-Flo DC. After filling, they were subjected to a light intensity of 1250 mWcm(-2) with a light-emitting diode light through their open ends for 20 seconds. The resulting specimens were stored in a light-proof box at 37°C, and the Knoop hardness gradients of each polymerized material were measured after 0.5 hour, 24 hours, and 120 hours. The surface readings were obtained in 1-mm intervals at 1 mm to 10 mm away from the open ends. The collected data were analyzed by 2-way ANOVA and the Student-Newman-Keuls test (α=.05). RESULTS: Before the Knoop hardness numbers of the 6 materials became stable, they decreased gradually in depth at each time point (P<.001). However, the depths at which they became stable differed. The Knoop hardness numbers of Luxa Core and Core-Flo DC reached stability at a depth of 3 mm, Para Core at 4 mm, and Clearfil DC Core, Multi Core Flow, and Gradia Core at 5 mm. Additionally, at 120 hours after exposure, the ratios of the Knoop hardness numbers at a depth of 5 mm to those at 1 mm were 63.08% for Luxa Core, 70.48% for Clearfil DC Core, 81.38% for Para Core, 80.49% for Gradia Core, 86.30% for Multi Core Flow, and 96.28% for Core-Flo DC. CONCLUSIONS: In simulated root canals, the flowable composite resin foundation materials tested had better polymerization under dual polymerizing than under chemical polymerizing, and their chemical-polymerized capabilities could determine the definitive polymerization depth.

Comparison of mandibular buccal shelf morphology between adolescents and adults with different vertical patterns using CBCT.

OBJECTIVE: This retrospective study aimed to analyze the anatomical structure of the mandibular buccal shelf (MBS) in adolescents and adults with different vertical patterns to determine the optimal location for miniscrew insertion in orthodontic treatment. METHODS: Cone-beam computed tomography (CBCT) scans of 230 patients were utilized for measurements. The morphology and thickness of alveolar bone at the MBS were measured. Two-way ANOVA and regression analysis were conducted to analyze the influencing factors on alveolar bone and cortical bone thickness. RESULTS: Age had a significant effect on alveolar bone thickness (level I: F = 62.449, level II: F = 18.86, p < 0.001), cortical bone thickness (level II: F = 18.86, p < 0.001), alveolar bone tilt (F = 6.267, p = 0.013), and second molar tilt (F = 6.693, p = 0.01). Different vertical patterns also influenced alveolar bone thickness (level I: F = 20.950, level II: F = 28.470, p < 0.001), cortical bone thickness (level I: F = 23.911, level II: F = 23.370, p < 0.001), and alveolar bone tilt (F = 27.046, p < 0.001). As age increased, the alveolar bone thickness at level I decreased by 0.096 mm and at level II decreased by 0.073 mm. Conversely, the thickness of alveolar bone at level I and level II increased by 0.06 mm and 0.075 mm, respectively. The cortical bone thickness at level I and level II increased by 0.024 mm and 0.29 mm, respectively. However, the alveolar bone thickness decreased by 0.931 mm and 1.545 mm at level I and level II, and the cortical bone thickness decreased by 0.542 mm and 0.640 mm at level I and level II, respectively. CONCLUSION: Age, different vertical patterns, alveolar bone inclination, and different shapes of MBS significantly affected the thickness of alveolar bone and cortical bone in the MBS area. Notably, only alveolar bone thickness and cortical bone thickness at level II were affected by age and different vertical patterns simultaneously. These findings can provide valuable insights for orthodontic practitioners in selecting the most suitable location for miniscrew insertion during treatment planning.

Construction of a Ni(OH)2@CaTiO3 heterostructure on a Ti implant for enhanced osseointegration through NIR photoactivated bacterial inactivation and microenvironment optimization.

Desirable antibacterial and osseointegration abilities are essentially important for long-term survival of a Ti-orthopedic implant. Herein, a near-infrared light (NIR) excited antibacterial platform with excellent osseointegration composed of perovskite calcium titanate/nickel hydroxide on a Ti implant (Ni(OH)2@CaTiO3/Ti) was designed and successfully fabricated. The construction of the heterostructure efficiently separated the photogenerated electron-hole pairs to produce sufficient reactive oxygen species (ROS), which enabled the photoactivated bacterial inactivation (PBI) of Ti implants. The results showed that the surface-modified Ti implant displayed remarkable antibacterial ability with bacterial inhibition rates of 95.5% for E. coli and 93.8% for S. aureus under NIR excitation. Also, the intervention of Ni(OH)2 could create a slightly alkaline surface on the Ti implant, which synchronized with Ca-rich CaTiO3 to regulate the osteogenic microenvironment in favor of the adhesion, proliferation and differentiation of MC3T3-E1 cells as well as the up-regulation of osteogenesis-related gene expressions. The in vivo implantation experiments further confirmed that the heterostructured coating prominently accelerated the formation of new bone and promoted the osseointegration of Ti implants. Our work may provide a novel concept for improving the antibacterial and osseointegration abilities of Ti implants in orthopedic and dental applications.

Bioconjugation of luminescent silicon quantum dots for selective uptake by cancer cells.

Conventional quantum dots have great potential in cancer-related imaging and diagnostic applications; however, these applications are limited by concerns about the inherent toxicity of their core materials (e.g., cadmium, lead). Virtually all imaging applications require conjugation of the imaging agent to a biologically active molecule to achieve selective uptake or binding. Here, we report a study of biocompatible silicon quantum dots covalently attached to biomolecules including lysine, folate, antimesothelin, and transferrin. The particles possess desirable physical properties, surface chemistry, and optical properties. Folate- and antimesothelin-conjugated silicon quantum dots show selective uptake into Panc-1 cells. This study contributes to the preclinical evaluation of silicon quantum dots and further demonstrates their potential as an imaging agent for cancer applications.

Multimodal imaging probes based on Gd-DOTA conjugated quantum dot nanomicelles.

Recently, multimodal nanoparticles integrating dual- or tri-imaging modalities into a single hybrid nanosystem have attracted plenty of attention in biomedical research. Here, we report the fabrication of two types of multimodal micelle-encapsulated nanoparticles, which were systematically characterized and thoroughly evaluated in terms of their imaging potential and biocompatibility. Optical and magnetic resonance (MR) imaging probes were integrated by conjugating DOTA-gadolinium (Gd) derivative to quantum dot based nanomicelles. Two amphiphilic block copolymer micelles, amine-terminated mPEG-phospholipid and amine-modified Pluronic F127, were chosen as the capping agents because of their excellent biocompatibility and ability to prevent opsonization and prolong circulation time in vivo. Owing to their different hydrophobic-hydrophilic structure, the micellar aggregates exhibited different sizes and protection of core QDs. This work revealed the differences between these nanomicelles in terms of the stability over a wide range of pH, along with their cytotoxicity and the capacity for chelating gadolinium, thus providing a useful guideline for tailor-making multimodal nanoparticles for specific biomedical applications.

Bioconjugated PLGA-4-arm-PEG branched polymeric nanoparticles as novel tumor targeting carriers.

In this study, we have developed a novel carrier, micelle-type bioconjugated PLGA-4-arm-PEG branched polymeric nanoparticles (NPs), for the detection and treatment of pancreatic cancer. These NPs contained 4-arm-PEG as corona, and PLGA as core, the particle surface was conjugated with cyclo(arginine-glycine-aspartate) (cRGD) as ligand for in vivo tumor targeting. The hydrodynamic size of the NPs was determined to be 150-180 nm and the critical micellar concentration (CMC) was estimated to be 10.5 mg l( - 1). Our in vitro study shows that these NPs by themselves had negligible cytotoxicity to human pancreatic cancer (Panc-1) and human glioblastoma (U87) cell lines. Near infrared (NIR) microscopy and flow cytometry demonstrated that the cRGD conjugated PLGA-4-arm-PEG polymeric NPs were taken up more efficiently by U87MG glioma cells, over-expressing the α(v)ß(3) integrin, when compared with the non-targeted NPs. Whole body imaging showed that the cRGD conjugated PLGA-4-arm-PEG branched polymeric NPs had the highest accumulation in the pancreatic tumor site of mice at 48 h post-injection. Physical, hematological, and pathological assays indicated low in vivo toxicity of this NP formulation. These studies on the ability of these bioconjugated PLGA-4-arm-PEG polymeric NPs suggest that the prepared polymeric NPs may serve as a promising platform for detection and targeted drug delivery for pancreatic cancer.

Fatigue Performance and Stress Distribution of Endodontically Treated Premolars Restored with Direct Bulk-fill Resin Composites.

PURPOSE: To investigate the fatigue performance and stress distribution of endodontically treated maxillary premolars with occlusal (O), mesio-occlusal (MO), or mesio-occluso-distal (MOD) cavities filled directly with bulk-fill composite. Materials and Methods: Besides the intact teeth (control group), sixty sound maxillary first premolars, standardized by size and morphology, were subjected to root canal treatment and randomly allocated to three groups throughout cavity preparations (O/MO/MOD). All cavities were restored with a bulk-fill composite (Tetric N-Ceram Bulk fill) and universal adhesive (Tetric N-Bond Universal) using etch-and-rinse mode. Half of the specimens of each group underwent 20,000 thermocycles (5°C-55°C). All specimens were subjected to a 50-N load perpendicular to their buccal bevels on the palatal cusps for 1,200,000 cycles. The survival curve and fracture mode were analyzed by log-rank and Fisher's exact tests, respectively. Finite element analysis (FEA) was conducted to simulate the working condition of premolars with O/MO/MOD cavities. The von Mises stress and the first principal stress were calculated for three FEA models. Results: Premolars with O cavity restorations exhibited better stress distributions than did those with MO and MOD cavity restorations. Compared to the intact premolars, no significant difference was detected in the fatigue performance of O/MO/MOD restorations, regardless of whether they underwent thermocycling. Only one specimen presented unrestorable fracture, while the rest of the fractured premolars were restorable. Conclusion: The cavity design of endodontic premolars restored with a bulk-fill composite has no influence on the stress distribution or fatigue survival, with a biomechanical performance similar to that of an intact tooth.

Biocompatible PEGylated gold nanorods as colored contrast agents for targeted in vivo cancer applications.

In this contribution, we report the use of a PEGylated gold nanorods formulation as a colored dye for tumor labeling in vivo. We have demonstrated that the nanorod-targeted tumor site can be easily differentiated from the background tissues by the 'naked eye' without the need of sophisticated imaging instruments. In addition to tumor labeling, we have also performed in vivo toxicity and biodistribution studies of PEGylated gold nanorods in vivo by using BALB/c mice as the model. In vivo toxicity studies indicated no mortality or adverse effects or weight changes in BALB/c mice treated with PEGylated gold nanorods. This finding will provide useful guidelines in the future development of diagnostic probes for cancer diagnosis, optically guided tumor surgery, and lymph node mapping applications.

Enhancing the targeting ability of nanoparticles via protected copolymers.

It is important to maintain the balance between therapeutic efficiency and cytotoxicity when using nanomaterials for biomedical applications. Here, we propose a new method (i.e., non-covalent coating of protected copolymers onto the nanoparticle surface) to enhance the active targeting of nanoparticles to the cancer cells by combining the dissipative particle dynamics simulation and in vitro experiments. When coating the protected copolymer onto the nanoparticle surface, the uptake efficiency could be greatly altered due to the competition between the copolymer-ligand interaction and the receptor-ligand interaction-the non-covalent coating is more efficient than the covalent coating. Furthermore, the effect of the physicochemical properties of the protected copolymer on the targeting ability of nanoparticles was also investigated. This study offers useful insight into the optimal design of nanocarriers in biomedicine.

A new species and two new combinations of Monolophus (Zingiberaceae) from Indo-Burma.

Monolophus odontochilus Y.H.Tan & H.B.Ding, a new species from Northern Myanmar, is described and illustrated. The new species is morphologically similar to M. linearis, but differs by having elliptic to oblong leaves (vs. linear-lanceolate to lanceolate), bilobed ligules (vs. entire), purely white corolla (vs. pinkish white), semi-orbicular crenate labellum (vs. trilobed). In addition, a diagnostic key to the new species of Monolophus and its closely related non-yellow flowered species is provided. New combinations are proposed here for Caulokaempferia phokhamii Picheans. & Douangde. and C. wongsuwaniae Picheans. & Douangde. from Laos.

VEGF-PLGA controlled-release microspheres enhanced angiogenesis in encephalomyosynangiosis-based chronic cerebral hypoperfusion.

Treatments enhancing angiogenesis for chronic cerebral hypoperfusion (CCH) are still in the research stage. Although encephalomyosynangiosis (EMS) is a common indirect anastomosis for the treatment of CCH, the effectiveness to promote angiogenesis is not satisfactory. Vascular endothelial growth factors (VEGF) is a cytokine found to specifically act directly on vascular endothelial cells, promote neovascularization, and enhance capillary permeability. However, the short half life and unstable property of VEGF underlies the need to explore available delivery system. In this study, poly (lactide-co-glycolide) (PLGA) was used to prepare VEGF controlled-release microspheres. In vitro and in vivo analysis of release kinetics showed that the microspheres could release VEGF continuously within 30 days. Then, modified chronic cerebral hypoperfusion rat model was established by ligation of bilateral internal carotid artery and one vertebral artery. At 14 days after ischemia, the EMS and the VEGF microspheres injection were performed. At 30 days after the injection, the result of Morris water maze displayed that combinating VEGF microspheres and EMS significantly ameliorated cognitive deficit after ischemia. We observed that combinating VEGF microspheres and EMS could further significantly increase cerebral blood flow. We speculated that this enhancement of cerebral blood flow was attributed to more angiogenesis induced by combination of VEGF microspheres and EMS, which verified by more collateral circulation with cerebral angiography and higher expression of CD31 or α-SMA. Our study demonstrated that combinating VEGF-PLGA controlled-release microspheres could significantly promote angiogenesis in EMS-based CCH rats model, providing new ideas for clinical treatment of CCH.

Biocompatible near-infrared quantum dots as ultrasensitive probes for long-term in vivo imaging applications.

A facile synthesis method to produce monodisperse, biocompatible, lysine crosslinked mercaptoundecanoic acid (MUA) CdSe(0.25)Te(0.75)/CdS near-infrared (NIR) quantum dots and use them as probes to study their long term in vivo distribution, clearance, and toxicity is presented. Large signal enhancements are demonstrated by these quantum dots, which enables their use as efficient and sensitive probes for live-animal imaging. An important finding is that mice intravenously injected with approximately 10.5 mg kg(-1) of NIR QDs survive for more than three months without any apparent adverse effect to their health. Furthermore, it is determined that there is a significant reduction in the number of the QDs in the liver and spleen three months post injection. In addition, histological analysis of heart, kidney, liver, spleen, and lung tissue indicates that there are no acute toxic effects from these lysine cross-linked MUA NIR QDs. This study suggests that these NIR QDs can be potentially used for long-term targeted imaging and therapy studies in vivo.

Synthesis, characterization and transfection of a novel folate-targeted multipolymeric nanoparticles for gene delivery.

Novel folate-conjugated biodegradable multipolymeric nanoparticles (NPs) were constructed and evaluated for potential use in gene delivery to human cervical carcinomas Hela cells, which overexpressed folate receptors. Folate-poly(ethylene glycol)-poly(D: , L: -lactic-co-glycolic acid) (PELGA-F) was synthesized and collaborated with poly-L: -lysine (PLL) to form polymer-polycationic peptide-DNA (PPD) NPs. Fluorescein sodium and polylysine-condensed DNA (PD) were encapsulated in both PELGA nanoparticles (PELGA-NPs) and folate modified nanoparticles (PELGA-F-NPs), which were prepared by a modified solvent extraction/evaporation method. Effects of the folate conjugation and PLL introduction on the uptake of NPs was qualified by fluorescent invert microscopy and quantified by spectrofluorometric measurement, while effect on the gene expression was measured by X-gal staining and luciferase assay, both using Hela cells as an in vitro model. Results showed that cellular uptake of NPs was enhanced by folate modification, but had no difference after PLL encapsulation. In transfection tests, increased gene expression also confirmed the different functions of folate and PLL introduction. It is feasible that folate-linked multipolymeric NPs should be an efficient targeted carrier for gene delivery.

Demonstration of three injection methods for the analysis of extrinsic and intrinsic blood supply of the peripheral nerve.

BACKGROUND: The use of free vascularized nerve grafts requires an intimate and accurate knowledge of the blood supply of peripheral nerve. This study was designed to compare the advantages and disadvantages of three methods employed to reveal the blood supply of the peripheral nerve, and to provide morphological basis for vascularized nerve grafts. METHODS: The blood supply of brachial plexus and its main branches (ulnar, median, radial, musculocutaneous and axillary nerve) were observed using three vascular injection techniques: three specimens were injected with red latex through the thoracic aorta; two side specimens were injected with a Chinese ink solution, through the subclavian artery, for diaphanization and histology; one fresh cadaver was injected with the gelatin-lead oxide mixture through the femoral artery for radiography. RESULTS: The blood supply of the brachial plexus and its main branches was well examined using the three different vascular injection techniques. Perfusion with red latex exposed the extrinsic blood supply. Diaphanization and histology showed the intrinsic blood supply, while gelatin-lead oxide injection technique interactively displayed both the intrinsic and extrinsic blood supply to the peripheral nerve. CONCLUSION: The standard method for the study of the extrinsic blood supply to the peripheral nerve is the red latex perfusion; diaphanization and histology are very suitable to study the intrinsic blood supply of the peripheral nerve; while gelatin-lead oxide technique is the standard for visualization of the integral topography of the blood supply of the peripheral nerve.