Visualization of the dentogingival junction using micro-plastination technique.

Plastination has revolutionized the field of anatomy and research by providing biosecurity and enabling the long-term preservation of biological material, ranging from entire bodies to individual organs and even micron sections. The dentogingival junction (DGJ) consists of both epithelial and connective tissues that are closely related to the tooth's mineralized tissues. Cutting-grinding techniques are commonly used to visualize DGJ histology. These techniques exclude enamel from preparations and focus on visualizing hard or soft tissues. To improve the micro-anatomical and histological study of this region, we applied micro-plastination technique to obtain micro-thin slices below 150 µm thick from human and animal samples. The DGJ microanatomy was visualized by applying histological stains to the micro-plastinated slices, highlighting the technique's endogenous autofluorescence capacity identifying periodontal tissues, including dentin, enamel, cementoenamel junction, dentinal tubules, connective tissue, and collagen. Based on our results, we confirm that micro-plastination is a useful technique for visualizing anatomical regions that are difficult to access, such as the DGJ. Micro-plastination can be used as an alternative technique, providing a new approach for its application in anatomical and morphological research protocols.

Effect of Experimental Bleaching Gels With Enzymes on Composite and Enamel.

INTRODUCTION AND AIMS: Potential secondary or toxic effects of peroxide-based whitening gels have driven the search for alternative methods that use natural compounds with gentle action on tooth enamel that provide remineralizing benefits. METHODS: This study introduces four innovative experimental whitening gels (GC, G1, G3, G4) formulated with enzymes (Bromelaine and Papaine) and natural extracts, along with SiO2. The efficacy of these gels was tested on nanohybrid dental composite (EsCOM100, Spident Company) and dental enamel stained with coffee and natural juice (Tedi) over 10 days. The structural changes in samples before and after bleaching were examined using scanning electron microscopy and atomic force microscopy. Additionally, cytotoxicity tests were conducted on the gels using mesenchymal stem cells from human dental pulp (dMSC) and human keratinocytes (HaCaT). Antibacterial activity was assessed on five strains (Streptococcus mutans. Porphyromonas gingivalis; Enterococcus faecalis; Escherichia coli; Staphylococcus aureus). RESULTS: Coffee and natural juice stains significantly increase the roughness of composite and enamel surfaces by forming deposits. The enzymatic action of bromelain and papain effectively disorganizes and removes these clusters, significantly reducing surface roughness. CONCLUSION: Notably, the gel containing papain and nanostructured SiO2 proved to be the most effective in removing coffee stains from both composite surfaces and enamel. On the other hand, the gel with bromelain and nanostructured SiO2 was the most efficient in removing natural juice stains. The absence of SiO2 in the experimental gels slightly decreased the enzymes' effectiveness in stain removal. The antibacterial activity observed in the experimental gels is attributed solely to the enzymatic compounds.

Asphalt dust influence on the distribution of polycyclic aromatic hydrocarbon in crankcase oil stains of a concrete, covered parking structure.

Used crankcase oil is an important source of environmental polycyclic aromatic hydrocarbons (PAHs). Here, we use gas chromatography-mass spectrometry (GC-MS) to measure and compare the concentration of PAHs, including alkylated PAHs, in used oil against new and old oil stains and parking dust collected from a concrete, covered, open parking structure to understand the distribution of PAH in crankcase oil stains. PAH concentration in used oils ranges from 606 ng/mg to 1,592 ng/mg. The PAH distribution in used oil does not match that observed in parking oils stains, parking background, or parking dust. A comparison with PAH distributions in traffic related dusts extracted from the literature and dust collected from a neighboring open asphalt-paved parking suggests that covered parking dust includes substantial contributions from asphalt-paved parking dust, road dust, and/or coal tar dust. The parking dust is the most concentrated source of PAHs in the covered parking structure (PAHs up to 4,371 ng/mg), a small contribution of which can alter the distribution of PAHs in oil stains. Even with this contribution, we were able to observe a significant decrease of the ratio of low molecular weight PAHs to high molecular weight PAHs, and a significant increase in values of the phenanthrene/anthracene and fluoranthene/(fluoranthene + pyrene) ratios when oil stains age, suggesting biodegradation is an active attenuation process in covered, open parking structures.

Use of 3D printing technology for custom bolus fabrication in the management of palmar or plantar fibromatosis with radiotherapy: A retrospective case series.

PURPOSE: Palmar or plantar fibromatosis is a benign fibroproliferative disorder affecting the fascia of the hands or feet. Management involves surgery, typically reserved for cases where progression limits function. Retrospective series demonstrate that radiation therapy (RT) can stabilize the disease course in many patients and improve symptoms in some cases. RT techniques vary between the use of electrons and superficial or orthovoltage photons and often require lead cutouts or custom boluses. We present a new approach demonstrating the implementation and effectiveness of three-dimensional (3D)-printed bolus material in patients receiving RT for fibromatosis. MATERIALS AND METHODS: A total of 3 patients, one with plantar and two with palmar fibromatosis, were treated with radiation using 3D-printed boluses over the past year. Bolus's design was based on computed tomography (CT) imaging data. Palmar patients were treated with a single en-face electron field, with a two-part accessory as a bolus and an immobilization device encasing the hand. The plantar case required 6MV photons delivered with a Volumetric Modulated Arc Therapy (VMAT) technique to cover the deeper target volume adequately. Dose and fractionation were based on guidelines from the Royal College of Radiologists in the United Kingdom. CT was used to assess printed shape and density accuracy. RESULTS: The mean deviations in shape between the printed bolus pieces and their designs were all less than 0.4 mm. The differences in mean Hounsefield units (HU) between the printed boluses and their expected values were between 7 and 44 HU. No significant issues were encountered when applying the bolus to patients. The thermoluminescent dosimeters (TLD) used demonstrated dose accuracy to within TLD precision (5 %). CONCLUSIONS: 3D printing bolus technology represents a novel approach to treating fibromatosis with radiation. It offers superior dosimetry through the reduction of air gaps and by permitting custom bolus thickness. Also, it simplifies clinical set-up by acting as an immobilization device and a visual aid for daily field placement.

Histochemistry for Molecular Imaging in Nanomedicine.

All the nanotechnological devices designed for medical purposes have to deal with the common requirement of facing the complexity of a living organism. Therefore, the development of these nanoconstructs must involve the study of their structural and functional interactions and the effects on cells, tissues, and organs, to ensure both effectiveness and safety. To this aim, imaging techniques proved to be extremely valuable not only to visualize the nanoparticles in the biological environment but also to detect the morphological and molecular modifications they have induced. In particular, histochemistry is a long-established science able to provide molecular information on cell and tissue components in situ, bringing together the potential of biomolecular analysis and imaging. The present review article aims at offering an overview of the various histochemical techniques used to explore the impact of novel nanoproducts as therapeutic, reconstructive and diagnostic tools on biological systems. It is evident that histochemistry has been playing a leading role in nanomedical research, being largely applied to single cells, tissue slices and even living animals.

Cytologic Findings and Ancillary Tests Results of Sclerosing Pneumocytoma: Our Institutional Experience.

INTRODUCTION: Sclerosing pneumocytoma (SP) is a rare benign tumor and a potential diagnostic pitfall. Our aim was to review the cytologic features of our surgically diagnosed SP cases including the clinical, immunohistochemical and available molecular findings. MATERIALS AND METHODS: A computerized search from 2013 to 2020 for surgical cases with corresponding cytology specimens diagnosed as SP was performed. The clinical data, cytology, and surgical specimens were collated for analysis. RESULTS: Six cytology specimens were collected. All were female (mean age = 35). Three have incidental lung nodules and three with cough. Cytologic findings showed variable architectural pattern (papillary, solid, singly scattered, acinar/rosette-like) and cellular heterogeneity (surface, stromal, epithelioid, plasmacytoid cells). Atypia was identified in 4/6 cases. The original cytology diagnoses were negative = 1, SP = 2 and adenocarcinoma = 3. The latter diagnoses were amended to SP after review of the surgical specimens. The three false positive cases on review have cytologic features mimicking adenocarcinoma. Immunohistochemical stains showed tumor cells (surface and stromal) were positive for TTF-1, and EMA with only the surface cells positive for pancytokeratin and Napsin A. Though two cases sent for molecular testing were negative for AKT1 or CTNNB1 exon 3 mutation, our panel did not evaluate AKT1 exon 4. CONCLUSIONS: SP is a diagnostic pitfall with 50% initially misdiagnosed as adenocarcinoma. Integrating the clinical/radiologic findings, cytologic features, and performance of immunohistochemistry on cell block are helpful in avoiding misdiagnosis. Molecular testing for recurrent mutations, if present, could be helpful for diagnosis and possible therapy options. However, routinely used molecular testing may not always capture relevant molecular markers for SP.

A Case Report of a Mysterious Mycetoma.

Mycetoma, a chronic subcutaneous infection caused by bacterial or fungal species from soil and water, presents a diagnostic challenge due to its rarity and diverse clinical manifestations. Predominantly affecting male workers in endemic regions, mycetoma typically manifests as painless swelling evolving into purulent lesions with draining sinuses in the extremities. Although historically uncommon in regions like North America, rising immigration and international travel have led to an increased prevalence, necessitating heightened clinical suspicion. Early diagnosis is crucial to prevent severe complications such as limb loss and septicemia. This case report details the diagnosis and management of chronic actinomycetoma due to Nocardia spp. in a Guatemalan immigrant landscaper and emphasizes the importance of comprehensive understanding and timely intervention in mycetoma cases.

Immunophenotypic, genetic, and clinical characterization of adult T-cell leukemia/lymphoma: A single tertiary care center experience in the United States.

OBJECTIVES: Adult T-cell leukemia/lymphoma (ATLL) is an aggressive mature T-cell neoplasm caused by human T-cell lymphotropic virus type 1 (HTLV-1). Its most common immunophenotype is CD4+/CD7-/CD25+, although unusual immunophenotypes can occur and may lead to misdiagnosis. METHODS: The immunophenotypes, cytogenetics, molecular features, clinical presentations, treatment, and prognosis of 131 patients with ATLL were retrospectively studied in a large tertiary medical center in the United States. RESULTS: All cases showed loss of CD7 expression. While 82.4% of cases demonstrated CD4+, 17.6% exhibited unusual phenotypes, including CD4+/CD8+ (6.9%), CD4-/CD8- (2.3%), CD5- (3.1%), CD2-, and CD3-. The most common cytogenetics abnormalities included polysomy 3 (34.6%), translocation 1 (23.1%), and abnormalities found on chromosome 11 (30.8%) and chromosome 14 (26.9%). The common gene mutations identified by the next-generation sequencing study were TP53 (16.7%), TBL1XR1 (16.7%), EP300 (14.3%), and NOTCH1 (14.3%). TBL1XR1 mutation is associated with genetic instabilities. There was no significant difference between the clinical presentations of these 2 groups. CONCLUSIONS: Adult T-cell leukemia/lymphoma exhibits versatile immunophenotypic, cytogenetic, and molecular features. Simultaneous involvement of blood, lymph nodes, and other organs, along with hypercalcemia in a patient from an endemic area, necessitates HTLV-1 testing to avoid underdiagnosis of this dismal disease that might need aggressive chemotherapy followed by bone marrow transplant.

Does Sunlight Affect the Quality for Purposes of DNA Analysis of Blood Stain Evidence Collected from Different Surfaces?

The aim of this study was to investigate the effect of sunlight on the degradation of DNA samples taken from blood stains from different types of surfaces. A blood sample obtained from a single male donor was placed on seven different surfaces (galvanized sheet, iron rod, newspaper, white printer paper, glass, soil, and ceramic panel). Samples were kept, during a 4-week summer period, in a room, but next to an open window. Every 7 days, 1 mm2 of blood sample was collected from each substrate and stored in labeled tube for later analysis. DNA was extracted with the Chelex method, amplified using AmpFISTRTM MinifilerTM Plus Amplification Kit, and quantified using a QuantifilerTM Human DNA Quantification kit. After 7 days of sun exposure, the highest DNA concentration was determined to be from the sample from a galvanized sheet stain, followed by, in order of decreasing concentration, the ceramic panel, glass, newspaper, iron rod, and white printer paper surface. As expected, the DNA concentration from all samples decreased as the sunlight exposure time progressed. The results obtained after the amplification in the MiniFilerTM system were in correlation with the DNA concentrations measured by the qPCR method for all samples, except for the glass, soil, and white printer paper samples. The obtained data show that DNA degradation is correlated to the length of sunlight exposure and to the type of surface the samples are collected from. A negative qPCR result does not mean negative PCR amplification in the STR system; therefore, both methods should be applied when analyzing forensic samples collected from trace evidence.

Dynamic pathology and pathogen distribution of the yeast Metschnikowia bicuspidata in the Chinese mitten crab (Eriocheir sinensis).

In this study, the "milky disease" model of Eriocheir sinhensis was constructed via intramuscular injection with the pathogenic yeast Metschnikowia bicuspidata. The dynamic pathological changes of E. sinensis after injection were elucidated with two staining methods (haemotoxylin-eosin and alcian blue periodic acid-Schiff) and fluorescence in situ hybridization technology. Anatomical observation revealed three stages of the "milky disease": no clinical signs (1-4 days after infection), the appearance of signs of disease (5-7 days), and significant liquefaction (10 days). Histological observation also revealed three stages of the disease: yeast diffusion (1-2 days after infection), yeast slow development (3-4 days), and yeast rapid proliferation (5 days). And FISH technique was suitable for the early detection of infection with M. bicuspidata in E. sinensis. We found that M. bicuspidata spread to the whole body of the crab through the haemolymph and developed into fungal septicaemia. These results elucidate the systemic pathological characteristics of "milky disease" in E. sinensis and suggest the pathogenic mechanism of M. bicuspidata.

Histomorphometric Study of the Tunics of Ductus Arteriosus in Human Fetal Cadavers Using the ImageJ Software.

INTRODUCTION: The ductus arteriosus (DA) connects the left pulmonary artery with the aorta during fetal life. Although it connects two elastic arteries, histological studies have shown that it is a muscular artery. There are very few studies on the histomorphometry of human fetal cadaveric DA. There are few studies on the changes in the tunics of the DA at various stages of fetal development. The present study aimed to observe the histomorphometric features of DA and its histological variations according to the gestational age of the fetus. METHODS: The study sample was DA dissected from 34 fetal cadavers of different gestational ages and stained with standard hematoxylin and eosin staining (H&E). We studied the structure of DA under a light microscope. We used ImageJ software (National Institutes of Health, Bethesda, Maryland) to measure the thickness of all three layers of the DA wall. RESULTS: The thickness of the DA wall was directly proportional to the gestational age of the fetus. In each trimester, we observed distinct histological changes in the tunics. CONCLUSION: The formation of multiple intimal mounds and the increase in intimal thickness observed during the last trimester are responsible for the closure of the ductus after birth. Elastosis is associated with patent DA. The disappearance of elastosis at a later gestational age ensures the closure of the ductus.

Cydnidae Pigmentation on Palms Resembling Black-Ink Stains.

We report a case of a 26-year-old male, a student by profession, who noticed black-ink-like stains over palms not removed with soap and water on relocating to a new flat during monsoons. The patient had no history of trauma, contact with chemicals, or drug intake. Sudden onset during monsoons, absence of symptoms, characteristic clinical and dermoscopic findings, and spontaneous resolution within 10 days led to a diagnosis of cydnidae pigmentation.

A causal association between lipid-lowering medications and rotator cuff syndrome: a drug-targeted mendelian randomization study.

Background: Previous research has suggested that dyslipidemia may be a risk factor for rotator cuff syndrome (RCS), and lipid-lowering drugs may aid in its treatment, though conclusions have not been definitive. Mendelian randomization is a statistical method that explores the causal relationships between exposure factors and diseases. It overcomes the confounding issues inherent in traditional observational studies, thereby providing more reliable causal inferences. We employed this method to investigate whether hyperlipidemia is a risk factor for rotator cuff syndrome and whether lipid-lowering drugs can effectively treat this condition. Methods: Genetic variations linked to lipid traits low-density lipoprotein cholesterol (LDL-C), triglyceride (TG), and total cholesterol (TC) were acquired from the UK Biobank and the Global Lipids Genetics Consortium (GLGC). Data on genetic variation in rotator cuff syndrome were obtained from FinnGen, including 24,061 patients and 275,212 controls. In the next step, we carried out two-sample Mendelian randomization analyses to determine whether lipid traits correlate with rotator cuff syndrome risk. Additionally, we performed drug-target Mendelian randomization (MR) analyses on 10 drug targets related to rotator cuff syndrome. For the drug targets that showed significant results, further analysis was done using Summary-data-based Mendelian Randomization (SMR) and colocalization techniques. We performed a mediation analysis to identify potential mediators between HMG-CoA reductase (HMGCR) and RCS. Results: No causative link was established between these lipid traits and rotator cuff syndrome. However, a significant association has been identified where HMGCR inhibition corresponds to a reduced risk of rotator cuff disease (OR = 0.68, [95% CI, 0.56-0.83], p = 1.510 × 10-4). Additionally, enhanced expression of HMGCR in muscle tissues is also linked to a decreased risk of rotator cuff syndrome (OR = 0.88, [95% CI, 0.76-0.99], p = 0.03). Body mass index (BMI) mediated 22.97% of the total effect of HMGCR on RCS. Conclusion: This study does not support low-density LDL-C, TG, and TC as risk factors for rotator cuff syndrome. HMGCR represents a potential pharmaceutical target for preventing and treating rotator cuff syndrome. The protective action of statins on the rotator cuff syndrome might not be associated with their lipid-lowering properties.

Evaluation of Forensic Luminol in Detection of Blood Stains in Instruments Following Dental Treatment.

Background Saliva and blood, being biological materials with a high potential for infectious transmission in dental environments, pose significant risks to dental professionals, assistants, and patients alike. Therefore, practitioners must adopt stringent security measures to ensure patient care, considering all parties as potential carriers of microorganisms capable of causing infectious diseases. Currently, various methods of disinfection and sterilization are employed to maintain the aseptic chain effectively. Having reliable methods for detecting substances in liquids, particularly body fluids, is crucial and highly convenient. Luminol, a chemiluminescent agent widely used in forensic science for detecting minute traces of blood that are invisible to the naked eye, presents itself as a valuable tool. Blood, a major bodily fluid often present in instruments following dental procedures, underscores the importance of its detection. Hence, in this study, luminol was utilized to detect blood traces in dental instruments following dental treatment, both before and after sterilization or disinfection. Objective Blood and saliva splashes, together with highly contagious aerosols, are always a part of dental procedures. The objective of the current study is to detect traces of blood stains on face shields, surgical instruments, and endodontic files using luminol before and after sterilization. Materials and methods Sample size calculation was done with G*Power software (Version 3.1.9.4, Düsseldorf, Germany), and a total of 30 instruments were selected for the study. In the present study, a total of 30 items were collected and utilized, including 14 instruments used after implant placement, 12 endodontic files employed after root canal treatment, and four face shields utilized during these procedures. Meanwhile, a freshly prepared luminol solution was applied to these instruments, and they were viewed in a dark environment both before and after sterilization procedures. Luminescence generated by luminol was observed in the instruments, indicative of the presence of blood not visible to the naked eye. Statistical analysis for both groups was done with IBM SPSS Statistics for Windows, Version 16.0 (Released 2007; SPSS Inc., Chicago, IL, USA). Intragroup comparison was done using the Friedman test, and intergroup comparison was done using the Wilcoxon signed-rank test. Results Blood stains and chemiluminescence were visualized in two out of 10 endodontic files (one #15 K-file and #20 K-files) and two out of four face shields. The intragroup comparison was done using the Friedman test, and it was found to be statistically significant (p < 0.05). Intergroup comparison was done using the Wilcoxon signed-rank test and was found to be statistically insignificant (p > 0.05). Conclusion Following sterilization and disinfection, there were no visual blood stains or chemiluminescence. Therefore, luminol was found to be effective in detecting blood stains in endodontic files, surgical instruments, and face shields, as well as in validating the sterilization and disinfection processes. Hence, sterilization in dentistry stands as a critical measure to guarantee patient safety, halt the dissemination of infections, and uphold exemplary clinical care standards.

Clinical Observation on the Therapeutic Effect of Port-Wine Stains with Intravenous Injection of Hematoporphyrin Monomethyl Ether (HMME).

Background: Hematoporphyrin monomethyl ether (HMME) is a promising photosensitizer for photodynamic therapy (PDT) and has found wide application in the treatment of port-wine stains (PWS). Objective: This study aims to observe and analyze the clinical efficacy and safety of HMME-PDT in the treatment of PWS patients. It also aims to evaluate the usefulness of color Doppler flow imaging (CDFI), an ultrasound technique for detecting blood flow in skin lesions, in assessing clinical efficacy. Methods: Thirty-three patients with PWS underwent HMME-PDT at our dermatology outpatient clinic between January 2019 and March 2020. Data on patient demographics, lesion location, lesion type (pink, purple, nodular thickening), treatment frequency, and pre- and post-treatment images were collected and retrospectively analyzed. CDFI was performed on three patients. Results: All patients received intravenous HMME and underwent irradiation with 532 nm green LED light. Of these, 5 patients received 1 session of HMME-PDT, 14 received 2 sessions, 9 received 3 sessions and the remaining 5 patients received more than 3 sessions. Of the 33 patients, 9 were cured (27.27%), 10 showed improvement (30.30%), 11 experienced a reduction in symptoms (33.33%), and 3 showed no significant improvement (9.09%). Most patients reported local pain and oedema, and no systemic adverse effects were observed. Clinical efficacy correlated with lesion type and total number of treatment sessions. CDFI appears to be an excellent technique for assessing clinical efficacy. Conclusion: HMME-PDT is a safe and effective method for the treatment of PWS. CDFI examination appears to be a promising assessment tool. However, further validation with larger sample sizes is warranted.

Utility and performance of cell blocks in cerebrospinal fluid cytology: Experience at two teaching hospitals.

BACKGROUND: Cytology cell blocks (CBs) are not routinely made for cerebrospinal fluid (CSF) specimens. The goal of this study was to identify when CSF CB preparation improves diagnostic performance. MATERIALS AND METHODS: Under institutional review board approval, a retrospective review of CSF cytology cases was conducted at a tertiary university-based hospital and an affiliated county hospital. Patient history, CSF volume, final diagnosis, use of stains, and whether the CB was contributory was determined from the cytopathology report. CSF nucleated cell count data was obtained from the medical record. RESULTS: A total of 69 CSF specimens with CBs from January 2006 to March 2023 were identified from 61 patients. The median CSF volume was 8 mL (interquartile range, 4-13 mL; range, 1-800 mL), with immunohistochemical stains performed on 29 (42%) cases. Per cytology report, CB was contributory in 23 cases (33%), not contributory in 34 cases (49%), and not discussed in 12 cases (17%). The median volume was 8 mL for cases in which CB was contributory, not contributory, or not discussed. There was no difference in average nucleated cell counts between cases in which CB was contributory versus not contributory (73.9 vs. 40.0, p = .175). CONCLUSIONS: CBs for CSF samples were contributory in a subset (33%) of cases. The authors were unable to identify any specific pre-analytic factors, including specimen volume and average nucleated cell counts, for cases in which CB was contributory. Further evaluation is needed to identify if there are scenarios in which CSF CBs should be routinely prepared.

Photodynamic therapy for the treatment of port-wine stains in phakomatosis pigmentovascularis.

BACKGROUND: Phakomatosis pigmentovascularis (PPV) is a rare congenital syndrome. Only a few studies have reported the treatment of PPV, including a case using photodynamic therapy (PDT) to treat PPV-associated port-wine stains (PWS). OBJECTIVE: To investigating the efficacy and adverse effects of hemoporfin-PDT in PPV-associated PWS. METHODS: The efficacy and adverse effects in patients with PPV who underwent two sessions of hemoporfin-PDT from January 2019 to December 2022 were retrospectively analyzed. RESULTS: Twenty patients were included (13 females, 7 males, age range: 2-31 years; mean: 8.20 ± 8.92 years). Two, nine, seven, and two patients had PPV types Ia, IIa, IIb, and IIIa, respectively. After two treatments, the visual evaluation indicated the color of the PWS in 4, 5, 6, and 5 patients showed poor, fair, good, and excellent improvements, respectively. The combined good and excellent improvement rates in patients with PWS and pigmentary nevus overlapping in the same treatment area and in patients with PWS in the treatment areas only were 33.3% versus 87.5%, respectively, and were significantly different (p = 0.02). Minor side effects, such as edema, scabbing, hyperpigmentation, and blistering, were observed in some patients after PDT. CONCLUSION: Hemoporfin-PDT is an effective treatment for PPV-associated PWS. Patients with PWS and pigmentary nevus overlapping in the same treatment area showed poorer efficacy than patients with PWS in the treatment areas only.

[Rapamycin and HPPH Co-Loaded Nanodrug Delivered via Dissolvable Microneedles to Treat Port-Wine Stains]. / å ±è½½é›·å¸•éœ‰ç´ å’Œå ‰å ‹æ´›çš„çº³ç±³è¯ç‰©å¤åˆå¯æº¶è§£å¾®é’ˆæ²»ç–—é²œçº¢æ–‘ç—£çš„å®žéªŒç ”ç©¶.

Objective: Port-wine stains are a kind of dermatological disease of congenital capillary malformation. Based on the biological characteristics of port-wine stains and the advantages of microneedle transdermal administration, we intend to construct a nanodrug co-loaded with rapamycin (RPM), an anti-angiogenesis drug, and photochlor (HPPH), a photosensitizer, and integrate the nanodrug with dissolvable microneedles (MN) to achieve anti-angiogenesis and photodynamic combination therapy for port-wine stains. Methods: First, RPM and HPPH co-loaded nanoparticles (RPM-HPPH NP) were prepared by the emulsification solvent-volatilization method, and its ability to generate reactive oxygen species (ROS) was investigated under 660 nm laser irradiation. Mouse hemangioendothelioma endothelial cells (EOMA) were used as the subjects of the study. The cellular uptake behaviors were examined by fluorescence microscopy and flow cytometry. The cytotoxicity effects of RPM-HPPH NP with or without 660 nm laser irradiation on EOMA cells were examined by MTT assays (with free RPM serving as the control). Then, hyaluronic acid (HA) dissolvable microneedles loaded with RPM-HPPH NP (RPM-HPPH NP@HA MN) were obtained by compounding the nanodrug with HA dissolvable microneedle system through the molding method. The morphological characteristics and mechanical properties of RPM-HPPH NP@HA MN were investigated by scanning electron microscope and electronic universal testing machine. The penetration ability of RPM-HPPH NP@HA MN on the skin of nude mice was evaluated by trypan blue staining and H&E staining experiment. Results: The RPM-HPPH NP prepared in the study had a particle size of 150 nm and generated large amounts of ROS under laser irradiation. At the cellular level, RPM-HPPH NP was taken up by EOMA cells in a time-dependent manner. The cytotoxicity of RPM-HPPH NP was higher than that of free RPM with or without laser irradiation. Under laser irradiation, RPM-HPPH NP exhibited stronger cytotoxic effects and the difference was statistically significant (P<0.05). The height of the needle tip of RPM-HPPH NP@HA MN was 600 µm and the mechanical property of a single needle was 0.75048 N. Trypan blue staining and HE staining showed that pressing on the microneedles could produce pores on the skin surface and penetration of the stratum corneum. Conclusion: RPM-HPPH NP@HA MN can deliver RPM-HPPH NP percutaneously to the lesion tissue and realize the synergistic treatment of port-wine stains with anti-angiogenic therapy and photodynamic therapy, providing a new strategy for the construction of nanodrug-loaded microneedle delivery system and the clinical treatment of port-wine stains.