TGF-ß loaded exosome enhances ischemic wound healing in vitro and in vivo.

Rationale: With over seven million infections and $25 billion treatment cost, chronic ischemic wounds are one of the most serious complications in the United States. The controlled release of bioactive factor enriched exosome from finbrin gel was a promising strategy to promote wound healing. Methods: To address this unsolved problem, we developed clinical-grade platelets exosome product (PEP), which was incorporate with injectable surgical fibrin sealant (TISSEEL), to promote chronic wound healing and complete skin regeneration. The PEP characterization stimulated cellular activities and in vivo rabbit ischemic wound healing capacity of TISSEEL-PEP were performed and analyzed. Results: PEP, enriched with transforming growth factor beta (TGF-ß), possessed exosomal characteristics including exosome size, morphology, and typical markers including CD63, CD9, and ALG-2-interacting protein X (Alix). In vitro, PEP significantly promoted cell proliferation, migration, tube formation, as well as skin organoid formation. Topical treatment of ischemic wounds with TISSEEL-PEP promoted full-thickness healing with the reacquisition of hair follicles and sebaceous glands. Superior to untreated and TISSEEL-only treated controls, TISSEEL-PEP drove cutaneous healing associated with collagen synthesis and restoration of dermal architecture. Furthermore, PEP promoted epithelial and vascular cell activity enhancing angiogenesis to restore blood flow and mature skin function. Transcriptome deconvolution of TISSEEL-PEP versus TISSEEL-only treated wounds prioritized regenerative pathways encompassing neovascularization, matrix remodeling and tissue growth. Conclusion: This room-temperature stable, lyophilized exosome product is thus capable of delivering a bioactive transforming growth factor beta to drive regenerative events.

Ethyl Alcohol Versus Botulinum Toxin A: A Comparative Study of the Visual and Histopathological Outcomes in the Rabbit Anterior Auricular Muscle Model.

BACKGROUND: Botulinum toxin has long been known for its paralytic effects at the neuromuscular junction. Although it has been widely used for vascular and nervous tissues, there has been no study of the aesthetic effects of the application of ethanol to muscle tissues to date. OBJECTIVE: The authors aimed to demonstrate the effects of the application of ethanol to muscle tissues after an intramuscular injection and to compare the effects of botulinum toxin A (BTA) and ethanol. METHODS AND MATERIALS: A total of 28 rabbits were divided into 4 groups (n = 7 each). Botulinum toxin A (5 units) and different concentrations of ethanol (5 cc) were injected into the left and right anterior auricular muscles of all rabbits, respectively. Ear ptosis was assessed, and histopathological examination was performed after all rabbits were euthanized in the eighth week. RESULTS: Muscle function was affected earlier in ethanol-treated ears than in botulinum-treated ears; however, the ptotic effect lasted for a significantly shorter duration in ethanol-injected ears than in BTA-applied ears. CONCLUSION: Ethanol can block muscle function reversibly and can serve as an alternative to BTA, particularly when rapid results are desirable.

Malignant (necrotizing) externa otitis: the experience of a single hyperbaric centre.

INTRODUCTION: Malignant otitis externa (MOE) is a potentially life-threatening infection of the soft tissues of the external ear, quickly spreading to involve the periosteum and bone of the skull base. Treatment includes antibiotics and eventually surgery. Hyperbaric oxygen treatment (HBOT) has been proposed as an adjunctive therapy. However, in the tenth consensus conference, this disease was considered as a non-indication for HBOT. The aim of this study was to evaluate the effectiveness of HBOT in MOE treatment. METHODS: Retrospective and observational study was conducted of patients with MOE treated in our centre. Staging of the disease was made according to the clinicopathological classification system. RESULTS: From March 1998 to November 2016, 16 patients were referred. 6% patients were on stage 1 of the disease at the time they were referred, 20% in stage 2, 7% in stage 3a, 13% in stage 3b and 53% in stage 4. Seven (43.75%) patients had VII nerve palsy and three (18.75%) patients had multiple nerve palsy. Average length of symptoms of disease was 5 months (maximum 11 months). Average number of sessions was 33 and the length of hospitalization prior to HBOT (median 90 days) was significantly longer than the time between beginning HBOT and cure (p = 0.028, Wilcoxon signed rank test). There were no fatalities due to MOE and all patients were considered free of disease after HBOT. CONCLUSION: HBOT was well tolerated and revealed to be a helpful adjuvant treatment in MOE. According to our data, HBOT should be considered for patients who failed conventional treatments and in severe cases.

Drug Repurposing by Simulating Flow Through Protein-Protein Interaction Networks.

As drug development is extremely expensive, the identification of novel indications for in-market drugs is financially attractive. Multiple algorithms are used to support such drug repurposing, but highly reliable methods combining simulation of intracellular networks and machine learning are currently not available. We developed an algorithm that simulates drug effects on the flow of information through protein-protein interaction networks, and used support vector machine to identify potentially effective drugs in our model disease, psoriasis. Using this method, we screened about 1,500 marketed and investigational substances, identified 51 drugs that were potentially effective, and selected three of them for experimental confirmation. All drugs inhibited tumor necrosis factor alpha-induced nuclear factor kappa B activity in vitro, suggesting they might be effective for treating psoriasis in humans. Additionally, these drugs significantly inhibited imiquimod-induced ear thickening and inflammation in the mouse model of the disease. All results suggest high prediction performance for the algorithm.

Current management of necrotising otitis externa in the UK: survey of 221 UK otolaryngologists.

INTRODUCTION: Necrotising otitis externa (NOE) is a rare infection causing skull base osteomyelitis. The evidence regarding NOE consists mostly of case series. Hence, there is a limited evidence base to guide decision-making. This survey aims to report the experience of 221 otolaryngologists in this condition. MATERIALS AND METHODS: Internet survey administered to the membership of the British Association of Otorhinolaryngology - Head and Neck Surgery (ENT UK). RESULTS: Respondents' detailed replies on diagnosis, treatment and follow up are presented. One third of respondents reported increasing incidence of NOE. Over 80% diagnosed NOE based on pre-existing risk factors, severe pain, non-resolution of infection and CT scan. Most respondents managed NOE with intravenous antibiotics (90%) and blood sugar control (82%). There was less agreement in certain aspects of management including the role of surgery and the nature and duration of follow up. CONCLUSIONS: Our survey provides a picture of NOE management in the UK. While there is consensus in some aspects of NOE management, other aspects attract widely differing answers. This may reflect the lack of strong evidence in the literature. Future work should aim to address this.

Skin permeation profile and anti-inflammatory effect of anemonin extracted from weilingxian.

The aim of this study was to evaluate the skin permeability of anemonin, which was extracted from the Chinese herb weilingxian, and its potency of relieving the inflammation caused by rheumatoid arthritis (RA). To optimize the formulation, the solubility of anemonin in water and selected concentration of ethanol-water vehicles was determined. The effect of ethanol on the permeation of anemonin through human skin was then studied. Additionally, the influences of hydroxypropyl methylcellulose E50 (HPMC) and Carbomer 934 in different concentrations on the permeation of drug were investigated. Finally, the anti-inflammatory effect of the optimized formulation was assessed by murine model of xylene-induced ear edema. The results showed that the solubility and transdermal permeation of anemonin in ethanol-water vehicles linearly depended on the ethanol concentration. The combination of 30% ethanol and 3% Azone had a synergistic enhancement effect and was therefore selected for gel preparation. The 0.14% anemonin gel prepared with 1% HPMC exhibited the highest transdermal flux. The xylene-induced ear edema inhibitory rate of the optimized formulation was 48.85%. The results indicated that transdermal administration of anemonin is a potential modality for combating inflammation caused by RA.

Synthesis and pharmacological evaluation of novel limonin derivatives as anti-inflammatory and analgesic agents with high water solubility.

A novel series of water-soluble derivatives of limonin were synthesized by introducing various tertiary amines onto the C (7)-position of limonin. Ten target compounds were characterized and screened for their anti-inflammatory and analgesic activity in vivo. Compound 3c exhibited the strongest analgesic and anti-inflammatory activity among the limonin and its derivatives tested; its analgesic activity is more potent than that of aspirin and its anti-inflammatory activity is stronger than that of naproxen.

Strain-dependent effects of the histamine H4 receptor antagonist JNJ7777120 in a murine model of acute skin inflammation.

The effects of the histamine H(4) receptor antagonist JNJ7777120 were evaluated in a model of acute skin inflammation induced by local application of croton oil. The influence of strain on the effect of JNJ7777120 was investigated in four different mouse strains (CD-1, NMRI, BALB/c and C57BL/6J). In CD-1 mice, JNJ777720 (30-100 mg/kg subcutaneously, s.c.) exerted a dose-dependent inhibition of croton oil-induced ear inflammation and polymorphonuclear leucocyte infiltration, as confirmed by histological evaluation of ear tissues. JNJ7777120 (30-100 mg/kg) did not reduce ear oedema in NMRI, BALB/c or C57BL/6J mice. The positive control, dexamethasone (2 mg/kg s.c.) induced significant anti-inflammatory effects only in CD-1 and NMRI mice. In these strains, also the histamine H(1) -receptor blocker pyrilamine (30 mg/kg s.c.) significantly reduced ear oedema at 2 h after croton oil challenge, being as effective as JNJ7777120 in CD-1 mice. Taken together, these data demonstrate that the H(4) receptor antagonist JNJ7777120 may reduce acute croton oil-induced skin inflammation as effectively as H(1) receptor blockade. However, present experiments evidenced for the first time marked strain-related differences in the JNJ7777120 pharmacological activity, which have to be carefully considered when using this ligand to characterize histamine H(4) receptor functions in murine models and translating preclinical data to clinical human settings.

Increase of therapeutic activity of doxorubicin by long circulating liposomes in combination with curcumin.

In this study, doxorubicin (DOX)-loaded long circulating liposomes combined with curcumin (CUR) (DOX-CUR-LCLs) were successfully prepared as a novel formulation for cancer treatment. The particle size and distribution, zeta potential, drug loading capacity, and entrapment efficiency (EE) of the preparation were characterized. The in vitro anti-tumor activities of DOX-CUR-LCLs and DOX-LCLs against A549 cells were then evaluated and compared with that of free DOX. Cytotoxicity evaluation showed that DOX-CUR-LCLs had a significantly higher antitumor activity than other DOX preparations. These results suggest that novel DOX-CUR-LCLs, combination of DOX and CUR administered in long-circulating liposomes, could improve antitumor activity.

Inhibitory effects of essential oil from rhizomes of Ligusticum chuanxiong on hypertrophic scarring in the rabbit ear model.

CONTEXT: Hypertrophic scarring, a common proliferative disorder of dermal fibroblasts, results from an overproduction of collagen and excessive deposition of extracellular matrix. Although the treatment with surgical excisions or steroid hormones can modify the symptoms, numerous treatment-related complications have also been established. OBJECTIVE: To investigate the effects of essential oil (EO) from rhizomes of Ligusticum chuanxiong Hort. (Umbelliferae) on hypertrophic scarring in a rabbit ear model. MATERIALS AND METHODS: A rabbit ear model of hypertrophic scarring was established. EO (5, 10, and 20%) was applied once daily to the scars for 22 days. After 28 days of post-wounding, excision of scars was respectively performed for both histological examination and assays of the levels of collagen I, collagen III, matrix metalloproteinase-1 (MMP-1), and transforming growth factor beta 1 (TGF-ß1). The scar elevation index (SEI) was also determined. RESULTS: After 22 days of treatment with indicated concentrations of EO, hypertrophic scarring was significantly inhibited in the rabbit ears. The levels of TGF-ß1, collagen I, and collagen III evidently decreased and MMP-1 level markedly increased in the scar tissue. SEI was also significantly reduced. Immunohistochemical findings exhibited significant amelioration of the scar tissue. DISCUSSION AND CONCLUSION: EO suppresses hypertrophic scarring in the rabbit ear model and is a probably effective cure for human hypertrophic scarring.

Effect of pedunculagin investigated by non-invasive evaluation on atopic-like dermatitis in NC/Nga mice.

BACKGROUND/PURPOSE: Atopic dermatitis (AD) is a chronic relapsing inflammatory skin disorder that is becoming increasingly prevalent. Experimental animal models have been an indispensable tool for studying its pathological mechanisms and for in vivo testing of novel therapeutic approaches. AD-like lesions can be induced experimentally in NC/Nga mice. Pedunculagin, an ellagitannin purified from the Manchurian alder, Alnus hirsuta var. microphylla, Betulaceae, is a novel immunomodulator. To evaluate the effect of pedunculagin for AD-like lesions in NC/Nga mice, using clinical and non-invasive methods. METHODS: AD-like lesions were induced in NC/Nga mice using 2,4,6-trinitrochlorobenzene (TNCB). A cream containing 0.1% or 0.5% pedunculagin was applied to the positive treatment group, and the base cream without pedunculagin was applied to the negative treatment group. The control group did not receive any kind of topical agents. We evaluated the therapeutic efficacy of pedunculagin for AD by statistical evaluation of the clinical severity score using non-invasive biomedical engineering tools before treatment, and 1 day, 3 days, 1 week, 2 weeks and 4 weeks afterwards. RESULTS: An AD-like skin rash was successfully induced using TNCB in NC/Nga mice. The group receiving higher concentrations of pedunculagin showed faster and greater improvement. CONCLUSION: Our results suggest that remedies made from natural materials like pedunculagin are now showing promise for medical applications, and many new studies are expected to explore this potential.

Water-related occupations and diet in two Roman coastal communities (Italy, first to third century AD): correlation between stable carbon and nitrogen isotope values and auricular exostosis prevalence.

The reconstruction of dietary patterns in the two Roman imperial age coastal communities of Portus and Velia (I-III AD) by means of stable isotope analysis of bone remains has exposed a certain degree of heterogeneity between and within the two samples. Results do not correlate with any discernible mortuary practices at either site, which might have pointed to differential social status. The present study tests the hypothesis of a possible connection between dietary habits and occupational activities in the two communities. Among skeletal markers of occupation, external auricular exostosis (EAE) has proved to be very informative. Clinical and retrospective epidemiological surveys have revealed a strong positive correlation between EAE development and habitual exposure to cold water. In this study, we show that there is a high rate of occurrence of EAE among adult males in both skeletal samples (21.1% in Portus and 35.3% in Velia). Further, there is a statistically significant higher prevalence of EAE among those individuals at Velia with very high nitrogen isotopic values. This points to fishing (coastal, low-water fishing) as the sea-related occupation most responsible for the onset of the ear pathology. For Portus, where the consumption of foods from sea and river seems to be more widespread through the population, and where the scenario of seaport and fluvial activities was much more complex than in Velia, a close correlation between EAE and fish consumption by fishermen is less easy to establish.

Minimally invasive ear reshaping with a 1450-nm diode laser using cryogen spray cooling in New Zealand white rabbits.

BACKGROUND: Otoplasty is the current standard of care for treating prominent ears, a psychologically and sometimes functionally disabling disorder. The technically demanding procedure carries many risks such as poor aesthetic outcome, need for revision surgery, and need for general anesthesia. This study investigates the use of laser irradiation combined with cryogen skin cooling and stenting to reshape cartilage in the ears of New Zealand white rabbits. METHODS: In this prospective, randomized, internally controlled animal study, the right ears of 9 rabbits were mechanically deformed with a jig and then irradiated with a 1450-nm diode laser combined with cryogen skin cooling (14 J/pulse with cryogen spray for 33 milliseconds per cycle and a 6-mm spot size). The left ear served as the control. The ears were splinted for 1, 3, or 4 weeks. The rabbits were then given a lethal dose of intravenous pentobarbital, and the splints were removed and ears examined and photographed. Light and confocal microscopy were performed on the specimens. RESULTS: Shape change was observed in all 9 treated rabbit ears, while none of the control ears (stenting alone) showed significant change. Qualitatively, reshaped ears were stiffer after 4 weeks of splinting than after 1 or 3 weeks. None of the rabbits showed evidence of skin injury nor did they show signs of postprocedural pain. Findings from histologic analysis in the treated areas showed evidence of an expanded chondrocyte population in the region of laser irradiation, along with some perichondrial thickening and some fibrosis of the deep dermis. Confocal microscopy revealed minimal cellular death at 1 week and none thereafter. CONCLUSIONS: Cartilage reshaping using laser energy can be performed safely transcutaneously using cryogen spray cooling in rabbits. This animal model has similarity to human ears with regard to skin and cartilage thickness and is a stepping stone toward developing minimally invasive laser auricle reshaping in humans.

Non-invasive 3D facial analysis and surface electromyography during functional pre-orthodontic therapy: a preliminary report.

OBJECTIVES: Functional orthodontic devices can modify oral function thus permitting more adequate growth processes. The assessment of their effects should include both facial morphology and muscle function. This preliminary study investigated whether a preformed functional orthodontic device could induce variations in facial morphology and function along with correction of oral dysfunction in a group of orthodontic patients in the mixed and early permanent dentitions. MATERIAL AND METHODS: The three-dimensional coordinates of 50 facial landmarks (forehead, eyes, nose, cheeks, mouth, jaw and ears) were collected in 10 orthodontic male patients aged 8-13 years, and in 89 healthy reference boys of the same age. Soft tissue facial angles, distances, and ratios were computed. Surface electromyography of the masseter and temporalis muscles was performed, and standardized symmetry, muscular torque and activity were calculated. Soft-tissue facial modifications were analyzed non-invasively before and after a 6-month treatment with a functional device. Comparisons were made with z-scores and paired Student's t-tests. RESULTS: The 6-month treatment stimulated mandibular growth in the anterior and inferior directions, with significant variations in three-dimensional facial divergence and facial convexity. The modifications were larger in the patients than in reference children. In several occasions, the discrepancies relative to the norm became not significant after treatment. No significant variations in standardized muscular activity were found. CONCLUSIONS: Preliminary results showed that the continuous and correct use of the functional device induced measurable intraoral (dental arches) and extraoral (face) morphological modifications. The device did not modify the functional equilibrium of the masticatory muscles.

Non-invasive 3D facial analysis and surface electromyography during functional pre-orthodontic therapy: a preliminary report

OBJECTIVES: Functional orthodontic devices can modify oral function thus permitting more adequate growth processes. The assessment of their effects should include both facial morphology and muscle function. This preliminary study investigated whether a preformed functional orthodontic device could induce variations in facial morphology and function along with correction of oral dysfunction in a group of orthodontic patients in the mixed and early permanent dentitions. MATERIAL AND METHODS: The three-dimensional coordinates of 50 facial landmarks (forehead, eyes, nose, cheeks, mouth, jaw and ears) were collected in 10 orthodontic male patients aged 8-13 years, and in 89 healthy reference boys of the same age. Soft tissue facial angles, distances, and ratios were computed. Surface electromyography of the masseter and temporalis muscles was performed, and standardized symmetry, muscular torque and activity were calculated. Soft-tissue facial modifications were analyzed non-invasively before and after a 6-month treatment with a functional device. Comparisons were made with z-scores and paired Student's t-tests. RESULTS: The 6-month treatment stimulated mandibular growth in the anterior and inferior directions, with significant variations in three-dimensional facial divergence and facial convexity. The modifications were larger in the patients than in reference children. In several occasions, the discrepancies relative to the norm became not significant after treatment. No significant variations in standardized muscular activity were found. CONCLUSIONS: Preliminary results showed that the continuous and correct use of the functional device induced measurable intraoral (dental arches) and extraoral (face) morphological modifications. The device did not modify the functional equilibrium of the masticatory muscles.

Anti-inflammatory effects of ethanolic extract and alkamides-derived from Heliopsis longipes roots.

ETHNOPHARMACOLOGICAL RELEVANCE: Heliopsis longipes (A. Gray) Blake (Asteraceae) is a broadly used species in the Mexican, Central and South American Traditional Medicine for its anaesthetic, analgesic, anti-inflammatory and anti-ulcerative properties. The ethanolic extract contains alkamides, mainly affinin (spilanthol). This family of compounds exerts an in vitro inhibitory action on the cyclooxygenase and lipoxygenase enzymes. AIM OF THE STUDY: The present study approaches the anti-inflammatory effect of the extract and its main bioactive component affinin and derived isobutyl-decanamide. MATERIALS AND METHODS: The anti-inflammatory effect was evaluated through the mouse ear oedema test by means of two irritating agents, arachidonic acid (AA) and phorbol myristate acetate (PMA). RESULTS: Heliopsis longipes, affinin and isobutyl-decanamide displayed a marked anti-inflammatory effect on the AA model with ED(50)=0.8, 1.2 and 0.9 mg/ear, respectively. Nimesulide (1 mg/ear) was used as a reference drug. In PMA model, the extract and two alkamides also showed a dose-dependent anti-inflammatory effect with ED(50)=2.0, 1.3 and 1.1 mg/ear, respectively. Indomethacin (3 mg/ear) was used as reference drug. CONCLUSIONS: These results could represent an important contribution to explain the anti-inflammatory ethnobotanical effects reported for Heliopsis longipes and other species containing affinin (spilanthol). For the first time the topical anti-inflammatory effects of Heliopsis longipes, affinin and isobutyl-decanamide were studied.

Endogenous ochronosis.

Endogenous ochronosis or alkaptonuria is a rare, autosomal recessive disease of tyrosine metabolism that is caused by a deficiency of the enzyme homogentisic acid oxidase. The disease results in the accumulation and deposition of homogentisic acid in the cartilage, eyelids, forehead, cheeks, axillae, genital region, buccal mucosa, larynx, tympanic membranes, and tendons. The disease generally presents in adults with arthritis and skin abnormalities; occasionally, involvement of other organs may be seen. A 49-year-old man was referred to our clinic with verrucous lesions on his hands. On physical examination, caviar-like ochronotic papules were found around his eyes and the helix cartilage of his ears, and on the dorsa of both hands. There were brown macules on the sclera (Osler's sign). The patient had arthritis and nephrolithiasis, and a sample of his urine darkened upon standing. Histopathological examination showed deposition of ochronotic pigment. High-dose ascorbic acid was given, and the patient showed improvement on follow-up examination 6 months later.

Anti-inflammatory effects of essential oil isolated from the buds of Cleistocalyx operculatus (Roxb.) Merr and Perry.

Cleistocalyx operculatus (Roxb.) Merr and Perry buds (Myrtaceae) are widely used in folk medicine for the treatment of gastric ailments as well as an antiseptic agent in China, Vietnam and some other tropical countries. However, to be clinically useful, more scientific data are needed. In the present study, we investigated the in vitro and in vivo anti-inflammatory activities of the essential oil of the C. operculatus buds (EO-CO). In the applied tests, EO-CO significantly inhibited lipopolysaccharide (LPS)-induced secretion of pro-inflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta), in RAW 264.7 cells, a mouse macrophage-like cell line. Also the mRNA expression of TNF-alpha and IL-1beta was suppressed by treatment with EO-CO in LPS-stimulated RAW 264.7 cells. Moreover, reporter gene analysis revealed that the EO-CO significantly blocked LPS-induced transcriptional activation of NF-kappaB in RAW 264.7 cells. Nuclear translocation of p65 subunit was also suppressed by EO-CO treatment. In addition, EO-CO inhibited phorbol ester-induced increase in ear swelling and skin water content in BALB/c mice. These results suggest that EO-CO might exert an anti-inflammatory effect by suppressing the expression of pro-inflammatory cytokines which is mediated, at least in part, by blocking NF-kappaB activation.

Anti-inflammatory and analgesic activities of Edgeworthia chrysantha and its effective chemical constituents.

The barks and roots of Edgeworthia chrysantha LINDL., which have been used as the folk medicine "Zhu shima" in southern China due to their detumescence and acesodyne effects, were investigated for their anti-inflammatory and analgesic activities using a xylene-induced ear edema assay in mice and Freund's complete adjuvant-induced paw edema as inflammation models, and the acetic acid-induced writhing test as an analgesic model. Fractions effective in terms of anti-inflammatory and analgesic activities were obtained from E. chrysantha. The chloroform-soluble fraction (CHF) showed significant anti-inflammatory (p<0.01-0.001) and analgesic (p<0.01) effects. On further purification by silica gel, three major coumarins, edgeworin (EdN), edgeworosides A and C (EdeA and EdeC), were isolated from the chloroform fraction and both anti-inflammatory and analgesic activities were evaluated. EdN and EdeA had anti-inflammatory (p<0.05-0.01) and analgesic (p<0.001) effects, while EdeC only showed an analgesic effect. The results of this study thus demonstrated that the coumarins EdN, EdeA and EdeC in this plant may be active constituents that contribute to the anti-inflammatory and analgesic effects.