Mitigation of Equine Recurrent Uveitis Through Topical Suppressor of Cytokine Signaling-1 Mimetic Peptide: Open Label Safety and Efficacy Pilot Study

Equine recurrent uveitis (ERU) is a painful and debilitating autoimmune disease, and represents the only spontaneous model of human recurrent uveitis (RU). Despite the efficacy of existing treatments, RU remains a leading cause of visual handicap in horses and humans. Cytokines, which utilize Janus kinase 2 (Jak2) for signaling, drive the inflammatory processes in ERU that promote blindness. Notably, suppressor of signaling-1 (SOCS1), which naturally limits the activation of Jak2 through binding interactions, is often deficient in autoimmune disease patients. Significantly, we previously showed that topical administration of a SOCS1 peptide mimic (SOCS1-KIR) mitigated induced rodent uveitis. In this pilot study, we test the potential to translate the therapeutic efficacy observed in experimental rodent uveitis to equine patient disease. Through bioinformatics and peptide binding assays we demonstrate putative binding of the SOCS1-KIR peptide to equine Jak2. We also show that topical, or intravitreal injection of SOCS1-KIR was well tolerated within the equine eye through physical and ophthalmic examinations. Finally, we show that topical SOCS1-KIR administration was associated with significant clinical ERU improvement. Together, these results provide a scientific rationale, and supporting experimental evidence for the therapeutic use of a SOCS1 mimetic peptide in RU.

Comparative Pharmacokinetic Studies of Marketed and Microsponges Gel Loaded with Diclofenac Diethylamine in Rabbits

Oral administration of the non-steroidal anti-inflammatory drug, Diclofenac Diethylamine (DDEA) is often associated with gastrointestinal ulcers, bleeding and extensive first-pass hepatic metabolism. As an alternative to oral administration, formulated microsponges-based gel of DDEA was developed for topical administration, to quantify diclofenac diethylamine in plasma of rabbits for this a sensitive Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method was developed using carbamazepine as Internal standard (IS) and DDEA (pure drug) was provoked on Hypersil RP C18 column (250mm × 4.6mm 5µm) using a mobile phase mixture of potassium dihydrogen buffer pH 2.5 and acetonitrile in the ratio of 30:70 v/v at an isocratic flow rate of 1mL/min. The drug peak area was detection and found at 276nm. The retention time of DDEA was found to be 5.3 min. The calibration curve was linear over the concentration range of 50-750ng/ml of DDEA. This method was accurate for quantitative estimation of the drug from the marketed gel and optimized microsponge gel. The main of investigation is to compare pharmacokinetic profile of diclofenac diethylamine in pharmaceutical dosage forms (marketed gel and microsponges gel) using WinNonlin software version 8.1.

Cryptomphalus aspersa Eggs Extract Potentiates Human Epidermal Stem Cell Regeneration and Amplification

Modern life and extended life expectancy have prompted the search for natural compounds alleviating skin aging. Evidence supports the beneficial effects on skin integrity and health from the topical administration of preparations of the mollusc Cryptomphalus aspersa eggs extract (IFC-CAF®) and suggests these effects are partly derived from an impact on skin renewal and repair mechanisms. The objective was to dissect in vitro the specific impact of IFC-CAF® on different parameters related to the regenerative potential, differentiation phenotype and exhaustion of skin stem cells. A prominent impact of IFC-CAF® was the induction of stratification and differentiated phenotypes from skin stem cells. IFC-CAF® slowed down the cell cycle at the keratinocyte DNA repair phase and, decelerated proliferation. However, it preserved the proliferative potential of the stem cells. IFC-CAF® reduced the DNA damage marker, γH2AX, and induced the expression of the transcription factor p53. These features correlated with significant protection in telomere shortening upon replicative exhaustion. Thus, IFC-CAF® helps maintain orderly cell cycling and differentiation, thus potentiating DNA repair and integrity. Our observations support the regenerative and repair capacity of IFC-CAF® on skin, through the improved mobilization and ordered differentiation of keratinocyte precursors and the enhancement of genome surveillance and repair mechanisms that counteract aging.

FORMULATION, CHARACTERIZATION, AND DETERMINATION OF THE DIFFUSION RATE STUDY OF ANTIOXIDANT SERUM CONTAINING ASTAXANTHIN NANOEMULSION

Objective: Astaxanthin is one of the natural carotenoids with strong antioxidant characteristics which is widely used in skin care. The aim of this study was developed to formulate and characterize the antioxidant serum containing astaxanthin nanoemulsion and the diffusion rate studies using diffusion Franz method. Methods: Astaxanthin nanoemulsion (As-NE) was prepared by using the self-nanoemulsifying method, followed by incorporation into serum preparation with the using carbomer as a gelling agent. Evaluation of serum As-NE was performed by physical, chemical characterizations and diffusion assay. Stability study was carried out in both accelerated (temperature of 40±2 °C/75±5%RH) and non-accelerated (at ambient temperature) conditions. Results: These results suggest that antioxidant serum As-NE had good physical and chemical characteristics that are suitable for topical administration. Conclusion: For the study of diffusion and stability under different storage conditions, it was proven that antioxidant serum As-NE form was packed in a carbomer as a gelling agent that could enhance the stability and diffusion rate of the astaxanthin.

Topical Administration of Drugs Incorporated in Carriers Containing Phospholipid Soft Vesicles for the Treatment of Skin Medical Conditions

This review focuses on the improved topical treatment of various medical skin conditions by the use of drugs delivered from carriers containing phospholipid soft vesicles. Topical drug delivery has many advantages over other ways of administration, having increased patient compliance, avoiding the first-pass effect following oral drug administration or not requesting multiple doses administration. However, the skin barrier prevents the access of the applied drug, affecting its therapeutic activity. Carriers containing phospholipid soft vesicles are a new approach to enhance drug delivery into the skin and to improve the treatment outcome. These vesicles contain molecules that have the property to fluidize the phospholipid bilayers generating the soft vesicle and allowing it to penetrate into the deep skin layers. Ethosomes, glycerosomes and transethosomes are soft vesicles containing ethanol, glycerol or a mixture of ethanol and a surfactant, respectively. We review a large number of publications on the research carried out in vitro, in vivo in animal models and in humans in clinical studies, with compositions containing various active molecules for treatment of skin medical conditions including skin infections, skin inflammation, psoriasis, skin cancer, acne vulgaris, hair loss, psoriasis and skin aging.

Topical Administration of an Ointment Prepared From Satureja sahendica Essential Oil Accelerated Infected Full-Thickness Wound Healing by Modulating Inflammatory Response in a Mouse Model

Introduction. Satureja sahendica has antibacterial and anti-inflammatory properties that can have beneficial effects for decreasing inflammation in infected wounds. Objective. This study was conducted to evaluate the effects of an ointment prepared from S sahendica essential oil (SSO) on an infected wound model in BALB/c mice. Materials and Methods. One full-thickness excisional skin wound was surgically created per animal and inoculated with 5 × 107 colony-forming units of Pseudomonas aeruginosa and Staphylococcus aureus. Following induction of the wound, the mice (N = 90) were treated with soft yellow paraffin (negative control, n = 18), mupirocin (positive control, n = 18) and 1%, 2%, and 4% SSO (n = 18 in each of the 3 groups). To determine the effect of the treatments on healing of an infected wound, the following factors were assessed: rate of the wound area, tissue bacterial count, histopathology, collagen biosynthesis, immunohistochemistry, and the expressions of insulin-like growth factor (IGF)-1, fibroblast growth factor (FGF)-2, vascular endothelial growth factor (VEGF), interleukin (IL)-1ß, IL-4, transforming growth factor beta (TGF-ß), and chemokine (CXC motif) ligand 1 (CXCL-1) on days 3, 7, and 14 after induction of the wound. Results. Topical administration of SSO shortened the inflammatory phase, accelerated cellular proliferation, and increased fibroblast distribution per 1 mm2, collagen deposition, and rapid reepithelialization in comparison with control animals (P <.05). The messenger RNA levels of IGF-1, IL-10, FGF-2, VEGF, TGF-ß1, and CXCL-1 were remarkably increased, and IL-1ß level decreased (P <.05) in the treated animals compared with the control group (P <.05). The immunohistochemical analyses showed topical administration of SSO increased collagen biosynthesis in the treated group (P <.05). Conclusions. Topical administration of SSO shows evidence of accelerating wound healing by upregulating the expression of IGF-1, IL-10, FGF-2, VEGF, TGF-ß, and CXCL-1; shortening the inflammatory stage; and promoting the proliferative phase.

Confocal Raman Spectroscopic Imaging for Evaluation of Distribution of Nano-Formulated Hydrophobic Active Cosmetic Ingredients in Hydrophilic Films

Film-forming systems are highly relevant to the topical administration of active ingredients (AI) to the body. Enhanced contact with the skin can increase the efficacy of delivery and penetration during prolonged exposure. However, after the evaporation of volatile solvents to form a thin film, the distribution of the ingredient should remain homogenous in order to ensure the effectiveness of the formula. This is especially critical for the use of hydrophobic molecules that have poor solubility in hydrophilic films. In order to address this concern, hydroxyphenethyl esters (PHE) of Punica granatum seed oil were prepared as a nanosuspension stabilised by poloxamers (NanoPHE). NanoPHE was then added to a formulation containing polyvinyl alcohol (PVA) as a film forming agent, Glycerol as a plasticiser and an antimicrobial agent, SepicideTM HB. Despite their reliability, reference methods such as high-performance liquid chromatography are increasingly challenged due to the need for consumables and solvents, which is contrary to current concerns about green industry in the cosmetics field. Moreover, such methods fail to provide spatially resolved chemical information. In order to investigate the distribution of ingredients in the dried film, Confocal Raman imaging (CRI) coupled to Non-negatively Constrained Least Squares (NCLS) analysis was used. The reconstructed heat maps from a range of films containing systematically varying PHE concentrations highlighted the changes in spectral contribution from each of the ingredients. First, using NCLS scores it was demonstrated that the distributions of PVA, Glycerol, SepicideTM HB and PHE were homogenous, with respective relative standard deviations (RSD) of 3.33%, 2.48%, 2.72% and 6.27%. Second, the respective relationships between ingredient concentrations in the films and their Raman responses, and the spectral abundance were established. Finally, a model for absolute quantification for PHE was be constructed using the percentage of spectral abundance. The prepared %w/w concentrations regressed against predicted %w/w concentrations, displaying high correlation (R2 = 0.995), while the Root Mean Squared Error (0.0869% w/w PHE) confirmed the precision of the analysis. The mean percent relative error of 3.75% indicates the accuracy to which the concentration in dried films could be determined, further supporting the suitability of CRI for analysis of composite solid film matrix. Ultimately, it was demonstrated that nanoformulation of hydrophobic PHE provides homogenous distribution in PVA based film-forming systems independent of the concentration of NanoPHE used in the formula.

Investigation of fluorescein stain–based tear film breakup time test reliability in dogs in a clinical setting

OBJECTIVE To determine intra- and interobserver reliability of a fluorescein stain–based tear film breakup time (TFBUT) test as performed in a clinical environment with and without administration of a topical anesthetic. ANIMALS 21 privately owned dogs. PROCEDURES A randomized study design was used. Two independent observers that commonly perform the TFBUT test in clinical practice read the same description of TFBUT. Observers performed TFBUT testing for each dog before and after topical administration of 0.5% proparacaine solution in 4 testing periods with a 1-hour interval between periods. Intraclass correlation coefficient (ICC) analysis was used to assess inter- and intraobserver test reliability. Linear mixed models were used to assess the main effects of testing period, observer, eye, and presence of ophthalmic disorders and their interactions on TFBUT. RESULTS Mean TFBUT measurements performed by observer 1 and observer 2 were 5.9 seconds and 8.6 seconds, respectively, when adjusted for other effects in the model. Intraobserver ICC was poor for one observer and moderate for the other. Interobserver ICC was poor without use of topical anesthetic and slightly lower when anesthetic was used. Observer and testing period were each significantly associated with TFBUT; the measurements decreased and were more variable after multiple applications of fluorescein stain and proparacaine. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested tear film stability is negatively affected by topical administration of 0.5% proparacaine solution and repeated applications of fluorescein stain. The TFBUT test as performed in this study had poor to moderate reliability.

Effects of Vibration Characteristics on the Atomization Performance in the Medical Piezoelectric Atomization Device Induced by Intra-Hole Fluctuation

Oral inhalation of aerosolized drugs has be widely applied in healing the affected body organs including lesions of the throat and lungs and it is more efficient than those conventional therapies, such as intravenous drip, intramuscular injection and external topical administration in the aspects of the dosage reduction and side effects of drugs. Nevertheless, the traditional atomization devices always exhibit many drawbacks. For example, non-uniformed atomization particle distribution, the instability of transient atomization quantity and difficulties in precise energy control would seriously restrict an extensive use of atomization inhalation therapy. In this study, the principle of intra-hole fluctuation phenomenon occurred in the hole is fully explained, and the produced volume change is also estimated. Additionally, the mathematical expression of the atomization rate of the atomizing device is well established. The mechanism of the micro-pump is further clarified, and the influence of the vibration characteristics of the atomizing film on the atomization behavior is analyzed theoretically. The curves of sweep frequency against the velocity and amplitude of the piezoelectric vibrator are obtained by the Doppler laser vibrometer, and the corresponding mode shapes of the resonance point are achieved. The influence of vibration characteristics on atomization rate, atomization height and atomization particle size are also verified by experiments, respectively. Both the experimental results and theoretical calculation are expected to provide a guidance for the design of this kind of atomization device in the future.