Collagenase Activity Research Articles

P.776 Targeting of an F-BAR domain protein to the synaptic periactive zone ensures uniform size of synaptic vesicles

Osteoarthritis is a degenerative disease of the joint, affecting over 30 million people in the US1. A key characteristic of OA is chondrocyte hypertrophy, characterized by chondrocyte changes to a more rounded and osteoblastic phenotype, characterized by increased IL-6 and IL-8 secretion2. While there are no cures for OA, treatments focus on mitigating pain and inflammation, the two main symptoms of OA. However, the analgesics, NSAIDS and corticosteroids commonly used, do not target regeneration and have negative side effects. Local anesthetics (LA) can be used as a pain management alternative but are usually short lasting and therefore, not suited for chronic conditions such as OA. Our engineered sustained release local anesthetic construct successfully delivers bupivacaine for an extended period of time3-5. This study is designed to evaluate the effect of the LA system on chondrocytes in an inflammatory OA-like environment.Chondrocytes were cultured with bolus, liposomal, or construct LA and either untreated or treated with TNF-α and IL-1α for 24 hrs, 48 hrs, or 96 hrs. Chondrocyte viability, interleukin-8 (IL-8), interleukin-6 (IL-6), collagenase activity and proteoglycan deposition were assessed.In the presence of the engineered construct, the chondrocytes retained viability and regenerative function. Moreover, the construct allowed for higher initial doses to be used, which promoted more regeneration and decreased inflammation without compromising cellular viability.The construct promotes a less hypertrophic chondrocyte environment while promoting a more anti-inflammatory environment. These two factors are consistent with a less OA progressive environment when using the engineered construct, compared to bolus LA.

P.774 Olfactory reference syndrome as a prodrome of Alzheimer disease

Osteoarthritis is a degenerative disease of the joint, affecting over 30 million people in the US1. A key characteristic of OA is chondrocyte hypertrophy, characterized by chondrocyte changes to a more rounded and osteoblastic phenotype, characterized by increased IL-6 and IL-8 secretion2. While there are no cures for OA, treatments focus on mitigating pain and inflammation, the two main symptoms of OA. However, the analgesics, NSAIDS and corticosteroids commonly used, do not target regeneration and have negative side effects. Local anesthetics (LA) can be used as a pain management alternative but are usually short lasting and therefore, not suited for chronic conditions such as OA. Our engineered sustained release local anesthetic construct successfully delivers bupivacaine for an extended period of time3-5. This study is designed to evaluate the effect of the LA system on chondrocytes in an inflammatory OA-like environment.Chondrocytes were cultured with bolus, liposomal, or construct LA and either untreated or treated with TNF-α and IL-1α for 24 hrs, 48 hrs, or 96 hrs. Chondrocyte viability, interleukin-8 (IL-8), interleukin-6 (IL-6), collagenase activity and proteoglycan deposition were assessed.In the presence of the engineered construct, the chondrocytes retained viability and regenerative function. Moreover, the construct allowed for higher initial doses to be used, which promoted more regeneration and decreased inflammation without compromising cellular viability.The construct promotes a less hypertrophic chondrocyte environment while promoting a more anti-inflammatory environment. These two factors are consistent with a less OA progressive environment when using the engineered construct, compared to bolus LA.

Characterization of collagenase found in the nonpathogenic bacterium Lysinibacillus sphaericus VN3.

High collagenolytic activity has been detected in pathogenic bacteria. Collagenase plays an essential role in the invasion step in animals and humans. In this study, we characterized collagenase found in the nonpathogenic bacterium Lysinibacillus sphaericus VN3, which was isolated from soil in Vietnam. The collagenase activity of the purified enzyme was strongly inhibited by Cu2+, but it was significantly increased by Zn2+. The purified enzyme with a molecular mass of approximately 110 kDa exhibited collagenolytic, gelatinolytic, and caseinolytic activity. The kinetic studies showed that this enzyme had greater hydrolyzing activity toward collagen and gelatin compared with casein. Based on the ratio V max/K m, collagen is likely to be the best substrate among three proteins. We found that this collagenase could digest small pieces of bovine skin and tendon into a collagen solution. Interestingly, at pH 6.0-8.0, the soluble collagen could form a collagen membrane, which is useful as a wound-healing biomaterial.

층층갈고리둥굴레 (Polygonatum sibiricum Redoute) 뿌리줄기와 잎의 증숙 시간과 증숙 온도에 따른 생리활성 변이 연구

Background: The aim of this study was to analyze the total phenol and total flavonoid contents and antioxidant activity of steam-treated leaves and rhizomes of Polygonatum sibiricum Redoute. In addition, we aimed to confirm their potential use as cosmetic materials by investigating their anti-aging and skin-whitening activity.<BR>Methods and Results: The leaves and rhizomes of P. sibiricum were treated with steam at different temperatures for different durations, and the antioxidant activity (DPPH and ABTS radical scavenging activity) and total phenol and total flavonoid contents of each sample were tested. The steam temperature and treatment duration siginificantly affected the antioxidant activity and, total phenol and total flavonoid content of the leaves and rhizome of P. sibiricum. Treating the P. sibiricum samples with steam at 120℃ for 12 h, yielded higher total phenol and total flavonoid contents. Comparatively, the samples treated with steam at 120℃ for 12 to 24 h showed significantly higher antioxidant activity. Further, the steamed samples of P. sibiricum demonstrated collagenase and tyrosinase inhibition activity, which indicated their anti-aging and skin-whitening properties. The samples steamed at 120℃ for 12 h, exhibited higher collagenase and tyrosinase inhibition activity.<BR>Conclusions: Leaves and rhizomes of P. sibiricum steamed at 120℃ for 12 h, showed highest antioxidant activity and, total phenol and total flavonoid contents than all other samples. Our results indicate the potential of using P. sibiricum as a cosmetic material by confirming its excellent antiaging and whitening activity.

Dieckol, an algae-derived phenolic compound, suppresses airborne particulate matter-induced skin aging by inhibiting the expressions of pro-inflammatory cytokines and matrix metalloproteinases through regulating NF-κB, AP-1, and MAPKs signaling pathways

Exposure to particulate matter causes skin aging. In the present study, we investigated the effect of an algae-derived phenolic compound, dieckol (DK), against Chinese particulate matter (CPM)-stimulated aging in vitro in human dermal fibroblasts (HDF cells) and in vivo in zebrafish. DK effectively protected HDF cells against CPM-induced oxidative stress by scavenging intracellular reactive oxygen species. Moreover, DK significantly improved collagen synthesis and inhibited intracellular collagenase activity in CPM-stimulated HDF cells. In addition, DK remarkably reduced the expression of pro-inflammatory cytokines and matrix metalloproteinases via regulating the nuclear factor kappa B, activator protein 1, and mitogen-activated protein kinases signaling pathways in CPM-stimulated HDF cells. Furthermore, the in vivo test results demonstrated that DK effectively improved the survival rate of CPM-stimulated zebrafish via suppressing oxidative stress and inflammatory response. In conclusion, this study suggests that DK is a potential anti-aging compound that can be used as a therapeutic agent to improve CPM-induced skin aging, or as an ingredient to develop a cosmetic or medicine in the cosmeceutical and pharmaceutical industries.

Efeitos anti-idade dos óleos essenciais de lavanda, alecrim e melaleuca em fibroblastos dérmicos humanos

Introdução: O envelhecimento cutâneo é um processo multifatorial que tem grande importância social. Ao longo dos anos, o poder de renovação e de combate aos radicais livres da pele diminui, acarretando numa barreira cutânea menos efetiva e com maiores danos a derme. A indústria cosmética tem desenvolvido diversos produtos para desacelerar o processo desse envelhecimento cutâneo, de forma mais efetiva e menos agressiva ao ser humano e ao meio ambiente, por meio do uso de ativos de origem vegetal. Dentre esses ativos, os óleos essenciais têm se destacado pela sua diversidade de bioativos. Objetivos: Neste trabalho, objetivou-se avaliar a influência dos óleos essenciais de lavanda, alecrim e melaleuca sobre a proliferação, expressão de metaloproteinases e a síntese de colágeno e de ácido hialurônico, além da atividade de colagenase em fibroblastos dérmicos humanos. Resultados: Os óleos essenciais de lavanda e de alecrim mostraram potencial efeito no combate as alterações cutâneas causadas no processo de envelhecimento, como promover o aumento significativo da síntese de ácido hialurônico (p&lt;0,05) e, no caso do óleo essencial de lavanda, promover o aumento significativo de colágeno total (p&lt;0,05). Além disso, os óleos essenciais de lavanda e alecrim, reduziram significativamente (p&lt;0,05) a atividade da enzima colagenase. Por fim, os resultados demonstraram que os óleos essenciais de lavanda, alecrim reduziram significativamente (p&lt;0,05) a secreção de MMP-1, MMP-2 e MMP-9. Conclusão: Os resultados obtidos mostraram redução na expressão de MMPs, aumento na proliferação de fibroblastos e estímulo na síntese de colágeno e de ácido hialurônico. Sugerimos que os óleos essenciais de alecrim e lavanda possam ser em formulações cosméticas destinadas à prevenção do envelhecimento cutâneo.

Optimization of the Antioxidant Activities of Mixtures of Melastomataceae Leaves Species (M. malabathricum Linn Smith, M. decemfidum, and M. hirta) Using a Simplex Centroid Design and Their Anti-Collagenase and Elastase Properties

In this study, a simplex-centroid mixture design (SCMD) approach was used to select the optimal proportions of three different Melastomataceae leaves species (M. malabathricum, M. decemfidum, and M. hirta) extracts to determine the optimum antioxidant activities of total phenolic compound (TPC) and ABTS (2, 2′-azino-bis (ethylbenzthiazoline-6-sulfonic acid)) radical-scavenging activities. Twelve experimental designs were set up, consisting of points that were equally weighted mixtures of 0 to 1 components, which were pure blends (1, 0, …, 0), binary blends (1/2, 1/2, 0, …, 0), tertiary blends (1/3, 1/3, 1/3, 0, …, 0), and a control point. The in vitro anti-elastase and collagenase activities were evaluated in order to determine the anti-aging efficacy of the optimized mixture extracts. A high-performance liquid chromatography (HPLC) analysis was employed to identify the flavonoid content (rutin and quercetin) present in the optimized mixture extracts. The results showed that the best proportions of the optimum phenolic compounds and ABTS activity corresponded to 30%, 40%, and 30% of M. malabathricum, M. hirta, and M. decemfidum leaf extracts, respectively. The in vitro anti-collagenase and elastase activity evaluation of the optimized mixture extracts showed 70% inhibition against both elastase and collagenase enzymes. The HPLC analysis revealed two flavonoids (rutin and quercetin) at retention time 7.770 and 8.769 min, respectively, in the mixture extracts. This study suggests the potential exploitation of mixtures of Melastomataceae leaves (M. malabathricum, M. decemfidum, and M. hirta) as cosmetic ingredients for antioxidant and anti-wrinkles applications.

Intralesional injection of the calcium channel blocker Verapamil in Peyronie's disease: A critical review.

To assess the effectiveness of an intralesional injection of verapamil in men with Peyronie's disease (PD). The data provided in the current review are based on a thorough review of the available original articles on PD retrieved with a systematic literature search using PubMed- Medline, and the Cochrane Central Register of Controlled Trials, up to December 2019, to identify studies dealing with Peyronie's disease and its treatment. Included were only original articles, that we thoroughly evaluated. We searched for the primary and secondary terms of: "Peyronie's disease," "Penile curvature," "Erectile dysfunction," "Verapamil and Peyronie's disease," "Calcium channel blocker," and "Intralesional injection." The initial search of the databases yielded a total of 1240 studies (PubMed: 1058; Cochrane: 182), as of December 2019. Seventy studies were removed due to duplication. Further 986 studies were removed due to not being in English (except for one study by Arena F. for which we got a translation form Italian), being about animal experimentations, not being full-text, and not being clinical trials. Likewise, studies not referring at all to verapamil were excluded (148). From the remaining 36 full-text articles we focused on 13 studies which met the inclusion criteria, mainly being deemed relevant to the context of this study. Calcium channel blockers have been shown in both in vitro and in vivo studies to inhibit the synthesis and secretion of extracellular matrix molecules, as well as to increase collagenase activity. Patients with localised plaque are the best candidates for intralesional injections of verapamil. The beneficial effects of intralesional verapamil are apparent within the first three months. For patients who respond to treatment, the injections should be continued for six months. Patients who fail to respond to intralesional verapamil or whose angulation is greater than 30° at presentation should be considered candidates for surgery. Injection of verapamil is clinically safe for patients with Peyronie's disease, and it appears to induce a rapid, beneficial effect in patients for the reduction of plaque size. Intralesional verapamil injection for Peyronie's disease could reduce pain, decrease penile curvature, and improve sexual function.

Evaluation Biological Activities of Diphylleia grayi Extract as a Natural Cosmetic Ingredient

Purpose: In this study, we investigated the biological activities of Diphylleia grayi extract as a natural cosmetic ingredient. Methods: The antioxidant activities of Diphylleia grayi extract was measured through 2,2-diphenyl-1-(2,4,6-trinitrophenyl)- hydrazinyl (DPPH), 2,2’-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) scavenging assay and Fluorescence recovery after photobleaching (FRAP) assay. We further investigated whether Diphylleia grayi extract could reduce the intracellular reactive oxygen species (ROS) using dichlorofluorescin diacetate (DCF-DA) fluorescence. To investigate the whitening effect, we conducted tyrosinase inhibition assay and L-DOPA oxidation inhibition assay. Furthermore, we performed collagenase inhibition assay to evaluate the anti-wrinkle activity of Diphylleia grayi extract. Results: Diphylleia grayi extract showed strong antioxidant activities on DPPH, ABTS radical scavenging assay. Furthermore, Diphylleia grayi extract decreased the intracellular ROS on skin normal cells. Diphylleia grayi extract has also inhibition activities of tyrosinase, L-DOPA oxidation and collagenase. Conclusion: Therefore, Diphylleia grayi extract was identified as a promising natural cosmetic ingredient which has antioxidant, whitening and anti-wrinkle properties.

Skin Protective Activity of LactoSporin-the Extracellular Metabolite from Bacillus Coagulans MTCC 5856

Background: Probiotics and their products are increasingly used in skincare in recent years. Postbiotics are defined as any substance derived through the metabolic activity of a probiotic microorganism, which exerts a direct or indirect beneficial effect on the host. The extracellular metabolites of probiotic bacteria have antimicrobial activities, protect against acne, and improve skin condition. We studied skin protective activities of the extracellular metabolite (LactoSporin) of a spore-forming probiotic Bacillus coagulans MTCC 5856 in vitro. Methods: LactoSporin was evaluated for antioxidant activity by free radical scavenging activity and reactive oxygen quenching activity in human dermal fibroblast cells. Protection of fibroblasts from UV-induced apoptosis and cell death was studied by flow cytometry and neutral red uptake assays. Enzyme inhibition assays were carried out for collagenase, Elastase, and Hyaluronidase. Gene expression studies were carried out using polymerase chain reaction. Results: LactoSporin showed antioxidant activity and was found to protect skin cells from UV-induced apoptosis and cell death. LactoSporin inhibited collagenase, elastase, and hyaluronidase activity and upregulated the expression of hyaluronan synthase, transforming growth factor and epidermal growth factor, which are associated with extracellular matrix integrity. Conclusions: These results suggest LactoSporin is a skin protective postbiotic with wide application in cosmetic formulations.

Changes in DNA methylation accompany changes in gene expression during chondrocyte hypertrophic differentiation in vitro.

During osteoarthritis (OA), articular chondrocytes undergo phenotypic changes that resemble developmental patterns characteristic of growth plate chondrocytes. These phenotypic alterations lead to a hypertrophy-like phenotype characterized by altered production of extracellular matrix constituents and increased collagenase activity, which, in turn, results in cartilage destruction in OA disease. Recent studies have shown that the phenotypic instability and dysregulated gene expression in OA are associated with changes in DNA methylation patterns. Subsequent efforts have aimed to identify changes in DNA methylation with functional impact in OA disease, to potentially uncover therapeutic targets. Here, we paired an in vitro 3D/pellet culture system that mimics chondrocyte hypertrophy with RNA sequencing (RNA-Seq) and enhanced reduced representation of bisulfite sequencing (ERRBS) to identify transcriptomic and epigenomic changes in murine primary articular chondrocytes undergoing hypertrophy-like differentiation. We identified hypertrophy-associated changes in DNA methylation patterns in vitro. Integration of RNA-Seq and ERRBS datasets identified associations between changes in methylation and gene expression. Our integrative analyses showed that hypertrophic differentiation of articular chondrocytes is accompanied by transcriptomic and epigenomic changes in vitro. We believe that our integrative approaches have the potential to uncover new targets for therapeutic intervention.

Comparison of antiaging, anti-melanogenesis effects, and active components of Raspberry (Rubus occidentalis L.) extracts according to maturity.

In this study, we carried out a comparative evaluation of antiaging and anti-melanogenesis activities of raspberry extracts (Rubus occidentalis L.) according to their stage of ripening (uRo: unripe raspberry, Ro: ripe raspberry), and analyzed the active component (ellagic acid) present in these extracts. Our results showed higher inhibitory effects of the uRo extract in terms of elastase and collagenase activities than Ro extract. In the CCD-986sk cells, uRo extract significantly inhibited MMP-1 activity by 18% and increased the rate of type 1 pro-collagen synthesis by 25%. Besides, treatment with uRo extract significantly inhibited α-melanocyte-stimulating hormone-induced melanin synthesis and tyrosinase activity in B16F10 mouse melanoma cells. Overall, uRo was a more potent mediator of antiaging and anti-melanogenesis effects than Ro extract. Further analysis showed that the functional effects of uRo could be attributed to its 18.5 times higher ellagic acid content than that in Ro extract. PRACTICAL APPLICATIONS: This study reported the differential effect of the raspberry extracts depending on their stage of ripening. To the best of our knowledge, this was the first study to report the antiaging, anti-wrinkle, and anti-pigmentation effects of the uRo extracts. We showed that the extracts from the uRo have an overall better antiaging and skin-whitening effect than ripe ones. The effects were attributed to high ellagic acid content in uRo. We believed that our study makes a significant contribution to the literature because the outcome of the study has both, cosmetic as well as therapeutic implications.

Collagenase and Melanogenesis Inhibitory Effects of Perilla Frutescens Pomace Extract and Its Efficacy in Topical Cosmetic Formulations

In previous studies, Perilla frutescens pomace was shown to contain bioactive phenolic compounds and good anti-oxidative activity. However, reports about collagenase activity and melanogenesis inhibitory effects of P. frutescens pomace are limited. This study aimed to evaluate the bioactivity of P. frutescens pomace extract and incorporate the extract into a cosmetic formulation for evaluating its effects on collagenase and melanogenesis inhibition on human skin. The P. frutescens seeds after an oil pressing process were extracted with ethanol (70% v/v) in order to examine the remaining phytochemical compounds, the bioactivity in pomace perilla, and its efficacy as a skincare product. In this study, total phenolic and total flavonoid contents of P. frutescens seed extract (PFSE) were determined using spectrophotometry. The free radical scavenging activity was determined with 2, 2′-diphenyl-1-picrylhydrazyl (DPPH) radicals and 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) ABTS radicals. Additionally, the effects on collagenase activity, melanin content, and alpha-Melanocyte stimulating hormone (α-MSH) on the viability of cultured B16F10 melanoma cells were investigated. Skin irritation and efficacy of PFSE cream for skin elasticity and skin color were also clinically evaluated. The total phenolic content with gallic acid equivalents (GAE) value and total flavonoids content with catechin equivalents (CE) value were, respectively, 92.79 ± 1.19 and 56.02 ± 2.83 mg/g. Furthermore, PFSE significantly inhibited the collagenase activity (p &lt; 0.001) at the concentration of 400 µg/mL (82 ± 3.23%). These results clearly demonstrated the anti-melanogenic effects on B16F10 cells without causing any cytotoxicity or death. Although there was a slight improvement in skin elasticity in the 4th week compared to the previous week, the 4th week melanin content of the skin significantly decreased from the beginning (p &lt; 0.05) without any irritations. In conclusion, PFSE could be cosmetically considered as a key ingredient that effectively lessens the effects of skin aging and skin hyperpigmentation disorders.

Wound healing of the pelvic floor concerning pelvic organ prolapse – What do we know?

O prolapso de órgão pélvico (POP) é resultado do enfraquecimento dos tecidos de sustentação e de suspensão dessa região, incluindo os músculos levantadores do ânus, a fáscia endopélvica, os ligamentos e a parede vaginal. Sabe-se que os fatores de risco mais importantes para o POP, como gravidez, parto vaginal e aumento da pressão intra-abdominal, causam lesão nos tecidos do assoalho pélvico. Assim, é possível que um processo de cicatrização imperfeito desses tecidos possa afetar sua homeostase e contribuir para o desenvolvimento do POP. Dessa forma, o objetivo deste artigo é revisar a fisiologia da cicatrização de tecidos do assoalho pélvico e discutir os fatores que interferem nesse processo. Para isso, foi feita uma revisão da literatura na base de dados do MEDLINE até 2017. Percebeu-se que, assim como na pele, a cicatrização das lesões do assoalho pélvico ocorre em quatro fases (hemostasia, inflamação, proliferação e remodelação), porém a duração de cada fase é maior nas diferentes estruturas do assoalho pélvico. Além disso, a pressão constante no assoalho pélvico está associada ao aumento da atividade da colagenase, afetando negativamente a cicatrização, enquanto o estrogênio melhora as propriedades mecânicas do tecido estirado e parece ser benéfico para a cicatrização das lesões na parede vaginal. De acordo com a literatura, os músculos, nervos, ligamentos ou parede vaginal danificados não recuperam totalmente suas características prévias. Desse modo, conclui-se que uma cicatrização imperfeita no assoalho pélvico resulta em tecidos com composição e propriedades mecânicas alteradas, o que pode levar à incidência ou progressão do POP.

Influence of TEGDMA monomer on MMP-2, MMP-8, and MMP-9 production and collagenase activity in pulp cells

ObjectivesResin-based composites may leach monomers such as triethylene-glycol dimethacrylate (TEGDMA), which could contribute to intrapulpal inflammation. The aim of this investigation was to examine whether various concentrations of TEGDMA are able to influence dentally relevant Matrix metalloproteinase (MMP)-2, MMP-8, and MMP-9 production, total collagenase/gelatinase activity in pulp cells, and suggest possible signaling mechanisms.Materials and methodsPulp cells were cultured, followed by a 1-day exposure to sublethal TEGDMA concentrations (0.1, 0.2, and 0.75 mM). Total MMP activity was measured by an EnzCheck total collagenase/gelatinase assay, while the production of specific MMPs and the relative changes of phosphorylated, i.e., activated signaling protein levels of extracellular signal-regulated kinase (ERK)1/2, p38, c-Jun N-terminal kinase (JNK) were identified by western blot. Immunocytochemistry image data was also plotted and analyzed to see whether TEGDMA could possibly alter MMP production.ResultsAn increase in activated MMP-2, MMP-8, and MMP-9 production as well as total collagenase activity was seen after a 24-h exposure to the abovementioned TEGDMA concentrations. Increase was most substantial at 0.1 (P = 0.002) and 0.2 mM (P = 0.0381). Concurrent p-ERK, p-p38, and p-JNK elevations were also detected.ConclusionsResults suggest that monomers such as TEGDMA, leached from resin-based restorative materials, activate and induce the production of dentally relevant MMPs in pulp cells. Activation of ERK1/2, p38, or JNK and MMP increase may play a role in and/or can be part of a broader stress response.Clinical relevanceInduction of MMP production and activity may further be components in the mechanisms of intrapulpal monomer toxicity.

Study of detection method for collagenase enzyme reaction using two dimensional electrochemical sensor

AbstractThis paper addresses the assay of collagenase activity using Light Addressable Amperometric Sensor (LAAS). The redox current of LAAS was reduced by deposited collagen layers and recovered by collagenase solution. The collagen layers on the LAAS is considered to be broken down by collagenase. We expect an application of LAAS to activity evaluation.

In vivo bioactivity assessment on Epilobium species: A particular focus on Epilobium angustifolium and its components on enzymes connected with the healing process

Ethnopharmacological relevanceEpilobium species are generally known as “Yakı Otu” in Turkey, which mens “plaster herb” in English. Young shoots of Epilobium angustifolium L., Epilobium stevenii Boiss., and Epilobium hirsutum L. are consumed as salad or meal. These species have been used as a poultice for the treatment of mouth wounds in traditional medicine. An ointment prepared from leaves is used for skin disorders in children. Aim of the studyWe aimed to evaluate the ethnopharmacological use of Epilobium angustifolium, E. stevenii, and E. hirsutum by using in vivo and in vitro experimental models, and to identify the active wound-healer compound(s) and to explain the probable mechanism of the wound-healing activity. Materials and methodsEvaluation of wound healing effects of plant extracts was performed in rats and mice by linear incision and circular excision wound models. Determination of total phenolic constituents and antioxidant capacities, which are known to promote the wound healing process, were carried out through Folin-Ciocalteau method and 2,2 Diphenyl 1 picrylhydrazyl (DPPH) scavenging assay as well as determination of total antioxidant status (TAS) and total oxidant status (TOS) on the treated tissues. The active ethyl acetate (EtOAc) sub-extract of E. angustifolium was fractionated by different chromatographic separation techniques. The structures of isolated compounds were elucidated via detailed analyzes (NMR and LC/MS). In addition, in vitro collagenase, hyaluronidase, and elastase enzymes inhibitory activity tests were performed on the isolated compounds to discover the activation pathways of the samples. ResultsAmong the methanol (MeOH) extracts, E. angustifolium had the highest wound healing activity. Among the sub-extracts, EtOAc showed the highest wound healing activity. Thus, EtOAc sub-extract was subjected to chromatography to isolate the active compounds. Five known flavonoids namely hyperoside (quercetin-3-O-β-D-galactoside) (1), kaempferol (2), kaempferol-3-O-α-L-rhamno pyranoside (3), quercetin-3-O-α-L-rhamno pyranoside (4), and quercetin-3-O-α-L-arabino pyranoside (5) were isolated from the EtOAc sub-extract of E. angustifolium. In vitro tests showed that hyperoside could be the compound responsible for the wound-healing activity by its significant anti-hyaluronidase, anti-collagenase, and antioxidant activities. ConclusionThe EtOAc sub-extract of the aerial part of Epilobium angustifolium displayed remarkable wound-healing activity with anti-hyaluronidase, anti-collagenase, and antioxidant activities. Hyperoside was detected as the primary active compound of the aerial parts. According to the results, we suggest that EtOAc sub-extract of E. angustifolium and hyperoside may be a potent nominee to be used for the improvement of a wound-healing agent.

Thymocid®, a Standardized Black Cumin (Nigella sativa) Seed Extract, Modulates Collagen Cross-Linking, Collagenase and Elastase Activities, and Melanogenesis in Murine B16F10 Melanoma Cells.

Black cumin (Nigella sativa) seed extract has been shown to improve dermatological conditions, yet its beneficial effects for skin are not fully elucidated. Herein, Thymocid®, a chemically standardized black cumin seed extract, was investigated for its cosmeceutical potential including anti-aging properties associated with modulation of glycation, collagen cross-linking, and collagenase and elastase activities, as well as antimelanogenic effect in murine melanoma B16F10 cells. Thymocid® (50, 100, and 300 µg/mL) inhibited the formation of advanced glycation end-products (by 16.7–70.7%), collagen cross-linking (by 45.1–93.3%), collagenase activity (by 10.4–92.4%), and elastases activities (type I and III by 25.3–75.4% and 36.0–91.1%, respectively). In addition, Thymocid® (2.5–20 µg/mL) decreased melanin content in B16F10 cells by 42.5–61.6% and reduced cellular tyrosinase activity by 20.9% (at 20 µg/mL). Furthermore, Thymocid® (20 µg/mL for 72 h) markedly suppressed the mRNA expression levels of melanogenesis-related genes including microphthalmia-associated transcription factor (MITF), tyrosinase-related protein 1 (TYRP1), and TYRP2 to 78.9%, 0.3%, and 0.2%, respectively. Thymocid® (10 µg/mL) also suppressed the protein expression levels of MITF (by 15.2%) and TYRP1 (by 97.7%). Findings from this study support the anti-aging and antimelanogenic potential of Thymocid® as a bioactive cosmeceutical ingredient for skin care products.

Intralesion Triamcinolon Injection Therapy on Esophageal Stricture as Side Effect of Long Term Use Biphosphonate

Systemic Lupus Erythematous (SLE) is a chronic inflammatory disease that affect almost any organ system. Patient with SLE is at risk of osteoporosis. Biphosphonate is one of osteoporosis treatments. However, esophageal stricture (ES) caused by continuous esophageal inflammation as a result of long-term use Biphosphonate can occur. Intralesion Triamcinolone Injection (ITI) becomes one of ES therapy by inhibiting inflammatory response to injury and decrease subsequent collagen formation. ITI inhibits transcription of matrix protein genes, including fibronectin and procollagen. It also reduces the synthesis of α2-macroglobulin, an inhibitor of collagenase activity. A 43 year old woman was diagnosed with SLE. She was treated with Risedronic Acid (bisphosphonate class) 35 mg once a week. After 8 months, she started complaining difficulty to swallow. Symptom was getting worse 6 months later. One-third distal ES with inflammation process was found with endoscopy. ITI 150 mg was given in inflammation area. She was scheduled to follow up after 7 days for re-endoscopy. Endoscopy showed that inflammation process had been improved. Six months later, she had again complained difficulty to swallow and we found that she still consumed Risedonic Acid. ITI 150 mg was given because inflammation and ES was found with endoscopy. After 7 days, endoscopy showed that there was no ES and inflammation process had been improved. She was advised to stop Risedronic Acid consumption.

Evaluation of antioxidant and anti-collagenase activity of Rosa damascena L. flower petal and receptacle extract

Background: Rosa damascena L. is the most notable species of the Rosaceae family in the world, and has been used in food, cosmetics, and the pharmaceutical industry. Bioactive compounds in this flower are known to have several activities, such as antioxidant, antimicrobial, and anti-inflammatory. In this study, the antioxidant and collagenase inhibitory activities of R. damascena L. petal and receptacle extracts were evaluated. Methods: Ethanolic extraction of R. damascena L. petals (EERP) and R. damascena L. receptacles (EERR) were obtained, and bioactive compounds (flavonoids, phenolics, alkaloids, steroids, tannins, terpenoids, and triterpenoids) were classified by phytochemical screening. Antioxidant activities were analyzed by Ferric Reducing Antioxidant Power (FRAP) assay, while anti-collagenase analysis was examined through the inhibition of collagenase. Results: Phytochemical test revealed the presence of flavonoids, phenolics, alkaloids, steroids, triterpenoids, triterpenes, and tannin. EERP showed higher FRAP activity (164.23 ± 1.34 μM Fe(II)) than EERR (12.85 ± 6.19 μM Fe(II)). EERP also had higher inhibitory activity of collagenase (IC50 = 115.48±1.78 µM/mL) compared to EERR (IC50 = 141.96±6.13 µM/mL). Conclusions: R. damascena L. petal and receptacle ethanol extracts contain several components, such as phenolics, flavonoids, alkaloids, tannins, terpenes, triterpenoids, and steroids. These extracts exhibit antioxidant activity and collagenase inhibition. R. damascena L. petal extract showed higher antioxidant activity through FRAP assay and inhibitory activity of collagenase than R. damascena L. receptacle extract.