Collagenase Inhibitory Activity Research Articles

Anti-aging potential and anti-tyrosinase activity of three Hypericum species with focus on phytochemical composition by LC–MS/MS

Hypericum species are one of the most commonly used medicinal plants all over the world and traditional uses of the Hypericum species are mainly characterized by external applications for the treatment of dermatological disorders. The purpose of this study was to investigate the chemical compounds with anti-aging and anti-tyrosinase activities of Hypericum perforatum L., H. calycinum L., and H. confertum Choisy from Turkey and to bring innovation for further studies on developing formulations with Hypericum species for topical applications. In this study, chemical compositions of methanol extracts prepared from flowering aerial parts of the following species collected from Uludağ, H. calycinum, H. confertum, and H. perforatum were investigated by LC–MS/MS. Nine metabolites, p-coumaric acid, gallic acid, quinic acid, chlorogenic acid, malic acid, protocatechuic acid, rutin, quercitrin, and isoquercitrin were identified in all analyzed species. To investigate the anti-aging and skin-whitening properties, enzyme inhibition activity of the three species were tested against, collagenase, elastase, hyaluronidase, and tyrosinase enzymes. H. calycinum appeared the most active in all enzyme inhibition activity assays which is mainly due to its rich phenolic content. The methanol extract of H. calycinum showed the highest collagenase, elastase and hyaluronidase inhibitory activities with IC50 values of 51.24 ± 1.33 μg/mL, 55.77 ± 1.32 μg/mL and 22.17 ± 0.02 μg/mL, respectively. Additionally, H. calycinum emerged as the most potential tyrosinase inhibitor among these species with 54.30 ± 0.49% at 200 μg/mL concentration. The most abundant compounds determined in H. calycinum methanol extract are, chlorogenic acid (2595.15 ± 0.179 μg/g), quercitrin (1805.9 ± 0.98 μg/g), quinic acid (885.50 ± 0.073 μg/g) and isoquercitrin (346.16 ± 0.046 μg/g), respectively. Our study demonstrates that all three extracts, especially H. calycinum extract are potential agents to use in cosmeceuticals for anti-aging and skin-whitening purposes.

Comparison of Antioxidant and Anti-collagenase Activity of Genistein and Epicatechin

Background : Skin aging is a complex natural process caused by both intrinsic or genetically programmed aging and extrinsic aging caused by environmental factors, such as free radical. The use of antioxidant and anti-collagenase to prevent the aging proses has been known. Natural compounds from plants are one of the sources of antioxidant and anti-collagenase that has ability to prevent aging. Genistein and Epicatechin are the major phenolic compound found in G. max. Objectives : This research was to evaluate the antiaging potential of genistein and epicatechin through antioxidant activity assay (ABTS-reducing activity assay) and collagenase inhibitory activity assay. Methods : Antioxidant analysis of Genistein and Epicatechin was performed by 2,2’- azinobis-3-ethylbenzo-thiazoline-6-sulfonic acid (ABTS) reducing activity Assay. Antiaging assay was conducted through inhibitory of collagenase, one of important enzyme in aging process. Results : ABTS- reducing activity assay showed that both compounds had great ABTS-reducing activity in which epicatechin had better activity than genistein. Epicatechin had low value of IC50 ABTS-reducing activity around 14.39 µg/ml better than genistein with IC50 about 43.17 µg/ml. In terms of collagenase inhibitory activity assay, epicatechin had lower value of IC50 (9.08+-3.46 ug/ml), better than genistein (98.74+-4.25 ug/ml). Conclusion : Epicatechin had higher antioxidant and anti- collagenase activity compared to Genistein.

Proanthocyanidin Dimers and Trimers from Vitis vinifera Provide Diverse Structural Motifs for the Evaluation of Dentin Biomodification.

Aimed at exploring the dentin biomodification potential of proanthocyanidins (PACs) for the development of dental biomaterials, this study reports the phytochemical and dental evaluation of nine B-type PACs from grape seed extract (GSE). Out of seven isolated dimers (1-7), four new compounds (2, 3, 5, and 6) involved relatively rare ent-catechin or ent-epicatechin monomeric flavan-3-ol units. Low-temperature NMR analyses conducted along with phloroglucinolysis and electronic circular dichroism enabled unequivocal structural characterization and stereochemical assignment. Additionally, one known (8) and one new (9) B-type trimer were characterized. Differential 13C NMR chemical shifts (Δδ) were used to determine the absolute configuration of 9, relative to the dimers 1 and 2 as the possible constituent subunits. Compared to the dimers, the trimers showed superior dentin biomodification properties. The dimers, 1-7, exhibited pronounced differences in their collagenase inhibitory activity, while enhancing dentin stiffness comparably. This suggests that PAC structural features such as the degree of polymerization, relative and absolute configuration have a differential influence on enhancement of dentin biomechanical and biostability. As mechanical enhancement to dentin and resistance to proteolytic biodegradation are both essential properties functional and stable dentin substrate, the structurally closely related PACs suggest a new metric, the dentin biomodification potential (DBMP) that may rationalize both properties.

Static hydrothermal processing and fractionation for production of a collagen peptide with anti-oxidative and anti-aging properties

Abstract Collagen, in a form suitable for human consumption, can be obtained from the large amounts of fish skin via protein hydrolysis to produce low-molecular-weight substances with enhanced bioavailability and function. To optimize this method, we investigated the synergistic effect of elevated temperature (150–250 °C) and pressure (350–3900 kPa) of a hydrothermal process on the hydrolytic ability and characteristics of collagen hydrolysates. Elevated temperature and pressure of the hydrothermal process increased the levels of free amino groups and lower-molecular-weight collagen hydrolysates, particularly at 210 °C and 2100 kPa. The resulting hydrolysates were fractionated by ultrafiltration membranes of different molecular weight cutoff and evaluated for their antioxidant (ABTS radical scavenging activity and reducing power) and anti-aging (tyrosinase and collagenase inhibition) activities. The

Antioxidative, Anti-Inflammatory, and Anti-Aging Properties of Mycosporine-Like Amino Acids: Molecular and Cellular Mechanisms in the Protection of Skin-Aging.

Prolonged exposure to ultraviolet (UV) radiation causes photoaging of the skin and induces a number of disorders, including sunburn, fine and coarse wrinkles, and skin cancer risk. Therefore, the application of sunscreen has gained much attention to reduce the harmful effects of UV irradiation on our skin. Recently, there has been a growing demand for the replacement of chemical sunscreens with natural UV-absorbing compounds. Mycosporine-like amino acids (MAAs), promising alternative natural UV-absorbing compounds, are a group of widely distributed, low molecular-weight, water-soluble molecules that can absorb UV radiation and disperse the absorbed energy as heat, without generating reactive oxygen species (ROS). More than 30 MAAs have been characterized, from a variety of organisms. In addition to their UV-absorbing properties, there is substantial evidence that MAAs have the potential to protect against skin aging, including antioxidative activity, anti-inflammatory activity, inhibition of protein-glycation, and inhibition of collagenase activity. This review will provide an overview of MAAs, as potential anti-aging ingredients, beginning with their structure, before moving on to discuss the most recent experimental observations, including the molecular and cellular mechanisms through which MAAs might protect the skin. In particular, we focus on the potential anti-aging activity of mycosporine-2-glycine (M2G).

Anti-Oxidative and Anti-Aging Activities of Porcine By-Product Collagen Hydrolysates Produced by Commercial Proteases: Effect of Hydrolysis and Ultrafiltration.

To investigate methods for improving the processing of porcine waste, porcine skin was hydrolyzed using different commercially available proteases (Alcalase, Flavorzyme, Neutrase, Bromeline, Protamex, and Papain) under several optimal conditions. Following enzymatic hydrolysis, the collagen hydrolysates (CHs) were fractionated by molecular weight (3 kDa) via membrane ultrafiltration. The CHs were analyzed for physical properties (pH, protein recovery, free amino group content, molecular weight distribution, and amino composition) as well as for functional properties (antioxidant activities and anti-aging activities). Among the CHs, CHs hydrolyzed by Alcalase (CH-Alcalase) exhibited the highest degree of hydrolysis compared to other CHs. Both “CH-Alcalase” and “CH-Alcalase < 3 kDa” fractions showed a considerably high antioxidant activity and collagenase inhibition activity. Therefore, resulting bioactives have potential for development as antioxidants and anti-aging ingredients in the food, cosmetics, and pharmaceuticals, from animal by-products.

Collagenase inhibition by water-pepper (Polygonum hydropiper L.) sprout extract

Introduction: Collagenase plays an important role in the degradation of dermal matrix proteins leading to wrinkle formation. The objectives of this study were to evaluate the inhibitory effect of water-pepper (Polygonum hydropiper L.) sprout extract on the activity of collagenase and to identify the inhibitory compounds.Methods: Collagenase inhibitory activity was measured by spectrophotometric assay. Activity-guided fractionation was performed using liquid-liquid extraction of water and n-butanol and Diaion HP-20 column chromatography, followed by high-performance liquid chromatography (HPLC) fraction collection.Results: A methanolic extract of water-pepper sprout inhibited collagenase activity in a concentration-dependent manner with an IC50 value of 156.7 μg/mL. Collagenase inhibitory activity (IC50 = 23.5 μg/mL) was found in 50% methanol eluate from the HP-20 column chromatography of the n-butanol soluble fraction. The active compound (IC50 = 1.9 μg/mL) in the eluate was isolated by HPLC and identified as quercetin-3-O-galactoside (hyperoside) from comparing retention time, UV-Vis absorption, and mass spectra with those of the standard. Lineweaver-Burk plots revealed that hyperoside was an uncompetitive inhibitor against collagenase. Hyperoside was also the most abundant flavonoid present in the methanolic extract.Conclusion: These results suggest that water-pepper sprouts could be beneficial as a natural source of collagenase inhibitor which might be used for the treatment of skin aging.

Free Radical Scavenging and Anticollagenase Properties of Sargachromanol I from Myagropsis myagroides

The present study investigated the free radical scavenging activity and antiwrinkle properties of methanol extract, hexane, chloroform, ethyl acetate, and butanol partition fractions of Myagropsis myagroides. The methanol extract showed higher free radical scavenging (65.46 % at 0.5 mg/ml) and collagenase inhibitory activity (29.64 % at 1000 μg/ml) than the water extract (16.68 % at 0.5 mg/ml and < 5 % at 1000 μg/ml). The hexane fraction exhibited strong free radical scavenging activity (96.36 % at 0.5 mg/ml) and collagenase inhibitory activity (IC50 of 712.21 μg/ml). The hexane fraction was subjected to silica gel column chromatography, Sephadex LH-20 column chromatography, octadecyl silica Sepak cartridge, and preparative high performance liquid chromatography. Sargachromaol I was isolated, which showed strong free radical scavenging activity (95.82 % at 0.1 mg/ml) and collagenase inhibitory activity (IC50 value of 54.01 μg/ml). The potential value in preventing skin aging could be confirmed through effects of extract from Myagropsis myagroides on free radical scavenging activity and collagenase inhibitory properties.

신품종 그린볼(Green ball; Malus pumila Mill.) 사과의 기능성

The peel of newly bred Green ball apple was extracted using water and ethanol. Water and 70% ethanol extract showed high phenolic contents of 5.30 and 8.31 mg/g, respectively. The water and ethanol extracts of Green ball apple peel showed 88.26% and 100% DPPH radical scavenging activity, and 99.27% and 99.09% ABTS radical scavenging activity at phenolic concentration of 100 μg/mL, respectively. The water and ethanol extract of Green ball apple peel showed antioxidant protection factor of 1.43 PF and 1.21 PF, respectively. The water and ethanol extracts showed 60.29% and 75.92% anti-oxidative effect on thiobarbituric acid reactive substances (TBARS) at phenolic concentration of 100 μg/mL. Hence, Green ball apple peel extract can be considered a potential anti-oxidant with anti-aging properties. The water and ethanol extracts 6.45% and 20.78% hyaluronidase inhibition, respectively, at phenolic concentration of 200 μg/mL. The water extract showed 4.55% and 40.06% elastase inhibition and collagenase inhibition, while the ethanol extract showed 63.38% and 80.07% inhibition, respectively. Green ball apple peel was found to exhibit anti-oxidation activity, as well as hyaluronidase, elastase and collagenase inhibitory activities. Therefore, Green ball apple peel can be considered a potential source for new functional foods.

Evaluation of collagen type I scaffolds including gelatin-collagen microparticles and Aloe vera in a model of full-thickness skin wound.

Research on collagen type I scaffolds with Aloe vera is sparse. The aim of this work was to develop collagen type I scaffolds with gelatin-collagen microparticles and loaded with a dispersion of A. vera, to assess their performance as grafting material for healing of skin wounds. Scaffolds were evaluated in a Cavia porcellus model with full-thickness skin wound and compared with wounds healed by secondary intention (controls). Animals grafted with scaffolds without A. vera and their control wounds were also included in the study. Evaluation of enzymatic degradation and percentage of the scaffolds' free amino groups-as an indirect assessment of their cross-linking-were also carried out because A. vera contains compounds which affect their stability. We found that dispersions of lyophilized A. vera extract loaded on scaffolds do not have cytotoxic potential, and they decrease collagenase degradation of scaffolds in the range of 0.1 to 0.3% w/v in a dose-dependent manner. Only the A. vera dispersion with the highest concentration (0.3% w/v) decreased the percentage of free amino groups, which are the ones involved in the cross-link of collagen fibers. This finding suggests that cross-linking is not the mechanism by which the tested dispersions stabilize the scaffolds. Preclinical, histochemical, and histomorphometric analyses of repaired wound tissue indicate that loading collagen type I scaffolds, including microparticles of gelatin-collagen, with A. vera in the concentrations tested does not improve wound healing. Low biodegradability of the tested scaffolds caused by the inhibition of collagenase activity might account for these results.

Effect of angiotensin II receptor blocker on TGF-β1, MMP-1, and collagen type I and type III concentration in New Zealand rabbit urethral stricture model.

IntroductionUrethral stricture is a disease with a high recurrence rate. Angiotensin II via AT1 receptor increases collagen formation through its effects on TGF-β1 and inhibition of collagenase activity. In this study, we evaluated the antifibrotic effect of angiotensin II receptor blocker on urethral stricture formation by creating a urethral stricture model in a male rabbit.Material and methodsThirty three male adult rabbits were separated into 3 groups (control, treatment, and sham). Group I consisted of 15 rabbits with urethral stricture that did not undergo any treatment, group II consisted of 15 rabbits with urethral stricture that were treated with a daily dose of 15 mg/kg losartan, given orally. Group III consisted of 3 rabbits with normal urethra and without any treatment. After 1, 2, and 4 weeks, the urethral tissues were collected, processed, and examined for TGF-β1, MMP-1, collagen type I, and collagen type III using enzyme-linked immunosorbent assay. Data were analyzed using 2-way analysis of variance using SPSS version 20.0.ResultsUrethral TGF-β1 concentration in the treatment group was significantly lower during the 2nd and 4th week of observation (p<0.0001), MMP-1 was significantly higher in the 1st, 2nd, and 4th week of observation (p<0.0001), collagen type I was significantly lower during the 2nd (p=0.001) and 4th week (p<0.0001), and collagen type III concentration was significantly lower in the 2nd and 4th week of observation (p<0.0001).ConclusionAngiotensin II receptor blocker could limit the progression of urethral stricture. The mechanism may be related to the AT1 blockage that leads to a decrease in TGF-β1 concentration, eventually resulting in lower collagen concentration due to increased MMP-1 activity.

Investigation of Hyaluronidase, Collagenase and Elastase Inhibitory Potentials and Comparative Evaluation of the Antimicrobial, Antioxidant and Homeostatic Activities of Two Natural Polysaccharides

The aim of this study was to investigate the hyaluronidase, collagenase and elastase inhibitory effects, which play important role for wound healing, together with the antibacterial, antioxidant and homeostatic activities of tragacanth gum (TG) and locust bean gum (LBG). The antimicrobial activities were tested against four bacteria and the antioxidant activities were estimated by the 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydrogen peroxide (H2O2) radical scavenging and β-carotene bleaching assays. Homeostatic effect was evaluated with the Prothrombin Time (PT) and Activated Partial Thromboplastin Time (aPTT) test parameters. The wound healing potentials were determined with the inhibition of hyaluronidase, collagenase and elastase enzymes. The TG showed antibacterial activity against Pseudomonas aeruginosa ATCC27853 and Escherichia coli ATCC25922. The results showed that TG and LBG possessed antioxidant properties including DPPH scavenging (21.0% and 17.6%, respectively) and H2O2 radical scavenging (59.4% and 79.0%, respectively) activities. The polysaccharides displayed significantly reducing PT and aPTT results. Between the two tested polysaccharides LBG showed significant hyaluronidase and collagenase inhibition activity at 10 mg/mL concentration. These findings show that these natural polysaccharides can be used to support of wound healing.

Identification of a Collagenase-Inhibiting Flavonoid from Alchemilla vulgaris Using NMR-Based Metabolomics.

This paper describes the use of 1H NMR profiling and chemometrics in order to facilitate the selection of medicinal plants as potential sources of collagenase inhibitors. A total of 49plants with reported ethnobotanical uses, such as the healing of wounds and burns, treatment of skin-related diseases, rheumatism, arthritis, and bone diseases, were initially chosen as potential candidates. The in vitro collagenase inhibitory activity of hydroalcoholic extracts of these plants was tested. Moreover, their phytochemical profiles were analyzed by 1H NMR and combined with the inhibitory activity data by an orthogonal partial least squares model. The results showed a correlation between the bioactivity and the concentration of phenolics, including flavonoids, phenylpropanoids, and tannins, in the extracts. Considering the eventual false-positive effect on the bioactivity given by tannins, a tannin removal procedure was performed on the most active extracts. After this procedure, Alchemilla vulgaris was the most persistently active, proving to owe its activity to compounds other than tannins. Thus, this plant was selected as the most promising and further investigated through bioassay-guided fractionation, which resulted in the isolation of a flavonoid, quercetin-3-O-β-glucuronide, as confirmed by NMR and HRMS spectra. This compound showed not only a higher activity than other flavonoids with the same aglycone moiety, but was also higher than doxycycline (positive control), the only Federal Drug Administration-approved collagenase inhibitor. The approach employed in this study, namely the integration of metabolomics and bioactivity-guided fractionation, showed great potential as a tool for plant selection and identification of bioactive compounds in natural product research.

Antioxidant and Cosmeceutical Activities of Agarum cribrosum Phlorotannin Extracted by Ultrasound Treatment

An ultrasonic process for the effective extraction of phlorotannins from Agarum cribrosum with high total phlorotannin content (TPC) was optimized using response surface methodology (RSM). Optimal conditions for ultrasound extraction consisted of temperature: 30°C; time: 5.75 h; ratio of solvent to solid: 35 mL/g, where the TPC was 19.59 mg PGE/g. The crude extract was further partitioned using hexane, dichloromethane, ethyl acetate (EtOAc), and water, in that order. The EtOAc fraction with the highest TPC exhibited the strongest radical-scavenging activity against DPPH, ABTS, hydrogen peroxide, and reducing power, which was similar to that of ascorbic acid. Furthermore, the IC50 values of the EtOAc fraction against collagenase, elastase, and tyrosinase inhibitory activity were 29.63, 16.13, and 6.17 μg/mL, respectively, whereas epigallocatechin gallate (EGCG), EGCG, and kojic acid, the counterpart positive controls, were 68.90, 23.34, and 43.29 μg/mL, respectively. The EtOAc fraction was further fractionated using Sephadex LH-20 column chromatography, which yielded two major and four minor subfractions. The structure of the SF5 and SF6 subfractions were further determined through HPLC-DAD-QMS analyses and 1H and 13C NMR, in which only trifuhalol A was identified. The SF5 and SF6 subfractions exhibited significantly higher radical-scavenging activity against DPPH and reducing power than the EtOAc fraction, but no statistically significant differences were detected for ABTS, hydrogen peroxide, collagenase, elastase, and tyrosinase inhibitory activity. Based on the results, we propose that A. cribrosum phlorotannins could be a good source for natural antioxidants and cosmeceutical.

초임계 유체로 추출된 산삼 부정 배양근 오일의 효모균 처리에 의한 비사포닌계 지방산 함량 변화 및 Collagenase 및 Elastase 저해 활성 증대

Background: To obtain useful cosmetic resources, this study aimed to determine the non-saponin fatty acid and inhibitory activities of collagenase and elastase by treatment of Saccharomyces cerevisiae in supercritical fluid extracted oil of the adventitious root culture of wild mountain ginseng.BR Methods and Results: We performed supercritical fluid extraction at various conditions such as pressure, temperature, time, and use of co-solvents, unlike the n-hexane extraction for the adventitious roots culture of wild mountain ginseng. The non-saponin-fatty acid obtained from the oil of the adventitious roots culture was incresed by treatment with S. cerevisiae. The supercritical fluid extraction was conducted using gas chromatography. Non-saponin-fatty acid content, in the oil of adventitious roots culture of wild mountain ginseng treated with S. cerevisiae for 2 days were three times higher than that in the control. In addition, the oil of the adventitious roots culture treated with S. cerevisiae was investigated for the anti-wrinkle effect by using collagenase and elastase. The oil of adventitious roots culture treated with S. cerevisiae exhibited higher collagenase and elastase inhibitory activities than those in the control.BR Conclusions: Supercritical fluid extracted oil of the adventitious roots culture of wild mountain ginseng treated with S. cerevisiae was found to have decreased ratio of saturated fatty acids and incresed ratio and content of unsaturated fatty acids increased. Furthermore, it showed anti-wrinkle effects in vitro.

Antioxidants, anticollagenase and antielastase potentials of ethanolic extract of ripe sesoot (Garcinia picrorrhiza Miq.) fruit as antiaging

Introduction: Nitric oxide (NO) is a free radical that belongs to reactive nitrogen species (RNS). The excess amount of NO in body generates physical changing on skin as a consequence of alteration in connective tissue through formation of lipid peroxides, cell content, and enzymes. These free radicals induce damage to extracellular matrix (ECM) and are responsible in reducing skin elasticity. Antioxidants possess significant role in delaying aging process by scavenging free radicals and preventing collagenase and elastase enzymes activities. This study aimed to evaluate antioxidants, anticollagenase and antielastase potentials of ethanolic extract of ripe sesoot (Garcinia picrorrhiza Miq.) fruit (GpKar) as antiaging remedy. Methods: Antioxidant activity was performed by NO scavenging activity assay, while anti-aging activity was performed through inhibitory effects of collagenase and elastase activities. Results: In antioxidant activity, GpKar had lower NO scavenging activity (IC50=1530.34 μg/mL) compared to xanthone (IC50=85.40 μg/mL). In collagenase inhibitory activity, GpKar also had lower inhibition collagenase activity (IC50= 1169.31 μg/mL) compared to xanthone (IC50= 286.32 μg/mL). In elastase inhibitory activity, GpKar had lower inhibition elastase activity (IC50= 152.93 μg/mL) compared to xanthone (IC50= 21.26 μg/mL). Conclusion: In summary, GpKar and its compounds possess antioxidant, anticollagenase, and antielastase activities for antiaging, and might be beneficial in these subjects.

3,5,6,7,8,3',4'-Heptamethoxyflavone, a Citrus Flavonoid, Inhibits Collagenase Activity and Induces Type I Procollagen Synthesis in HDFn Cells.

Citrus fruits contain various types of flavonoids with powerful anti-aging and photoprotective effects on the skin, and have thus been attracting attention as potential, efficacious skincare agents. Here, we aimed to investigate the chemical composition of Citrus unshiu and its protective effects on photoaging. We isolated and identified a bioactive compound, 3,5,6,7,8,3′,4′-heptamethoxyflavone (HMF), from C. unshiu peels using ethanol extraction and hexane fractionation. HMF inhibited collagenase activity and increased type I procollagen content in UV-induced human dermal fibroblast neonatal (HDFn) cells. HMF also suppressed the expression of matrix metalloproteinases 1 (MMP-1) and induced the expression of type I procollagen protein in UV-induced HDFn cells. Additionally, HMF inhibited ultraviolet B (UVB)-induced phosphorylation of the mitogen-activated protein kinases (MAPK) cascade signaling components—ERK, JNK, and c-Jun—which are involved in the induction of MMP-1 expression. Furthermore, HMF affected the TGF-β/Smad signaling pathway, which is involved in the regulation of type I procollagen expression. In particular, HMF induced Smad3 protein expression and suppressed Smad7 protein expression in UV-induced HDFn cells in a dose-dependent manner. These findings suggest a role for Citrus unshiu in the preparation of skincare products in future.

Anti-aging potential of extracts from Sclerocarya birrea (A. Rich.) Hochst and its chemical profiling by UPLC-Q-TOF-MS

BackgroundDegradation of components of the extracellular matrix such as elastin and collagen by elastase and collagenase accelerates skin aging. Phytochemicals that inhibit the activity of these enzymes can be developed as anti-aging ingredients. In this study, an investigation of the anti-aging properties of Sclerocarya birrea (A. Rich.) Hochst (Marula) extracts was conducted in vitro with the aim of developing chemically characterized anti-aging ingredients.MethodsMarula stems, leaves and fruits were extracted using methanol:dichloromethane (DCM) (1:1). The stems were later extracted using acetone, ethanol, methanol:DCM (1:1) and sequentially using hexane, DCM, ethyl acetate and methanol. The stem ethanol extract was defatted and concentrated. Elastase and collagenase inhibition activities of these extracts and Marula oil were determined using spectrophotometric methods. The chemical profile of the ethanolic stem extract was developed using Ultra-performance-liquid chromatography quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) with MassLynx software. Pure standards were used to confirm the identity of major compounds and were screened for anti-elastase and anti-collagenase activity.ResultsMarula stems extracts were the most active as they exhibited anti-elastase activity comparable to that of elafin (> 88%) and anti-collagenase activity as potent as EDTA (> 76%). The leaf extract had moderate anti-elastase activity (54%) but was inactive agains collagenase. Marula fruits and oil exhibited limited activity in both assays. The ethanolic extract of Marula stems was the most suitable based on its acceptability to the cosmetic industry and its anti-collagenase activity (99%). Defatting and concentration improved its antiaging activity and lowered the colour intensity. Six compounds have been tentatively identified in the chemical profile of the ethanolic extract of Marula stems of which four; quinic acid, catechin, epigallocatechin gallate and epicatechin gallate have been confirmed using pure standards. Epigallocatechin gallate and epicatechin gallate were as potent (p < 0.05) as EDTA at 5 μg/ml in the anti-collagenase assay.ConclusionsThe ethanolic extract of Marula stems can be developed into an anti-aging ingredient as it exhibited very good in vitro anti-aging activity and its chemical profile has been developed. Epicatechin gallate and epigallocatechin gallate contribute to the anti-aging activity of Marula stem ethanol extract.

Reduced synovial inflammation and inhibition of matrix metalloproteinases explicates anti-osteoarthritis activity of polyherbal formulations.

OBJECTIVES:Current osteoarthritis (OA) research experiences an incline toward Ayurveda to attain a complete cure without notable adverse effects. Ayurveda uses natural products, which are known to perform the multi-faceted role, a much demanding approach for OA management. However, lack of scientific evidence is a major drawback hindering their wider use. The present work investigated the anti-arthritic potential of Ashwagandharishta, Balarishta, Dashmoolarishta, and Triphala-extract to establish molecular-evidence for their clinical use.MATERIALS AND METHODS:Rabbit synoviocytes were induced using interleukin-1 beta (IL-1 β) and lipopolysaccharide (LPS) separately and were further treated with study formulations to test anti-inflammatory and anti-oxidant potential, using nitric oxide (NO) and malondialdehyde (MDA) assays. Collagenase inhibition activity was estimated with N-(3-[2-Furyl] acryloyl)-Leu-Gly-Pro-Ala (FALGPA)-substrate and gelatinase spot assays. Data were analyzed with GraphPad Prism using one-way ANOVA followed by Bonferroni's multiple comparison.RESULTS:The study formulations were effective against synovitis, oxidative-stress, and inhibiting collagenase. They caused NO reduction in selected concentrations. DA showed the maximum NO decline of 0.02 ± 0 and 0.97 ± 0.62 μM/ml with IL-1 β and LPS induction at 5 and 20 μg/ml concentrations, respectively. Estimated by FALGPA assay, increasing collagenase inhibition was observed as the function of concentration. All formulations showed a significant MDA decline, in dose-dependent manner.CONCLUSION:We assessed the anti-OA efficacy of conventionally prescribed Ayurvedic drugs using relevant biochemical assays. The studied formulations revealed potential to restrain synovitis, cartilage degeneration and to reduce oxidative stress, and the signature OA features. With established molecular authenticity, Ayurvedic drugs can offer a safer and affordable therapeutic option for OA.

Polyphenols extracted from red grape pomace by a surfactant based method show enhanced collagenase and elastase inhibitory activity

AbstractBACKGROUNDThe aim of this study is to separate polyphenols from grape pomace using a surfactant‐based separation, colloidal gas aphrons (CGA) and to investigate their inhibitory activity against skin relevant enzymes, collagenase and elastase. Ethanolic (EE) and hot water crude extracts (HWE) were produced first and then the CGA generated using TWEEN20 were applied resulting in polyphenols enriched fractions (CGA‐EE and CGA‐HWE, ethanol and hot water extracts derived fractions, respectively).RESULTSBoth crude extracts inhibited the enzymes in a dose‐dependent manner, however, further extraction by CGA led to fractions with higher inhibitory efficiency against collagenase. Although gallic acid was the main component of the CGA‐HWE, others such as kaempferol must have contributed to its potency which was more than six times that of gallic acid. The CGA‐EE was found to be about four times more efficient than its crude extract and more than six times more efficient than gallic acid for collagenase inhibition; quercetin was the major polyphenol in this fraction.CONCLUSIONIt is evident that ethanol and hot water extraction processes led to different polyphenols composition and thus different inhibitory activity against collagenase and elastase. Further separation with CGA increased the inhibitory potency of both extracts against collagenase. Overall the results here showed the potential for application of CGA fractions from grape extracts in cosmetics. © 2017 Society of Chemical Industry