Pensak Jantrawut

Anti-inflammatory activity of gel containing novel elastic niosomes entrapped with diclofenac diethylammonium

The objective of this study was to develop a novel elastic bilayer vesicle entrapped with the non-steroidal anti-inflammatory drug (NSAID), diclofenac diethylammonium (DCFD) for topical use. Eighteen bilayer vesicular formulations composing of DPPC or Tween 61 or Span 60 mixed with cholesterol (at 1:1, 3:7 and 1:1 molar ratios, respectively) and ethanol at 0-25% (v/v), by chloroform film method with sonication were developed. The elastic Tween 61 niosomes which gave no sedimentation, no layer separation, unchanged particle sizes (about 200 nm) were selected to entrap DCFD. The entrapment efficiency of the drug in the conventional and elastic Tween 61 niosomes was 65 and 93%, respectively. At least 87% of DCFD determined by HPLC remained in elastic Tween 61 niosomes when kept at 4, 27 and 45 degrees C for 3 months. The deformability index values of the elastic niosomes were 13.76 and 3.44 times higher than the conventional niosomes entrapped and not entrapped with the drug, respectively, indicating the higher flexibility of the elastic vesicle especially, when entrapped with the drug. Transdermal absorption through excised rat skin was performed by vertical Franz diffusion cell at 32+/-2 degrees C for 6h. Gel containing elastic niosomes exhibited fluxes of DCFD in the stratum corneum (SC), deeper skin layer (viable epidermis and dermis, VED) and receiver chamber at 191.27+/-9.52, 16.97+/-2.77 and 3.76+/-0.54 microg/(cm2 h), whereas the commercial emulgel, containing an equivalent DCFD, gave 60.84+/-13.63, 7.33+/-1.70 and 0.14+/-0.01 microg/(cm2 h), respectively. The in vivo anti-inflammatory activity was evaluated by ethyl phenylpropiolate (EPP)-induced rat ear edema (n=3). DCFD entrapped in the developed elastic niosomes and incorporated in gel gave the same ear edema inhibition percentages of 23.81% at 30 min, but 2 and 9 times more inhibition percentages at 45 and 60 min than the commercial emulgel, respectively. This result has not only demonstrated the enhancement of transdermal absorption through rat skin, but also the in vivo anti-inflammatory effect of DCFD when entrapped in the developed novel elastic Tween 61 niosomes, as well.

In vitro anti-aging activities of Terminalia chebula gall extract

Context: The Thai Lanna region has its own folklores and wisdoms in various fields such as traditional medicines. The galls of Terminalia chebula Retz. (Combretaceae) frequently appear in many Thai Lanna medicinal plant recipes for promoting longevity. Objectives: To investigate the in vitro anti-aging activities of the extracts from 15 plants including T. chebula gall selected from the Thai medicinal plant recipes that have been traditionally used for longevity. Materials and methods: The plant extracts were prepared by four extraction methods including hot (HW) and cold (CW) aqueous processes and hot (HM) and cold (CM) methanol processes. These extracts were tested for antioxidative and tyrosinase inhibition activity as well as the proliferative and MMP-2 inhibition activity on early aging human skin fibroblasts in order to evaluate their in vitro anti-aging activity. Results: At 0.1u2009mg/mL, the CW extract of T. chebula gall exhibited the highest DPPH radical scavenging activity with scavenging of 84.64%u2009±u20092.22%, whereas ascorbic acid, α-tocopherol and butylated hydroxyl toluene gave 96.50%u2009±u20090.1%, 35.74%u2009±u20090.2% and 27.43%u2009±u20090.1%, respectively. The CW extract of T. chebula gall indicated the highest stimulation index (SI) on normal human fibroblast proliferation of 1.441 which was more active than ascorbic acid (SI 1.21). This extract has also demonstrated MMP-2 inhibition on fibroblasts determined by zymography 1.37 times more potent than ascorbic acid. Discussion and conclusion: This study has confirmed the traditional use of T. chebula gall in many Thai medicinal plant recipes for longevity which will be beneficial for further development of anti-aging products.

Biological Activities of Phenolic Compounds and Triterpenoids from the Galls of Terminalia chebula

Nine phenolic compounds, including two phenolic carboxylic acids, 1 and 2, seven hydrolyzable tannins, 3–9, eight triterpenoids, including four oleanane‐type triterpene acids, 10–13, and four of their glucosides, 14–17, isolated from a MeOH extract of the gall of Terminalia chebula Retz. (myrobalan tree; Combretaceae), were evaluated for their inhibitory activities against melanogenesis in B16 melanoma cells induced by α‐melanocyte‐stimulating hormone (α‐MSH), against the Epsteinuf8ffBarr virus early antigen (EBV‐EA) activation induced by 12‐O‐tetradecanoylphorbol 13‐acetate (TPA) in Raji cells, and against TPA‐induced inflammation in mice. Their 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical‐scavenging activities and cytotoxic activities against four human cancer cell lines were also evaluated. Compounds 6–9 and 12 exhibited potent inhibitory activities against melanogenesis (39.3–66.3% melanin content) with low toxicity to the cells (74.5–105.9% cell viability) at a concentration of 10u2005μM. Western‐blot analysis revealed that isoterchebulin (8) reduced the protein levels of MITF (=microphtalmia‐associated transcription factor), tyrosinase, and TRP‐1 (=tyrosine‐related protein 1), mostly in a concentration‐dependent manner. Eight triterpenoids, 10–17, showed potent inhibitory effects on EBV‐EA induction with the IC50 values in the range of 269–363u2005mol ratio/32u2005pmol TPA, while these compounds exhibited no DPPH scavenging activities (IC50>100u2005μM). On the other hand, the nine phenolic compounds, 1–9, exhibited potent radical‐scavenging activities (IC50 1.4–10.9u2005μM) with weak inhibitory effects on EBV‐EA induction (IC50 460–518u2005mol ratio/32u2005pmol TPA). The tannin 6 and seven triterpenoids, 10–16, have been shown to inhibit TPA‐induced inflammation (1u2005μg/ear) in mice with the ID50 values in the range of 0.06–0.33u2005μmol/ear. Arjungenin (10) exhibited inhibitory effect on skin‐tumor promotion in an in vivo two‐stage mouse‐skin carcinogenesis test based on 7,12‐dimethylbenz[a]anthracene (DMBA) as initiator and with TPA as promoter. Compounds 1, 2, 4, 5, 7–9, 12, and 13, against HL60 cell line, compounds 1 and 4, against AZ521 cell line, and compounds 1, 11, and 12, against SK‐BR‐3 cell line, showed moderate cytotoxic activities (IC50 13.9–73.2u2005μM).

Biological activities of phenolic compounds isolated from galls of Terminalia chebula Retz. (Combretaceae)

The aqueous extract of galls from Terminalia chebula Retz. (Combretaceae) was fractionated on Diaion and refractionated on octadecyl silica column. Six phenolic compounds were isolated and identified as gallic acid (1), punicalagin (2), isoterchebulin (3), 1,3,6-tri-O-galloyl-β-D-glucopyranose (4), chebulagic acid (5) and chebulinic acid (6). All of the compounds showed stronger 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and melanin inhibitory activities than ascorbic acid, butylated hydroxytoluene, α-tocopherol, arbutin and kojic acid, the reference compounds. Gallic acid (1) exhibited inhibitory activity against nitric oxide production in lipopolysaccharide-activated macrophages. However, all isolated compounds exhibited less activity than the reference compounds in mushroom tyrosinase inhibition and human tumour cytotoxicity assays. This study has demonstrated that the phenolic compounds isolated from galls of T. chebula might contribute significantly due to their antioxidant and whitening activities.

In vitro and in vivo skin anti-aging evaluation of gel containing niosomes loaded with a semi-purified fraction containing gallic acid from Terminalia chebula galls

Context: The galls of Terminalia chebula Retz. (Combretaceae) frequently appear in many Thai Lanna medicinal plant recipes for promotion of longevity. Objective: The objective of this study was to evaluate the skin anti-aging of gel containing niosomes loaded with a semi-purified fraction containing gallic acid from T. chebula galls. Method: The semi-purified fraction containing phenolic compounds including gallic acid isolated from T. chebula galls loaded in non-elastic or elastic niosomes, and its developed gel, were evaluated for rabbit skin irritation by the closed patch test and skin anti-aging in human volunteers by measuring skin elasticity and roughness. Results: Gel containing the fraction unloaded (SS) or loaded in non-elastic (SN) or elastic (SE) niosomes and gallic acid loaded in non-elastic (GN) or elastic (GE) niosomes showed no skin irritation, whereas the unloaded gallic acid (GS) gave the irritation in rabbit’s skin by the closed patch test. The % parameter changes of skin elastic recovery and skin elastic extension when applied with SN and SE gels were +28.73 and +32.57; −21.25 and −22.63%, respectively. SN and SE gel also showed a significant decrease of the maximum and average roughness values with the parameter changes of −29.43 and −32.38; −39.47 and −35.28%, respectively. Conclusion: The semi-purified fraction loaded in niosomes indicated not only higher chemical stability of gallic acid containing in the fraction, but also more in vivo anti-aging activities than the unloaded fraction when incorporated in gel.

Influence of low methoxyl pectin gel textures and in vitro release of rutin from calcium pectinate beads

This study described the preparation and characterization of low methoxyl pectin (LMP) gels and beads for controlled release applications. The rheological characterization of the various formulations was proposed. Then the mechanical and morphological characterizations of beads were determined. Finally, the controlled release studies taking rutin as a model drug was evaluated. The results showed that Youngs modulus values of non-amidated LMP gels decrease when adding up to 15% sorbitol. Calcium pectinate beads loaded with rutin are about 600 μm, oblong shaped with dense matrix. Beads containing sodium bicarbonate showed about 80% lower rutin encapsulation efficiency by increasing the pH of the cross-linking solution. The rutin loaded in non-amidated pectinate beads containing 15% sorbitol showed the best release efficiency and swelling behavior. Therefore, the gel texture affects the release rate of the active compounds encapsulated in calcium pectinate beads and can be used as a parameter to modulate drug release.

Transdermal absorption enhancement of gel containing elastic niosomes loaded with gallic acid from Terminalia chebula galls

Objectives: To investigate transdermal absorption enhancement of gel containing elastic niosomes loaded with gallic acid in the semipurified fraction isolated from Terminalia chebula Retz. (Combretaceae) galls. Materials and methods: Nonelastic and elastic niosomes loaded with gallic acid in pure form or in the semipurified fraction were developed. Rat skin permeation by vertical Franz diffusion cells of gallic acid from various gel formulations containing elastic niosomes loaded with gallic acid or the semipurified fraction was performed. Results: Elastic and nonelastic niosomes loaded with gallic acid or the semipurified fraction exhibited the mixture of unilamellar and multilamellar structures with negative zeta potential values and in the size range of 200–400u2009nm. Both loaded elastic and nonelastic niosomes showed good physical and chemical stability for 3 months. The percentages remaining of gallic acid in nonelastic were slightly higher than in elastic niosomes. Both elastic and nonelastic niosomes retarded rat skin permeation of the loaded pure gallic acid, while enhanced the loaded gallic acid in the semipurified fraction. However, elastic niosomes exhibited higher percentages of gallic acid through rat skin than the nonelastic niosomes. Discussion and conclusion: This study has demonstrated the potential of niosomes, especially elastic niosomes, for the enhancement of chemical stability and rat skin transdermal absorption of gallic acid in the semipurified fraction from T. chebula galls, which will be beneficial for topical antiaging application.

In vitro anti-proliferative activity on colon cancer cell line (HT-29) of Thai medicinal plants selected from Thai/Lanna medicinal plant recipe database "MANOSROI III".

ETHNOPHARMACOLOGICAL RELEVANCEnThai/Lanna region has its own folklore wisdoms including the traditional medicinal plant recipes. Thai/Lanna medicinal plant recipe database MANOSROI III has been developed by Prof. Dr. Jiradej Manosroi. It consists of over 200,000 recipes for all diseases including cancer. To investigate the anti-proliferative and apoptotic activities on human colon cancer cell line (HT-29) as well as the cancer cell selectivity of the methanolic extracts (MEs) and fractions of the 23 selected plants from the MANOSROI III database.nnnMATERIALS AND METHODSnThe 23 selected plants were extracted with methanol under reflux and evaluated for their anti-proliferative activity by sulforhodamine B assay. The 5 plants (Gloriosa superba, Caesalpinia sappan, Fibraurea tinctoria, Ventilago denticulata and Psophocarpus tetragonolobus) with potent anti-proliferative activity were fractionated by liquid-liquid partition to give 4 fractions including each hexane (HF), methanol-water (MF), n-butanol (BF) and water (WF) fractions. They were tested for anti-proliferative activity and cancer cell selectivity. The ME and fractions of G. superba which showed potent anti-proliferative activity were further examined for morphological changes and apoptotic activities by acridine orange (AO)/ethidium bromide (EB) staining.nnnRESULTSnThe ME of G. superba root showed active with the highest anti-proliferative activity at 9.17 and 1.58 folds of cisplatin and doxorubicin, respectively. After liquid-liquid partition, HF of V. denticulata, MFs of F. tinctoria, V. denticulata and BF of P. tetragonolobus showed higher anti-proliferative activities than their MEs. The MF of G. superba indicated the highest anti-proliferative activity at 7.73 and 1.34 folds of cisplatin and doxorubicin, respectively, but only 0.86 fold of its ME. The ME and HF, MF and BF of G. superba and MF of F. tinctoria demonstrated high cancer cell selectivity. At 50 µg/ml, ME, HF, MF and BF of G. superba demonstrated higher apoptotic activities than the two standard drugs.nnnCONCLUSIONSnThis present study has not only confirmed the traditional use of the Thai/Lanna medicinal plant recipes for cancer treatments, but also the potential of the selected plant, G. superba for the further development as a modern anti-cancer drug.

Anticancer activities of the extract from Longkong (Lansium domesticum) young fruits

Context: “Longkong”(Lansium domesticum Corr., Family: Meliaceae) is a fruit found in the south of Thailand. This plant has been used in traditional medicines. Objectives: To investigate the antiproliferative activities and the phytoconstituents of Longkong extracts. Materials and methods: Cytotoxicity and apoptotic activity of 48 extracts were tested using the SRB assay and acridine orange (AO)/ethidium bromide (EB) staining, respectively. The extracts which gave the highest anticancer activity were selected to prepare the semipurified extracts and analysis for the constituents by gas chromatography–mass spectrometry (GC/MS). Results: The highest percentage yield (59.38%) was from the cold water extract of Longkong ripe fruits (RFWC). The highest total phenolic and flavonoid contents were observed in cold and hot methanol extract of Longkong stalks (STMC and STMH). The hot and cold chloroform young Longkong fruit extracts (YFCH and YFCC) exhibited a cytotoxic effect (IC50 < 1 mg/mL) against cancer cells. For apoptotic induction, YFCH demonstrated the highest activity against KB of 13.84 ± 4.21% at 0.5 mg/mL which was 0.88 and 1.35 times of cisplatin and 5-FU, respectively, while apoptotic cells in HT-29 were 8.68 ± 1.85% at 5 mg/mL, which was 0.61 and 1.43 times of cisplatin and 5-FU, respectively. YFCC showed the highest apoptotic effect against KB cells at 10.70 ± 2.15% at 0.5 mg/mL, which was 0.68 and 1.07 times of cisplatin and 5-FU, respectively. The major phytoconstituents in YFCH were hexadecanoic acid (11.53%) and ethyl oleate (10.58%). Discussion and conclusion: The crude extracts of Longkong showed anticancer activities and may provide new lead compounds for the development of anticancer products.

Effect of Plasticizer Type on Tensile Property and In Vitro Indomethacin Release of Thin Films Based on Low-Methoxyl Pectin

This study developed the interests of low-methoxyl pectin (LMP) together with plasticizers for the preparation of elastic thin films. The effect of different plasticizer types (glycerol: Gly; sorbitol: Sor; propylene glycol: PG; and polyethylene glycol 300: PEG 300) and concentrations (20–40% w/w) on mechanical and thermal properties of LMP films as well as on in vitro release of indomethacin were evaluated. Without any plasticizer, a brittle LMP film with low tensile strength and % elongation at break was obtained. Addition of plasticizers from 20% to 40% caused reduction in the tensile strength and Young’s modulus values, whereas percent elongation was increased. Forty percent Gly-plasticized and PG-plasticized films were selected to deliver indomethacin in comparison with non-plasticized film. No significant difference in indomethacin release profiles was displayed between the films. The analysis of indomethacin release model indicated that more than one drug release mechanism from the film formulation was involved and possibly the combination of both diffusion and erosion. Even though indomethacin incorporated in non-plasticized film showed similar release profile, Gly or PG should be added to enhanced film flexibility and decrease film brittleness.