Effectiveness of Ya-Samarn-Phlae in diabetic wound healing: Evidence from in vitro studies and a multicenter randomized controlled clinical trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Ya-Samarn-Phlae (YaSP) has traditionally been widely used in southern Thailand for treating chronic and infected wounds, including diabetic foot ulcers. However, there are only a limited number of clinical studies supporting the use of this polyherbal formulation. Therefore, the present work aims to provide clinical evidence to support the application of YaSP, prepared according to a standardized traditional procedure (T-YaSP). Additionally, its potential chemical markers and wound healing-related biological activities were examined. MATERIALS AND METHODS: The in vitro wound healing-related biological activities of YaSP ethanol extract and T-YaSP, including antibacterial activity against Staphylococcus epidermidis, inhibition and eradication of staphylococcal biofilm, anti-inflammatory effects, and enhancement of human dermal fibroblast migration in scratch wounds, were examined using well-established protocols. The chemical profiles of the ethanol extract of YaSP and T-YaSP were compared, and with promising chemical markers, arecoline, alpha-mangostin, and curcumin were selected and quantified using the HPLC method. A prospective, multicenter, randomized, controlled, parallel-group study was conducted over 12 weeks to evaluate the efficacy of the YaSP solution as an adjunct therapy, combined with standard wound care, for diabetic ulcers compared to standard treatment. RESULTS: The YaSP extract reduces NO production and can scavenge NO radicals in LPS-induced RAW 264.7 macrophage cells. Additionally, in a scratch assay, this extract and one of its herbal components, Curcuma longa, enhance the migration of human dermal fibroblasts. T-YaSP, containing 2.412 ± 0.002 mg/g of arecoline, 2.399 ± 0.005 mg/g of curcumin, and 0.017 ± 0.000 mg/g of α-mangostin, has shown the ability to inhibit the development and eradicate the mature biofilm of S. epidermidis. The use of T-YaSP as an adjunct therapy led to a significantly higher proportion of patients achieving healing within six weeks compared to the standard treatment group (36%/9 patients vs. 4%/1 patient; p = 0.013). After 12 weeks, 19 out of 25 patients in the T-YaSP group experienced complete healing, whereas only four patients in the standard treatment group achieved complete wound healing (76% in the T-YaSP group vs. 16% in the control group; p < 0.001). CONCLUSION: The results presented here represent the first randomized controlled trial to demonstrate the effectiveness of the traditional polyherbal solution, T-YaSP, which exhibits a wide range of wound healing-related activities. Utilizing T-YaSP as an adjunctive treatment resulted in a significant improvement in the number of type 2 diabetic patients achieving complete healing. However, to explore and utilize YaSP further, conducting a double-blind, randomized controlled trial with a larger population is necessary.

Potential Galactagogue Effect of Tri-Than-Thip Remedy on Milk Synthesis in Lactating Rats: Impact on Milk Production-Related Parameters.

Herbal galactagogues have been widely used as a treatment for postpartum hypogalactia due to the potential side effects associated with pharmacological therapy. Tri-Than-Thip (Tri-TT) is a Thai herbal medicine remedy that contains three main components: Cassia fistula, Pithecellobium dulce, and Ficus benjamina. These components are believed to have properties that contribute to milk production. However, despite the traditional use of Tri-TT, there is a lack of academic evidence supporting its efficacy in enhancing milk production. Therefore, the purpose of this study was to investigate the effect of Tri-TT on milk production and determine if it has a galactagogue effect. The weight suckle weight model was used to determine total milk production in lactating rats, while histological analysis was performed to assess the alveolar diameter of the mammary gland. The findings of this study revealed a significant increase in total milk production among lactating rats treated with 500 mg/kg of Tri-TT, compared to the control group. Furthermore, both the Tri-TT and Domperidone-treated groups exhibited a larger alveolar diameter of the mammary gland in comparison to the control group. In summary, these findings provide supportive evidence for the galactagogue activity of Tri-TT. The observed enhancement in milk production may be associated with Tri-TT could potentially be attributed to its ability to widen the alveolar diameter of the mammary gland, thereby facilitating increased milk volume.

Evaluation of Chemical Compositions and the Antioxidant and Cytotoxic Properties of the Aqueous Extract of Tri-Yannarose Recipe (Areca catechu, Azadirachta indica, and Tinospora crispa).

Tri-Yannarose is a Thai traditional herbal medicine formula composed of Areca catechu, Azadirachta indica, and Tinospora crispa. It possesses antipyretic, diuretic, expectorant, and appetite-stimulating effects. This study aimed to evaluate the antioxidant activities, cytotoxicity, and chemical constituents of an aqueous extract following a Tri-Yannarose recipe and its plant ingredients. The phytochemical analysis was performed using LC-QTOF-MS. Antioxidant activities were determined using DPPH, ABTS, TPC, TFC, FRAP, NBT, MCA, and ORAC assays. Cytotoxicity was investigated using a methyl thiazol tetrazolium (MTT) assay. In addition, the relationship between the chemical composition of Tri-Yannarose and antioxidant activities was investigated by examining the structure-activity relationship (SAR). The results of the LC-QTOF-MS analysis revealed trigonelline, succinic acid, citric acid, and other chemical constituents. The aqueous extract of the recipe showed significant scavenging effects against ABTS and DPPH radicals, with IC50 values of 1054.843 ± 151.330 and 747.210 ± 44.173 µg/mL, respectively. The TPC of the recipe was 92.685 mg of gallic acid equivalent/g of extract and the TFC was 14.160 mg of catechin equivalent/g of extract. All extracts demonstrated lower toxicity in the Vero cell line according to the MTT assay. In addition, the SAR analysis indicated that prenyl arabinosyl-(1-6)-glucoside and quinic acid were the primary antioxidant compounds in the Tri-Yannarose extract. In conclusion, this study demonstrates that Tri-Yannarose and its plant ingredients have potent antioxidant activities with low toxicity. These results support the application of the Tri-Yannarose recipe for the management of a range of disorders related to oxidative stress.

Repeated 28-Day Oral Toxicological Study and Gastroprotective Effects of Nigella sativa L. Oil (Shuhada) against Ethanol-Induced Gastric Mucosal Injury in Rats.

Nigella sativa L. and black seeds are traditionally used for cooking and medicinal purposes in Arab and other countries. Although N. sativa seed extract has many known biological effects, the biological effects of cold-pressed N. sativa oil are poorly understood. Therefore, the objective of this study was to investigate the gastroprotective effects and subacute oral toxicity of black seed oil (BSO) in an animal model. The gastroprotective effects of oral BSO (50% and 100%; 1 mg/kg) were tested using acute experimental models of ethanol-induced gastric ulcers. Gross and histological gastric lesions, ulcerated gastric areas, ulcer index score, percentage of inhibition rate, gastric juice pH, and gastric wall mucus were all evaluated. The subacute toxicity of BSO and its thymoquinone (TQ) content were also examined. The results indicated that the administration of BSO exerted gastroprotective effects by increasing the gastric wall mucus and decreasing gastric juice acidity. In the subacute toxicity test, the animals behaved normally, and their weight and water and food intake did not show significant variations. High-performance liquid chromatography detected 7.3 mg/mL TQ in BSO. These findings suggest that BSO may be a safe therapeutic drug for preventing gastric ulcers.

Potential applications of Rhodomyrtus tomentosa leaf extract as natural anti-staphylococcal additive in food systems: Efficacy and in vivo safety evaluation.

This work aimed to explore the potential use of Rhodomyrtus tomentosa ethanol leaf extract (RTEL) as an alternative food preservative agent for controlling the growth of Staphylococcus aureus. Antibacterial activities against food-isolated S. aureus were performed using disc diffusion and broth microdilution assays, followed by evaluating in vivo subacute oral toxicity of the extract. Salad dressing was used as a food model to study bactericidal properties and consumer acceptability. RTEL remarkably inhibited S. aureus with minimum inhibitory concentrations (MICs) ranging from 7.81-62.5 µg/mL. Repeated oral doses (5, 50, and 300 mg/kg RTEL) for 28 days did not affect any of the measured toxicity parameters. The no-observed-adverse-effect-level (NOAEL) of RTEL was noted as more than 300 mg/kg body weight/day. The utilization of RTEL (12.5 mg/mL) in the vinaigrette salad dressing did not affect the consumer acceptability of the product, remarkably killed the pathogen within 3-9 h of exposure. The results indicated that RTEL is safe and effective as a natural anti-staphylococcal controlling agent that could be utilized in food systems. Further work is required on the effects of enterotoxin production, an important virulence factor of S. aureus responsible for food-borne disease.

LC-QTOF-MS Characterization, Antioxidant Activity, and In Vitro Toxicity of Medicinal Plants from the Tri-Than-Thip Remedy.

BACKGROUND: The world population has exhibited increased trust in folk medicine, including Thai folk medicine, for the treatment of various illnesses. However, the comparative antioxidant and cytoprotective activities against oxidative damage of Tri-Than-Thip (Tri-TT), a Thai folk remedy, have not been reported. OBJECTIVES: The purpose of this study was to evaluate the antioxidant capacities of Tri-TT and its herbal constituents, Cassia fistula, Pithecellobium dulce, and Ficus benjamina. METHODS: Extracts were obtained from Tri-TT and its herbal constituents. The free radical scavenging activities, cytotoxicity, ferric-reducing antioxidant power (FRAP), metal chelating activities, total phenolic compound (TPC) contents, and total flavonoid (TF) contents of Tri-TT extract were investigated, and qualitative analysis of the chemical composition of Tri-TT extract was performed by LC-QTOF-MS. RESULTS: Tri-TT extract exhibited remarkable scavenging activities toward DPPH, ABTS, and superoxide anion radicals, with IC50 values of 0.081 ± 0.00, 0.021 ± 0.00, and 0.205 ± 0.057 mg/mL, respectively. The oxygen radical antioxidant capacity (ORAC) and FRAP values of Tri-TT were 6.499 ± 0.67 µM TE/g extract and 1,919.71 ± 63.14 mM FeSO4/mg sample, respectively. P. dulce had the highest scavenging activities and antioxidant capacity followed by C. fistula and F. benjamina. The TPC and TF content of Tri-TT extract were 287.87 mg equivalence/g extract and 225.62 mg catechin equivalent/g extract, respectively. The highest TPC was obtained from P. dulce, and TF content was observed in C. fistula. Using LC-QTOF-MS, a total of 25 compounds were tentatively identified in Tri-TT, including polyphenols such as luteolin, gallic acid, baicalein, apigenin, epicatechin, and ellagic acid. In addition, Tri-TT extract demonstrated nontoxicity (cell viability >90%) to Vero cells at the highest tested concentration of 80 µg/mL. CONCLUSION: This study demonstrated that the Tri-TT remedy is a promising candidate as a natural source of antioxidant activity, suggesting that the polyphenol content of plants may contribute to antioxidant activities.

Gastroprotective and Antioxidative Effects of the Traditional Thai Polyherbal Formula Phy-Blica-D against Ethanol-Induced Gastric Ulcers in Rats.

Phy-Blica-D is a traditional Thai polyherbal formula that has reduced oxidative stress in non-communicable diseases. However, evidence supporting the gastroprotective effects of Phy-Blica-D has not been previously reported. Therefore, this study aimed to evaluate the gastroprotective effects of Phy-Blica-D against gastric ulcers in rats and investigate the potential underlying mechanism. To estimate the possible mechanisms of action, we examined the levels of oxidative stress markers, such as reactive oxygen species (ROS) and malondialdehyde (MDA), as well as antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD), and glutathione (GSH). According to our results, rats treated with only 80% ethanol (vehicle group) exhibited significant increases in their ulcer area and ulcer index (UI). Moreover, the levels of ROS and MDA markedly increased in the vehicle group compared with the normal control group. Daily oral administration of Phy-Blica-D (500 and 1000 mg/kg) for 7 days not only significantly decreased the ulcer area and UI, but also remarkably decreased the ROS and MDA levels in gastric tissue. Gastric ulcers induced by ethanol had significantly decreased antioxidant enzyme activities (CAT and SOD) and non-enzymatic antioxidant (GSH), whereas pretreatment with Phy-Blica-D significantly improved the activities of CAT, SOD, and GSH. Moreover, after exposure to ethanol, the rats exhibited a significantly increased level of inducible nitric oxide synthase (iNOS), which was reduced after treatment with Phy-Blica-D. These findings suggest that Phy-Blica-D potentially exerts its gastroprotective effects by suppressing oxidative stress and stimulating antioxidant enzymes, which is one of the causes of destruction of cell membranes, and it is involved in the pathogenesis of acute gastric ulcers induced by ethanol.

Effects of a traditional Thai polyherbal medicine 'Ya-Samarn-Phlae' as a natural anti-biofilm agent against Pseudomonas aeruginosa.

Ya-Samarn-Phlae (YaSP) is a traditional Thai polyherbal formula for the treatment of chronic wounds. Although its ethanol extract has been proven to possess several wound-related biological activities, there is no scientific information available for the infused oil of YaSP, which is its traditionally prepared form. This present study therefore aimed to evaluate the efficacy of different infused oils obtained from either fresh or oven-dried herbal parts of YaSP (F-YaSP and D-YaSP) against biofilms of Pseudomonas aeruginosa, which reside in chronic wounds. Its main active herbal component, Garcinia mangostana (F-GM and D-GM), as well as α-mangostin were also tested in this study. All infused oils significantly inhibited the biofilm formation of P. aeruginosa with a percentage of reduction ranging from 50 to 90%. Visualization of the inhibition of biofilm development was confirmed using scanning electron and atomic force microscopes. All tested agents resulted in a reduction in the mean average roughness of the biofilm, whereas only treating with D-YaSP, D-GM, and α-mangostin led to a decrease in both peak height and peak-valley height. MTT reduction assays revealed that the metabolic activity of P. aeruginosa mature biofilms decreased considerably up to 50% after only 3 h of incubation and after only 9 h of exposure to D-YaSP. Confocal laser scanning micrographs illustrated that a maximum biofilm eradication was found when treated with the extracts for 3 h, whereas the biomass, the average thickness, maximum thickness, and the surface to volume ratio of the treated biofilm was reduced after up to 18 h of contact time. It can be concluded that D-YaSP can effectively inhibit biofilm formation and eradicate mature biofilms of P. aeruginosa. It should be noted that G. mangostana and α-mangostin contribute in YaSP as principle active agents for anti-biofilm efficacy.