Enhancement of cryopreserved rooster semen and fertility potential after oral administration of Thai ginger (Kaempferia parviflora) extract in Thai native chickens.

Objective: Semen cryopreservation is an effective method of preserving genetic material, particularly in native chicken breeds facing a substantial decline. In this study, we evaluated the quality of frozen/thawed rooster semen treated with different concentrations of oral administrations of black ginger (Kaempferia parviflora: KP) extract and determined its fertility. Methods: Thirty-two Thai native roosters (Pradu Hang Dum, 42 weeks old) were used in this study. The treatments were classified into four groups according to the concentration of KP extract administered to the roosters: 0, 100, 150, and 200 mg/kg body weight. The quality of fresh semen was analyzed before cryopreservation. Post-thaw sperm quality and fertility potential were determined. Also, lipid peroxidation was determined. Results: The results showed that sperm concentration and movement increased in roosters treated with 200 mg/kg of KP extract (P<0.05). The malondialdehyde (MDA) in the rooster receiving 200 mg/kg KP extract was lower than that in the other but had an insignificant difference within the KP treatment groups (P>0.05). The highest MDA levels were observed in the control group (P<0.05). The percentage of motile sperm (total motility and progressive motility) after semen thawing was higher in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (P<0.05). MDA levels decreased significantly in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (P<0.05). Fertility and hatchability were greater in the KP150 and KP200 groups than in the KP100 and control groups (P<0.05). Conclusion: The optimal amount of KP extract influencing initial sperm quality was determined to be 200 mg/kg. However, 150 mg/kg was the optimal low dosage of KP extract administration that maintained sperm quality and fertility following semen cryopreservation.

The effect of Thai ginger (Kaempferia parviflora) extract orally administration on sperm production, semen preservation, and fertility in Thai native chickens under heat stress.

Thai indigenous roosters are exposed to unsuitable temperatures and humidity, resulting in a lower reproductive potential. Kaempferia parviflora (KP) extract containing methoxyflavones was fed to roosters to improve their reproductive performance. Thirty-two Thai native roosters were orally administered KP extract at 300, 450, and 600 mg, calculated according to their average body weight, for at least 14 d before semen collection and continued supplementation until the end of the experiment. The nonsupplemented group served as the control. Fresh semen in terms of semen volume, sperm concentration, mass movement score, and sperm viability were evaluated. Semen preservation at 5°C and fertility test were examined for total motility (MOT), progressive motility (PMOT), sperm viability, and lipid peroxidation up to 48 h of storage. Testosterone concentrations and testicular function were also determined. The results showed that the highest sperm concentration and sperm motility of fresh semen were observed in KP extract at 600 mg (P < 0.001). KP extract at 600 mg resulted in higher sperm viability than the control and KP extract at 300 mg (P < 0.05), but was not different from KP at 450 mg (P > 0.05). The highest MOT, PMOT, and viability were found in the roosters that received 600 mg oral KP extract (P < 0.05), while those of the roosters that received oral KP extract 300 mg and the control were the lowest (P < 0.05) at all storage times. Lipid peroxidation was significantly lower in the KP extract up to 24 h (P < 0.05). The fertility and hatchability of the KP extract at 600 mg at T48 showed a minor decrease compared to the control at T0. These results might be inferred as a result of good spermatogenesis, as revealed by the results of histological examination and testosterone activity. In summary, oral administration of 600 mg KP extract improved sperm production and successfully preserved rooster semen for a long duration of up to 48 h of storage.

Formulation and In Vitro Evaluation of Mucoadhesive Sustained Release Gels of Phytoestrogen Diarylheptanoids from Curcuma comosa for Vaginal Delivery.

Diarylheptanoids (DAs) characterized by a 1,7-diphenylheptane structural skeleton are considered a novel class of phytoestrogens. The DAs available in Curcuma comosa Roxb. (C. comosa) extract demonstrated significant estrogenic activities both in vitro and in vivo. This study aimed to develop and comprehensively evaluate a mucoadhesive vaginal gel for the sustained release of DAs. Different mucoadhesive polymers as gelling agents were investigated. C. comosa ethanolic crude extract was used as a source of DAs. All C. comosa gels were light brown homogeneous with pH within 4.4-4.6. Their flow behaviors were pseudoplastic with a flow behavior index of 0.18-0.38. The viscosity at a low shear rate varied from 6.2 to 335.4 Pa·s. Their mechanical and extrudability properties were associated well with rheological properties. Polycarbophil (PCP):hydroxypropyl methylcellulose (HPMC) blends had a higher mucoadhesiveness to porcine vaginal mucosa than those of PCP-based or HPMC-based gels. All C. comosa gels exhibited a sustained, zero-order DA release pattern over 72 h. Korsmeyer and Peppas equation fitting indicated a non-Fickian, case II transport release mechanism. C. comosa gels had good physical and chemical stability under low-temperature storage for up to 12 months. PCP:HPMC-based mucoadhesive gels could be a proper delivery system for vaginal administration of DAs.

Supplementation of Kaempferia parviflora Extract Enhances Physical Fitness and Modulates Parameters of Heart Rate Variability in Adolescent Student-Athletes: A Randomized, Double-Blind, Placebo-Controlled Clinical Study.

This randomized double-blind controlled study aimed to investigate the effects of a standardized Kaempferia parviflora (KP) extract on the physical fitness and heart rate variability (HRV) parameters in adolescent sport school students. 194 male students were recruited and randomized into two groups (n = 97), matched by age and sports. The KP-treated group received KP extract capsules at a dose of 360 mg/day and the control group received placebo capsules, continuously for 12 weeks. Physical fitness performance and HRV parameters were monitored with blood biochemical analysis for product safety. KP extract significantly increased the right-hand grip strength, the back-leg strength and maximal oxygen consumption (VO2 max) and decreased the time used for 50-meter sprint test without changing the sit-and-reach test and the 40 yard technical test. For HRV parameters, KP extract significantly increased standard deviation of normal to normal intervals (SDNN), square root of the mean of square of successive normal to normal interval differences (RMSSD) and high frequency (HF) norm, without changing low frequency (LF) norm and LF/HF ratio. The increase in stress resistance and decrease in stress index were found in the KP-treated group, without changing the autonomic nervous system (ANS) activity and balance. Blood biochemical analysis showed normal values of all participants. This data indicates the safety and positive effects of KP on muscle strength, endurance and speed, but not on the flexibility and agility. The modulatory effects of KP extract on HRV parameters suggest its anti-stress effects and would encourage the application in a sport training and exercise.

Curcuma comosa loaded transfersomal gel for transdermal application: formulation, in vitro and in vivo evaluation.

RATIONAL: Diarylheptanoids, extracted from the Curcuma comosa (CC) rhizome, have been reported to exhibit estrogenic activity. However, oral administration of the extract showed a short half-life. OBJECTIVES: This study aimed to formulate and to investigate the potential of transfersomal gels for the transport of phytoestrogenic diarylheptanoids across the skin into the blood circulation. MATERIALS AND METHODS: The transfersomes were developed and optimized for their compositions including sources of phospholipid (egg yolk and soybean), types of edge activators (polysorbate 80, sorbitan oleate 80, and sodium cholate), and concentrations of CC extract (10-60 mg). The optimal formulation was further incorporated into Carbopol® Ultrez 21 gel and evaluated for in vitro release, permeation, and in vivo absorption. RESULTS: The optimal transfersomes containing 10% of polysorbate 80 were selected due to high drug entrapment efficiency and a small diameter. The release kinetic of transfersomal gels followed a zero model. The maximum permeation flux through porcine ear skin was 1.38 ± 0.25 µg/cm2/h for (4E, 6E)-1, 7-diphenylhepta-4, 6-dien-3-ol, and 0.40 ± 0.11 µg/cm2/h for (6E)-1, 7-diphenylhept-6-en-3-ol. Results of the in vivo pharmacokinetics study in rats showed that transfersomal gel provided a maximum concentration of 219.71 ± 4.05 ng/ml and prolonged plasma concentration of diarylheptanoids for over 12 h. There was no significant variation found in the physical characteristics including viscosity, pH, and size after six months of storage at room temperature (30 ± 1 °C) and high temperature (40 ± 1 °C). CONCLUSIONS: The obtained data suggested that the developed transfersomal gel of CC extract should be beneficial for improving the delivery of phytoestrogenic diarylheptanoids.

Development and evaluation of topical films containing phytoestrogenic diaryheptanoids from Curcuma comosa extract.

RATIONAL: Phytoestrogens have been found to delay signs of skin aging in post-menopausal women, in a way similar to the effects of estrogens. Diarylheptanoids from a rhizome of traditional Thai herb named Curcuma comosa is considered to be a novel class of phytoestrogens. OBJECTIVES: The aims of this study were to prepare effective topical films using mixed types and vary ratios of hydrophobic (Eudragit RL, Eudragit RS, and Eudragit NE) and hydrophilic polymer (hydroxylpropyl methycellulose, HPMC) with Transcutol as a permeation enhancer for delivery of diarylheptanoids to improve signs of skin aging in post-menopausal women. MATERIAL AND METHODS: Topical films were characterized for their physical and mechanical properties. In vitro release, skin permeation and accumulation were evaluated using Franz diffusion cell and the concentrations of diarylheptanoids were determined using high-performance liquid chromatography. RESULTS: The combined formulations between HPMC and Eudragit NE showed the satisfactory physical and mechanical properties, and also provided the highest amount of drug released compared to Eudragit RL and Eudragit RS. When the proportion of HPMC amount in the polymer matrix increased, the cumulative drug release also increased (HPMC: Eudragit NE 6:4 > 5:5 > 4:6). Moreover, they provided a high accumulation of diarylheptanoids within skin when using transcutol as a permeation enhancer. CONCLUSION: The obtained data provided the skin permeation and accumulation behavior of diarylheptanoids, indicating the feasibility of a skin delivery of the C. comosa extract. The developed films might be topically used as an alternative therapy for protection of skin aging in peri and post-menopausal women.

Effects of Vehicles and Enhancers on the Skin Permeation of Phytoestrogenic Diarylheptanoids from Curcuma comosa.

Curcuma comosa (C. comosa) is widely used in traditional medicine as a dietary supplement for health promotion in postmenopausal women in Thailand. It contains several diarylheptanoids, which are considered to be a novel class of phytoestrogens. However, the diarylheptanoids isolated from the plant rhizome are shown to have low oral bioavailability and faster elimination characteristics. The aim of this study was to investigate the permeation behavior of the active compounds of diarylheptanoids. The effects of binary vehicle systems and permeation enhancers on diarylheptanoids permeation and accumulation within the skin were studied using side-by-side diffusion cells through the porcine ear skin. Among the tested binary vehicle systems, the ethanol/water vehicle appeared to be the most effective system for diarylheptanoids permeation with the highest flux and shortest lag time. The presence of transcutol in the vehicle system significantly increased diarylheptanoid's permeation and accumulation within the skin in a concentration-dependent manner. Although the presence of terpenes in formulation decreased the flux of diarylheptanoids, it raised the amount of diarylheptanoids retained within the skin substantially. Based on the feasibility of diarylheptanoid permeation, C. comosa extract should be further developed into an effective transdermal product for health benefits and hormone replacement therapy.

A monolithic drug-in-adhesive patch of methoxyflavones from Kaempferia parviflora: in vitro and in vivo evaluation.

The aim of this study was to design and develop a suitable monolithic drug-in-adhesive type patch of methoxyflavones from Kaempferia parviflora (K. parviflora) using acrylic polymer Durotak(®) 87-2852. The absence of interaction between components in K. parviflora extract and the adhesive polymer was confirmed by attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR). Thirteen K. parviflora patches with different extract loading and permeation enhancers were prepared by the solvent evaporation technique. All formulations showed good physicochemical properties, good stability and satisfactory adhesive properties. The effect of K. parviflora loading and permeation enhancers on methoxyflavones transport across porcine ear skin was also evaluated. The permeation of methoxyflavones increased with the amount of K. parviflora. Among the permeation enhancers investigated, oleic acid increased permeation flux of total methoxyflavones by 1.25 fold compared to the control; whereas menthol shortened the lag time. When oleic acid and menthol were combined, the maximum flux of methoxyflavones and shortest lag time were observed, suggesting a synergistic effect of menthol with oleic acid. The optimal patch formulation contained 15% K. parviflora, 3% oleic acid and 3% of menthol, and this was evaluated via an in vivo pharmacokinetic study using rats. The maximum plasma drug concentration (Cmax) of total methoxyflavones was 218.08ng/ml with Tmax at 8h. The concentrations of methoxyflavones in plasma continued to increase until the end of the experiment, indicating a sustained release into the systemic circulation.

Transdermal permeation of Kaempferia parviflora methoxyflavones from isopropyl myristate-based vehicles.

Kaempferia parviflora (K. parviflora) rhizomes have long been used in traditional folk medicines and as general health-promoting agents. Several biological activities of K. parviflora, especially its anti-inflammatory effect, are due to its major constituents, methoxyflavones. However, the oral bioavailability of these methoxyflavones has been shown to be low. The aim of this study was to investigate the permeation behaviors of K. parviflora methoxyflavones from isopropyl myristate (IPM)-based vehicles. We studied the effects of ethanol and propylene glycol (PG) as the hydrophilic, solvent-type vehicles as well as fatty acids as the permeation enhancers. A permeation experiment was performed in vitro, using side-by-side diffusion cells through the full thickness of pig ear skin. The solubility and permeation of methoxyflavones were able to be modified by choice and ratio of vehicles. The ethanol/IPM vehicle was shown to be more effective in enhancing the solubility and permeation of methoxyflavones when compared to the PG/IPM vehicle. Regarding an optimal balance between solubility or affinity to vehicle and skin to vehicle partition coefficient, the ethanol/IPM vehicle in the ratio of 1:9 maximized the flux. Among the investigated fatty acids, oleic acid showed the greatest enhancing effect on the permeation of methoxyflavones, indicating that saturated fatty acids are less effective than unsaturated fatty acids. Long chain fatty acids increased diffusion coefficient parameter and shortened the lag time. The number of carbon atoms and double bonds of fatty acids did not show direct relation to the profile of permeation of methoxyflavones.