The aloe vera (Aloe vera) plant has a long history of traditional therapeutic topical use. Its leaf and inner leaf juices are also ingested in various forms. Aloe vera leaf juice is made from macerated aloe leaves, and when it is intended for human ingestion, it is purified of latex constituents via a charcoal filtration process known as decolorization. Several publications have shown immunomodulatory effects, as well as antibacterial, antifungal and anti-parasitic properties of aloe vera inner leaf juice, but limited information is available regarding these effects for aloe vera leaf juice after the decolorization process. The aim of this preliminary study was to compare biologically-relevant antioxidant and immune-modulatory effects of aloe vera inner leaf and decolorized leaf preparations using a variety of in vitro techniques.
Objective: To evaluate a blend of two natural ingredients on immune parameters relevant for their current topical use and potential support of microcirculation in skin tissue.
Materials and methods: A blend (BL) of Aloe vera-based Nerium oleander extract (NAE-8i, oleandrin-free) and hydrolyzed water-soluble egg membrane (WSEM) was applied to human whole-blood cultures for 24 hours, with each separate ingredient serving as a control. Immune-cell subsets were analyzed for expression levels of the activation markers CD69 and CD25. Culture supernatants were analyzed for cytokines, chemokines, and immunoregulating peptides.
Results: BL increased CD69 expression on lymphocytes, monocytes, and CD3–CD56+ natural killer cells, and CD25 expression on natural killer cells. The number of CD69+CD25+ lymphocytes increased in cultures treated with BL and the separate ingredients. BL triggered production of multiple cytokines and chemokines, where CC chemokines MIP1α and MIP3α, as well as cytokines involved in wound healing – Groα, Groβ, ENA78, and fractalkine – reached levels manyfold above treatment with either NAE-8i or WSEM alone.
Conclusion: Data on BL showed that WSEM strongly enhanced NAE-8i’s effects on immunoactivation in vitro. This has potential relevance for support of immunity in skin tissue, including antibacterial and antiviral defense mechanisms, wrinkle reduction, and wound care.
Objective: The aim of this study was to evaluate the effects of water-soluble egg membrane (WSEM) on wrinkle reduction in a clinical pilot study and to elucidate specific mechanisms of action using primary human immune and dermal cell-based bioassays.
Methods: To evaluate the effects of topical application of WSEM (8%) on human skin, an open-label 8-week study was performed involving 20 healthy females between the age of 45 years and 65 years. High-resolution photography and digital analysis were used to evaluate the wrinkle depth in the facial skin areas beside the eye (crow’s feet). WSEM was tested for total antioxidant capacity and effects on the formation of reactive oxygen species by human polymorphonuclear cells. Human keratinocytes (HaCaT cells) were used for quantitative polymerase chain reaction analysis of the antioxidant response element genes Nqo1, Gclm, Gclc, and Hmox1. Evaluation of effects on human primary dermal fibroblasts in vitro included cellular viability and production of the matrix components collagen and elastin.
Results: Topical use of a WSEM-containing facial cream for 8 weeks resulted in a significant reduction of wrinkle depth (P<0.05). WSEM contained antioxidants and reduced the formation of reactive oxygen species by inflammatory cells in vitro. Despite lack of a quantifiable effect on Nrf2, WSEM induced the gene expression of downstream Nqo1, Gclm, Gclc, and Hmox1 in human keratinocytes. Human dermal fibroblasts treated with WSEM produced more collagen and elastin than untreated cells or cells treated with dbcAMP control. The increase in collagen production was statistically significant (P<0.05).
Conclusion: The topical use of WSEM on facial skin significantly reduced the wrinkle depth. The underlying mechanisms of this effect may be related to protection from free radical damage at the cellular level and induction of several antioxidant response elements, combined with stimulation of human dermal fibroblasts to secrete high levels of matrix components.
Objective: The objective of this study is to evaluate the effects of consumption of nattokinase on hypertension in a North American hypertensive population with associated genetic, dietary, and lifestyle factors. This is in extension of, and contrast to, previous studies on Asian populations.
Materials and methods: A randomized, double-blind, placebo-controlled, parallel-arm clinical study was performed to evaluate nattokinase (NSK-SD), a fermented soy extract nattō from which vitamin K2 has been removed. Based on the results from previous studies on Asian populations, 79 subjects were enrolled upon screening for elevated blood pressure (BP; systolic BP ≥130 or diastolic BP ≥90 mmHg) who consumed placebo or 100 mg nattokinase/d for the 8-week study duration. Blood collections were performed at baseline and 8 weeks for testing plasma renin activity, von Willebrand factor (vWF), and platelet factor-4. Seventy-four people completed the study with good compliance.
Results: Consumption of nattokinase was associated with a reduction in both systolic and diastolic BP. The reduction in systolic BP was seen for both sexes but was more robust in males consuming nattokinase. The average reduction in diastolic BP in the nattokinase group from 87 mmHg to 84 mmHg was statistically significant when compared to that in the group consuming placebo, where the average diastolic BP remained constant at 87 mmHg (P<0.05), and reached a high level of significance for males consuming nattokinase, where the average diastolic BP dropped from 86 mmHg to 81 mmHg (P<0.006). A decrease in vWF was seen in the female population consuming nattokinase (P<0.1). In the subpopulation with low plasma renin activity levels at baseline (<0.29 ng/mL/h), an increase was seen for 66% of the people after 8-week consumption of nattokinase (P<0.1), in contrast to only 8% in the placebo group.
Conclusion: The data suggest that nattokinase consumption in a North American population is associated with beneficial changes to BP in a hypertensive population, indicating sex-specific mechanisms of action of nattokinase’s effect on vWF and hypertension.
Objectives: The aim of this study was to evaluate the effects of consumption of an aqueous cyanophyta extract (ACE) from Arthrospira platensis on chronic pain in humans, in two clinical pilot studies.
Design and interventions: The two pilot studies each involved 12 subjects experiencing chronic pain. The initial study followed an open-label 4-week study design involving consumption of 1 g ACE per day. A subsequent placebo-controlled, single-blind, crossover study involved consumption of 500 mg ACE, 250 mg ACE, or 0 mg ACE (placebo) per day for 1-week duration, separated by 1-week washout period.
Subjects: Adult subjects of both sexes, with chronic joint-related pain for at least 6 months prior to enrollment, were recruited after obtaining written informed consent.
Outcome measures: Visual analog scales were used to score pain at rest and during physical activity for each person’s primary and secondary areas of chronic pain. An activities of daily living questionnaire was used to collect data on physical functioning.
Results: The data showed rapid reduction of chronic pain in people consuming ACE, where the reduction in pain scores for each person’s primary pain area reached a high level of statistical significance after 2 weeks of consumption (P<0.01), both when at rest and when being physically active. Secondary pain areas when physically active showed highly significant improvements within 1 week of consumption of 1 g/d (P<0.001) and borderline significant improvements within 1 week of consuming 500 mg/d (P<0.065) and 250 mg/d (P<0.05). This was accompanied by an increased ability to perform daily activities (P<0.05). A small but significant weight loss was observed during the 4-week study, as the average body mass index dropped from 31.4 to 29.4 (P<0.01).
Conclusion: Consumption of ACE was associated with reduction of chronic pain, as well as a dose-dependent increased ability to perform activities of daily living.
Objective: The goal for this study was to evaluate the effects of daily consumption of Puer tea extract (PTE) on body weight, body-fat composition, and lipid profile in a non-Asian population in the absence of dietary restrictions.
Materials and methods: A randomized, double-blind, placebo-controlled study design was used. A total of 59 overweight or mildly obese subjects were enrolled upon screening to confirm fasting cholesterol level at or above 220 mg/dL (5.7 mmol/dL). After giving informed consent, subjects were randomized to consume PTE (3 g/day) or placebo for 20 weeks. At baseline and at 4-week intervals, blood lipids, C-reactive protein, and fasting blood glucose were evaluated. A dual-energy X-ray absorptiometry scan was performed at baseline and at study exit to evaluate changes to body composition. Appetite and physical and mental energy were scored at each visit using visual analog scales (0–100).
Results: Consumption of PTE was associated with statistically significant weight loss when compared to placebo (P<0.05). Fat loss was seen for arms, legs, and the gynoid region (hip/belly), as well as for total fat mass. The fat reduction reached significance on within-group analysis, but did not reach between-group significance. Consumption of PTE was associated with improvements to lipid profile, including a mild reduction in cholesterol and the cholesterol:high-density lipoprotein ratio after only 4 weeks, as well as a reduction in triglycerides and very small-density lipoproteins, where average blood levels reached normal range at 8 weeks and remained within normal range for the duration of the study (P<0.08). No significant changes between the PTE group and the placebo group were seen for fasting glucose or C-reactive protein. A transient reduction in appetite was seen in the PTE group when compared to placebo (P<0.1).
Conclusion: The results from this clinical study showed that the daily consumption of PTE was associated with significant weight loss, reduced body mass index, and an improved lipid profile.
The Nopal cactus grows widespread in many parts of the world, and in some areas is considered a noxious weed. In addition to the use of the stems and leaves (also called pads or cladodes) and fruits (prickly pears) as food, Nopal has a long use in traditional folk medicine. The fruit shares some constituents with the stems, and contains additional unique polyphenols and betalain pigments with antioxidant and anti-inflammatory activities. A randomized, double-blind, controlled, parallel-arm human study was performed to compare consumption of Nopal fruit juice (NFJ) to control (apricot) juice. Serum C-reactive protein (CRP) and uric acid were measured at baseline and after 8 and 12 weeks. Reduced CRP levels were seen in both the NFJ and control groups. The 21.0% reduction at 8 weeks in the NFJ group was significantly more robust than the 6.6% reduction in the control group (P <.05). The reduced CRP level within the NFJ group was highly significant at 8 weeks (P <.0001), and remained reduced by 10.6% at 12 weeks compared to baseline (P <.065). The CRP reduction was not significant within the control group. There was no significant difference in uric acid levels between the groups at either 8 or 12 weeks. The 5.2% increase in uric acid levels within the control group was highly significant at 8 weeks (P <.0003), remaining significant at 12 weeks (P <.04). In contrast, a transient 3.6% increase in uric acid in the NFJ group at 8 weeks (P <.02) returned almost back to baseline levels at 12 weeks. Consumption of NFJ was associated with significant reduction in the CRP inflammatory biomarker while maintaining uric acid well within healthy range.
Objective: Aloe has been used for the treatments of various ailments dating back almost 6000 years. There are more than 450 species of aloe coming from various parts of Africa and South America, and from the island of Madagascar that contains unique species endemic to the island. One such species is Aloe macroclada that has been used for centuries by the local residents as a remedy for a wide variety of ailments. We investigated whether the mechanism of action behind the wide-ranging health benefits of A. macroclada could be mobilization of bone marrow stem cells.
Methods: A. macroclada was prepared into small spherical pellets by Malagasy healers using traditional methods of fabrication. The traditional dose of three pellets was fed to 4 volunteers and the number of circulating stem cells was quantified 1, 2 and 3 hours after consumption using flow-cytometry.
Results: The usual dose and preparation of A. macroclada traditionally used in Madagascar triggered a significant increase (up to 53%) in the number of circulating CD45dim CD34+ and CD34+ CD133+ stem cells within 2 hours of consumption. This increase lasted more than 3 hours and was significant after 120 and 180 minutes of consumption.
Conclusion: Consumption of A. macroclada has been credited with significant improvements in a wide variety of heath conditions. This data suggest that stem cell mobilization may be an important mechanism of action behind the health benefits of A. macroclada.
Objective: The goal for this study was to evaluate the effects of an Aloe vera-based Nerium oleander extract (NAE-8®), compared to an extract of A. vera gel alone (ALOE), and to an aqueous extract of N. oleander (AQ-NOE) in bioassays pertaining to dermatologic potential with respect to antioxidant protection, anti-inflammatory effects, and cytokine profiles in vitro.
Methods: Cellular antioxidant protection was evaluated in three separate bioassays: The cellular antioxidant protection of erythrocytes (CAP-e) assay, protection of cellular viability and prevention of apoptosis, and protection of intracellular reduced glutathione levels, where the last two assays were performed using human primary dermal fibroblasts. Reduction of intracellular formation of reactive oxygen species (ROS) was tested using polymorphonuclear cells in the absence and presence of oxidative stress. Changes to cytokine and chemokine profiles when whole blood cells and human primary dermal fibroblasts were exposed to test products were determined using a 40-plex Luminex array as a method for exploring the potential cross-talk between circulating and skin-resident cells.
Results: The NAE-8® provided significantly better antioxidant protection in the CAP-e bioassay than AQ-NOE. NAE-8® and AQ-NOE both protected cellular viability and intracellular reduced glutathione, and reduced the ROS formation significantly when compared to control cells, both under inflamed and neutral culture conditions. ALOE showed minimal effect in these bioassays. In contrast to the NAE-8®, the AQ-NOE showed induction of inflammation in the whole blood cultures, as evidenced by the high induction of CD69 expression and secretion of a number of inflammatory cytokines. The treatment of dermal fibroblasts with NAE-8® resulted in selective secretion of cytokines involved in collagen and hyaluronan production as well as re-epithelialization during wound healing.
Conclusion: NAE-8®, a novel component of a commercial cosmetic product, showed beneficial antioxidant protection in several cellular models, without the induction of leukocyte activation and secretion of inflammatory cytokines. The biological efficacy of NAE-8® was unique from both ALOE and AQ-NOE.
The goal for this work was to characterize basic biological properties of a novel Arthrospira platensis-based aqueous cyanophyta extract (ACE), enriched in the known anti-inflammatory cyclooxygenase-2 (COX-2) inhibitor phycocyanin (PC), but also containing a high level of non-PC bioactive compounds. Antioxidant properties were tested in parallel in the Folin-Ciocalteu assay (chemical antioxidant capacity) and in the cellular antioxidantprotection (CAP-e) bioassay, where both the PC and the non-PC fractions contributed to the antioxidant capacity and CAP of ACE. In contrast to the COX-2 inhibition seen in the presence of PC, the inhibition of enzymatic activity of the inflammatory mediator Lipoxygenase was associated specifically with the non-PC fraction of ACE. Inhibition of formation of reactive oxygen species (ROS) was evaluated using polymorphonuclear cells from healthy human donors. The inhibition of ROS formation was seen for both the PC and non-PC fractions, with ACE showing the most robust effect. The effects of PC, non-PC, and ACE on clotting and clot lysing was tested using a modified Euglobulin fibrinolytic assay in vitro. In the presence of PC, non-PC, and ACE, the time for clot formation and lysis was not affected; however, the clots were significantly more robust. This effect was statistically significant (p<.05) at doses between 125-500 μg/mL, and returned to baseline at lower doses. Both PC and the non-PC fraction contributed to the antioxidant properties and anti-inflammatory effects, without a negative impact on blood clotting in vitro. This suggests a potential benefit for the consumable ACE extract in assisting the reduction of inflammatory conditions.