Rodney Alexandre Ferreira Rodrigues

Evaluation of wound healing properties of Arrabidaea chica Verlot extract.

ETHNOPHARMACOLOGICAL RELEVANCE Arrabidaea chica Verlot. (Bignoniaceae), popularly known as Crajiru, has been traditionally used as wound healing agent. AIM OF THE STUDY Investigate in vitro and in vivo healing properties of Arrabidaea chica leaves extract (AC). MATERIALS AND METHODS AC was evaluated in vitro in fibroblast growth stimulation (0.25-250 microg/mL) and collagen production stimulation (250 microg/mL) assays. Allantoin (0.25-250 microg/mL) and vitamin C (25 microg/mL) were used as controls respectively. DPPH and Folin-Ciocalteau assays were used for antioxidant evaluation, using trolox (0.25-250 microg/mL) as reference antioxidant. To study wound healing properties in rats, AC (100mg/mL, 200 microL/wound/day) was topically administered during 10 days and wound area was evaluated every day. Allantoin (100mg/mL, 200 microL/wound/day) was used as standard drug. After treatment, wound sites were removed for histopathological analysis and total collagen determination. RESULTS AC stimulated fibroblast growth in a concentration dependent way (EC50=30 microg/mL), increased in vitro collagen production and demonstrated moderate antioxidant capacity. In vivo, AC reduced wound size in 96%, whereas saline group showed only 36% wound healing. CONCLUSION AC efficiency seems to involve fibroblast growing stimulus and collagen synthesis both in vitro and in vivo, beyond moderate scavenging activity, corroborating Crajiru folk use.

Liposomes incorporating essential oil of Brazilian cherry (Eugenia uniflora L.): Characterization of aqueous dispersions and lyophilized formulations

Multilamellar liposomes incorporating essential oil of Brazilian cherry (Eugenia uniflora L.) leaves were produced by dry film hydration. Gas chromatography demonstrated the compounds found in the essential oil were effectively incorporated in the aqueous dispersions of liposomes. Differential scanning calorimetry analyses revealed the incorporation of the essential oil did not cause phase separation in the membrane structure; the gel-liquid crystalline transition temperature (main transition) remained the same despite the higher heterogeneity indicated by the transition peak broadening. Different cryoprotectors (sucrose and trehalose) were added to the liposomal formulations to be tested in their ability to protect the liposomal structure during the lyophilization. The morphological aspect of the lyophilized powders analysed by scanning electron microscopy showed significant differences among the samples with and without cryoprotectors. Fourier-transform infrared spectroscopy indicated the cryoprotectors interacted effectively with the polar heads of phospholipids in the bilayer. In terms of water absorption, trehalose was identified as a much more effective protector agent against it than sucrose. The cryoprotectors showed different degrees of effectiveness of preservation of the liposomal structure when the rehydration assays of lyophilized liposomes were carried out, as particle size measurements indicated a moderate process of fusion when the formulations with sucrose were rehydrated.

Furanoditerpenes from Pterodon pubescens benth with selective in vitro anticancer activity for prostate cell line

Activity guided fractionation of Pterodon pubescens Benth. methylene chloride-soluble fraction afforded novel 6α-acetoxi 7β-hydroxy-vouacapan 1 and four known diterpene furans 2, 3, 4, 5. The compounds were evaluated for in vitro cytotoxic activities against human normal cells and tumour cell lines UACC-62 (melanoma), MCF-7 (breast), NCI-H460 (lung, non-small cells), OVCAR-03 (ovarian), PC-3 (prostate), HT-29 (colon), 786-0 (renal), K562 (leukemia) and NCI-ADR/RES (ovarian expressing phenotype multiple drugs resistance). Results were expressed by three concentration dependent parameters GI50 (concentration that produces 50% growth inhibition), TGI (concentration that produces total growth inhibition or cytostatic effect) and LC50 (concentration that produces -50% growth, a cytotoxicity parameter). Also, in vitro cytotoxicity was evaluated against 3T3 cell line (mouse embryonic fibroblasts). Antiproliferative properties of compounds 1, 4 and 5 are herein reported for the first time. These compounds showed selectivity in a concentration-dependent way against human PC-3. Compound 1 demonstrated selectivity 26 fold more potent than the positive control, doxorubicin, for PC-3 (prostrate) cell line based on GI50 values, causing cytostatic effect (TGI value) at a concentration fifteen times less than positive control. Moreover comparison of 50% lethal concentration (LC50 value) with positive control (doxorubicin) suggested that compound 1 was less toxic.

Structural and biochemical alterations during the healing process of tendons treated with Aloe vera

AIMS The tendon is composed of highly organized collagen fibers that form a complex supramolecular structure. After lesions, the organization and composition of the tendon are not completely restored. Our purpose was to evaluate if the application of Aloe vera improves tendon healing, considering the effectiveness in the stimulus of collagen synthesis. MAIN METHODS The calcaneal tendon of male Wistar rats was partially transected with subsequent topical application of A. vera ointment at the injury. The animals were separated into groups with tendons treated with the A. vera extract for 7days and excised on the 7th, 14th and 21st days after surgery; control rats received only ointment base without plant extract. KEY FINDINGS Morphological analysis using polarization microscopy showed that the entire tendon undergoes a remodeling process, with disorganized collagen fibers by days 7 and 14 in plant-treated and non-treated groups and with a higher birefringence in tendons of the plant-treated group on the 21st day. A higher concentration of hydroxyproline was found in plant-treated tendons on days 7 and 14 compared with their controls. Western blots showed lower amounts of type I collagen in the plant-treated group on day 14 compared with the control. MMP-9 diminished 14days after lesion and the active isoform of MMP-2 increased on day 21 in plant-treated groups. SIGNIFICANCE The present study indicates a beneficial effect of A. vera in the tissue reorganization in the transected region of the tendon 21days after injury and is supported by an increase of active MMP-2.

Geranylgeraniol and 6α,7β-dihydroxyvouacapan-17β-oate methyl ester isolated from Pterodon pubescens Benth.: Further investigation on the antinociceptive mechanisms of action.

The crude alcoholic extracts obtained from Pterodon pubescens Benth. seeds are widely used in Brazilian folk medicine as anti-inflammatory, analgesic, anti-rheumatic tonics and depurative preparations. We previously demonstrated the antinociceptive activity on writhing capsaicin, glutamate, and hot-plate tests of two compounds isolated from P. pubescens: geranylgeraniol (C1) and 6α,7β-dihydroxyvouacapan-17β-oate methyl ester (C2). This work is a continuation of the previous study investigating the possible mechanisms of action for compounds C1 and C2, and the differences between them. The present study demonstrated that when administered intraperitoneally (i.p.): i), compounds C1 and C2 produced significant anti-allodynic activity during the acute phase of the Complete Freunds Adjuvant (CFA)-induced persistent pain model; ii) compound C1 produced significant anti-hypernociception activity in the carrageenan-induced pain model; iii) compound C2 presented a significant loss of activity after p-chlorophenylalanine methyl ester hydrochloride (PCPA) [5-HT synthesis inhibitor] treatment, suggesting that the mechanisms of action could be related to either the synthesis or release of serotonin; iv) compound C1 presented a significant loss of activity after ondansetron (5-HT(3) receptor antagonist) treatment suggesting activity upon 5-HT(3) serotonin receptors; v) compound C1 presented a significant loss of activity after efaroxan (mixed I(1) imidazoline/α(2)-adrenoceptor antagonist) treatment suggesting the participation of this compound upon imidazoline I(1) receptors; and vi) both compounds C1 and C2 did not appear to exert their activity via 5-HT(1A), 5-HT(2A), imidazoline I(2), α(2)-adrenoceptor, nitric oxide, GABA(A), acetylcholine muscarinic, and nicotinic receptors when evaluated in acetic acid-induced nociception.

Otimização do processo de extração e isolamento do antimalárico artemisinina a partir de Artemisia annua L.

Malaria is still one of the major diseases in the world, causing physical and economic problems in tropical regions. Artemisinin (Qinghaosu), a natural compound identified in Artemisia annua L. , is an effective drug mainly against cerebral malaria. The action of this drug is immediate and parasitaemia in the treatment of drug-resistant malaria is rapidily reduced, justifying the industrial production of artemisinin. This article focuses on the industrial production of this potent antimalarial drug, including strategies for enhancing yield using inexpensive and easy steps.

Desenvolvimento e validação de metodologia analítica por CLAE-IR para determinação de artemisinina em Artemisia annua L

The aim of this work was to develop and validate an analytical methodology for determination of artemisinin used as antimalaric. The method was based on high performace liquid chromatography, using a CN column with mobile phase composed of methanol : H2O 50:50 (V/V). The results showed that the method presented linearity from 50 to 1500 µg/mL. It was considered selective, accurate, precise according to the specific resolution from ANVISA, the Brazilian regulatory agency.

In vitro activity of Artemisia annua L (Asteraceae) extracts against Rhipicephalus (Boophilus) microplus.

The activity of plant extracts on parasites may indicate groups of substances that are potentially useful for controlling Rhipicephalus (Boophilus) microplus. The aim of the present study was to investigate the in vitro action of Artemisia annua extracts on this tick. The concentrations of the sesquiterpene lactones artemisinin and deoxyartemisinin present in plant extracts were quantified via high-performance liquid chromatography. Four extracts produced from the concentrated crude extract (CCE) were evaluated on larvae using the impregnated paper method, with readings after 24 hours of incubation. The engorged females were immersed in the CCE and in its four derived extracts for five minutes, with incubation for subsequent analysis of biological parameters. The extracts were not effective on the larvae at the concentrations tested (3.1 to 50 mg.mL(-1)). The CCE showed greater efficacy on engorged females (EC(50) of 130.6 mg.mL(-1) and EC(90) of 302.9 mg.mL(-1)) than did the derived extracts. These results tend to confirm that the action of artemisinin on engorged females of R. (B.) microplus is conditional to their blood intake. In this case, in vitro methods would be inadequate for effective evaluation of the action of A. annua on R. (B.) microplus.

Anticancer and Anti-Inflammatory Activities of a Standardized Dichloromethane Extract from Piper umbellatum L. Leaves.

Despite the advances in anticancer drug discovery field, the worldwide cancer incidence is remarkable, highlighting the need for new therapies focusing on both cancer cell and its microenvironment. The tumor microenvironment offers multiple targets for cancer therapy, including inflammation. Nowadays, almost 75% of the anticancer agents used in chemotherapy are derived from natural products, and plants are an important source of new promising therapies. Continuing our research on Piper umbellatum species, here we describe the anticancer (in vitro antiproliferative activity and in vivo Ehrlich solid tumor model) and anti-inflammatory (carrageenan-induced paw edema and peritonitis models) activities of a standardized dichloromethane extract (SDE) from P. umbellatum leaves, containing 23.9% of 4-nerolidylcatechol. SDE showed in vitro and in vivo antiproliferative activity, reducing Ehrlich solid tumor growth by 38.7 and 52.2% when doses of 200 and 400 mg/kg, respectively, were administered daily by oral route. Daily treatments did not produce signals of toxicity. SDE also reduced paw edema and leukocyte migration on carrageenan-induced inflammation models, suggesting that the anticancer activity of SDE from Piper umbellatum leaves could involve antiproliferative and anti-inflammatory effects. These findings highlight P. umbellatum as a source of compounds against cancer and inflammation.

Effect of Calendula Officinalis Cream on Achilles Tendon Healing

In recent years, the scientific community has undertaken research on plant extracts, searching for compounds with pharmacological activities that can be used in diverse fields of medicine. Calendula officinalis L. is known to have antioxidant, anti‐inflammatory, antibacterial, and wound healing properties when used to treat skin burns. Therefore, the purpose of this study was to analyze the effects of C. officinalis on the initial phase of Achilles tendon healing. Wistar rats were separated in three groups: Calendula (Cal)—rats with a transected tendon were treated with topical applications of C. officinalis cream and then euthanized 7 days after injury; Control (C)—rats were treated with only vehicle after transection; and Normal (N)—rats without tenotomy. Higher concentrations of hydroxyproline (an indicator of total collagen) and non‐collagenous proteins were observed in the Cal group in relation to the C group. Zymography showed no difference in the amount of the isoforms of metalloproteinase‐2 and of metalloproteinase‐9, between C and Cal groups. Polarization microscopy images analysis showed that the Cal group presented a slightly higher birefringence compared with the C group. In sections of tendons stained with toluidine blue, the transected groups presented higher metachromasy as compared with the N group. Immunocytochemistry analysis for chondroitin‐6‐sulfate showed no difference between the C and Cal groups. In conclusion, the topical application of C. officinalis after tendon transection increases the concentrations of collagen and non‐collagenous proteins, as well as the collagen organization in the initial phase of healing. Anat Rec, 298:428–435, 2015.