Ethyl gallate isolated from phenol-enriched fraction of Caesalpinia mimosoides Lam. Promotes cutaneous wound healing: a scientific validation through bioassay-guided fractionation.

The tender shoots of Caesalpinia mimosoides Lam. are used ethnomedically by the traditional healers of Uttara Kannada district, Karnataka (India) for the treatment of wounds. The current study was aimed at exploring phenol-enriched fraction (PEF) of crude ethanol extract of tender shoots to isolate and characterize the most active bio-constituent through bioassay-guided fractionation procedure. The successive fractionation and sub-fractionation of PEF, followed by in vitro scratch wound, antimicrobial, and antioxidant activities, yielded a highly active natural antioxidant compound ethyl gallate (EG). In vitro wound healing potentiality of EG was evidenced by a significantly higher percentage of cell migration in L929 fibroblast cells (97.98 ± 0.46% at 3.81 µg/ml concentration) compared to a positive control group (98.44 ± 0.36%) at the 48th hour of incubation. A significantly higher rate of wound contraction (98.72 ± 0.41%), an elevated tensile strength of the incised wound (1,154.60 ± 1.42 g/mm2), and increased quantity of connective tissue elements were observed in the granulation tissues of the 1% EG ointment treated animal group on the 15th post-wounding day. The accelerated wound healing activity of 1% EG was also exhibited by histopathological examinations through Hematoxylin and Eosin, Masson's trichome, and Toluidine blue-stained sections. Significant up-regulation of enzymatic and non-enzymatic antioxidant contents (reduced glutathione, superoxide dismutase, and catalase) and down-regulation of oxidative stress marker (lipid peroxidation) clearly indicates the effective granular antioxidant activity of 1% EG in preventing oxidative damage to the skin tissues. Further, in vitro antimicrobial and antioxidant activities of EG supports the positive correlation with its enhanced wound-healing activity. Moreover, molecular docking and dynamics for 100 ns revealed the stable binding of EG with cyclooxygenase-2 (-6.2 kcal/mol) and matrix metalloproteinase-9 (-4.6 kcal/mol) and unstable binding with tumor necrosis factor-α (-7.2 kcal/mol), suggesting the potential applicability of EG in inflammation and wound treatment.

Attenuation of dermal wounds through topical application of ointment containing phenol enriched fraction of Caesalpinia mimosoides Lam.

Caesalpinia mimosoides Lam. is one of the important medicinal plants used by the traditional healers of Uttara Kannada district, Karnataka (India) for treating wounds. In our previous study ethanol extract of the plant was evaluated for its wound healing activity. In continuation, the present study was aimed to evaluate the phenol enriched fraction (PEF) of ethanol extract for wound healing activity along with its antioxidant, anti-inflammatory and antimicrobial properties. The potent wound healing activity of PEF was evidenced by observation of increased rate of cell migration in L929, 3T3L1 and L6 cells (92.59 ± 1.53%, 98.42 ± 0.82% and 96.63 ± 0.61% respectively) at 7.81 µg/ml doses in assays carried out in vitro. Significantly enhanced rate of wound contraction (97.92 ± 0.41%), tensile strength (973.67 ± 4.43 g/mm2), hydroxyproline (31.31 ± 0.64 mg/g) and hexosamine (8.30 ± 0.47 mg/g) contents were observed on 15th post wounding day in 5% PEF treated animals. The enzymatic and non-enzymatic cellular antioxidants (superoxide dismutase, catalase and reduced glutathione) were upregulated (15.89 ± 0.17 U/mg, 48.30 ± 4.60 U/mg and 4.04 ± 0.12 µg/g respectively) with the administration of 5% PEF. The significant antimicrobial, antioxidant and anti-inflammatory activities support the positive correlation of PEF with its enhanced wound healing activity. PEF contains expressive amounts of total phenolic and total flavonoid contents (578.28 ± 2.30 mg GAE/g and 270.76 ± 2.52 mg QE/g). Of the various chemicals displayed in RP-UFLC-DAD analysis of PEF, gallic acid (68.08 µg/mg) and ethyl gallate (255.91 µg/mg) were predominant. The results indicate that PEF has great potential for the topical management of open wounds.

Evaluation of wound healing property of Caesalpinia mimosoides Lam.

ETHNOPHARMACOLOGICAL RELEVANCE: Caesalpinia mimosoides Lam. is one of the important traditional folk medicinal plants in the treatment of skin diseases and wounds used by healers of Uttara Kannada district of Karnataka state (India). However scientific validation of documented traditional knowledge related to medicinal plants is an important path in current scenario to fulfill the increasing demand of herbal medicine. AIM OF THE STUDY: The study was carried out to evaluate the claimed uses of Caesalpinia mimosoides using antimicrobial, wound healing and antioxidant activities followed by detection of possible active bio-constituents. MATERIALS AND METHODS: Extracts prepared by hot percolation method were subjected to preliminary phytochemical analysis followed by antimicrobial activity using MIC assay. In vivo wound healing activity was evaluated by circular excision and linear incision wound models. The extract with significant antimicrobial and wound healing activity was investigated for antioxidant capacity using DPPH, nitric oxide, antilipid peroxidation and total antioxidant activity methods. Total phenolic and flavonoid contents were also determined by Folin-Ciocalteu, Swain and Hillis methods. Possible bio-active constituents were identified by GC-MS technique. RP-UFLC-DAD analysis was carried out to quantify ethyl gallate and gallic acid in the plant extract. RESULTS: Preliminary phytochemical analysis showed positive results for ethanol and aqueous extracts for all the chemical constituents. The ethanol extract proved potent antimicrobial activity against both bacterial and fungal skin pathogens compared to other extracts. The efficacy of topical application of potent ethanol extract and traditionally used aqueous extracts was evidenced by the complete re-epithelization of the epidermal layer with increased percentage of wound contraction in a shorter period. However, aqueous extract failed to perform a consistent effect in the histopathological assessment. Ethanol extract showed effective scavenging activity against DPPH and nitric oxide free radicals with an expressive amount of phenolic and moderate concentration of flavonoid contents. Ethyl gallate and gallic acid were found to be the probable bio-active compounds evidenced by GCMS and RP-UFLC-DAD analysis. CONCLUSION: The study revealed the significant antimicrobial, wound healing and antioxidant activities of tender parts of C. mimosoides and proved the traditional folklore knowledge.

Ethnomedicinal plants to cure skin diseases-an account of the traditional knowledge in the coastal parts of Central Western Ghats, Karnataka, India.

ETHNOPHARMACOLOGICAL RELEVANCE: Documentation of ethnomedicinal knowledge pertaining to the treatment of different types of skin diseases from the Central Western Ghats of India, a rich habitat of different ethnic communities. MATERIALS AND METHODS: Frequent field surveys were carried out to invent the 'key informants' in the treatment of skin diseases in the study area. The information was collected through semi-structured open ended interviews with questionnaire in their local Kannada language. All medicinal plants recorded for the treatment of skin diseases were photographed in the field; voucher specimens were made subsequently and are deposited in the Herbarium, P.G. Department of Botany, Karnatak University, Dharwad. The information such as botanical name, status, family, vernacular name, habit and habitat, analysis like percentage of parts used, percentage of drug preparations, use value (UV), informants consensus factor (ICF), fidelity level (FL) and correlation between UV and use mention (Np) of the plants are provided. RESULTS: In all, 48 informants were interviewed. Amongst which 38 were the 'key informants' who gave the information exclusively about the treatment of skin diseases. Among 102 plant species collected, seven species are endemic to India and eleven species have their nativity outside India. Twelve species could be considered as new claims for skin diseases as their use has not been mentioned in Ayurveda or any other research articles surveyed. Of all the drug formulations, paste is the most preferred method (50%) followed by oil extraction (18.89%), juice (14.44%), ash (4.44%) etc. The highest UV is for Pongamia pinnata, Naregamia alata, Randia dumetorum and Girardinia diversifolia (1.50 each). The treatment for different types of skin diseases by the herbal healers are classified into 13 categories, out of which ringworm scored the highest ICF value. Similarly, the 100% FL value scored was in the order of 10 plants for boils, 4 plants for different types of sore, 2 plants for ringworm, intertrigo, eczema and pruritus respectively. As Caesalpinia mimosoides and Basella alba in the treatment of boils, Hygrophila schulli for inter trigo, Cissus discolor for ringworm, Mammea suriga for eczema scored high FL and high Np value, they can be considered as important species. CONCLUSIONS: The documentation and data analysis of the ethnomedicinal knowledge in the coastal regions of Uttara Kannada district of Central Western Ghats have provided the information about important plants in the treatment of different types of skin diseases. Further scientific analysis of such plants may provide novel compounds for the treatment of skin diseases.

Ethnomedicinal practices in different communities of Uttara Kannada district of Karnataka for treatment of wounds.

ETHNOPHARMACOLOGICAL RELEVANCE: Uttara Kannada district is located in the heart of the Western Ghats, one of the biodiversity hotspots, in Karnataka state of India. The thick evergreen forests are home to several ethnic communities. The study was under taken for documentation and analysis of ethnomedicinal plants in the treatment of wounds. MATERIALS AND METHODS: Field trips were made in Uttara Kannada district to identify the key informants. The collection of information was through semi-structured open ended interviews with a questionnaire. The questionnaire was designed to obtain the information about their experience in the field of treatment, number of patients treated per week, knowledge about the medicinal plants, vernacular names, parts of the plants used and other ingredients added during the drug formulations. Plants mentioned for treatment were photographed in the field, cuttings of the samples were taken and voucher specimens are deposited in the herbarium of P.G. Department of Botany, Karnatak University, Dharwad. The information such as botanical name, status, family, vernacular name habit and habitat, statistical analysis like percentage of parts used, Use value (UV) and Informants Consensus Factor (ICF) are provided. RESULTS: Present study resulted in recording 106 medicinal plant species of 55 families and 86 formulations to treat different types of wounds by 44 key informants. Among the 106 plants recorded four species are endemic to India and 22 species have the nativity outside India. Rest of the species have nativity both in India and elsewhere. The highest number of species belonged to Apocynaceae and Rubiaceae (6 species each). Trees are used more often (35.84%), followed by herbs (28.30%), shrubs (23.58%), climbers (11.32) and parasites (0.80%). Leaves are the major part of the plants used in the formulations (28.57%). The highest Use value is for Calycopteris floribunda (1.80), followed by Rauvolfia serpentina and Achyranthes aspera (1.67). The different types of wounds treated by traditional healers are classified into 15 categories and the highest ICF scored is for the burns (0.66). CONCLUSIONS: Ethnomedicinal survey in Uttara Kannada district of Karnataka revealed uses of 106 plants in traditional practices for curing various types of wounds. The statistical analysis confirmed high degree of sharing the knowledge amongst 44 key informants. Information about the largest number of remedies was obtained from the Havyaka Brahmin ethnic community which has strong Sanskrit background.