Fabrication and evaluation of a halal medicated hydrogel film from standardized Centella Asiatica for wound healing

Ethnopharmacological relevance: Asiaticoside is claimed as a bioactive compound capable of wound healing. In order to ensure that the pharmacological activity of the extract is traceable and measurable, the present study attempted to evaluate the bioactivity of rich fractionated extract of asiaticos...

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Main Authors: Abdul Aziz, Huwaida, Bakhtiar, M. Taher, Mandal, Uttam Kumar, Md. Jaffri, Juliana, Wan Sulaiman, Wan Mohd. Azizi, Darnis, Deny Susanti
Format: Monograph
Language:English
Published: 2016
Subjects:
Online Access:http://irep.iium.edu.my/53195/
http://irep.iium.edu.my/53195/1/53195_FABRICATION%20AND%20EVALUATION%20OF%20A%20HALAL.pdf
_version_ 1848784194235269120
author Abdul Aziz, Huwaida
Bakhtiar, M. Taher
Mandal, Uttam Kumar
Md. Jaffri, Juliana
Wan Sulaiman, Wan Mohd. Azizi
Darnis, Deny Susanti
author_facet Abdul Aziz, Huwaida
Bakhtiar, M. Taher
Mandal, Uttam Kumar
Md. Jaffri, Juliana
Wan Sulaiman, Wan Mohd. Azizi
Darnis, Deny Susanti
author_sort Abdul Aziz, Huwaida
building IIUM Repository
collection Online Access
description Ethnopharmacological relevance: Asiaticoside is claimed as a bioactive compound capable of wound healing. In order to ensure that the pharmacological activity of the extract is traceable and measurable, the present study attempted to evaluate the bioactivity of rich fractionated extract of asiaticoside. Aim of the study: The current study evaluates the wound healing efficacy via in vitro scratch assay and in vivo circular wound excision model. Materials and methods: The ethanol extract was fractionated into seven fractions via vacuum liquid chromatography. The compound of interest in the fractions was qualitatively identified using thin layer chromatography and the positive fraction containing asiaticoside was further quantified using reverse-phase HPLC. The asiaticoside rich fraction was subjected to: i) colorimetric MTT (methylthiazoltetrazolium) cytotoxicity assay following incubation with human dermal fibroblast (HDF) and human dermal keratinocyte (HaCaT); ii) in vitro 12-well plate scratch assay (using HDF and HaCaT cells); iii) topically apply(40, 10, and 2.5%, w/w) on in vivo circular wound excision of rabbits. Data on wound contraction, epithelization period, hydroxyproline content and histophatological analysis was collected from in vivo study. Results: The result showed that the methanol fraction of extract contained about 2.4% of asiaticoside. Based on the results of colorimetric MTT (methylthiazoltetrazolium) cytotoxicity assay, both HDF and HaCaT showed significant stimulation upon application of the methanolic fraction of extract at concentrations of 100 µg/mL and 0.19 µg/mL. The methanol fraction showed almost no toxicity effect at the concentrations tested since their IC50 could not be determined in concentrations ranging from 100 µg/mL to 0.19 µg/mL. Since all the concentrations tested allowed for more than 90% cell viability, the concentrations chosen for the scratch assay were randomly chosen and designated as highest (100 µg/mL), medium (6 µg/mL), and lowest (0.2 µg/mL) concentrations. In the scratch assay, methanol fraction of extract with concentration of 0.2 µg/mL and 100 µg/mL showed significant effect on HDF and HaCaT compared to the positive control (p< 0.05). In vivo, it was shown that the methanol fraction of the extract induced collagen synthesis. Histopathology data also concluded that dose-dependent effect of the tested extract as a wound healer was present. Conclusions: Taken together, recent findings suggest that methanol fraction of C. asiatica demonstrated remarkable polyvalent activity, thus has potential as an effective wound healer. In conclusion, the claim on the presence of wound healing properties in C. asiatica had been well supported based on the results obtained in this study.
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institution International Islamic University Malaysia
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spelling iium-531952018-03-01T08:41:48Z http://irep.iium.edu.my/53195/ Fabrication and evaluation of a halal medicated hydrogel film from standardized Centella Asiatica for wound healing Abdul Aziz, Huwaida Bakhtiar, M. Taher Mandal, Uttam Kumar Md. Jaffri, Juliana Wan Sulaiman, Wan Mohd. Azizi Darnis, Deny Susanti RS Pharmacy and materia medica Ethnopharmacological relevance: Asiaticoside is claimed as a bioactive compound capable of wound healing. In order to ensure that the pharmacological activity of the extract is traceable and measurable, the present study attempted to evaluate the bioactivity of rich fractionated extract of asiaticoside. Aim of the study: The current study evaluates the wound healing efficacy via in vitro scratch assay and in vivo circular wound excision model. Materials and methods: The ethanol extract was fractionated into seven fractions via vacuum liquid chromatography. The compound of interest in the fractions was qualitatively identified using thin layer chromatography and the positive fraction containing asiaticoside was further quantified using reverse-phase HPLC. The asiaticoside rich fraction was subjected to: i) colorimetric MTT (methylthiazoltetrazolium) cytotoxicity assay following incubation with human dermal fibroblast (HDF) and human dermal keratinocyte (HaCaT); ii) in vitro 12-well plate scratch assay (using HDF and HaCaT cells); iii) topically apply(40, 10, and 2.5%, w/w) on in vivo circular wound excision of rabbits. Data on wound contraction, epithelization period, hydroxyproline content and histophatological analysis was collected from in vivo study. Results: The result showed that the methanol fraction of extract contained about 2.4% of asiaticoside. Based on the results of colorimetric MTT (methylthiazoltetrazolium) cytotoxicity assay, both HDF and HaCaT showed significant stimulation upon application of the methanolic fraction of extract at concentrations of 100 µg/mL and 0.19 µg/mL. The methanol fraction showed almost no toxicity effect at the concentrations tested since their IC50 could not be determined in concentrations ranging from 100 µg/mL to 0.19 µg/mL. Since all the concentrations tested allowed for more than 90% cell viability, the concentrations chosen for the scratch assay were randomly chosen and designated as highest (100 µg/mL), medium (6 µg/mL), and lowest (0.2 µg/mL) concentrations. In the scratch assay, methanol fraction of extract with concentration of 0.2 µg/mL and 100 µg/mL showed significant effect on HDF and HaCaT compared to the positive control (p< 0.05). In vivo, it was shown that the methanol fraction of the extract induced collagen synthesis. Histopathology data also concluded that dose-dependent effect of the tested extract as a wound healer was present. Conclusions: Taken together, recent findings suggest that methanol fraction of C. asiatica demonstrated remarkable polyvalent activity, thus has potential as an effective wound healer. In conclusion, the claim on the presence of wound healing properties in C. asiatica had been well supported based on the results obtained in this study. 2016-11-28 Monograph NonPeerReviewed application/pdf en http://irep.iium.edu.my/53195/1/53195_FABRICATION%20AND%20EVALUATION%20OF%20A%20HALAL.pdf Abdul Aziz, Huwaida and Bakhtiar, M. Taher and Mandal, Uttam Kumar and Md. Jaffri, Juliana and Wan Sulaiman, Wan Mohd. Azizi and Darnis, Deny Susanti (2016) Fabrication and evaluation of a halal medicated hydrogel film from standardized Centella Asiatica for wound healing. Project Report. UNSPECIFIED, Kuala Lumpur. (Unpublished)
spellingShingle RS Pharmacy and materia medica
Abdul Aziz, Huwaida
Bakhtiar, M. Taher
Mandal, Uttam Kumar
Md. Jaffri, Juliana
Wan Sulaiman, Wan Mohd. Azizi
Darnis, Deny Susanti
Fabrication and evaluation of a halal medicated hydrogel film from standardized Centella Asiatica for wound healing
title Fabrication and evaluation of a halal medicated hydrogel film from standardized Centella Asiatica for wound healing
title_full Fabrication and evaluation of a halal medicated hydrogel film from standardized Centella Asiatica for wound healing
title_fullStr Fabrication and evaluation of a halal medicated hydrogel film from standardized Centella Asiatica for wound healing
title_full_unstemmed Fabrication and evaluation of a halal medicated hydrogel film from standardized Centella Asiatica for wound healing
title_short Fabrication and evaluation of a halal medicated hydrogel film from standardized Centella Asiatica for wound healing
title_sort fabrication and evaluation of a halal medicated hydrogel film from standardized centella asiatica for wound healing
topic RS Pharmacy and materia medica
url http://irep.iium.edu.my/53195/
http://irep.iium.edu.my/53195/1/53195_FABRICATION%20AND%20EVALUATION%20OF%20A%20HALAL.pdf