scholarly journals Evaluation of Eye Irritation Potential of Aqueous Leaf Extract of Achyranthes aspera by In Vitro and In Vivo Method

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Gajanan Rajpal Deshmukh ◽  
Kuntrapakam Hema Kumar ◽  
Poojari Venkata Suresh Reddy ◽  
Boddapati Srinivasa Rao ◽  
Chirumamilla Venkata Satish Kumar
Keyword(s):  
Leaf Extract ◽  
Chorioallantoic Membrane ◽  
Negative Control ◽  
Eye Irritation ◽  
Achyranthes Aspera ◽  
Hen’S Egg ◽  
Irritation Test ◽  
Aqueous Leaf Extract

The present paper is an attempt to investigate the eye irritation potential of aqueous leaf extract of Achyranthes aspera by in vitro, Hen's Egg Chorioallantoic Membrane Test (HET-CAM) and in vivo acute eye irritation test in rabbits. The irritation score (IS) obtained after treatment of the extract on HET-CAM is 0.07 and that of in rabbits is 0.55, Which does not comes under either category 1 or 2 as per the harmonized integrated classification system. The aqueous extract of Achyranthes aspera showed no eye irritation properties both in vitro and in vivo methods when compared with negative control whereas positive controls showed eye irritation potential.

HET-CAM test for determining the possible eye irritancy of pesticides

2010 ◽  
Vol 58 (3) ◽  
pp. 369-377 ◽  
Author(s):  
Péter Budai ◽  
József Lehel ◽  
Judit Tavaszi ◽  
Éva Kormos
Keyword(s):  
Screening Method ◽  
Chorioallantoic Membrane ◽  
Alternative Methods ◽  
Eye Irritation ◽  
Parallel Data ◽  
Hen’S Egg ◽  
Irritation Test ◽  
Veterinary Products

Agrochemicals and veterinary products must undergo numerous toxicological tests before registration. The use of animals in these studies is a controversial issue. The Draize eye irritation test is one of the most criticised methods because of the injuries inflicted on the test animals. Several in vitro methods have been used to investigate the toxicity of potential eye irritants with a view to replacing in vivo eye irritation testing. One of these alternative methods is the Hen’s Egg Test — Chorioallantoic Membrane (HET-CAM) test. In the present studies comparative screening was performed with a set of agrochemicals to establish parallel data on in vitro (HET-CAM) and in vivo (Draize) results. The examined materials were: Totril (ioxynil), Omite 57 E (propargit), Actellic 50 EC (pyrimiphos-methyl), Stomp 330 EC (pendimethalin), Mospilan 3 EC (acetamiprid), Alirox 80 EC (EPTC), Match 050 EC (lufenuron), Nonit (dioctyl sulphosuccinate sodium), Perenal (haloxyfop-R methyl ester), Pyrinex 48 EC (chlorpyrifos). These experiments showed good correlation between results obtained by the HET-CAM test and those of the Draize rabbit eye test in most cases. The present form of the HET-CAM test can be proposed as a pre-screening method for the determination of eye irritative potential, therefore the number of test animals can be reduced and/or experimental animals can be replaced.

Hen Egg Chorioallantoic Membrane Bioassay: An In Vitro Alternative to Draize Eye Irritation Test for Pesticide Screening

2008 ◽  
Vol 27 (6) ◽  
pp. 449-453 ◽  
Author(s):  
A. S. Kishore ◽  
P. A. Surekha ◽  
P. V. R. Sekhar ◽  
A. Srinivas ◽  
P. Balakrishna Murthy
Keyword(s):  
Chorioallantoic Membrane ◽  
Laboratory Animals ◽  
Test Compound ◽  
Eye Irritation ◽  
Test Chemical ◽  
Draize Test ◽  
Irritation Test ◽  
Hen Egg

As an alternative to the standard Draize eye irritation test, the potential irritancy of compounds was evaluated by observing adverse changes that occur in chorioallantoic membrane CAM) of the hen egg (HECAM) after exposure to a test chemical placed directly on the CAM. The occurrence of hemorrhage, coagulation, and lysis in response to a test compound is the basis for employing this technique to evaluate its potential for in vivo damage to mucous membrane, in particular the eye. Irritancy is scored according to the severity and speed at which damage occurs. In the present study, five different classes of pesticides were screened for irritation potential. There was good correlation between the HECAM assay and the in vivo Draize eye irritation test. The proposed HECAM assay, which reduces the requirement for laboratory animals, could be a painless alternative to the Draize test.

scholarly journals Investigating the ocular toxicity potential and therapeutic efficiency of in situ gel nanoemulsion formulations of brinzolamide

2020 ◽  
Vol 9 (4) ◽  
pp. 578-587
Author(s):  
Sima Talaei ◽  
Mohammad Mehdi Mahboobian ◽  
Mojdeh Mohammadi
Keyword(s):  
Intraocular Pressure ◽  
Membrane Surface ◽  
Chorioallantoic Membrane ◽  
In Situ Gel ◽  
Toxicity Potential ◽  
Vessel Injury ◽  
Hen’S Egg

Abstract Glaucoma is an ocular disease i.e. more common in older adults with elevated intraocular pressure and a serious threat to vision if it is not controlled. Due to the limitations regarding the conventional form of brinzolamide (Azopt®), two optimum formulations of in situ gel nanoemulsion were developed. To ensure the safety and efficacy of developed formulations for ocular drug delivery, the current study was designed. MTT assay was carried out on the human retinal pigmentation epithelial cells. To investigate the irritation potential of the chosen formulations, hen’s egg test-chorioallantoic membrane as a borderline test between in vivo and in vitro methods has been done. The modified Draize method was utilized to evaluate eye tolerance against the selected formulations. Intraocular pressure was measured by applying the prepared formulations to the eyes of normotensive albino rabbits in order to assess the therapeutic efficacy. Based on MTT test, cell viability for NE-2 at 0.1% and NE-1 at 0.1 and 0.5% concentrations was acceptable. The results of the hen’s egg test-chorioallantoic membrane test indicated no sign of vessel injury on the chorioallantoic membrane surface for both formulations. Also, during 24 h, both formulations were well-tolerated by rabbit eyes. The pharmacodynamics effects of formulations had no difference or were even higher than that of suspension in case of adding lower concentration (0.5%) of brinzolamide to the formulations. With regard to the results of the mentioned methods, our advanced formulations were effective, safe, and well-tolerated, thus can be introduced as an appropriate vehicle for ocular delivery of brinzolamide.

scholarly journals GREEN TEA EXTRACT IN AN EYELASH GROWTH ENHANCER GEL FORMULATION: STABILITY TEST, EYE IRRITATION TEST, AND HUMAN EYELASH GROWTH ACTIVITY

2017 ◽  
Vol 10 (6) ◽  
pp. 243
Author(s):  
Indra Syahputra Roes Lie ◽  
Joshita Djajadisastra ◽  
Fadlina Chany Saputri
Keyword(s):  
Green Tea ◽  
Chorioallantoic Membrane ◽  
Test Group ◽  
Green Tea Extract ◽  
Stability Test ◽  
Eye Irritation ◽  
Before And After ◽  
Irritation Test ◽  
Tea Extract

Objective: To formulate a green tea extract (GTE), which is often used as a hair growth product, to produce an eyelash gel with good stability, effectiveness, and safety for growing eyelashes.Methods: GTE was formulated into a gel. A stability test was performed at a high temperature (40±2°C), room temperature (25±2°C), low temperature (4±2°C), and a cycling temperature. An in vitro hen’s egg test-chorioallantoic membrane assay was performed to evaluate potential eye irritation. An eyelash growth test was conducted by length measurement using an eyelash ruler before and after 2 mo of application in human volunteers. Results: The GTE gel was stable in storage at high, room, and low temperatures and at cycling temperatures and did not cause eye irritation. Eyelashes grew significantly more in the test group than in the placebo group after 2 mo of application (p<0.05). Conclusion: GTE gel provides a new, safe, and effective option for growing natural eyelashes.

scholarly journals In vitro Antioxidant and In vivo Hepatocurative and Nephrocurative Activities of Aqueous Leaf Extract of Newbouldia laevis in Albino Rats

2021 ◽  
pp. 114-125
Author(s):  
Mohammed A. Sulaiman ◽  
Mahmoud S. Jada ◽  
Augustine Elizabeth ◽  
Abubakar Umar Modibbo
Keyword(s):  
Oral Administration ◽  
Leaf Extract ◽  
Kidney Diseases ◽  
Albino Rats ◽  
Protein Levels ◽  
Ameliorative Effect ◽  
Liver And Kidney ◽  
Aqueous Leaf Extract

The in vitro antioxidant activity and in vivo hepatocurative and nephrocurative potential of Newbouldia laevis aqueous leaf extract (NLALE) was evaluated. The study used 30 male, albino rats (Rattus norvegicus) weighing 180 ± 20 g, of which 25 were intoxicated by oral administration of a single dose of diclofenac (100 mg/kg b. wt.). Animals were treated by oral administration of silymarin (200 mg/kg b. wt.), furosemide (1.5 mg/kg b. wt.) and NLALE (200 mg/kg and 400 mg/kg b. wt.) for seven consecutive days before animals were sacrificed on the 8th day and serum/plasma was analyzed for biochemical markers of hepatotoxicity and nephrotoxicity. Phytochemical screening of NLALE revealed the presence of alkaloids, flavonoids, glycosides, phenols, saponins, steroids and tannins. The extract scavenged DPPH radical, reduced Fe3+ and inhibited TBARs in comparable manner to ascorbic acid in vitro. NLALE also attenuated diclofenac-induced liver and kidney intoxication as indicated by the significantly (p<0.05) reduced levels of serum biomarkers of hepatotoxicity: ALT, AST, bilirubin, but increased total protein levels and nephrotoxicity: urea, creatinine, Na+ and K+. The observed effects are dose dependent as the 400 mg/kg b. wt. appeared to be more potent than the 200 mg/kg b. wt. dose. It may be concluded from this study that Newbouldia laevis leaf has ameliorative effect against diclofenac-induced hepatotoxicity and nephrotoxicity probably through antioxidative mechanism and the curative claim and the folkloric use of the plant in the treatment of liver and kidney diseases have been scientifically validated

scholarly journals Suppression of inflammatory mediators by aqueous leaf extract of Crotalaria verrucosa: in vivo and in vitro analysis

2020 ◽  
Vol 9 (12) ◽  
pp. 1897
Author(s):  
Mohammad Mustakim Billah ◽  
Abir Huzaifa ◽  
M. Abdul Kader Khan ◽  
Nusrat Jahan Vabna ◽  
Kashfia Nawrin ◽  
...  
Keyword(s):  
Leaf Extract ◽  
Protein Denaturation ◽  
Inflammatory Activity ◽  
Medicinal Value ◽  
Aqueous Leaf Extract ◽  
Anti Inflammatory ◽  
Vitro Analysis ◽  
In Vitro Analysis

Background: Crotalaria verrucosa is a traditional plant frequently prescribed by the tribes for its medicinal value against inflammation. The present study was designed to investigate the scientific basis for medicinal value in inflammation by in vivo and in vitro analysis.Methods: Anti-inflammatory activity of the plant’s leaf was evaluated by two in vivo methods - carrageenan induced rat paw edema and xylene induced mice ear edema. Moreover, in vitro analysis was performed through heat induced hemolysis and heat induced protein denaturation methods.Results: The inflammation produced by carrageenan and xylene were effectively suppressed by the aqueous leaf extract of C. verrucosa (CVAQ) at 600 mg/kg body weight which was comparable to the standards. In heat induced hemolysis test the extract was able to inhibit the lysis up to 70% at 500 µg/ml whereas in heat induced protein denaturation test it reduces the percentage till 69% at the same concentration.Conclusions: The findings suggested that CVAQ possess moderate to high anti-inflammatory activity when applied in low to high concentrated doses. However, the study can only conclude from this basic evaluation that the extract needs to be further investigated for identifying the potential compound which contributed to such medicinal value of the plant.

Validation and Regulatory Acceptance of Alternatives

1999 ◽  
Vol 8 (1) ◽  
pp. 73-79 ◽  
Author(s):  
RICHARD N. HILL ◽  
WILLIAM S. STOKES
Keyword(s):  
Ad Hoc ◽  
Toxicity Testing ◽  
Test Method ◽  
Regulatory Agencies ◽  
Eye Irritation ◽  
Current Test ◽  
Irritation Test ◽  
Test Guideline

For years there was no focus within the U.S. federal government for alternatives to animal toxicity testing. Questions coming to regulatory agencies fell upon individuals to address in the best way they could. Given this void, the ad hoc Interagency Regulatory Alternatives Group was founded by staff in a number of federal agencies in the late 1980s to coalesce efforts in the field. The group sponsored two international workshops on eye irritation, the first making proposals for change in the current test method in rabbits, the second reviewing available data on in vitro alternatives. The result has been that the Organization for Economic Cooperation and Development (OECD) is considering revision of the in vivo eye irritation test guideline to incorporate a number of the workshop deliberations. However, movement of the in vitro eye irritation alternatives has been disappointing; attempts to determine their practical testing significance have thus far been unrewarding.

Recent Progress in the Eye Irritation Test

1993 ◽  
Vol 9 (6) ◽  
pp. 1017-1025 ◽  
Author(s):  
Ih Chu ◽  
Peter Toft
Keyword(s):  
Animal Studies ◽  
In Vitro Tests ◽  
Animal Testing ◽  
Regulatory Requirements ◽  
Eye Irritation ◽  
Significant Development ◽  
In Vivo Test ◽  
Irritation Test

The rabbit eye irritation test based on the Draize method is required for the hazard assessment of chemicals and products that may come into contact with the eye. Due to the potential for the suffering of animals and subjectivity of the test, many modifications of the method have been made that involved a reduction in the number of animals and a refinement of techniques. Additionally, there has been significant development of in vitro alternatives. This paper reviews recent advances in the in vivo test and in vitro alternatives, as well as regulatory requirements. While the refinement of in vivo protocols has resulted in a reduction in the number and discomfort on animals, the development of in vitro alternatives could lead to an eventual replacement of animal studies. In view of the inherent simplicity of many in vitro methods, some of which comprise cell cultures, further research into the relevance/mechanism of effects is required. Batteries of in vitro tests, when properly validated, may be considered as replacements for animal testing.

scholarly journals Mechanism of Wound-Healing Activity ofHippophae rhamnoidesL. Leaf Extract in Experimental Burns

2011 ◽  
Vol 2011 ◽  
pp. 1-9 ◽  
Author(s):  
Nitin K. Upadhyay ◽  
Ratan Kumar ◽  
M. S. Siddiqui ◽  
Asheesh Gupta
Keyword(s):  
Wound Healing ◽  
Wound Repair ◽  
Leaf Extract ◽  
Lipid Peroxide ◽  
Enzymatic Antioxidants ◽  
Burn Wound ◽  
Burn Wounds ◽  
Aqueous Leaf Extract

The present investigation was undertaken to evaluate the healing efficacy of lyophilized aqueous leaf extract of Sea buckthorn (Hippophae rhamnoidesL., family Elaeagnaceae) (SBT) and to explore its possible mechanism of action on experimental burn wounds in rats. The SBT extract, at various concentrations, was applied topically, twice daily for 7 days. Treatment with silver sulfadiazine (SSD) ointment was used as reference control. The most effective concentration of the extract was found to be 5.0% (w/w) for burn wound healing and this was further used for detailed study. The SBT-treated group showed faster reduction in wound area in comparison with control and SSD-treated groups. The topical application of SBT increased collagen synthesis and stabilization at the wound site, as evidenced by increase in hydroxyproline, hexosamine levels and up-regulated expression of collagen type-III. The histological examinations and matrix metalloproteinases (MMP-2 and -9) expression also confirmed the healing efficacy of SBT leaf extract. Furthermore, there was significant increase in levels of endogenous enzymatic and non-enzymatic antioxidants and decrease in lipid peroxide levels in SBT-treated burn wound granulation tissue. The SBT also promoted angiogenesis as evidenced by anin vitrochick chorioallantoic membrane model andin vivoup-regulated vascular endothelial growth factor (VEGF) expression. The SBT leaf extract had no cytotoxic effect on BHK-21 cell line. In conclusion, SBT aqueous leaf extract possesses significant healing potential in burn wounds and has a positive influence on the different phases of wound repair.

scholarly journals Annona muricata L. extract decreases intestinal glucose absorption and improves glucose tolerance in normal and diabetic rats

2021 ◽  
Vol 10 (3) ◽  
pp. 359-366
Author(s):  
Ana María Guevara-Vásquez ◽  
Julio Víctor Campos-Florián ◽  
Jesús Haydee Dávila-Castillo
Keyword(s):  
Aqueous Extract ◽  
Leaf Extract ◽  
Glucose Absorption ◽  
Diabetic Rats ◽  
Annona Muricata ◽  
Antihyperglycemic Activity ◽  
Aqueous Leaf Extract ◽  
Intestinal Glucose Absorption

Introduction: Poorly controlled hyperglycemia causes numerous health complications. Postprandial hyperglycemia is an important indicator of diabetic status. The aim of this research was to evaluate the effect of Annona muricata L. extract on the in vitro intestinal glucose absorption in diabetic rats and in vivo antihyperglycemic activity in both normal and diabetic rats. Methods: Phytochemical screening of the aqueous extract from the leaves of A. muricata was carried out. Albino rats were randomly assigned into normal and diabetic groups. Each group was divided into three subgroups: control (vehicle), experimental (A. muricata), and standard (Metformin) groups, to determine antihyperglycemic activity at different times after glucose overload. The everted intestinal sac technique was used to study intestinal glucose absorption in diabetic rats. Results: Aqueous leaf extract of Peruvian A. muricata exhibited statistically significant (P < 0.05) in vivo antihyperglycemic activity in both normal and diabetic rats when compared to the control group. The magnitude of the effect was similar to metformin treatment. Moreover, the aqueous leaf extract of A. muricata significantly diminished in vitro intestinal glucose absorption, with a magnitude similar to metformin treatment. Phytochemical analysis of the aqueous extract revealed the presence of tannins, flavonoids, alkaloids, and leucoanthocyanidins, among others. Conclusion: This study reveals that A. muricata aqueous extract is able to reduce in vitro intestinal glucose absorption and improve oral glucose tolerance in rats.

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