Corneal Ulcer Caused by AcanthamoebaNCT07148752
Evaluation of Resveratrol and Its Loaded Nanoparticles on Acanthamoeba
An observational study of Corneal Ulcer Caused by Acanthamoeba, run by Assiut University.
Trial summary
Acanthamoeba is a free-living amoeba responsible for severe human infections. There are at least twenty-two different genotypes of Acanthamoeba (T1-T22). T4 is the most common virulent genotype, mainly Acanthamoeba castellani and Acanthamoeba polyphaga. Read more about this study
- Who is it for?
- Any age
- Conditions the study is for
- Corneal Ulcer Caused by Acanthamoeba
- How many people
- 50 participants
- Where
- No sites listed yet
- Study dates
- 15 Oct 2025to 1 Nov 2028
- How long will I take part?
- 4 Weeks
- Phase
- Observational
- Are there results?
- Not yet: the study is under way
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages Any age
- All sexes
- No healthy volunteers
- Child, Adult, Older adult
Study population: study participants will be enrolled form Ophthalmology department clinics
Who may be able to join
- Patients presented with keratitis to the outpatient clinic.
- Patients presented with corneal ulcers to the outpatient clinic.
Who cannot join
- Patients presented with corneal ulcers caused by pathogens rather than Acanthamoeba (bacterial, viral or fungal).
- Patients presented with other eye conditions that act as confounding variable as corneal perforation, severe dry eye, glaucoma, scleritis and active uveitis.
- Patients with history of previous ocular surgery or corneal transplant.
About this study
Official title: Evaluation of Amoebicidal Efficacy of Resveratrol and Resveratrol Loaded Nanoparticles on Acanthamoeba Keratitis Isolates
Acanthamoeba is a free-living amoeba responsible for severe human infections. There are at least twenty-two different genotypes of Acanthamoeba (T1-T22). T4 is the most common virulent genotype, mainly Acanthamoeba castellani and Acanthamoeba polyphaga.
Proper treatment still represents a challenge for healthcare professionals.So, we need novel therapeutic strategies, including newer drugs and/or drug delivery systems that can overcome the developing drug resistance as well as have minimal toxicity to human cells.
Read more
There is a new approach in the treatment of parasitic disease by using nanotechnology and natural compounds as plant extracts.
The current study aims to evaluate the therapeutic potential of resveratrol, both in its free form and when loaded on nanoparticles, against clinically isolated Acanthamoeba strains obtained from patients with persistent AK or corneal ulcers.
Read the detailed description
: 1-Specimen collection using: A- Corneal swabs. B- Corneal scraping and superficial keratectomy.
C- Contact lenses and contact lens solutions. 2-Culture: The collected samples will be cultivated on non-nutrient agar (NNA) plates seeded with heat-killed Escherichia coli.
3-Cultivation and maintenance of Acanthamoeba isolates in axenic culture: PYG media will be used in order to obtain a significant number of parasites with a minimum presence of bacteria.
4-Molecular analysis:Positive samples by culture will be subjected to conventional PCR technique for molecular characterization of Acanthamoeba at the genus level using genus-specific primers: Forward primer JDP1: F5'-GGC CCAGATCGTTTACCGTGAA-3'
- Reverse primer JDP2:R5' TCTCACAAGCTGCTAGGG AGTCA-3' 5- Sequencing and phylogenetic study: The amplified DNA will be purified, sequencing ASA.S1 region of the 18s rRNA gene will be performed. Following genotyping, the Basic Local Alignment Search Tool (BLAST) of the US National Center for Biotechnology Information (NCBI) will be used to identify similar Acanthamoeba sequences.
6-Experimental study: Preparation of Resveratrol (Plant extract), preparation and Characterization of nanoparticles and parasite material preparation from PYG axenic culture media will be done.
study groups: Group 1: Negative control: No drug is applied. Group 2: Positive control: Propamidine isethionate as a reference drug. Group 3: Resveratrol alone is applied. Group 4: Nanoparticles alone are applied. Group 5: Resveratrol loaded nanoparticles.
- Group 6: Propamidine isethionate and Resveratrol.
- Group 7: Propamidine isethionate and Resveratrol loaded nanoparticles.
I- In-vitro experimental study:
For assessment of applied drugs/extracts efficacy:
A- In vitro drug susceptibility testing :The Acanthamoeba isolates will be tested for their susceptibility to various drug groups serial dilutions. The number of cysts/trophozoites will be counted using a hemocytometer.
B-Viability measurements: Using trypan blue viability stain. C-Scanning electron microscopy (SEM) D-Host cells cytotoxicity evaluation using MTT assay on HeLa cell line: The MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) will be performed to determine the cell cytotoxicity of the tested extract on human cells that are represented by the HeLa cell line.
-Percentage of cell cytotoxicity will be calculated, and the acceptable limit of cytotoxicity will be taken as the viability cutoff of 60%; thus, a cytotoxicity of >40% is considered unacceptable.
II- In-vivo experimental study:
-Sample size calculation: The sample size is calculated to be 35 tested animals (5 in each group).
- Procedure:
- Induction of keratitis and confirmation of the infection
- Application of the tested drug (7 groups as previously discussed).
- Evaluation of treatment by:
- Clinical evaluation (examination and scoring of the tested animal's eyes).
- Histopathological evaluation of the animal's corneal tissue:
- Parasitological evaluation.
- Conditions
- Corneal Ulcer Caused by Acanthamoeba
What is being measured
The primary outcome is the main question the study is designed to answer.
Invitro evaluation of amoebicidal efficacy of resveratrol and resveratrol loaded nanoparticles against the isolated Acanthamoeba pathogenic strains from corneal ulcers patients in comparison to the reference drug through experimental study.
In-vitro experimental study:
A-Drug susceptibility testing :The Acanthamoeba isolates will be tested for their susceptibility to various drug groups serial dilutions. The number of cysts/trophozoites will be counted using a hemocytometer.
B-Viability measurements: Using trypan blue viability stain. C-Scanning electron microscopy (SEM).
D-Host cells cytotoxicity evaluation using MTT assay on HeLa cell line:
-Percentage of cell cytotoxicity will be calculated, and the acceptable limit of cytotoxicity will be taken as the viability cutoff of 60%; thus, a cytotoxicity of >40% is considered unacceptable.
Measured: 2025-2028
Invivo evaluation of amoebicidal efficacy of resveratrol and resveratrol loaded nanoparticles against the isolated Acanthamoeba pathogenic strains from corneal ulcers patients in comparison to the reference drug through experimental study.
II- In-vivo experimental study:
- The sample size is calculated to be 35 tested animals (5 in each group).
- Procedure:
Induction of keratitis and confirmation of the infection Application of the tested drug (7 groups as previously discussed).
-Evaluation of treatment by: Clinical evaluation (examination and scoring of the tested animal's eyes).
Histopathological evaluation of corneal tissue:
Parasitological evaluation.
Measured: 2025-2028
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted29 Aug 2025
- Study start15 Oct 2025 (estimated)
- Primary completion30 Oct 2028 (estimated)
- Study completion1 Nov 2028 (estimated)
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Other trials for Corneal Ulcer Caused by AcanthamoebaStudies of the same condition, open and under way.
- More trials run by Assiut UniversityThe sponsor's other registered studies.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Other
- Time perspective
- Prospective
- Enrolment
- 50 participants (estimated)
- Target duration
- 4 Weeks
Sponsor and investigators
- Lead sponsor
- Assiut University
- Sponsor type
- University, hospital or other
- Responsible party
- Principal investigator, Reham Hany Mohammed, Assiut University
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT07148752
- Sponsor's protocol ID
- Acanthamoeba new treatment
- Status last verified
- Aug 2025
- First submitted
- 11 Aug 2025
- Last updated
- 29 Aug 2025
- Results posted
- No
- Data monitoring committee
- Yes
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- No
Study at a glance
From the registry record, last updated by the sponsor on 29 Aug 2025. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Evaluation of Amoebicidal Efficacy of Resveratrol and Resveratrol Loaded Nanoparticles on Acanthamoeba Keratitis Isolates
- ClinicalTrials.gov ID
- NCT07148752
- Acronym / sponsor ID
- Acanthamoeba new treatment
- Conditions
- Corneal Ulcer Caused by Acanthamoeba
- Study type
- Observational
- Recruitment status
- Not yet recruiting
- Enrolment
- 50 participants (estimated)
- Lead sponsor
- Assiut UniversityUniversity, hospital or other
- Study start
- 15 Oct 2025 (estimated)
- Primary completion
- 30 Oct 2028 (estimated)
- Study completion
- 1 Nov 2028 (estimated)
- First posted
- 29 Aug 2025 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
Acanthamoeba is a free-living amoeba responsible for severe human infections. There are at least twenty-two different genotypes of Acanthamoeba (T1-T22). T4 is the most common virulent genotype, mainly Acanthamoeba castellani and Acanthamoeba polyphaga.
Proper treatment still represents a challenge for healthcare professionals.So, we need novel therapeutic strategies, including newer drugs and/or drug delivery systems that can overcome the developing drug resistance as well as have minimal toxicity to human cells.
There is a new approach in the treatment of parasitic disease by using nanotechnology and natural compounds as plant extracts.
The current study aims to evaluate the therapeutic potential of resveratrol, both in its free form and when loaded on nanoparticles, against clinically isolated Acanthamoeba strains obtained from patients with persistent AK or corneal ulcers.
Detailed description
: 1-Specimen collection using: A- Corneal swabs. B- Corneal scraping and superficial keratectomy.
C- Contact lenses and contact lens solutions. 2-Culture: The collected samples will be cultivated on non-nutrient agar (NNA) plates seeded with heat-killed Escherichia coli.
3-Cultivation and maintenance of Acanthamoeba isolates in axenic culture: PYG media will be used in order to obtain a significant number of parasites with a minimum presence of bacteria.
4-Molecular analysis:Positive samples by culture will be subjected to conventional PCR technique for molecular characterization of Acanthamoeba at the genus level using genus-specific primers: Forward primer JDP1: F5'-GGC CCAGATCGTTTACCGTGAA-3'
- Reverse primer JDP2:R5' TCTCACAAGCTGCTAGGG AGTCA-3' 5- Sequencing and phylogenetic study: The amplified DNA will be purified, sequencing ASA.S1 region of the 18s rRNA gene will be performed. Following genotyping, the Basic Local Alignment Search Tool (BLAST) of the US National Center for Biotechnology Information (NCBI) will be used to identify similar Acanthamoeba sequences.
6-Experimental study: Preparation of Resveratrol (Plant extract), preparation and Characterization of nanoparticles and parasite material preparation from PYG axenic culture media will be done.
study groups: Group 1: Negative control: No drug is applied. Group 2: Positive control: Propamidine isethionate as a reference drug. Group 3: Resveratrol alone is applied. Group 4: Nanoparticles alone are applied. Group 5: Resveratrol loaded nanoparticles.
- Group 6: Propamidine isethionate and Resveratrol.
- Group 7: Propamidine isethionate and Resveratrol loaded nanoparticles.
I- In-vitro experimental study:
For assessment of applied drugs/extracts efficacy:
A- In vitro drug susceptibility testing :The Acanthamoeba isolates will be tested for their susceptibility to various drug groups serial dilutions. The number of cysts/trophozoites will be counted using a hemocytometer.
B-Viability measurements: Using trypan blue viability stain. C-Scanning electron microscopy (SEM) D-Host cells cytotoxicity evaluation using MTT assay on HeLa cell line: The MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) will be performed to determine the cell cytotoxicity of the tested extract on human cells that are represented by the HeLa cell line.
-Percentage of cell cytotoxicity will be calculated, and the acceptable limit of cytotoxicity will be taken as the viability cutoff of 60%; thus, a cytotoxicity of >40% is considered unacceptable.
II- In-vivo experimental study:
-Sample size calculation: The sample size is calculated to be 35 tested animals (5 in each group).
- Procedure:
- Induction of keratitis and confirmation of the infection
- Application of the tested drug (7 groups as previously discussed).
- Evaluation of treatment by:
- Clinical evaluation (examination and scoring of the tested animal's eyes).
- Histopathological evaluation of the animal's corneal tissue:
- Parasitological evaluation.
Study design
- Observational model
- Other
- Time perspective
- Prospective
- Target follow-up
- 4 Weeks
Who may qualify
- Sex
- All
- Healthy volunteers
- Not accepted
- Age groups
- Child, Adult, Older adult
- Study population
- study participants will be enrolled form Ophthalmology department clinics
Key inclusion criteria (2 listed)
- Patients presented with keratitis to the outpatient clinic.
- Patients presented with corneal ulcers to the outpatient clinic.
Key exclusion criteria (3 listed)
- Patients presented with corneal ulcers caused by pathogens rather than Acanthamoeba (bacterial, viral or fungal).
- Patients presented with other eye conditions that act as confounding variable as corneal perforation, severe dry eye, glaucoma, scleritis and active uveitis.
- Patients with history of previous ocular surgery or corneal transplant.
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Primary outcome
Invitro evaluation of amoebicidal efficacy of resveratrol and resveratrol loaded nanoparticles against the isolated Acanthamoeba pathogenic strains from corneal ulcers patients in comparison to the reference drug through experimental study.
Time frame: 2025-2028
In-vitro experimental study: A-Drug susceptibility testing :The Acanthamoeba isolates will be tested for their susceptibility to various drug groups serial dilutions. The number of cysts/trophozoites will be counted using a hemocytometer. B-Viability measurements: Using trypan blue viability stain. C-Scanning electron microscopy (SEM). D-Host cells cytotoxicity evaluation using MTT assay on HeLa cell line: -Percentage of cell cytotoxicity will be calculated, and the acceptable limit of cytotoxicity will be taken as the viability cutoff of 60%; thus, a cytotoxicity of >40% is considered unacceptable.
Invivo evaluation of amoebicidal efficacy of resveratrol and resveratrol loaded nanoparticles against the isolated Acanthamoeba pathogenic strains from corneal ulcers patients in comparison to the reference drug through experimental study.
Time frame: 2025-2028
II- In-vivo experimental study: * The sample size is calculated to be 35 tested animals (5 in each group). * Procedure: Induction of keratitis and confirmation of the infection Application of the tested drug (7 groups as previously discussed). -Evaluation of treatment by: Clinical evaluation (examination and scoring of the tested animal's eyes). Histopathological evaluation of corneal tissue: Parasitological evaluation.
Results and safety
This study is still under way. Results are not available in the source record yet.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 29 Aug 2025.
- Individual participant dataSharing planned: No
Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part.
The record lists no contact. Use the official record to reach the sponsor.
Questions about this trial
What is this clinical trial studying?
An observational study of Corneal Ulcer Caused by Acanthamoeba, run by Assiut University.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 2 inclusion and 3 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: opening soon, as of the sponsor's last update on 29 Aug 2025. Status can change between updates, so confirm with the study contact before making plans.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
This study is still under way. Results are not available in the source record yet.
How can a doctor refer a patient or contact the research team?
The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Invitro evaluation of amoebicidal efficacy of resveratrol and resveratrol loaded nanoparticles against the isolated Acanthamoeba pathogenic strains from corneal ulcers patients in comparison to the reference drug through experimental study. (time frame: 2025-2028). The study lists 2 primary outcomes in all. It is what the study was designed to measure, not a finding.
Where can I read the original study information?
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 29 Aug 2025.