Ignite Creation Date:
2024-05-06 @ 8:08 PM
Last Modification Date:
2024-10-26 @ 3:21 PM
Study NCT ID:
NCT06260189
Status:
RECRUITING
Last Update Posted:
2024-02-15
First Post:
2024-02-07
Brief Title:
Assessment Of Serum And Tissue Levels Of Cold-Inducible RNA-Binding Protein In Patients With Lichen Planus
Sponsor:
Sohag University
Organization:
Sohag University
Study Overview
Official Title:
Assessment Of Serum And Tissue Levels Of Cold-Inducible RNA-Binding Protein In Patients With Lichen Planus
Status:
RECRUITING
Status Verified Date:
2024-02
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
Lichen planus LP is a chronic inflammatory mucocutaneous disease of unknown etiology
Pathogenesis of LP is not completely understood but its considered a T-cell-mediated autoimmune disease Cold-inducible RNA binding protein CIRP or CIRBP is a member of the glycine-rich RNA-binding protein family Recent studies proved that CIRP acts as a tumor promoter through its actions on different cellular proliferation levels Recently the role of the damage associated molecular proteins and cytokines was highlighted in the pathogenesis of many disorders including psoriasis alopecia areata vitiligo rheumatoid arthritis other autoimmune diseases as well as several types of cancer The aim of this study is to compare serum and tissue levels of CIRP in patients with LP and healthy controls
Pathogenesis of LP is not completely understood but its considered a T-cell-mediated autoimmune disease Cold-inducible RNA binding protein CIRP or CIRBP is a member of the glycine-rich RNA-binding protein family Recent studies proved that CIRP acts as a tumor promoter through its actions on different cellular proliferation levels Recently the role of the damage associated molecular proteins and cytokines was highlighted in the pathogenesis of many disorders including psoriasis alopecia areata vitiligo rheumatoid arthritis other autoimmune diseases as well as several types of cancer The aim of this study is to compare serum and tissue levels of CIRP in patients with LP and healthy controls
Detailed Description:
The aims of this study are
Assessment of serum and tissue levels of CIRP in patients with LP
To compare serum and tissue levels of CIRP in patients with LP and healthy controls
To compare CIRP levels according to LP severity A cross-sectional study will be conducted on 40 patients with LP and 40 age and gender matched healthy controls Diagnosis of LP will be clinically and will be confirmed by dermoscopy
Methods of the study
1 Evaluation of CIRP expression
Every participant in this study will be subjected to both blood sampling and skin biopsy taking in the same session then each sample will be processed separately and blindly at the Clinical Pathology and Histology Departments as following
Blood sampling
Three ml venous blood sample will be taken from every one of the two studied groups in a free anticoagulant tube enabling the blood to be coagulated After 30 minutes of leaving the tube at room temperature till coagulation then samples will be centrifuged at 3000 rpm for 15 min and the resultant serum will be stored at -80c until testing 22 Serum CIRP levels will be measured using an enzyme-linked immunosorbent assay analysis ELISA kit
Skin biopsies One skin biopsy will be taken with 3 mm punch from each patient and control These biopsies will be taken under local anesthesia from LP lesions and the same areas matched in control All biopsies will be submitted to Histology Department Sohag Faculty of Medicine They will be fixed in 10 neutral buffered formalin dehydrated in ascending grades of ethanol followed by immersion in xylene then impregnated in paraffin Several 5 micron 5um thick sections from each block will be taken One slide will be stained by hematoxylin and eosin HE for routine histopathological examination Other sections will be mounted on positive charged slides and stored at room temperature to be stained immunohistochemically
1 Histopathological assessment of HE-stained sections
Sections stained with HE will be evaluated under the light microscope to confirm the diagnosis of LP and to asses epidermal and dermal changes
2 Immunohistochemical IHC staining procedure The method used for immunostaining will be streptavidin-biotin amplified system Paraffin-embedded tissue sections will be deparaffinized in xylene rehydrated in a graded series of ethanol and incubated with 3 hydrogen peroxide Slides will be rinsed in phosphate - buffered saline PBS and then exposed to heat-induced epitope retrieval in citrate buffer solution Ph 6 for 20 minutes After cooling the slides will be incubated over night at room temperature with mouse monoclonal anti-CIRP antibody 01 ml concentrated and diluted by PBS in a dilution 175 Detection of immunoreactivity will be carried out using the Universal Labeled Streptavidin-Biotin system horseradish peroxidase Finally the reaction will be visualized by an appropriate substrate chromogen diaminobenzidine reagent The staining procedure will include the negative controls obtained by substitution of primary antibodies with phosphate-buffered saline 16
3 Interpretation of CIRP immunostaining
The CIRP immunostaining will be assessed separately and blindly in LP lesion and control cases Epidermal and dermal cells will be assigned positive by the presence of brownish coloration detected by DAB reaction either in the cytoplasm or nuclei in 1 stained cells 25
Assessment of serum and tissue levels of CIRP in patients with LP
To compare serum and tissue levels of CIRP in patients with LP and healthy controls
To compare CIRP levels according to LP severity A cross-sectional study will be conducted on 40 patients with LP and 40 age and gender matched healthy controls Diagnosis of LP will be clinically and will be confirmed by dermoscopy
Methods of the study
1 Evaluation of CIRP expression
Every participant in this study will be subjected to both blood sampling and skin biopsy taking in the same session then each sample will be processed separately and blindly at the Clinical Pathology and Histology Departments as following
Blood sampling
Three ml venous blood sample will be taken from every one of the two studied groups in a free anticoagulant tube enabling the blood to be coagulated After 30 minutes of leaving the tube at room temperature till coagulation then samples will be centrifuged at 3000 rpm for 15 min and the resultant serum will be stored at -80c until testing 22 Serum CIRP levels will be measured using an enzyme-linked immunosorbent assay analysis ELISA kit
Skin biopsies One skin biopsy will be taken with 3 mm punch from each patient and control These biopsies will be taken under local anesthesia from LP lesions and the same areas matched in control All biopsies will be submitted to Histology Department Sohag Faculty of Medicine They will be fixed in 10 neutral buffered formalin dehydrated in ascending grades of ethanol followed by immersion in xylene then impregnated in paraffin Several 5 micron 5um thick sections from each block will be taken One slide will be stained by hematoxylin and eosin HE for routine histopathological examination Other sections will be mounted on positive charged slides and stored at room temperature to be stained immunohistochemically
1 Histopathological assessment of HE-stained sections
Sections stained with HE will be evaluated under the light microscope to confirm the diagnosis of LP and to asses epidermal and dermal changes
2 Immunohistochemical IHC staining procedure The method used for immunostaining will be streptavidin-biotin amplified system Paraffin-embedded tissue sections will be deparaffinized in xylene rehydrated in a graded series of ethanol and incubated with 3 hydrogen peroxide Slides will be rinsed in phosphate - buffered saline PBS and then exposed to heat-induced epitope retrieval in citrate buffer solution Ph 6 for 20 minutes After cooling the slides will be incubated over night at room temperature with mouse monoclonal anti-CIRP antibody 01 ml concentrated and diluted by PBS in a dilution 175 Detection of immunoreactivity will be carried out using the Universal Labeled Streptavidin-Biotin system horseradish peroxidase Finally the reaction will be visualized by an appropriate substrate chromogen diaminobenzidine reagent The staining procedure will include the negative controls obtained by substitution of primary antibodies with phosphate-buffered saline 16
3 Interpretation of CIRP immunostaining
The CIRP immunostaining will be assessed separately and blindly in LP lesion and control cases Epidermal and dermal cells will be assigned positive by the presence of brownish coloration detected by DAB reaction either in the cytoplasm or nuclei in 1 stained cells 25
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None