Alterations in Mast Cell and Macrophage Infiltration, as Well as Micro Vessel Density

NCT07448896 | Recruiting

• 1 other identifier: Mast cells obesity
• Study Type: observational
• Target: 100
• Locations: 1 country
• Sites: 1

Brief Summary

Obesity is a global health problem that has reached epidemic proportions, affecting more than one billion people worldwide and significantly increasing the risk of multiple comorbidities, including type 2 diabetes, cardiovascular diseases, and cancer (World Health Organization, 2024). Increasing evidence suggests that chronic low-grade inflammation associated with obesity plays a critical role in the development of obesity-related malignancies, including gastric cancer. Adipose tissue dysfunction in obesity leads to the recruitment and activation of various immune cells, such as macrophages and mast cells, which contribute to a pro-inflammatory microenvironment through the release of cytokines, growth factors, and angiogenic mediators.

Conditions

Obesity

Keywords

gastric carcinogenesis; immunopathological changes; gastric mucosa

Outcome Measures

Primary Outcomes (5)

• Immunohistochemistry

  Compare the 2 groups: Adipose tissue macrophages (ATMs) was assessed by immunohistochemistry using a three-step biotin-avidin-peroxidase detection method. five-micrometer-thick serial sections were cut from formalin-fixed, paraffin-embedded gastric tissue of obese (GTO) and control normal tissue (NT) samples. Antigen retrieval was performed using a microwave oven (500 W for 10 minutes), followed by endogenous peroxidase blocking with a 3% hydrogen peroxide solution. Slides were incubated with the following primary antibodies for 1 hour at room temperature: * Anti-tryptase (clone AA1; Dako, Glostrup, Denmark; 1:100) for mast cell identification, * Anti-CD68 (clone KP1; Dako, Glostrup, Denmark; 1:100) for ATM detection, * Anti-CD31 (clone QB-END 10; Bio-Optica, Milan, Italy; 1:50) as a pan-endothelial marker to assess MVD.

  Baseline

• Immunohistochemistry

  Compare the 2 groups: Mast cells positive for tryptase (MCPT) was assessed by immunohistochemistry using a three-step biotin-avidin-peroxidase detection method. five-micrometer-thick serial sections were cut from formalin-fixed, paraffin-embedded gastric tissue of obese (GTO) and control normal tissue (NT) samples. Antigen retrieval was performed using a microwave oven (500 W for 10 minutes), followed by endogenous peroxidase blocking with a 3% hydrogen peroxide solution. Slides were incubated with the following primary antibodies for 1 hour at room temperature: * Anti-tryptase (clone AA1; Dako, Glostrup, Denmark; 1:100) for mast cell identification, * Anti-CD68 (clone KP1; Dako, Glostrup, Denmark; 1:100) for ATM detection, * Anti-CD31 (clone QB-END 10; Bio-Optica, Milan, Italy; 1:50) as a pan-endothelial marker to assess MVD.

  Baseline

• Immunohistochemistry

  Compare the 2 groups: Microvascular density (MVD) was assessed by immunohistochemistry using a three-step biotin-avidin-peroxidase detection method. five-micrometer-thick serial sections were cut from formalin-fixed, paraffin-embedded gastric tissue of obese (GTO) and control normal tissue (NT) samples. Antigen retrieval was performed using a microwave oven (500 W for 10 minutes), followed by endogenous peroxidase blocking with a 3% hydrogen peroxide solution. Slides were incubated with the following primary antibodies for 1 hour at room temperature: * Anti-tryptase (clone AA1; Dako, Glostrup, Denmark; 1:100) for mast cell identification, * Anti-CD68 (clone KP1; Dako, Glostrup, Denmark; 1:100) for ATM detection, * Anti-CD31 (clone QB-END 10; Bio-Optica, Milan, Italy; 1:50) as a pan-endothelial marker to assess MVD.

  Baseline

• Morphometric Analysis

  Compare the 2 groups: Quantitative assessment was performed using a light microscope. For GTO tissue section, five highly immunostained areas ("hot spots") were identified at low magnification. ATMs, MCPT, and MVD were then quantified at ×40 magnification. The mean value across five hot spots per marker was used for each sample. To evaluate mast cell degranulation, the presence of tryptase-positive granules in the extracellular matrix was examined. Degranulating mast cells were identified by the diffusion of immunoreactive granules outside the cell boundaries, indicating active release of tryptase. This extracellular localization of tryptase provided morphological evidence of mast cell activation and was considered a marker of tissue inflammation and remodeling.

  Baseline

• Morphometric analysis

  Compare the 2 groups: Quantitative assessment was performed using a light microscope. For NT tissue section, five highly immunostained areas ("hot spots") were identified at low magnification. ATMs, MCPT, and MVD were then quantified at ×40 magnification. The mean value across five hot spots per marker was used for each sample. To evaluate mast cell degranulation, the presence of tryptase-positive granules in the extracellular matrix was examined. Degranulating mast cells were identified by the diffusion of immunoreactive granules outside the cell boundaries, indicating active release of tryptase. This extracellular localization of tryptase provided morphological evidence of mast cell activation and was considered a marker of tissue inflammation and remodeling.

  Baseline

Study Arms (2)

patients with obesity undergoing bariatric surgery

Procedure: bariatric surgery laparoscopic sleeve gastrectomy

lean control patients undergoing endoscopic biopsy for benign or malignant gastric conditions

Procedure: lean control patients undergoing endoscopic biopsy for benign or malignant gastric conditions.

Interventions

lean control patients undergoing endoscopic biopsy for benign or malignant gastric conditions. PROCEDURE

Adipose tissue macrophages (ATMs), mast cells positive for tryptase (MCPT), and microvascular density (MVD) were assessed by immunohistochemistry. Quantitative assessment was performed using a light microscope. For each GTO and NT tissue section, five highly immunostained areas ("hot spots") were identified at low magnification.

lean control patients undergoing endoscopic biopsy for benign or malignant gastric conditions

bariatric surgery laparoscopic sleeve gastrectomy PROCEDURE

Adipose tissue macrophages (ATMs), mast cells positive for tryptase (MCPT), and microvascular density (MVD) were assessed by immunohistochemistry. Quantitative assessment was performed using a light microscope. For each GTO and NT tissue section, five highly immunostained areas ("hot spots") were identified at low magnification.

patients with obesity undergoing bariatric surgery

Eligibility Criteria

• Sex: all
• Healthy Volunteers: No
• Age Groups: Child (0-17), Adult (18-64), Older Adult (65+)
• Sampling Method: Probability Sample

Study Population

Fifty gastric tissue samples will be collected from patients with obesity undergoing bariatric surgery and fifty from lean control patients undergoing endoscopic biopsy for benign or malignant gastric conditions. All participants underwent preoperative evaluation including blood tests. All patients were evaluated by a multidisciplinary team consisting of a nutritionist, psychiatrist, endocrinologist, radiologist, anesthesiologist, and surgeon. Endocrinologic assessment excluded secondary causes of obesity (e.g., Cushing's syndrome, polycystic ovary syndrome). Only patients with a body mass index (BMI) \> 35 kg/m² were included. All surgical procedures were laparoscopic sleeve gastrectomies (LapSG).

You may qualify if:

• Adult patients undergoing bariatric surgery (laparoscopic sleeve gastrectomy).
• BMI \> 35 kg/m²
• All participants underwent preoperative evaluation, including blood tests and assessment by a multidisciplinary team (nutritionist, psychiatrist, endocrinologist, radiologist, anesthesiologist, and surgeon).

You may not qualify if:

• Patients with secondary causes of obesity, such as Cushing's syndrome or polycystic ovary syndrome (PCOS).
• Patients with malignant gastric conditions or previous gastric surgery.
• Patients with systemic inflammatory diseases, autoimmune disorders, or chronic infections that may influence immune cell infiltration.
• Patients with incomplete clinical data or poor-quality tissue samples.
• Patients taking anti-inflammatory, immunosuppressive, or corticosteroid therapy within the last 3 months before sampling.

Sponsors & Collaborators

• General Committee of Teaching Hospitals and Institutes, Egypt: lead

Study Sites (1)

The surgical department of Medical Research Institute Hospital, Alexandria University

Alexandria, Alexandria Governorate, 21531, Egypt

RECRUITING

MeSH Terms

Conditions

Obesity

Condition Hierarchy (Ancestors)

Overweight; Overnutrition; Nutrition Disorders; Nutritional and Metabolic Diseases; Body Weight; Signs and Symptoms; Pathological Conditions, Signs and Symptoms

Central Study Contacts

Mohamed H Ashour, PhD

CONTACT

00201002600970; dr.mhany@gmail.com

Study Design

• Study Type: observational
• Observational Model: COHORT
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER GOV
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Professor of General surgery

Study Record Dates

• First Submitted: February 20, 2026
• First Posted: March 4, 2026
• Study Start: April 15, 2026
• Primary Completion: May 1, 2026
• Study Completion: May 1, 2026
• Last Updated: April 29, 2026

Record last verified: 2026-04

Data Sharing

• IPD Sharing: Will share

Locations

EG (1)