Assessment of the Clinical, Cellular and Molecular Effects of Royalactin-Royal Jelly on the Healing of Wagner Grad II Diabetic Foot Ulcers. (ROYALACIN)

The aim of this study is to evaluate the clinical, cellular, and molecular effects of Royalactin as an adjunct to standard treatment, compared to standard treatment alone, in the treatment of Wagner grade II diabetic foot ulcers by assessing wound size, cell behavior (Epithelialization, polymorphonuclear neutrophils, fibroblasts, angiogenesis, and collagen), and growth factor expression. This is a prospective, randomized, double-blind, double-blind, experimental, longitudinal, comparative, controlled clinical trial.

Study Overview

• Status: Completed
• Conditions: Diabetic Foot Ulcer
• Intervention / Treatment: Drug: Royalactina; Drug: Placebo

Detailed Description

Materials and methods

Ethical Considerations The study was conducted in compliance with Good Clinical Practice (GCP) guidelines. All procedures adhered to the ethical standards of the Research and Bioethics Committee of the XXXXXXXXXX. the Declaration of Helsinki, and the General Health Law. Protocols were strictly followed, data were handled with rigorous confidentiality, and informed consent was obtained from all participants.

Inclusion Criteria Adult patients with Diabetes Mellitus undergoing treatment for foot and ankle wounds were included. Eligibility criteria required wounds larger than 4 cm², with more than 80% granulation tissue, involving skin, fat, or ligaments without bone involvement, and non-infected (Wagner Grade II). Additional requirements included a duration of more than 2 months, an ankle-brachial index (ABI) greater than 0.5, the signing of the informed consent form, and the ability to attend dressing changes three times per week for 4 weeks.

Experimental Agents and Randomization The FEMSA Biotechnology Center laboratory produced 20 tubes (50 ml) containing gold nanoparticles and emulsifiers. Ten tubes were formulated with Royalactin at 0.2 g/L, while the other 10 served as the control without the peptide. The laboratory randomized, numbered, and paired the tubes, providing a list for paired application.

Treatment Protocol Standardized dressing changes were performed three times per week for 4 weeks at the Chronic Wound, Diabetic Foot, and Hyperbaric Medicine Center of XXXXX. Each wound was first irrigated with sterile water and dried with sterile gauze. The wound was then divided (or the division was remarked) using black ink on the skin, and a 0.5 cm strip of Hypafix (BSN medical GmbH, ESSITY) was applied. A circular label was placed on the perilesional skin to identify the tube number (bottom), patient ID (top-right), and dressing change number (top-left).

Standard photographs were taken (Camera+ App, iPhone 15 Pro Max). The area was gently debrided three times with sterile gauze moistened with 0.12% Chlorhexidine (Clorhexi-Clean®, PISA). Ketanserin 2% gel (Sufrexal®, Janssen) was applied to both sides, followed by the application of the corresponding gel (Royalactin or placebo) to each designated wound section. Finally, a primary dressing of sodium carboxymethylcellulose hydrofiber and regenerated cellulose fiber (Aquacell extra®, Convatec) was applied, protected by a secondary adhesive border gauze dressing (Border gauze®, Medline).

Data Collection and Histological Analysis During the first and last dressing changes (sessions 1 and 13), photographs were taken to measure the area of each side in cm² (IMITO measure App, iPhone 15 Pro Max). Additionally, 3 mm punch biopsies of the granulation tissue were obtained from the distal edge of each side.

Biopsy samples were sectioned, labeled, and stained using Hematoxylin and Eosin (H&E), Masson's Trichrome, and immunohistochemistry (IHC) for the expression of:

• CD31 (Roche)
• Vascular Endothelial Growth Factor (VEGF) (Biocare)
• Epidermal Growth Factor Receptor (EGFR) (Cell Marque).

Statistical Analysis and Evaluation Results from sessions 1 and 13 were evaluated. Wound size was recorded in cm². Epithelialization, polymorphonuclear neutrophils (PMN), fibroblasts, angiogenesis, and collagen were scored according to a semi-quantitative histological scale for wound assessment. The percentage of tissue expressing VEGF and EGFR, as well as the intensity of expression (scored 0 to 3+), were reported separately (Figure 1 G-I). Data were compared at baseline and at four weeks, both as a total cohort and by comparing the Control vs. Royalactin groups; individual variables were also analyzed.

• Study Type: Interventional
• Enrollment (Actual): 10
• Phase: Phase 2

Contacts and Locations

Study Locations

• Mexico
  • Nuevo León
    • Monterrey, Nuevo León, Mexico, 64460
      • Facultad de medicina de la universidad autonoma de nuevo leon

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: No

Description

Inclusion Criteria:

- Confirmed diagnosis of Diabetes Mellitus according to the 2020 ADA guidelines.

• Stage II ulcer according to the Wagner classification for Diabetic Foot (deep ulcer, penetrating ligaments and muscles, without bone involvement or abscess formation).
• Ulcer located on any area of the foot or ankle.
• Ulcer with 80% granulation tissue.
• Ankle-brachial index greater than 0.5 (not considered severe or critical arterial disease).
• Negative stylet test.
• Ability to attend wound care and follow-up appointments three times a week for four weeks.
• Signed informed consent form.

Exclusion Criteria:

- History of hypersensitivity reactions to dermatological products.

• History of generalized dermatological pathology or pathology affecting the foot.
• Diagnosis of autoimmune disease.
• Local or systemic infection.
• Hypersensitivity to any of the components of the experimental product.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Double

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Active Comparator: Diabetic foot ulcers treated with Royalactin; Standardized wound care is performed 3 times a week with the application of Royalactin.	Drug: Royalactina; Standardized wound care is performed 3 times a week with application of Royalactin for 4 weeks
Placebo Comparator: Diabetic foot ulcers with placebo application; Standardized treatments are performed 3 times a week with placebo application.	Drug: Placebo; Standardized wound care is performed 3 times a week with placebo application for 4 weeks

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Ulcer measurement in square centimeters	A photograph is taken at the beginning and at 4 weeks and measured with a program in square centimeters.	4 weeks

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
The histological section of the ulcer margins at baseline and at 4 weeks is tabulated and compared	Biopsies are taken from the edges of the ulcer at baseline and at 4 weeks, and the epithelium, neutrophils, collagen, angiocenesis, and fibroblasts are tabulated using a semi-quantitative scale.	4 weeks

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
Measurement of VEGF and EGFR expression and intensity at baseline and 4 weeks	The percentage of granulation tissue expressing vascular endothelial growth factor (VEGF) and epithelial growth factor receptor (EGFR) was reported separately, as well as the intensity of their expression from 0 to 3 crosses.	4 weeks

Collaborators and Investigators

• Sponsor: Universidad Autonoma de Nuevo Leon
• Investigators: Principal Investigator: Jose L Elizondo, cirujano general, Sin afiliacion

Study record dates

Study Major Dates

• Study Start (Actual): September 9, 2022
• Primary Completion (Actual): September 30, 2025
• Study Completion (Actual): October 30, 2025

Study Registration Dates

• First Submitted: March 18, 2026
• First Submitted That Met QC Criteria: March 28, 2026
• First Posted (Actual): April 3, 2026

Study Record Updates

• Last Update Posted (Actual): April 3, 2026
• Last Update Submitted That Met QC Criteria: March 28, 2026
• Last Verified: March 1, 2026

More Information

Terms related to this study

• Keywords: DIABETIC FOOT DESEASE; ROYALACTINA; ROYAL JELLY
• Additional Relevant MeSH Terms: Endocrine System Diseases; Vascular Diseases; Cardiovascular Diseases; Diabetes Mellitus; Diabetic Angiopathies; Diabetes Complications; Skin Diseases; Skin Ulcer; Leg Ulcer; Diabetic Neuropathies; Foot Ulcer; Skin and Connective Tissue Diseases; Diabetic Foot
• Other Study ID Numbers: CM034/26092022; Without funding organization (Other Identifier: Without funding organization)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO
• IPD Plan Description: The informed consent form signed by the patients specifies that their personal data will not be shared.

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No