Effects of Chlorhexidine on Human Gingival Fibroblasts

NCT07669636 | Not Yet Recruiting

• 1 other identifier: Chx.01
• Study Type: observational
• Target: 3
• Locations: 0 countries
• Sites: N/A

Brief Summary

This prospective in vitro study aims to evaluate the effects of two chlorhexidine-based mouthwash formulations on the survival and function of human gingival fibroblasts (hGFs). Gingival fibroblasts obtained from discarded gingival tissue collected during routine dental procedures will be cultured and exposed to different concentrations of a conventional 0.2% chlorhexidine mouthwash and a 0.2% chlorhexidine mouthwash supplemented with N-acetylcysteine (NAC) and hyaluronic acid (HA). Cell viability, apoptosis, migration, and gene expression will be assessed at multiple time points using Trypan Blue exclusion assay, fluorescence-activated cell sorting (FACS), scratch assay, and quantitative real-time PCR. The study is designed to investigate the concentration-dependent effects of chlorhexidine on fibroblast viability and wound-healing-related functions and to determine whether the addition of NAC and HA may improve the biological compatibility of chlorhexidine-based formulations.

Conditions

HGF; Fibroblast; Chlorhexidine; Cytotoxicity; Biocompatibility

Outcome Measures

Primary Outcomes (1)

• Human Gingival Fibroblast Viability

  Cell viability of human gingival fibroblast cultures exposed in vitro to different concentrations of chlorhexidine-based mouthwash formulations will be assessed using Trypan Blue exclusion assay and live/dead analysis by fluorescence-activated cell sorting (FACS). Viability will be expressed as the proportion of viable cells compared with total counted cells.

  24 hours, 3 days and 7 days from the start of cell culture exposure

Study Arms (1)

Healthy Adult Donors

Human Gingival Fibroblast Cultures

Eligibility Criteria

• Age: 18 Years+
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Probability Sample

Study Population

The study population consists of three healthy adult donors undergoing routine dental surgical procedures at the Dental Unit of San Raffaele Hospital. Small fragments of discarded gingival tissue obtained during procedures such as tooth extraction or gingivectomy will be collected following informed consent. Human gingival fibroblasts will be isolated from these tissues and expanded in culture for in vitro experimental analyses.

You may qualify if:

• Male or female adults aged 18 years or older.
• Undergoing routine dental surgical procedures (e.g., tooth extraction or gingivectomy).
• Willing and able to provide written informed consent for the use of discarded gingival tissue for research purposes.
• Systemically healthy.

You may not qualify if:

• Presence of systemic diseases.
• Presence of relevant medical comorbidities.
• Current smoker.
• Refusal to provide informed consent for the use of discarded biological material.

Sponsors & Collaborators

• IRCCS San Raffaele: lead

Biospecimen

Retention: SAMPLES WITHOUT DNA

Prospective in vitro laboratory study using primary human gingival fibroblasts isolated from discarded gingival tissue obtained from healthy adult donors. Fibroblast cultures will be exposed to different concentrations of chlorhexidine-based mouthwash formulations and evaluated over time for viability, apoptosis, migration, and gene expression.

Central Study Contacts

Roberto Professor Roberto Rotundo, Associate Professor

CONTACT

0226432643; rotundo.roberto@hsr.it

Study Design

• Study Type: observational
• Observational Model: OTHER
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Associate Professor

Study Record Dates

• First Submitted: June 20, 2026
• First Posted: June 25, 2026
• Study Start: July 1, 2026
• Primary Completion (Estimated): October 1, 2026
• Study Completion (Estimated): November 1, 2026
• Last Updated: June 25, 2026

Record last verified: 2026-06