Effects of Standard PRP and Extracellular Vesicle-Enriched PRP on Human Gingival Fibroblasts

October 4, 2026 updated by: Kubra Aral, Inonu University

Comparative Evaluation of Standard Platelet-Rich Plasma (PRP) and Platelet-Derived Extracellular Vesicle-Rich Platelet-Rich Plasma (EV-rich PRP) on Human Gingival Fibroblast Function: An In Vitro Study

The goal of this study is to compare the biological effects of two blood-derived products, standard platelet-rich plasma (PRP) and extracellular vesicle-enriched PRP (EV-enriched PRP), on human gingival (gum) fibroblasts, the cells responsible for gum tissue repair. PRP is a concentrate of platelets prepared from a person's own blood and is used in periodontal and soft tissue regeneration because it releases growth factors. Extracellular vesicles (EVs) are nanometer-sized particles released by platelets that carry proteins and genetic material and may play an important role in tissue healing. No study has yet directly compared standard PRP with an EV-enriched formulation on gingival fibroblasts.

The main questions this study aims to answer are:

  • Does EV-enriched PRP increase gingival fibroblast proliferation more than standard PRP?
  • Does EV-enriched PRP accelerate gingival fibroblast migration (wound closure) more than standard PRP?
  • Do the two products differ in their effects on the expression of growth factor and collagen genes in gingival fibroblasts?

Six healthy, non-smoking adult volunteers aged 20 to 40 years will each give a single blood sample (two tubes, 18 mL in total) from an arm vein. No treatment will be given to the volunteers. Each volunteer's blood will be processed into both standard PRP and EV-enriched PRP, and both products will be tested in the laboratory on a commercially available human gingival fibroblast cell line.

Study Overview

Status

Status

Indicates the current recruitment status or the expanded access status.
Not yet recruiting

Conditions

Conditions

The disease, disorder, syndrome, illness, or injury that is being studied. On ClinicalTrials.gov, conditions may also include other health-related issues, such as lifespan, quality of life, and health risks.

Detailed Description

Study design: This is a laboratory-based study using blood donated by healthy volunteers, with a paired design. Blood from each of six healthy donors will be processed into both standard PRP and EV-enriched PRP, and each donor's PRP will be compared only with the same donor's EV-enriched PRP. Samples will not be pooled; each donor's samples will be analyzed separately. This design aims to minimize the influence of biological variability between donors on the comparison.

Blood collection and product preparation: Two tubes of peripheral venous blood (18 mL in total) will be collected from each volunteer. Standard PRP will be prepared using a commercially available, CE Class IIb certified, single-spin closed-system PRP kit (T-Lab, Turkey; 1150 RCF, 4 minutes). EV-enriched PRP will be prepared using the T-LAB Exomine kit (Turkey), in which Advanced PRX tubes with micro-lyzer blade technology activate platelets and induce EV release through mechanical stress. Stocks of PRP and EV-enriched PRP will be diluted in serum-free DMEM/F12 to final concentrations of 50%, 25%, 10%, 5%, 2.5%, and 1%.

Cell culture: Human gingival fibroblasts (HGF-1, ATCC) will be cultured in DMEM/F12 supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin, and 1% amphotericin B at 37°C and 5% CO2. Cells at passages 4 to 8 will be used in all experiments.

Cell proliferation assay: HGF-1 cells will be seeded in 96-well plates (2 x 10^3 cells/well) and, after overnight attachment, treated with a negative control (serum-free DMEM), a positive control (DMEM with 10% FBS), six PRP concentrations, and six EV-enriched PRP concentrations (14 groups in total, in technical triplicate). Absorbance will be measured spectrophotometrically at 570 nm at 24, 48, and 72 hours, and relative proliferation (%) will be calculated against the negative control.

Wound healing (scratch) assay: HGF-1 cells will be seeded in 24-well plates (8 x 10^4 cells/well). At 80% confluence a scratch will be created and cells will be treated using the same group layout. Images will be captured with a phase-contrast microscope at baseline, 24 hours, and 48 hours, and the percentage of cell-free area will be calculated with ImageJ software.

Gene expression analysis: After the most effective concentrations for proliferation and migration are identified, HGF-1 cells (5 x 10^4 cells/well, 24-well plates) will be treated with a control (DMEM with 10% FBS) or the optimal PRP and EV-enriched PRP concentrations. On days 3 and 7, expression of TGF-beta1, PDGF-A, PDGF-B, COL1A2, VEGF, and EGF will be analyzed by RT-qPCR, with beta-actin as the reference gene. Relative fold change will be calculated using the 2^-ddCt method.

Sample size: Sample size was based on the large effect sizes reported in the literature for the effects of platelet concentrates on gingival fibroblast function. With a paired t-test, alpha = 0.05, and an assumed effect size of d = 1.5, six donors provide 83% statistical power.

Statistical analysis: Normality will be assessed with the Shapiro-Wilk test. Paired comparisons between PRP and EV-enriched PRP will use the paired t-test or the Wilcoxon signed-rank test, depending on distribution. The combined effects of concentration and time will be evaluated with repeated-measures ANOVA (or the Friedman test) and appropriate post-hoc corrections. p < 0.05 will be considered statistically significant.

Hypothesis: EV-enriched PRP will show superior biological effects on gingival fibroblast proliferation and wound healing compared with conventional PRP.

Study Type

Study Type

Describes the nature of a clinical study. Study types include interventional studies (also called clinical trials), observational studies (including patient registries), and expanded access.
Observational

Enrollment (Estimated)

Enrollment

The number of participants in a clinical study. The "anticipated" enrollment is the target number of participants that the researchers need for the study.
6

Contacts and Locations

This section provides the contact details for those conducting the study, and information on where this study is being conducted.

Study Contact

Study Contact

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Study Locations

  • Turkey (Türkiye)
    • Battalgazi
      • Malatya, Battalgazi, Turkey (Türkiye), 44100
        • Inonu University Faculty of Dentistry

Participation Criteria

Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.

Eligibility Criteria

Eligibility Criteria

The key requirements that people who want to participate in a clinical study must meet or the characteristics they must have. Eligibility criteria consist of both inclusion criteria (which are required for a person to participate in the study) and exclusion criteria (which prevent a person from participating). Types of eligibility criteria include whether a study accepts healthy volunteers, has age or age group requirements, or is limited by sex.

Ages Eligible for Study

20 years to 40 years (Adult)

Accepts Healthy Volunteers

Yes

Genders Eligible for Study

All

Sampling Method

Non-Probability Sample

Study Population

Systemically healthy, non-smoking adult volunteers aged 20 to 40 years, recruited at Inonu University Faculty of Dentistry, Department of Periodontology (Malatya, Turkey). Individuals receiving treatment for any disease, pregnant or breastfeeding women, and individuals with limited capacity to give informed consent will not be included.

Description

Inclusion Criteria:

  • Age 20 to 40 years
  • Systemically healthy
  • Non-smoker
  • Not using anticoagulant medication
  • Willing to participate and able to provide written informed consent

Exclusion Criteria:

  • History of bleeding disorder or coagulation abnormality
  • Use of medications affecting platelet function (e.g., antiplatelet or anticoagulant agents)
  • Acute infection
  • Pregnancy or breastfeeding
  • Systemic disease (e.g., diabetes, autoimmune or hematological disease)
  • Age under 20 or over 40 years

Study Plan

This section provides details of the study plan, including how the study is designed and what the study is measuring.

How is the study designed?

Design Details

  • Observational Models: OTHER
  • Time Perspectives: PROSPECTIVE

Cohorts and Interventions

Group / Cohort

Group / Cohort

A group or subgroup of participants in an observational study that is assessed for biomedical or health outcomes.
Healthy Blood Donors

Six systemically healthy, non-smoking adult volunteers aged 20 to 40 years who are not using anticoagulant or antiplatelet medication. Each participant provides a single peripheral venous blood sample (two tubes, 18 mL in total); no treatment is administered to participants. From each donor's blood, both standard platelet-rich plasma (PRP) and extracellular vesicle-enriched PRP (EV-enriched PRP) are prepared and tested in vitro on human gingival fibroblasts (HGF-1), allowing a paired, within-donor comparison of the two products.

What is the study measuring?

Primary Outcome Measures

Primary Outcome Measures

In a clinical study's protocol, the planned outcome measure that is the most important for evaluating the effect of an intervention/treatment. Most clinical studies have one primary outcome measure, but some have more than one.
Outcome Measure
Measure Description
Time Frame
Gingival Fibroblast Proliferation
Time Frame: 24, 48, and 72 hours after treatment

Proliferation of human gingival fibroblasts (HGF-1) treated with standard PRP versus EV-enriched PRP at six concentrations (50%, 25%, 10%, 5%, 2.5%, 1%), measured spectrophotometrically at 570 nm and expressed as relative proliferation (%) compared with the negative control (serum-free DMEM). Each donor's PRP is compared with the same donor's EV-enriched PRP (paired comparison).
24, 48, and 72 hours after treatment
Gingival Fibroblast Wound Closure (Scratch Assay)
Time Frame: Baseline, 24 hours, and 48 hours after scratch

Migration of human gingival fibroblasts (HGF-1) in an in vitro scratch assay after treatment with standard PRP versus EV-enriched PRP at six concentrations, assessed by phase-contrast microscopy and expressed as the percentage of remaining cell-free area calculated with ImageJ software. Each donor's PRP is compared with the same donor's EV-enriched PRP (paired comparison).
Baseline, 24 hours, and 48 hours after scratch

Secondary Outcome Measures

Secondary Outcome Measures

In a clinical study's protocol, a planned outcome measure that is not as important as the primary outcome measure for evaluating the effect of an intervention but is still of interest. Most clinical studies have more than one secondary outcome measure.
Outcome Measure
Measure Description
Time Frame
Growth Factor and Collagen Gene Expression in Gingival Fibroblasts
Time Frame: Day 3 and day 7 after treatment

Relative mRNA expression of TGF-beta1, PDGF-A, PDGF-B, COL1A2, VEGF, and EGF in human gingival fibroblasts (HGF-1) treated with the optimal concentrations of standard PRP and EV-enriched PRP (selected from the proliferation and migration assays), compared with control (DMEM with 10% FBS). Expression will be measured by RT-qPCR with beta-actin as the reference gene and reported as relative fold change using the 2^-ddCt method.
Day 3 and day 7 after treatment

Collaborators and Investigators

This is where you will find people and organizations involved with this study.

Sponsor

Sponsor

The organization or person who initiates the study and who has authority and control over the study.
Inonu University

Publications and helpful links

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General Publications

Study record dates

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Study Major Dates

Study Start (Estimated)

Study Start

The actual date on which the first participant was enrolled in a clinical study. The "anticipated" study start date is the date that the researchers think will be the study start date.
October 11, 2026

Primary Completion (Estimated)

Primary Completion

The date on which the last participant in a clinical study was examined or received an intervention to collect final data for the primary outcome measure. Whether the clinical study ended according to the protocol or was terminated does not affect this date. For clinical studies with more than one primary outcome measure with different completion dates, this term refers to the date on which data collection is completed for all the primary outcome measures. The "anticipated" primary completion date is the date that the researchers think will be the primary completion date for the study.
November 18, 2026

Study Completion (Estimated)

Study Completion

The date on which the last participant in a clinical study was examined or received an intervention/treatment to collect final data for the primary outcome measures, secondary outcome measures, and adverse events (that is, the last participant's last visit). The "anticipated" study completion date is the date that the researchers think will be the study completion date.
November 18, 2026

Study Registration Dates

First Submitted

First Submitted

The date on which the study sponsor or investigator first submitted a study record to ClinicalTrials.gov. There is typically a delay of a few days between the first submitted date and the record's availability on ClinicalTrials.gov (the first posted date).
September 27, 2026

First Submitted That Met QC Criteria

First Submitted That Met QC Criteria

The date on which the study sponsor or investigator first submits a study record that is consistent with National Library of Medicine (NLM) quality control (QC) review criteria. The sponsor or investigator may need to revise and submit a study record one or more times before NLM's QC review criteria are met. It is the responsibility of the sponsor or investigator to ensure that the study record is consistent with the NLM QC review criteria.
September 27, 2026

First Posted (Actual)

First Posted

The date on which the study record was first available on ClinicalTrials.gov after National Library of Medicine (NLM) quality control (QC) review has concluded. There is typically a delay of a few days between the date the study sponsor or investigator submitted the study record and the first posted date.
October 2, 2026

Study Record Updates

Last Update Posted (Actual)

Last Update Posted

The most recent date on which changes to a study record were made available on ClinicalTrials.gov. There may be a delay between when the changes were submitted to ClinicalTrials.gov by the study's sponsor or investigator (the last update submitted date) and the last update posted date.
October 7, 2026

Last Update Submitted That Met QC Criteria

Last Update Submitted That Met QC Criteria

The most recent date on which the study sponsor or investigator submitted changes to a study record that are consistent with National Library of Medicine (NLM) quality control (QC) review criteria. It is the responsibility of the sponsor or investigator to ensure that the study record is consistent with the NLM QC review criteria.
October 4, 2026

Last Verified

Last Verified

The most recent date on which the study sponsor or investigator confirmed the information about a clinical study on ClinicalTrials.gov as accurate and current. If a study with a recruitment status of recruiting; not yet recruiting; or active, not recruiting has not been confirmed within the past 2 years, the study's recruitment status is shown as unknown.
October 1, 2026

More Information

Terms related to this study

Keywords

Other Study ID Numbers

Other Study ID Numbers

Identifiers or ID numbers other than the NCT number that are assigned to a clinical study by the study's sponsor, funders, or others. These numbers may include unique identifiers from other trial registries and National Institutes of Health grant numbers.
  • IU-2026 PRP

Plan for Individual participant data (IPD)

Plan to Share Individual Participant Data (IPD)?

NO

IPD Plan Description

Individual participant data will not be shared publicly because the study involves a small number of healthy donors and the data consist of in vitro laboratory measurements derived from donor blood products. Aggregated results will be reported in the resulting publication, and de-identified laboratory data may be made available by the corresponding author upon reasonable request.

Drug and device information, study documents

Studies a U.S. FDA-regulated drug product

No

Studies a U.S. FDA-regulated device product

No

This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.

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