Effect of Smoking on Growth Factors in Platelet-Rich Fibrin: Ex-Vivo Pilot Study

Summary

This protocol evaluates the effect of smoking on platelet-rich fibrin (PRF) by comparing growth factor release and blood parameters between smokers and non-smokers using a standardized and reproducible method.

Abstract

Platelet rich fibrin (PRF) and growth factors are routinely used to accelerate hard and soft tissue regeneration in periodontal surgical procedures, dental implant applications and oral surgery for the last 30 years in the field of dentistry. The aim of this study was to evaluate the effect of smoking on autologous growth factors within PRF. The study population consisted of 20 male participants (10 smokers/10 non-smokers) and 20 female participants (10 smokers/10 non-smokers). Each participant provided two tubes of venous blood (approximately 7 ml each). The first tube was used for a complete blood count (CBC). The second tube was centrifuged at 708 x g for 12 min, and the resulting PRF was analyzed using enzyme-linked immunosorbent assay (ELISA) to determine the levels of growth factors (Platelet-derived growth factor (PDGF), Transforming growth factor-beta (TGF-β), Vascular endothelial growth factor (VEGF), Fibroblast growth factor (FGF) and Insulin-like growth factor (IGF)). The results showed that smoking resulted in a decrease in mean platelet volume (MPV), platelet distribution width (PDW) and platelet larger cell ratio (P-LCR) values (p<0.05) in the whole population. In females, smoking led to a decrease in red blood cell (RBC), MPV, PDW and P-LCR levels, while in male subjects, it resulted in an increase in hematocrit (HCT) values (p<0.05). It was also found that smoking led to an increase in FGF and TGFβ-1 within PRF (p<0.05). Although current results showed a slight increase in certain growth factors in smokers, the clinical relevance of this finding should be investigated in further studies.

Introduction

The utilization of autologous platelet concentrates in various fields of dentistry began in the 1980s and 1990s. Since then, platelet concentrates and growth factors have been routinely employed in periodontal surgical procedures, dental implant applications, and oral surgery to accelerate the regeneration of hard and soft tissues¹. Platelets containing growth factors play crucial roles in cell migration, proliferation, differentiation, and angiogenesis, all of which are integral to tissue regeneration. Autologous platelet concentrates are produced by centrifuging venous blood at different speeds, with or without the use of thrombin and anticoa....

Protocol

This study was conducted at the Department of Periodontology, Faculty of Dentistry, Cukurova University, with the participation of 20 male and 20 female volunteers. All participants provided written informed consent after being informed about the purpose, methodology, and procedures of the study. Ethical approval was obtained from the Ethics Committee of the Faculty of Medicine, Cukurova University (Approval No: 125/54, Date: 16.09.2022). All procedures, including essential biosafety guidance for handling human blood and waste disposal instructions for biohazardous materials and chemical reagents, were performed in accordance with NIH-CDC guidelines^2....

Results

A total of 20 males (10 smokers and 10 non-smokers, aged between 18 and 25) and 20 females (10 smokers and 10 non-smokers, aged between 18 and 25) healthy participants were included in the study.

Evaluation of the Relationship Between Smoking and Blood Parameters
The comparison of blood parameters based on smoking status is presented in Table 1 and the correlation between the number of cigarettes smoked and blood parameters is shown in Table 2

Discussion

In this study, we aimed to evaluate the impact of smoking on autologous growth factors within Platelet-Rich Fibrin (PRF), a widely used material in dental surgeries to enhance wound healing. The findings indicated that smoking slightly affects blood parameters. In addition, the evaluation of the effects of smoking on the growth factors levels at different time points revealed that smoking influences the release profile of growth factors with these effects being correlated with the number of cigarettes smoked. Each of the.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Centrifuge (40° fixed-angle rotor)	N/A (Duo Centrifuge (Hettich))		708 x g, 12 min
ELISA Kit - FGF	Fine Test (Wuhan Fine Biotech)	EH0541
ELISA Kit - IGF-1	Fine Test (Wuhan Fine Biotech)	ER0030
ELISA Kit - PDGF	Fine Test (Wuhan Fine Biotech)	ER1240
ELISA Kit - TGFβ-1	Fine Test (Wuhan Fine Biotech)	EH0287
ELISA Kit - VEGF	Fine Test (Wuhan Fine Biotech)	EH0327
Eppendorf Tubes	N/A
Glass Blood Collection Tubes (10 mL)	N/A		No anticoagulant
HRP Solution	Included in ELISA kit
Micropipettes & Tips	N/A
Microplate Reader	N/A		450 nm
Orbital Shaker	N/A		Slow program, 72 h
Precision Scale	N/A
RPMI-1640 Cell Culture Medium	HyClon
Statistical analyses	IBM SPSS Statistics Version 20.0 (Armonk, NY: IBM Corp.)
Sterile Scissors	N/A
Stop Solution	Included in ELISA kit
Substrate Solution	Included in ELISA kit
Wash Buffer	Included in ELISA kit