Replica Study

First interlaboratory study in the world to replicate and validate high-profile studies on the cytotoxicity and inflammation induced by cigarette smoke and aerosol from electronic nicotine delivery systems (ENDS).

The Replica project team uses the most advanced tools and techniques and the most reliable methodologies to provide to the public up-to-date information on ENDS.

From 2019 to 2022, researchers of the Replica project have replicated the most important international studies in the field of Tobacco Harm Reduction, unequivocally demonstrating the reduced toxicity of electronic cigarettes (ECIG) and heated tobacco products (HTP) compared to conventional cigarettes. A new phase of research for Replica has started in 2023!

0 internal laboratories

Testing the latest generation of alternative products to conventional cigarettes

for one mission

To inform on the detrimental health effects of smoke, while accelerating efforts for the understanding, knowledge, and research capacity in the area of tobacco harm reduction and its role to end smoking, to better serve our worldwide communities.

Latest REPLICA findings were presented at the Asia-Pacific Conference, showing no significant impact of e-cigarette aerosols on cisplatin resistance in cancer cells—an important step in strengthening harm reduction science and academic dialogue in Indonesia

Stay up to date with the latest developments and research outcomes from the Replica Project at these upcoming international conventions:

📍 Bandung, Indonesia

📅 14 June 2025

🔗 Link 

📍 Athens, Greece

📅 14 September 2025

🔗 Welcome – EUROTOX Congress 2025

What is Replica

Replica project coordinates a multi-center research network of academic and affiliated laboratories worldwide.

Since 2019 the team has been conducting inter-laboratory studies comparing the latest generation of alternative products (i.e. ECIGs and HTPs) to traditional tobacco cigarettes, and replicating results from in vitro research published on high impact journals to assess validity of the original work put under scrutiny.

COEHAR SCIENTIFIC TEAM

Riccardo Polosa

CoEHAR Founder

Giovanni Li Volti

CoEHAR Director and Project Leader

Massimo Caruso

Co-project leader

Seven international laboratories

Italy

CoEHAR, University of Catania

Indonesia

Laboratorium Sentral,
Universitas Padjadjaran

Greece

NoSmoke Team,
Patras Science Park

Serbia

Center for Molecular Medicine and Stem Cell Research,
University of Kragujevac

USA

Temple University,

Oman

College of Medicine & Health Sciences,
Sultan Qaboos University

Russian Federation

Institute of Fundamental Medicine and Biology,
Kazan Federal University

Marocco

Faculté de Médecine
Université Mohammed 6 des Sciences de la Santé

Replica Goals

Non-reproducible studies result in economic damage and prevents innovation, can harm individuals and erode public trust in science.

Replica Project addresses the need to standardize research through reproducibility and provides reliable scientific data on ENDS.

Replicate
high-profile in vitro studies testing ENDS

Assess validity of the original work put under scrutiny

Inform the scientific academia and the public of the progress

Contribute to improve local and international THR policies

How we work

Our Know How

Replica project established fruitful partnerships with international universities and research centers. The activity of the network, although coordinated by the CoEHAR, was conducted independently. Knowledge sharing of the know–how with our international laboratory network is our priority. We have developed communication strategies of our technical expertise through multimedia methods.

Tools

Through a series of harmonization meetings, the team aligned their efforts and operated under standardized operating procedures. Utilizing innovative tools and equipment, the laboratories were able to establish seven specialized "smoking & vaping labs" compliant with ISO 3308 standards. These labs are equipped to follow both ISO and HCI smoking regimens—the latter simulating a more intense smoking topography—ensuring harmonized and reproducible results across the system.

Methodologies

The team investigated key biological effects—cytotoxicity, oxidative stress, mutagenicity, and genotoxicity—related to respiratory and cardiovascular health, and cancer. Standardized exposure protocols using tobacco smoke, e-cigarette (ECIG), and heated tobacco product (HTP) aerosols were applied. Biological endpoints were assessed with validated methodologies, including FDA-approved assays. To address reproducibility concerns in vitro, a robust approach using advanced high-throughput techniques like flow cytometry, real-time cell analysis, and high-content screening was employed, ensuring reliable and replicable results.

Our Working Process

The harmonization process required for all scientific documentation, materials, laboratories set up and experiments management is enriched by partner cultural diversity which prevents scientific bias or product stigma.

Commitment to pre-established design, methods and data analysis ensures value and credibility of scientific findings based on Reproducibility, while Multiple Methodology Comparison contributes to strengthen or debunk the original evidence.

Knowledge sharing of the leading center know-how with our international laboratory network is our priority. We have developed communication strategies of our technical expertise through multimedia methodologies.

Each project phase has been designed and managed following open science principles. Project result are shared in open access journals and repositories. Open education & training, inclusivity and open research characterize all our work.

HIGH-END EQUIPMENT

The smoking and vaping machines

LM1 smoking machine and LM4E vaping machine have been certified by Borgwaldt KC GmbH for instrument installation and calibration guarantee proper step by step functioning and puffing airflow. The LM4E vaping machine allows in line comparison between electronic cigarettes and HTPs (heating tobacco products). The separate LM1 smoking machine prevents contamination during testing between alternative tobacco products and reference cigarettes- representative of traditional tobacco products- allowing therefore proper reading of standard reference cigarettes. The tested product vapor or smoke is channeled in the system via a piston pump and delivered to either a trapping device for chemical analysis or directly delivered to an apparatus hosting human cells to assess biological endpoints.

Methodological gaps

Replica team identified and addressed critical gaps during replication of original studies and proposed solutions with recommendations to improve future research efforts.

New standards

Replica team evaluated the optimal standard condition for use and storage of samples, to create international reference standards.

The results

The team replicated and validated key findings from several influential in vitro studies in the field of tobacco harm reduction. Data generated independently across multiple laboratories consistently demonstrated reduced cytotoxicity of ENDS products compared to cigarette smoke, as well as lower in vitro impact on vascular-related markers associated with atherosclerosis, and reduced genotoxic and mutagenic responses.

Our latest publications

Uniting Global Efforts to Eradicate Smoking Within This Generation

Established in 2018 in Catania, Italy, the Center of Excellence for Harm Reduction (coEHAR) has forged partnerships with universities worldwide to conduct extensive research on Tobacco Harm Reduction. Aligned with our mission, our Indonesian partner advocates for similar research pursuits, aiming to expedite global initiatives aimed at mitigating health repercussions and fatalities linked to smoking. Together, we strive to achieve the ambitious objective of eradicating smoking on a global scale within this generation.

FAQ

Some false myths to dispel

Nicotine is an addictive substance and initiating any smoking and vaping activity is discouraged. ENDS contain many chemical compounds that are less harmful with respect to traditional cigarettes