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!



International 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.

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.


Riccardo Polosa

CoEHAR Founder


Giovanni Li Volti

CoEHAR Director and Project Leader


Massimo Caruso

Co-project leader

our network


Eight international laboratories


University of Catania


Laboratorium Sentral,
Universitas Padjadjaran


NoSmoke Team,
Patras Science Park


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


Temple University,


College of Medicine & Health Sciences,
Sultan Qaboos University

Russian Federation

Institute of Fundamental Medicine and Biology,
Kazan Federal University


Faculté de Médecine

Université Mohammed 6 des Sciences de la Santé

University of Kragujevac

The NOSMOKE Institute

Universitas Padjadjaran

Sultan Qaboos University

Sbarro Health Research Organization

Università degli Studi di Catania

Kazan University

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.

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.


The team, coordinated through several harmonization meeting, was able to operate using standardized operative procedures. The innovative tools and equipment inside the laboratories allowed to create seven “smoking & vaping labs” compliant with ISO3308 and capable to follow standard regimens such ISO and HCI, the latter intended to reproduce a more intense smoking topography.


The team focused on cytotoxicity, oxidation, mutagenicity and genotoxicity affecting respiratory and cardiovascular system as well as cancer disease. Standardized exposure to tobacco smoke or ECIGs and HTPs aerosol was followed by the evaluation of biological endpoints through a battery of diverse methodologies including FDA approved methodologies. Moreover, a well-rounded approach has been implemented to verify replicability of results through new high-throughput methodologies, including flow cytometry, real-time cell analysis and high-content screening analysis.

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.

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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 some gaps during the replication of the original works and accordingly found the solutions to recommend for new studies.

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 the result of some of the most relevant studies in the field of tobacco harm reduction: overall, the data obtained independently in different laboratories clearly confirm the reduced cytotoxicity of ENDS products compared with smoking, the reduced impact of ENDS on vascular system and consequently the onset of atherosclerosis, and the reduced genotoxicity and mutagenicity of ENDS.

Risk Reduction

Ecig is 95% less harmful than traditional cigarette


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