Imagine: Your collaborative research work is about to submitted for publication, but you are not convinced of its scientific validity. The lead author tells you: either you accept her interpretation of results and become co-author, or she kicks you off the paper. Her shady claims will pass peer review and be published, the scientific community as well as clinicians and patients will be misled. You can be part of it, with another paper decorating your CV, or you surrender your data and leave, but this paper is happening.
This is what happened to Jaywant Phopase, principal research engineer at the Linköping University (LiU) in Sweden, who now asks for your advice below. The university is known to readers of my site for the scandal around the fake professor and predatory conference organiser Ashutosh Tiwari. Incidentally, Phopase’s research was originally performed at the same Faculty of Science and Engineering (IFM), where Tiwari found support and protection by the former prefect Ulf Karlsson. Same Karlsson who allegedly used to bully Phopase, exactly because the latter raised a fuss about bad science being published and patients abroad being mistreated.
That science in question was led by former LiU professor May Griffith, then at LiU Faculty of Medicine and Health Sciences (FSM). Griffith’ research project was about artificial corneal implants, made as a composite of a chemical polymer and human collagen, manufactured at LiU and tested for biocompatibility on human subjects in Ukraine and India. For that she was found guilty of research misconduct by “repeated negligence”, in a LiU investigation from 2015 (see LiU press release and the Swedish-language report summary). Here is what was found by the three external investigators:
- Biocompatibility test operations with biosynthetic corneas have been conducted on blind patients, without adequate prior animal tests. One can only fantasise which benefit exactly the patients whose blindness was not caused by a corneal defect were promised from those corneal implants.
- Animal studies by the company Adlego Biomedical AB have not sufficiently advanced while human experiments in India and in Ukraine already were taking place: “these tests have not yet reached the level required to form the basis of an application to the regulatory authorities”.
- Research-grade porcine collagen was occasionally used to manufacture corneal implants for human tests in Ukraine. Not human collagen, and not medicinally approved quality, which brings severe risk of toxin contamination or infection. Investigation found “non-compliance with documentation procedures for the handling of chemicals”, “partly contradictory and irrelevant information” was filed by Griffith’ team.
LiU then assessed “whether the faculty’s decision will also lead to employment-related consequences for the researcher, such as a warning or a salary deduction”. Whatever they decided, Griffith is not at LiU anymore, but back in her home country Canada, as professor at the University of Montreal.
However, the results of those human experiments from India and Ukraine were published in a Nature-themed journal on January 31st 2018:
M. Mirazul Islam, Oleksiy Buznyk, Jagadesh C. Reddy, Nataliya Pasyechnikova, Emilio I. Alarcon, Sally Hayes, Philip Lewis, Per Fagerholm, Chaoliang He, Stanislav Iakymenko, Wenguang Liu, Keith M. Meek, Virender S. Sangwan & May Griffith
Biomaterials-enabled cornea regeneration in patients at high risk for rejection of donor tissue transplantation
npj Regenerative Medicine volume 3, Article number: 2 (2018) doi:10.1038/s41536-017-0038-8
The authors pretend that animal studies were performed before tests in patients, and of course all their materials are described as safe and certified: “European Medical Devices Directive MDD 93/42/ECC and its associated ISO standards were followed”. Never mind that the findings of LiU investigation say something different. Also, good for Griffith et al that the journal’s editors or reviewers never bothered to ask for ethical approvals for those clinical tests in Ukraine and India. “Written informed consent” from the patients, whatever that meant, sufficed here fully.
It is strange why LiU did not interfere with the journal to clarify the ethical shortcomings of that paper, maybe they might still do it. On the other hand, LiU coordinates a ongoing EU project, funded with €6mn to put those corneal implants into EU-wide clinical use (though Griffith is not part of it):
Advanced Regenerative and REStorative Therapies to combat corneal BLINDNESS
“ARREST BLINDNESS is therefore to develop and validate new regenerative-based therapies addressing a spectrum of blinding disorders of the cornea. These conditions either have no effective current treatments, depend on a scarce supply of donor tissue, or non-standardized methods are hindering validation of promising regenerative treatments. To achieve our objective, we will implant GMP-fabricated collagen-based bioengineered scaffolds to replace or regenerate the corneal stroma in cases of stromal thinning, scarring, dystrophy or trauma; deliver therapeutic epithelial stem and endothelial cells to the cornea to restore its transparency; deliver regenerative factors to promote neural growth and function; and actively maintain corneal immune privilege in high-risk situations by targeted therapeutic approaches to regress blood and lymphatic vessels.”
Now Griffith wants to publish a follow-up study. The corneal implants used in the clinical tests in Ukraine and India were not specially sterilised, though they were manufactured in sterile lab conditions. For a proper, i.e., ethically approved clinical trial one needs to ascertain sterility and the post-sterilisation stability of those collagen-based corneal implants. This is what Griffith announced to have achieved with electron beam irradiation, allegedly “without changing their physical, chemical or functional properties”. Only, regardless what Griffith’s tests in rabbits proved, that was not really the case in Phopase’s own lab analyses. Neither did Griffith ever ran a positive control for sterilisation efficiency by irradiating any actual dirty, bacteria contaminated corneal implants, as Phopase discusses below.
Appeal to scientific community for support to improve research results and oppose biased science
By Jaywant Phopase
I’m a researcher at Linköping University and would like to discuss some of my results from the manuscript which is supposed to be submitted soon. I have some concerns and doubts about our results and there is a serious disagreement about the conclusions between me and one of the senior authors, namely Professor May Griffith. I tried my best to discuss my views and concerns based on facts and my findings, however it’s not going anywhere. I therefore thought that I should reach out to the scientific community and get some help and expert comments.
The aim of this work was to investigate the effect of electron-beam (e-beam) irradiation on artificial corneal implants, which previously have been implanted into the eyes of blind humans to test biocompatibility. The material these corneal implants are made of, is called RHCIII-MPC: RHC being recombinant human collagen type III, MPC being 2-Methacryloyloxyethyl phosphorylcholine. We fabricated the RHCIII-MPC corneal implants in a clean lab and sealed them into sterile vials. These implants were then sent to Sterigenics, Denmark, and irradiated with three different doses of e-beam (17 kGy, 19kGy and 21 kGy). After e-beam irradiation, the implants were analyzed and compared with the control samples, which were unirradiated implants. The effect of e-beam irradiation on composition, structural changes and degradation rate of implants was evaluated using Fourier transformed infrared spectroscopy (FTIR) and biodegradation study, where irradiated and unirradiated implants were subjected to collagenase degradation. My observations were as follows.
FTIR analysis: The FTIR analysis of e-beam irradiated implants (all the doses, 17, 19 and 21 kGy) showed changes in the composition of implants. The amount of MPC polymer increased with increasing dose of e-beam irradiation. The peak intensity of MPC polymer increased with the increasing dose of e-beam. This also indicated the fact that these implants contained unreacted MPC monomer which got polymerized after e-beam irradiation. This also raised the concern about the use of these implants as they have been implanted into the humans without e-beam irradiation, meaning they likely contained unpolymerized MPC. We also observed the change in amide I and amide II bonds for collagen, indicating the changes in collagen structure. This is also in good agreement with the literature previously reported.
The biodegradation study showed that degradation rate of collagen in these implants increased significantly after e-beam irradiation and was much higher than in control unirradiated implants. The degradation rate slowed down after 12 hours because most of the collagen is degraded by that time and what remains is the MPC polymer which does not undergo degradation. The biodegradation study also confirmed the fact that the content of MPC polymer, the second polymer network in these implants, increased considerably after e-beam exposure in all the e-beam doses (17, 19 and 21 kGy). Furthermore, after exposing these implants to collagenase degradation for extended period, we got very stable thin film of MPC polymer from all the e-beam irradiated samples but not from the control samples (those without e-beam irradiation).We also confirmed that these films were nothing but the MPC polymer, by characterizing them using FTIR. These results are in good agreement with FTIR analysis described above. All this means, irradiation damaged the collagen structure and made it susceptible to enzymatic degradation. We also seeded human corneal epithelial cells (HCEC) on these MPC polymer films and found out that the cells were not able to attach or grow. Taken together, this means that irradiated RHCIII-MPC corneal implants are likely to quickly lose their collagen coating due to in vivo biodegradation and fail to be colonised by corneal epithelium when implanted into patients. They will basically lose their biocompatibility. These facts are quite crucial in my opinion and I’m insisting to include them in the manuscript. The senior author does not share the same view in this context and would like to exclude these observations and instead declare that low dose e-beam irradiation (17 kGy) has no significant effect on the RHCIII-MPC corneal implants.
Furthermore, the senior author has also included the following conclusion in our manuscript.
“Low dose e-beam irradiation (17 kGy) is an effective method of sterilization for RHCIII-MPC corneal implants.”
In my opinion this is clearly a misleading. The aim of our study was only to investigate the effect of e-beam irradiation on RHC-III-MPC implants. Therefore, we never validated if e-beam irradiation (any dose) was efficient to sterilize the contaminated/infected RHCIII-MPC corneal implants. It is very important to note that all the implants were fabricated in clean lab and were sealed into sterile vial. All the implants were sterile, before we sent them for e-beam irradiation. Therefore, it is no wonder that these implants were sterile after e-beam irradiation. I never understood, why so much of tax money was wasted to test the sterility of these implants after e-beam irradiation, when we knew that these implants were sterile before we subjected them to e-beam irradiation. We never performed any control study or even a single experiment to check if a 17 kGy dose is enough to kill any contamination present within these implants. I have also discussed this with few biologists as well with the person from Sterigenics, Denmark, who performed e-beam irradiation on our implants. He clearly indicated that the penetration depth of e-beam is limited, and it cannot penetrate thick materials. Therefore, we cannot claim that any of the e-beam doses we have used is enough to sterilize our implants, unless we validate it using infected samples (positive control). Moreover, the sterilization dose can also vary depending on the type of contaminants present in the implants. There are several articles about sterilization studies performed using different techniques, including e-beam irradiation which supports my argument above.
Based on my observations described above, I also have a concern regarding the long term performance and also use of these implants in highly inflamed corneal conditions (eg. chemical burn, HSV infection). The concentration of collagen-digesting enzymes (collagenases) is quite high during inflammation. Therefore, degradation of e-beam irradiated collagen implants will be higher than it was determined in past human experiments with unirradiated RHCIII-MPC corneal implants. The leftover MPC polymer network will then get in contact with the surrounding tissue and cells. As described above, cells were not able to attach to MPC polymer and I wonder what will happen in such scenario. MPC polymer is in fact known for its antifouling properties and is used to coat biosensor devices to avoid the attachment of proteins and cells on the surface. (http://www.whoi.edu/cms/files/Chae_07_36863.pdf).
I invite all the researchers and clinicians to help me with your critical comments and correct me, in case any of my observations/conclusions are inappropriate. Please do not hesitate to write me in case you need any more information or if you have any questions.
Thank you very much in advance for your support and help.
Jaywant Phopase, PhD, Docent
Email: jaywantphopase AT gmail DOT com
If you have an advice for Jaywant Phopase, please comment below.
Update 12.03.2018. The article in question has now been published, without Phopase:
Simpson F, Edin J, Islam MM, Buznyk O, Kozak ML, Liszka A, Merrett K, Gustafsson HP, Griffith M.
E-beam sterilization of recombinant human collagen-phosphorylcholine corneal implants for transplantation.
Ann Eye Sci 2018;3:AB085. doi: 10.21037/aes.2018.AB085
Update 13.03.2017. I received from LiU HR director, Pia Rundgren, an email in cc, where it looks like Linköping University blackmails me to remove Phopase’s institutional affiliations which I myself added to his guest post discussing his own work there. There seems to be an implicit threat of disciplinary consequences to Phopase should I not remove those affiliations:
As you must be well aware of by now, LiU has no intention of interfering with any statements made/information shared by you in your private capacity. And we have no opinion as to the content. But as an employee at LiU you are obligated to, when engaging in social media, at all times make sure that there is no confusion about whether or not your statements in this aspect are made by you as a representative of the University. We simply ask of you to abide to this obligation. How you chose to proceed to abide by this obligation is primarily your own choice.This is an obligation that applies to all employees of LiU. Neither the obligation, nor this communication is of any relevance to Mr Schneider.
Phopase asked me to comply with the order by LiU for his sake (hence the edited version above) and declared:
“Please kindly note that my intentions behind my article is purely scientific. There had been serious disagreement regarding the research results, which I believe are very serious and might be misused for clinical trails by the clinicians. Both the LiU Deans, (Prof. Nilssson and Prof. Södherholm) were involved in this discussions and I have informed both of them about my intentions that I will contact the scientific community to peer review my results and conclusions”.
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