How develop a Biological Evaluation: a case study

INTRODUCTION

Despite the numerous benefits of medical devices, all present some

degree of RISK even when used appropriately.

For this reason, risk assessments of medical devices must include a

Biological Evaluation Plan.

Such an evaluation plan is described in the ISO 10993 series of standards

for both Europe and the United States.

The purpose of Biological Evaluation, according to the ISO 10993

standards, is to estimate the biological risk in order for the medical
device to be considered biocompatible. The Biological Safety Evaluation
Plan (BEP), together with the Biological Evaluation Report (BER), can
help the manufacturer fully understand the device and its
biocompatibility. Moreover, these documents are useful for U.S. FDA
submission and/or CE marking.

Medical devices, must satisfy general safety and performance

requirements since contact with the human body can generate risks,

Moreover they must demonstrate that medical devices are safe, but
 How can they succeed in this task?

There is no standard pathway to follow. Manufacturers have to show

scientific proof of safety.

Chemical characterization: a case study

The new version of ISO 10993-1 emphasizes the importance of a

chemical characterization before any biological testing. Physicochemical
and morphological properties of a device and of its constituents should be
considered if they have an impact on biocompatibility.
Before performing any tests, especially in vivo, we need a better
understanding of the medical device by analyzing its composite materials,
manufacturing, sterilization and other post manufacturing processes.
On the basis of this preliminary evaluation, the intrinsic properties of a
medical device can be understood; so it will be possible to establish the
endpoints requiring further investigations through biological testing.

Chemical characterization and toxicological evaluation may provide

information related to long-term systemic effects; therefore they may
allow the waiver of some tests. Here an example of chemical
characterization of a real medical devices, a chemical characterization on
hydrophobic IOLs, as other equivalent hydrophobic intraocular lenses
available on the market. They are indicated for the visual correction of
aphakia:

Hydrophobic IOL
# Reference (component) Type of Materials % by weight Supplier
contact
Hydrophobic intraocular Direct contact 2-Ethoxyethyl /
1
lenses Methacrylate
2 Methyl Methacrylate /
Ethylene Glycol /
3
Dimethacrylate
2-(5-chloro-2H- /
benzotriazol-2-yl)-6-
4
(tert-butyl)-4-
ethenylphenol
4-(3-Vinylphenylazo) /
5
diphenylamine
N-Vinyl-N- /
6 methylacetamide

7 2-Phenyethyl /
Acrylate
 2,2'-azobis(2,4- /
dimethylvaleronitrile
8
)

9 Platinum Dioxide /

This kind of study, must be performed on medical devices before clinical

evaluation to evaluate:

- The interaction with the patient on the basis of the device’s nature
- Duration of the contact
- The part of the body involved

Certain classes of device require a more stringent chemical

characterization study. These studies, also known as device extractables
and leachables studies, follow ISO 10993-18 testing methods, and look at
the types and amounts of chemicals that may migrate from a device
during use. From this specific chemical profile, an overall risk assessment
of the safety of the device can be created.

Chemical characterization studies can be complex and are meant to

examine devices with a greater potential risk to users. Currently, the US
FDA requires a device chemical characterization, extractables and
leachables study with an associated toxicological risk assessment to be
performed on any device with the potential for cumulative systemic
 contact, long term implants would naturally be included, as would many
short-term yet repeated-use devices.

What are "extractables"?

In order to study the risk of materials extraction studies are performed;
generally using aggressive solvent conditions including acidic, basic,
organic and aqueous solvents and sohxlet or accelerated solvent
extraction.

What are "leachables"?

Leachables are chemical species that make their way into the product
under normal product, application or storage conditions.

In our device has been found a toxic substance using the leachables and
extractables test:

Final
Final Potential
propose Margin of Toxicological
Name CAS proposed patient daily
d TTC** safety concern
Number TE* [µg/day] intake
[µg/day] (MOS)
(µg/day)

PhenylethylAlcool 60-12-8 127 NA 3.36 >1 NO

50
Unknown
Probable
(probable
CAS 625‐06‐
aliphatic alcohols): NA 7.5 + 2.70 >1 NO
9;
18720‐66‐6;
589‐98‐0;
18720‐65‐5
 N-Acetyl-3.4- / NA 7.5 93.64 <1 YES
dehydroproline
Ciclobuthyltridecil / 1500 NA 15.06 >1 NO
Hexadecanol 36311-34- 5635 NA 2.60 >1 NO
Diphenylsuccinate 621-14-7 NA 150 51.42 >1 NO
3.5‐ 1138‐52‐9 NA 150 62.24 >1 NO
Octylphtalate 117‐84‐0 100 NA 14.22 >1 NO
Isopropylmyristat 110-27-0 5000 NA 15.42 >1 NO
2.6-bis(1.1- >1
dimethylethyl)-4- 6738-27-8 150 NA 12.02 NO
ethylidene-2-5- 0
Cyclohexadien-1- one
4.6-di(1.1-
616-55-7 125 NA 11.32 >1 NO
dimethylethyl)-2-
methyl-Phenol

The N-Acetyl-3.4-dehydroproline has been evaluated has a toxicological

concern. Then we decided to make a toxicological evaluation on that
substance, which is a derivate of main ingredient, generated during the
gamma irradiation process, as intermediate step of the manufacturing.

Toxicological evaluation
The toxicological evaluation was carried out in order to evaluate the
potential toxicological concerns of the extractable compounds from the
medical device “ADAGIO Hydrophobic preloaded IOLs”.
The toxicological evaluation is based on the results of chemical
characterization study. Extractable compounds analysed in the present
assessment were defined in document “ADAGIO Hydrophobic preloaded
IOLs residuals - TOXICOLOGICAL EVALUATION”, provided by the Sponsor.
In the technique used, GC/MS, the samples were extracted with acetone
(exhaustive extraction).
The extractable compound were toxicologically evaluated according to EN
ISO 10993-17:2009 and ISO/TS 21726:2019 therefore applying the
Threshold of Toxicological Concern (TTC) and Tolerable Exposure
concepts.
The potential patient daily intake (µg/day) was calculated considering 2
devices per patient as worst case scenario.
The comparison between the potential daily intake (µg/day) and the
TTC/TE (µg/day) was based on the assumption that the total release of
two devices identified in chemical study through the exhaustive
extraction occurs in one day (µg/item ~µg/day).
The amount of each organic extracted compound was compared with a
conservative Threshold of Toxicological Concern (TTC) value of 1.5 µg/day
(as described in ICH M7 and ISO/TS 21726:2019) which was considered
relevant for the device under evaluation since its contact in adult is long-
term (i.e. > 10 years to a lifetime).
Based on the data coming from the document “Hydrophobic preloaded
IOLs residuals - TOXICOLOGICAL EVALUATION”, combining the genotoxic
threshold, the clinical use of the medical device and the actual
toxicological evaluation, it can be concluded that in the sample “Shelf
Life T3 Accelerated” for the compound N‐Acetyl‐3,4‐ dehydroproline the
final proposed Tolerable Exposure (TE) value is lower than the potential
patient daily intake and therefore they could pose a toxicological
concern.

Compound SMILES Cramer Class Benigni/Bossa Proposed TTC

value (µg/person
rulebases
per day)
Negative for
genotoxic
carcinogenicity
N‐Acetyl‐3,4‐ CC(=O)N1CC=CC1
III 75
dehydroproline C(O)=O
Negative for
nongenotoxic
carcinogenicity

We can assume that the N-Acetyl-3.4-dehydroproline is a derivate of

main ingredient, generated during the gamma irradiation process, as
intermediate step of the manufacturing.
The possibility to use a more effective washing process in production
could be investigated.
However, the calculated Potential patient daily intake is higher than the
calculated TTC value, including the UF; therefore this leachable will trigger
additional biocompatibility tests.
The calculation of Potential patient daily intake of N-Acetyl-3.4-
dehydroproline was based on its SMILE methodology, an in silico tool
used when no toxicological data are available.
The obtained value is divided for 10 uncertainty factor (UF), as default
factor typically used for the general population in the absence of chemical
specific data, to extrapolate to the parenteral route of administration.
This methodology is a conservative approach, which include high safety
factor, but the potential toxicological properties should be then
investigated through the biocompatibility tests.

Biocompatibility Tests
Endpoints to consider are chosen based on ISO 10993-1 table A.1 and ISO
11979-5: 2006.
They represent the biological hazards that are associated with the device
under evaluation.
A summary of all main components, as well as the quantities present in
the final product, the maximal exposure dose and their toxicological
properties have been evaluated.
The type of contact and contact duration of the components of the
medical device PRELOADED HYRODPHOBIC IOL and patient exposure was
included in the risk assessment as part of the sequence of events, which
may lead to toxicological and biological harms for the patient. Considering
the type of contact and contact duration of the components of the
medical device PRELOADED HYRODPHOBIC IOL , its ingredients are well
below the toxicity threshold which could generate biocompatibility
concenrs. Therefore, no patient risk of exposure to medical device
PRELOADED HYRODPHOBIC IOL is expected.

CONCLUSION
The results of the in vitro and in vivo biocompatibility tests will be
evaluated in the final Biological Evaluation Report.

In conclusion we make a PLAN: the Biological Evaluation Plan, in order to

analize every single substance in the devices and understand which one
have a toxicological concern based on
- Type of contact with the patients
- Duration of the contact.

Based on the toxicological risk assessment we decide to perform some

biological tets, in order to demonstrate the safety of the devices.
The results of these tests are described in the Biological Evaluation
Report.