biomimetics

Article
Basic Research of Material Properties of
Mycelium-Based Composites
Hana Vašatko * , Lukas Gosch, Julian Jauk and Milena Stavric

Faculty of Architecture, Institute of Architecture and Media, Graz University of Technology, 8010 Graz, Austria;
[email protected] (L.G.); [email protected] (J.J.); [email protected] (M.S.)
* Correspondence: [email protected]

Abstract: The subject of this research is growing mycelium-based composites and exploring their
basic material properties. Since the building industry is responsible for a large amount of annual
CO2 emissions, rethinking building materials is an important task for future practices. Using such
composites is a carbon-neutral strategy that offers alternatives to conventional building materials. Yet,
in order to become competitive, their basic research is still needed. In order to create mycelium-based
composites, it was necessary to establish a sterile work environment and develop shaping procedures
for objects on a scale of architectural building elements. The composite material exhibited qualities
that make it suitable for compression-only structures, temporary assemblies, and acoustic and thermal
insulation. The methodology includes evaluating several substrates, focused on beech sawdust, with
two mycelium strains (Pleurotus ostreatus and Ganoderma lucidum), density calculations, compression
tests, three-point flexural tests and capillary water absorption. The results of this study are presented
through graphical and numerical values comparing material and mechanical properties. This study
established a database for succeeding investigations and for defining the potentials and limitations of
this material. Furthermore, future applications and relevant examinations have been addressed.
Citation: Vašatko, H.; Gosch, L.; Jauk,
Keywords: mycelium; growth; bio-composites; mechanical properties; architecture; materials science
J.; Stavric, M. Basic Research of
Material Properties of Mycelium-
Based Composites. Biomimetics 2022,
7, 51. https://doi.org/10.3390/
biomimetics7020051 1. Introduction

Academic Editors: Andrew

Contemporary problems, such as rapid population growth, the increased demand for
Adamatzky, Han A.B. Wösten and
food and housing, freshwater scarcity and inadequate waste management, are all influenced
Phil Ayres
by demographic, environmental and economic factors. The current linear economic model
relies on the extraction, transformation and disposition of raw materials once their life cycle
Received: 15 March 2022 ends. Meanwhile, the concept of circular economy is defined by the reuse of materials, with
Accepted: 19 April 2022 radical changes in the production itself taking place, thus preventing the accumulation of
Published: 21 April 2022
waste [1].
Publisher’s Note: MDPI stays neutral An additional fact driving this research is that embodied carbon emissions from the
with regard to jurisdictional claims in manufacturing of building materials and the construction sector account for 38% of annual
published maps and institutional affil- worldwide greenhouse gas emissions in the current world climate [2]. Out of the four raw
iations. material types currently extracted—minerals, ores, fossil fuels and biomass—the major
portion, totalling approximately 40%, finds its use in the construction and housing sector [3].
If the European energy strategy with net-zero emissions is to be realised by 2050 [4], it is
time to reconsider present design concepts as well as building elements and materials.
Copyright: © 2022 by the authors.
One way to accomplish this is by investigating new bio-based building materials.
Licensee MDPI, Basel, Switzerland.
In the last decade, bio-fabrication gained significance in architecture and has become an
This article is an open access article
integral part of sustainable building strategies. The production of mycelium-based com-
distributed under the terms and
posites is a low-energy and carbon-neutral process [5] that fits into the circular economy
conditions of the Creative Commons
and sustainable building concepts. The utilisation of mycelium-based composites is wide
Attribution (CC BY) license (https://
in terms of scale, functionality and application, as several architects, designers and enthu-
creativecommons.org/licenses/by/
4.0/).
siasts have begun to use them in their designs during the past decade. Furthermore, the

Biomimetics 2022, 7, 51. https://doi.org/10.3390/biomimetics7020051 https://www.mdpi.com/journal/biomimetics

Biomimetics 2022, 7, 51 2 of 16

composite has found its application as packaging material (Ecovative), acoustic insulation
(MOGU) and as temporary objects exhibited in a larger scale, such as MycoTree [6] and
the Growing Pavilion [7]. In comparison to building materials such as concrete or bricks,
mycelium-based composites are a new term in architecture. Hence, there is still a high
demand for basic research and testing in this respect. Several works have already pre-
sented some initial research by combining different substrates and mycelium strains and
subsequently elaborating on some of their mechanical properties [8–12], and morphology
and mechanics [13]. The work presented in this article extends the relevant references by
providing an overview of the basic material properties in very specific material combina-
tions, which include organic substrates, organic fibrous materials and inorganic materials.
The introduction of inorganic and fibrous materials as substrates—such as clay, sand and
soy silk fibres—contributes to the mentioned references. Since a wide variety of substrates
and mycelium strains are present, as well as several decisive factors during the production
process, the specification of the material properties is relevant. These preliminary experi-
ments introduce the concept of growing mycelium-based lignocellulosic products, whose
properties may subsequently be fine-tuned based on the material’s intended application.
The overarching purpose of this research is fabricating heterogeneous composites with
a defined material distribution, which will optimise the structural properties within one
geometry. This study offers the initial point of evaluating material properties that will be
used in these experiments.
Broadly speaking, mycelium is the vegetative part of mushrooms, which consists of
branching hyphae. Mycelial growth can be described as a hyphal penetration of a substrate,
which results in unifying it into one piece. A spore inoculated on a nutrient forms a tube
which experiences exponential non-photosynthetic growth [14]. Three growth phases can
be differentiated after inoculating a lignocellulosic substrate: (1) the lag phase (zero to
little population growth, the mycelium cells get used to their new environment), (2) the
exponential phase (if the conditions remain favourable, increase in biomass takes effect, as
well as the cell number—this is the optimal period for mycelial growth and continuation
of this for as long as possible is desirable) and (3) the stationary phase (the population
growth returns to zero, the fungal biomass remains constant and some fungal cells may
begin to perish) [15,16]. The oyster mushroom (Pleurotus ostreatus) mycelium was the most
frequently used species for this research, as this very common mushroom type shows
a high contamination resistance compared to other tested specimens and is capable of
consuming a variety of lignocellulosic substrates. Its by-products, water, carbon dioxide,
enzymes, alcohols and carbohydrates, serve as a nutritious foundation for other organisms
in nature [17].

2. Materials and Methods

The methods of this research were carried out through material experiments and
investigation of mechanical properties for an application as a building material. A series of
material mixtures was created, as well as samples with different geometries for material
testing experiments (Figure 1). The research presented here constitutes a database for
subsequent investigations conducted by the authors, as well for the overall understanding
of the potentials and limitations of the material. This initial study offers a comparison
between different material qualities and their fine-tuned versions according to the function
of their application.
Various organic substrates as well as inorganic additives were tested. However,
beech sawdust was the initial substrate for assessing all tested material properties due
to its availability in the local area, which minimises transportation and processing costs.
Additionally, bleached cellulose pulp was a substrate option, as it achieved solid results
and demonstrated no problems during inoculation and the growing phase, with very little
to no contamination. Obtaining large quantities of cellulose in a desired form, however,
proved to be challenging, leading to the choice of beech sawdust as the preferred substrate.
The mycelium grain spawn used for this research was bought from a local vendor. Other
 Biomimetics 2022, 7, 51 3 of 16

FOR PEER REVIEW 3 of 15

substrates were also tested on individual samples, e.g., sand and clay, in order to increase
the plasticity while comparing compressive strength.

(a) (b) (c)

Figure 1. Samples composed
Figure 1.ofSamples
various materials:
composed (a) cellulose
of various pulp
materials: and clay;
(a) cellulose (b)
pulp beech
and sawdust
clay; (b) and and
beech sawdust
clay; (c) oak sawdust. clay; (c) oak sawdust.
All samples were given abbreviations within a naming system, which is used through-
Various organic outsubstrates
this research: asPleurotus
well asostreatus
inorganic additives
mycelium were tested.
(PO), Ganoderma lucidum However,
mycelium (GL),
beech sawdust was beechthe initial substrate for assessing all tested material properties
sawdust (FS), oak sawdust (Q), bleached cellulose pulp (CS1), shredded duecardboard
to
its availability in the(SC),
localshredded newspaper
area, which (SN), cotton
minimises fibres (CO), soy
transportation andsilk fibres (SF), costs.
processing wheat bran
Ad- (WB),
straw (ST), burlap (B), clay (C) and sand (S). The nomenclature works in a way that the
ditionally, bleached cellulose pulp was a substrate option, as it achieved solid results and
abbreviation of the substrate is used firstly, followed by the abbreviation of the mycelium
demonstrated no problems
strain, and during
endinginoculation
with a sampleand the growing
number. phase, and
The abbreviations withthevery littlesequences
number to
no contamination. Obtaining
are connectedlarge quantities
with hyphens (e.g., of cellulose
FS-PO-01 is thein a sample
first desired form,sawdust
of beech however,inoculated
proved to be challenging, leading to the choice of beech sawdust as the preferred sub-
with oyster mushroom mycelium).
strate. The mycelium grain The spawn
following sections
used provide
for this a detailed
research wasdescription
bought fromof theaproduction
local vendor.procedure,
initial substrate exploration, together with the mechanical material testing and evaluation
Other substrates were also tested on individual samples, e.g., sand and clay, in order to
of basic material properties. The initial substrate exploration includes substrate compat-
increase the plasticity while
ibility andcomparing
investigatingcompressive strength.material testing includes evaluation of
density. The mechanical
All samples were given strength
compression abbreviations within aflexural
and a three-point naming test. system, which is the
Finally, determining usedcapillary
water absorption coefficient was conducted.
throughout this research: Pleurotus ostreatus mycelium (PO), Ganoderma lucidum mycelium
(GL), beech sawdust2.1. (FS), oak sawdust
Production Procedure(Q), bleached cellulose
of Mycelium-Based Compositespulp (CS1), shredded card-
board (SC), shredded newspaper
The starting(SN),
pointcotton fibres (CO),
for producing soy silk fibres
mycelium-based (SF), wheat
composites bran the
was preparing
(WB), straw (ST), burlap (B), clay (C) and sand (S). The nomenclature works in
substrate by soaking it in distilled water for 24 h. Afterwards, the excess water wasa way that
drained,
the abbreviation of theandsubstrate
the moisture is used
contentfirstly,
(MC) of followed by the
the substrate wasabbreviation
measured. Theofsubstrate
the myce- was then
lium strain, and endingput in with
a polypropylene
a sample microfilter
number. bag abbreviations andSacO2)
The(PP50/SEU4/V40-51, and sterilised
the number se- in a
pressure cooker at 121 ◦ C for 45 min to eliminate any competing microorganisms that might
quences are connected with hyphens (e.g., FS-PO-01 is the first sample of beech sawdust
hinder mycelial development. The moulds were made of perforated transparent foil with a
inoculated with oyster mushroom
thickness mycelium).
of 0.5 mm in order to produce samples for material testing, whose dimensions
The following sections
dependedprovide a detailed
on the conducted description
method. of the production
Once the sterilised substrate cooledprocedure,
down to room
temperature,
initial substrate exploration, mycelium
together withgrain
thespawn was homogeneously
mechanical distributed
material testing in microfilter bags
and evaluation
while working in a still-air box. The amount of mycelium used for inoculation was 10%
of basic material properties. The initial substrate exploration includes substrate compati-
of the weight of the sterilised substrate. The moulds were cleaned with rubbing alcohol
bility and investigating density.
(ethanol, The mechanical
70% solution) and filled with material testingsubstrate
the inoculated includes by evaluation
manual pressing. of The
compression strength and a bags
microfilter three-point
containingflexural
the closed test. Finally,
moulds were determining
sealed and stored thein capillary
an environment
◦
water absorption coefficient waslight
protected from conducted.
sources at temperatures ranging from 22 to 24 C. The initial phase of
development occurred in the moulds, followed by the second phase after unmoulding to
achieve growth of the outer protective skin of the sample. The duration of each growth
2.1. Production Procedure of Mycelium-Based Composites
phase is defined in the succeeding sections. Once the mycelium fully colonised the substrate,
The starting point for producing
dehydration mycelium-based
was initiated composites
in order to terminate was preparing the sub-
the growth.
strate by soaking it in distilled water for 24 h. Afterwards, the excess water was drained,
2.2. Initial Substrate Exploration
and the moisture content (MC) of the substrate was measured. The substrate was then put
Combinations of Pleurotus ostreatus and several substrates, including straw, beech
in a polypropylene microfilter
sawdust, wheat bagbran
(PP50/SEU4/V40-51, SacO2)
and bleached cellulose and sterilised
pulp (Figure in a pressure
2), were examined to evaluate
cooker at 121 °C for 45 min to eliminate any competing microorganisms that might hinder
mycelial development. The moulds were made of perforated transparent foil with a thick-
ness of 0.5 mm in order to produce samples for material testing, whose dimensions de-
pended on the conducted method. Once the sterilised substrate cooled down to room tem-
 2.2. Initial Substrate Exploration
Combinations of Pleurotus ostreatus and several substrates, including straw, beec
phase is defined in the succeeding sections. Once the mycelium fully colonised the sub-
sawdust,
strate,wheat bran was
dehydration andinitiated
bleached cellulose
in order pulp the
to terminate (Figure
growth. 2), were examined to evalua
breaking and shrinking, growth density, surface colour, and quality impression. The d
Biomimetics 2022, 7, 51 4 of 16
2.2. Initial
mensions Substrate
of the moulds Exploration
were 10 × 10 × 2 cm. Four samples were prepared for each su
strate type, with two of
Combinations of Pleurotus
them incorporating a piece
ostreatus and several of burlap
substrates, in the straw,
including centrebeech as addition
sawdust,
organic wheat
reinforcement. bran and bleached
The obtained cellulose
burlap pulp (Figure
fabriccolour, 2),
was woven were examined
fromimpression. to evaluate
jute and cut in squar
breaking and shrinking, growth density, surface and quality The
breaking and shrinking, growth density, surface colour, and quality impression. The di-
with dimensions
dimensions ofofthe 11moulds
× 11 cm and10had
were × 10a ×fabric
2 cm.weight of 180were
Four samples g/mprepared
2 . The individual
for each burla
mensions of the moulds were 10 × 10 × 2 cm. Four samples were prepared for each sub-
substrate
piecesstrate
were type,
larger with two of them incorporating a piece of burlap in the centre as additional
type, withthan
two ofthethem
moulds in ordera to
incorporating visually
piece of burlapassess
in the
organic reinforcement. The obtained burlap fabric was woven from jute and cut in squares
the continuation
centre as additional of myceli
growth on the
organic burlap. It The
reinforcement. wasobtained
used simultaneously
burlap for nutrition, since cutjute fibres are com
with dimensions of 11 × 11 cm and had afabric
fabric was woven
weight offrom
180 jute
g/mand2 . The in squares
individual
posedburlap
of lignocellulose
with dimensions of 11
pieces were largerand than
× 11 for decreasing
cm the moulds inthe
and had a fabric shrinking
weight
order of 180
to visually ofassess
g/m the composite.
2. The individual burlap
the continuation It was of stiff an
pieces
inelastic, were
and it larger
had athan the
moderate moulds in
moistureorder to visually
regain assess
[18]. Boththe continuation
burlap
mycelial growth on the burlap. It was used simultaneously for nutrition, since jute fibres and of
themycelial
substrate we
growth on
are composed the burlap. It was
of lignocellulose used simultaneously
and fordescribed
decreasingin for nutrition,
thetheshrinking since
of thejute fibres
composite. are com-
prepared according to the procedure previous section. TheItmoulds
was we
posed of lignocellulose
stiff and inelastic, and itand hadfor decreasing
a moderate the shrinking
moisture of the
regain [18]. composite.
Both burlap and It was
the stiff and
substrate
not covered
inelastic, withit had
and a lid, as increased
a moderate surface contact with and oxygen accelerates were myceli
were prepared according to the moisture
procedure regain [18].
described Both
in theburlap the substrate
previous section. The moulds
growth [11].
prepared
were not according
covered with to the procedure
a lid, as increaseddescribed
surfaceincontact
the previous section.accelerates
with oxygen The moulds were
mycelial
not covered
growth [11]. with a lid, as increased surface contact with oxygen accelerates mycelial
growth [11].

(a) (a) (b)(b) (c)(c) (d) (d)

FigureFigure
2. Substrate
Figure scale:
2.2.Substrate
Substrate (a)(a)
scale:
scale: straw;
(a)straw; (b)
straw;(b) wood
(b)wood chips;
woodchips;
chips;(c) (c) wheat
(c)wheat
wheat bran;bran;
bran; (d) (d) bleached
(d)bleached
bleached cellulosecellulose
cellulose pulp.
pulp. pulp.

Inaddition
additiontotosuccessfully
successfullygrown
grownsamples,
samples,aathorough
In
In addition to successfully grown samples,thorough
a thorough growthdocumentation
growth documentationwas
growth was
documentation w
obtained that
obtained that demonstrates changes of the growth patterns and speed of the used
changes of the growth patterns and speed of the used myce- mycelium
obtained
lium
that
strainstraindemonstrates
as well
as as itsas
well preferredchanges of
nutrition.
its preferred
the growth
The exponential
nutrition.
patterns
growth
The exponential
and speed
phase phase
growth
of the
[15,16][15,16] used
was visible
was
myc
lium visible
strain as well
after aafter
couple of as
days its preferred
only (Figure nutrition.
3). Since the The
moulds exponential
were made ofgrowth phase
transparent
a couple of days only (Figure 3). Since the moulds were made of transparent [15,16] w
plastic,
visiblethe growth
after
plastic, the on theof
a couple
growth oncovered
days sides (Figure
only
the covered was noticeably
sides was3). reduced.
Since
noticeably the After
moulds
reduced. 20were
After days, thethe
made
20 days, samples
of transpare
sam-
were
ples unmoulded,
were unmoulded, flipped upside-down
flipped upside-downand placed
and back
placed in the
back
plastic, the growth on the covered sides was noticeably reduced. After 20 days, microfilter
in the bag to
microfilter achieve
bag to the sam
homogeneous
achieveunmoulded, surface
homogeneousflippedgrowth on all
surface growth sides. After the
on all sides. unmoulding, the second growth phase
ples were
lasted for fivelasted
days. forFinally,
upside-down
the samples
andAfter the unmoulding,
placed back in the the second
microfilter bag
growth phase five days. Finally,were dried over
the samples werea heating source.
dried over The results
a heating source.of
achieve
The
homogeneous
these samples
results
surface
are analysed
of these samples in
growth
areSection
analysed3.1.on all sides. After the unmoulding, the secon
in Section 3.1.
growth phase lasted for five days. Finally, the samples were dried over a heating sourc
The results of these samples are analysed in Section 3.1.

(a) (b) (c)

Figure
Figure3.3.Close-up
Close-upofofthe
thegrowth
growthprocess
processofofoyster
oystermushroom
mushroommycelium
myceliumon
onbeech
beechsawdust:
sawdust:(a)
(a)after
after
three days; (b) after five days; (c) after 19 days.
three days; (b) after five days; (c) after 19 days.

2.3. Material Testing

(a) (b) (c)
Material testing included the evaluation of basic material properties and mechanical
Figurematerial
3. Close-up of the
testing. growth
The formerprocess of oyster
considered mushroomofmycelium
the calculation oncapillary
density and beech sawdust:
water (a) aft
absorption
three days; coefficient
(b) after (Figure
five days; 4c), and
(c) after the latter considered the compression strength and
19 days.
 2.3. Material Testing
Material testing included the evaluation of basic material properties and mechanical
material
Biomimetics 2022, 7, 51testing.
The former considered the calculation of density and capillary water ab-5 of 16
sorption coefficient (Figure 4c), and the latter considered the compression strength and
the three-point flexural test. The mechanical material testing was mostly executed at the
the three-point flexural test. The mechanical material testing was mostly executed at the
Institute of Technology and Testing of Building Materials, Graz University of Technology
Institute of Technology and Testing of Building Materials, Graz University of Technology
(Figure 4a,b). (Figure 4a,b).

(a) (b) (c)

Figure 4. Setup for mechanical
Figure 4. Setupmaterial testingmaterial
for mechanical and capillary water
testing and absorption:
capillary (a) compression
water absorption: (a) compression
strength;
strength; (b) three-point (b) three-point
flexural flexural test;
test; (c) capillary (c) capillary
water water absorption.
absorption.
The samples used to estimate the density, as well as all subsequent material tests,
The samples used to estimate
were prepared the density,
according as well outlined
to the procedures as all subsequent
in Section 2.1.material tests,
For this experiment,
were prepared according to the procedures outlined in Section 2.1. For this
the growth duration lasted for 14 days, i.e., until the substrate was fully colonised by experiment,
the growth duration mycelium.
lasted for The 14initial
days, growth phasethe
i.e., until lasting seven days
substrate wastookfullyplace in plasticby
colonised moulds
my- and
the second phase took place after unmoulding. Density
celium. The initial growth phase lasting seven days took place in plastic moulds and the was measured on samples grown
in dimensions of 10 × 10 × 10 cm. A drying cabinet was used for sample dehydration.
second phase took The
place after unmoulding. Density was measured on samples grown in
temperature in the cabinet was set to 40 ◦ C, and the drying process continued until
dimensions of 10 ×the 10samples
× 10 cm. A completely
were drying cabinet dry. The was used for sample
documentation of weight dehydration. The
loss and the calculation
temperature in theofcabinet
the averagewas water
set tocontent
40 °C,isand the drying
available process continued
in the supplementary until the(Table
documentation
S1). The dry.
samples were completely method Theof determining
documentation the water absorption
of weight losscoefficient
and thedue to capillaryof
calculation action
in hardened mortar [19] was used to determine the coefficient of water absorption of
the average water content is available in the supplementary documentation (Table S1).
mycelium-based composites. The detailed description of the procedure and its results is
The method of determining
provided in Sectionthe water 3.5. absorption coefficient due to capillary action in
hardened mortar [19] was Bothused to determine
compression strength andthe the
coefficient
three-pointofflexural
watertest absorption
were testedof onmyce-
a Shimadzu
AG-X plus testing machine (Figure 4a,b). The standard
lium-based composites. The detailed description of the procedure and its results is pro- used for these tests was EN 1015-
11 [20], whereas the loading rate was 10 N/mm 2 /sec. Compressive strength was measured
vided in Section 3.5.
on the same samples that were used for density calculations (10 × 10 × 10 cm). The
Both compression
failurestrength
criterion ofandthe the three-point
compression flexural
strength testindicated
tests was were testedby a dropon ofa measured
Shi-
madzu AG-X plus compressive
testing machine (Figure 4a,b). The standard used for these
force. This was caused by an abrupt deformation, which led to a loss of the tests was
EN 1015-11 [20], whereas the loading
overall integrity of the rate
grown wastest10 N/mmThe
samples. 2/sec. Compressive
samples strength
for the three-point was test
flexural
dimensions of 4 × 4 ×
measured on the same samples that were used for density calculations (10 × 10 × 10 cm). by a
had 16 cm and were placed on two linear bearings followed
centrally applied linear load on top of those (Figure 4b). Regarding flexural tests, the failure
The failure criterion of the compression strength tests was indicated by a drop of meas-
criterion was indicated by a force drop right after achieving the peak value, which was
ured compressive force.
causedThis by thewas causedofby
occurrence an abrupt
fractures in the deformation, whichand
sample. The graphical lednumerical
to a lossvalues
of of
the overall integrity of the
these grown are
experiments testpresented
samples. The samples
in Sections 3.3 and for
3.4. the three-point flexural
test had dimensions of 4 × 4 × 16 cm and were placed on two linear bearings followed by
3. Results
a centrally applied3.1.
linear load on top of those (Figure 4b). Regarding flexural tests, the
Substrate Selection
failure criterion was indicated by a force drop right after achieving the peak value, which
The first set of samples contained straw as a substrate. It was chopped into single pieces
was caused by theno occurrence
longer than 3ofcm fractures
in length andin the sample. into
hand-pressed Thethegraphical
moulds. The and MC numerical
of the substrate
values of these experiments
was 60.23%.are Thepresented
straw fragments in Sections 3.3 and
were visible after 3.4.
unmoulding and drying (Figure 5a,b).
Because the substrate had not been thoroughly compressed prior to inoculation, the airiness
3. Results in the material caused the samples to break. The shrinkage factor was negligible, yet the
growth speed was sufficient, as the mycelium expanded across the surface in five days.
3.1. Substrate Selection
The first set of samples contained straw as a substrate. It was chopped into single
pieces no longer than 3 cm in length and hand-pressed into the moulds. The MC of the
substrate was 60.23%. The straw fragments were visible after unmoulding and drying
(Figure 5a,b). Because the substrate had not been thoroughly compressed prior to inocu-
 smooth surface.
Beech sawdust was pressed manually into the moulds after inoculation but became
highly porous after the drying process. The MC of the substrate was 58%. Shrinkage was
10%, which is a reliable value for future use (Figure 5i–l). Similarly, as with all samples,
the density of the substrate particles was important for the stiffness of the dried product.
Biomimetics 2022, 7, 51
The outer layer of the samples was light brown, and the outer skin had not developed 6as of 16

uniformly as it did in the cellulose pulp samples.

(a) (b) (c) (d)

(e) (f) (g) (h)

(i) (j) (k) (l)

Figure 5. Samples
Figure of growth
5. Samples compatibility
of growth of Pleurotus
compatibility ostreatus
of Pleurotus with
ostreatus straw,
with wheat
straw, bran,
wheat bleached
bran, bleached
cellulose pulp and beech sawdust: (a) ST-PO-01; (b) ST-B-PO-03; (c) WB-B-PO-03; (d) WB-B-PO-04;
cellulose pulp and beech sawdust: (a) ST-PO-01; (b) ST-B-PO-03; (c) WB-B-PO-03; (d) WB-B-PO-
(e) CS1-PO-02; (f) CS1-PO-03; (g) CS1-B-PO-04; (h) CS1-B-PO-05; (i) FS-PO-20; (j) FS-PO-21; (k) FS-
04; (e) CS1-PO-02; (f) CS1-PO-03; (g) CS1-B-PO-04; (h) CS1-B-PO-05; (i) FS-PO-20; (j) FS-PO-21;
B-PO-22; (l) FS-B-PO-23.
(k) FS-B-PO-22; (l) FS-B-PO-23.

Using pure wheat bran as a substrate was not successful—the samples without burlap
became contaminated, while the other two with the burlap piece only retained their form as
the fabric kept them together. The MC of the substrate was 46.75%. The remaining samples
were fragile. As observed on the dark surface of the samples, the mycelium was hardly
visible (Figure 5c,d). However, using wheat bran as an additive would accelerate mycelium
growth [21], and it will thus be used for this purpose in further experiments.
Pieces of bleached cellulose pulp (2–6 mm diameter, MC 64.17%) were dispersed
in plastic moulds, and the mycelium grew entirely within. The substrate shrank by up
to 40% after drying (Figure 5e,f), making it exceedingly unpredictable in cases where
specified dimensions are to be achieved. The samples containing burlap shrank up to
5% (Figure 5g,h). Additionally, the samples became deformed during the drying process.
To anticipate or, at the very least, to decrease the considerable shrinkage, the cellulosic
substrate should be compressed firmly prior to inoculation. The samples had a white colour
and a smooth surface.
Beech sawdust was pressed manually into the moulds after inoculation but became
highly porous after the drying process. The MC of the substrate was 58%. Shrinkage was
10%, which is a reliable value for future use (Figure 5i–l). Similarly, as with all samples,
the density of the substrate particles was important for the stiffness of the dried product.
The outer layer of the samples was light brown, and the outer skin had not developed as
uniformly as it did in the cellulose pulp samples.

3.2. Density
The samples were slightly distorted after the drying process, which happened due to
the inconsistent pressure from the manual filling of the moulds and the standard shrinkage
Biomimetics 2022, 7, 51 7 of 16

factor. They were measured from edge to edge and in the centre of each side to obtain an
average side length, resulting in 24 measurements per sample. Consequently, the volume
was calculated followed by the density (Table 1). In addition to beech sawdust, further
density measurements of other substrates were carried out, such as bleached cellulose pulp,
soy silk fibres mixed with beech sawdust, cotton, cardboard, beech sawdust mixed with
sand, and beech sawdust mixed with clay.

Table 1. Density comparison (density of beech sawdust samples (FS) compared to various samples).

Name Average Side (cm) Volume (cm3 ) Weight (g) Density (g/cm3 )
FS-PO-01 9.26 794.67 215.00 0.27
FS-PO-02 9.29 802.20 220.00 0.27
FS-PO-03 9.42 836.12 210.16 0.25
FS-PO-04 9.41 832.80 212.52 0.26
FS-PO-05 9.35 817.40 209.67 0.26
FS-PO-06 9.42 835.01 209.28 0.25
FS-PO-average 9.36 819.70 212.77 0.26
CS1-PO-01 8.74 668.01 225.00 0.34
FS-SF-PO-01 8.96 718.92 170.00 0.24
FS-GL-01 9.30 804.36 205.00 0.25
SC-PO-01 8.93 710.93 297.00 0.42
S-FS-PO-01 9.73 919.75 215.00 0.23
S-FS-PO-02 9.47 848.38 429.00 0.51
CO-PO-01 8.82 685.35 151.00 0.22

3.3. Compression
3.3.1. Compression—Beech Sawdust
The samples whose growth was terminated after 14 days (FS-PO-05 and FS-PO-10)
showed similar compressive strength, as the ones that had three additional days to grow
(FS-PO-03 and FS-PO-07) (Table 2). The curves on the graph are defined by three stages:
the first showing mediocre endurance, the second being the weakest stage as the sample
softens, and finally, the recuperation phase, in which the curve grows more steeply than it
did previously (Figure 6). Results of this kind were to be expected, taking the porosity of
the material into account.

Table 2. Compression test of beech sawdust samples.

Name Maximum Force (N) Maximum Stress (MPa)

FS-PO-03 43,096.14 4.310
FS-PO-05 2822.56 0.280
FS-PO-07 42,275.38 4.230
FS-PO-10 11,166.73 1.120
average 24,840.20 2.490
Biomimetics 2022, 7, x FOR PEER REVIEW
Biomimetics 2022, 7, 51 8 of 16

Figure 6. Compression test of beech sawdust samples.

Figure 6. Compression test of

3.3.2. Compression—Various beech sawdust samples.
Samples
The various samples consisted of different combinations of organic and inorganic
3.3.2. Compression—Various
materials. Samples
A description of each sample is provided in the following text. The order of the
samples is defined by their composition similarities.
The various samples consisted of different combinations of organic and ino
FS-SF-PO-01—The sample consisted of horizontally stacked layers of soy silk fibres
materials.
between layersA description of each
of sawdust. Since the sample
sample wasis provided in the following
loaded perpendicularly text. The
to the layers of orde
samples is defined by their composition similarities.
soy silk fibres, it did not break apart as those made from sawdust had. The layers of fibres
behaved as elastic springs, adding
FS-SF-PO-01—The sample a certain flexibility
consisted to the sample. The
of horizontally ratio of
stacked soy fibres
layers of soy silk
to sawdust is 1:1, inoculated with 20% Pleurotus ostreatus.
between layers of sawdust. Since the sample was loaded perpendicularly to the la
CO-PO-01—Since cotton consists mostly (88–97%) of cellulose [22], this type of nutri-
soy
tionsilk
was fibres,
viable forit mycelial
did notgrowth.
break apart as those
The fibres used formade from had
the sample sawdust had. length
a significant The layers o
behaved
(5 cm), and asthe
elastic
task ofsprings, adding
homogenising a certain
them with the flexibility to the
mycelium grain sample.
spawn The ratio
thus proved to of soy
to sawdust is 1:1, inoculated with 20% Pleurotus ostreatus.
be challenging. The performance of this sample can be ranked between the one with soy
fibresCO-PO-01—Since
and sawdust, and thatcottonwith sawdust
consists only. CO-PO-01
mostly exhibited
(88–97%) ofacellulose
similar quality
[22],that
this type
of the sample with soy fibres, since it was compacted after testing and did not really break.
trition was viable for mycelial growth. The fibres used for the sample had a sign
SC-PO-01—The performance of this sample was high for withstanding compressive
length (5 cm),inoculating
forces. Before and the with
task 10%
of homogenising
Pleurotus ostreatus, them with thewas
the cardboard mycelium
soaked ingrain
hot spaw
proved
water and to later
be challenging.
torn into smallThe performance
pieces—35 of thisThe
mm in length. sample
samplecanwasbe ranked
found between t
to have
shrunk 11% when measured by the average side length and had a density of 0.42 g/cm 3,
with soy fibres and sawdust, and that with sawdust only. CO-PO-01 exhibited a
which contributed to the best compression strength results (Table 3), as the cardboard was
quality that of the sample with soy fibres, since it was compacted after testing and d
bound together tightly by the mycelium.
really break.
TableSC-PO-01—The performance
3. Compression test of various samples.of this sample was high for withstanding comp
forces. Before inoculating with 10% Pleurotus ostreatus, the cardboard was soaked
Name Maximum Force (N) Maximum Stress (MPa)
water and later torn into small pieces—35 mm in length. The sample was found t
FS-SF-PO-01 15,737.45 1.990
shrunk 11% when measured by the average side length and had a density of 0.42
C-FS-GL-01 4622.05 0.510
which contributed to the best compression strength results (Table 3), as the cardboa
CS1-PO-01 6429.34 0.850
bound together tightly by the mycelium.
FS-GL-01 6607.95 0.760
FS-GL-01—This blend consisted of the same sawdust type as the ones in the pr
SC-PO-01 20,975.92 2.650
section, but it was inoculated with Ganoderma lucidum. When compared to previ
S-FS-PO-02 2314.20 0.260
sults, it did not perform as well.
S-FS-PO-01 692.27
CS1-PO-01—The sample consisted of bleached cellulose0.090
pulp inoculated wi
CO-PO-01 6224.60 0.800
Pleurotus ostreatus. During the drying process, this sample shrunk significantly,
expected after testing the substrate compatibility. The sides are reduced in size to
(from the original 10 cm per side). The sample thus has a density of 0.34 g/cm3, w
higher than that of the average sample made from beech sawdust (0.26 g/cm3). The s
was brittle, similar to the C-FS-GL-01, but withstood greater force (Figure 7).
C-FS-GL-01—The mixture was made with one part of modelling clay to four p
sawdust; the composite was inoculated with 10% of Ganoderma lucidum grain spaw
 based on a sand to sawdust volume ratio of 1:4.

Table 3. Compression test of various samples.

Biomimetics 2022, 7, 51 Name Maximum Force (N) Maximum 9 ofStress

16 (M
FS-SF-PO-01 15,737.45 1.990
C-FS-GL-01 4622.05 0.510
FS-GL-01—This
CS1-PO-01 blend consisted of the same sawdust
6429.34 type as the ones in the previous
0.850
section, but it was inoculated with Ganoderma lucidum. When compared to previous results,
FS-GL-01
it did not perform as well. 6607.95 0.760
SC-PO-01 sample consisted of bleached
CS1-PO-01—The 20,975.92
cellulose pulp inoculated with 2.650
10%
Pleurotus S-FS-PO-02
ostreatus. During the drying process,2314.20
this sample shrunk significantly, as 0.260
was
expected after testing the substrate compatibility. The sides are reduced in size to 8.4 cm
(from theS-FS-PO-01 692.27
original 10 cm per side). The sample thus has a density of 0.34 g/cm3 , which0.090
is
3
higher thanCO-PO-01
that of the average sample made from 6224.60
beech sawdust (0.26 g/cm ). The sample0.800
was brittle, similar to the C-FS-GL-01, but withstood greater force (Figure 7).

Figure 7. Compression test of various samples.

FigureC-FS-GL-01—The
7. Compressionmixture
test of various
was madesamples.
with one part of modelling clay to four parts of
sawdust; the composite was inoculated with 10% of Ganoderma lucidum grain spawn. The
3.4. Three-Point
organic Flexural
portion was Test
also influenced by the density of the clay—the goal was to introduce
as much of the organic matter as possible in order to achieve a cohesive mycelial growth on
3.4.1. Three-Point Flexural Test—Beech Sawdust
the inside and as a result to enhance the properties of the composite. The sample showed
The
similar first setasof
brittleness samples consisted of six beech sawdust samples. The numbe
CS1-PO-01.
S-FS-PO-01 and S-FS-PO-02—The
played in the table below show a wide addition of sand to of
dispersion theresults
mixture(Table
did not 4),
adddespite
to its them
mechanical strength. The two samples are differentiated by the amount of sand in the
simultaneously inoculated and incubated for the same period of time, under the
mixture: S-FS-PO-01 had equal quantities of sand and sawdust, while S-FS-PO-02 was
conditions. A conclusion
based on a sand of whyratio
to sawdust volume theofresults
1:4. vary so much cannot be drawn to one s
factor. However, when comparing the curve from the sample with the highest resu
3.4. Three-Point Flexural Test
PO-13 (Figure 8) is compared to an average result from the cellulose pulp samples,
3.4.1. Three-Point Flexural Test—Beech Sawdust
are described in Section 3.4.2. A similar strain is not exhibited, whereby the sawdus
The first set of samples consisted of six beech sawdust samples. The numbers dis-
ples can bear only half of the force that cellulose samples can.
played in the table below show a wide dispersion of results (Table 4), despite them being
simultaneously inoculated and incubated for the same period of time, under the same
Table 4. Three-point
conditions. flexural
A conclusion test
of why theofresults
beechvary
sawdust samples.
so much cannot be drawn to one specific
factor. However, when comparing the curve from the sample with the highest result,
Name Maximum
FS-PO-13 Force
(Figure 8) is(N)
compared to Maximum Stress
an average result (MPa)
from Maximum
the cellulose Distance
pulp samples,
FS-PO-11 61.11 in Section 3.4.2. A similar strain
which are described 0.14324 3.68
is not exhibited, whereby the sawdust
samples can bear only half of the force that cellulose samples can.
FS-PO-12 32.54 0.07626 2.40
FS-PO-13 73.02 0.17114 3.88
FS-PO-14 33.35 0.07816 2.66
FS-PO-15 52.37 0.12275 2.67
FS-PO-16 39.59 0.09280 2.85
 Biomimetics 2022, 7, 51 10 of 16

Table 4. Three-point flexural test of beech sawdust samples.

Name Maximum Force (N) Maximum Stress (MPa) Maximum Distance (mm)
FS-PO-11 61.11 0.14324 3.68
FS-PO-12 32.54 0.07626 2.40
Biomimetics 2022, 7, x FOR PEER REVIEW FS-PO-13 73.02 0.17114 3.88
FS-PO-14 33.35 0.07816 2.66
FS-PO-15 52.37 0.12275 2.67

average FS-PO-16
48.66 39.59 0.09280
0.11406 2.85 3.02
average 48.66 0.11406 3.02

Figure 8. Three point flexural test of beech sawdust samples.

Figure 8. Three point flexural test of beech sawdust samples.
3.4.2. Three-Point Flexural Test—Various Samples
3.4.2.The
Three-Point Flexural
following section Test—Various
describes Samples tested in the same manner as
individual composites
the previous ones in Section
The following section3.4.1. The firstindividual
describes two samples, Q-GL-01 and
composites FS-GL-19,
tested in thewere
same man
inoculated simultaneously with the same mycelium strain, Ganoderma lucidum, and grown
the previous
under the sameones in Section
conditions 3.4.1.
for the same Theoffirst
period time.two
The samples,
substrate is Q-GL-01 and FS-GL-19
the sole distinction
inoculated simultaneously with the same mycelium strain, Ganoderma
between them, i.e., beech and oak sawdust. The sample inoculated with oak sawdust lucidum, and
under
shows the same conditions
significantly for(Table
better results the same
5). It isperiod of time.
important to noteThe
thatsubstrate is of
the particles the sole dist
oak
sawdust were smaller (1–2 mm) compared to beech sawdust (about 3
between them, i.e., beech and oak sawdust. The sample inoculated with oak samm). However, more
samples are needed in order to make viable conclusions. This result difference will be
shows significantly better results (Table 5). It is important to note that the particles
further explored in order to relate the substrate type and mycelium strain to their specific
sawdust
mechanicalwere smaller (1–2 mm) compared to beech sawdust (about 3 mm). Ho
properties.
more samples are needed in order to make viable conclusions. This result differen
Table
be 5. Three-point
further exploredflexural test of various
in order samples.
to relate the substrate type and mycelium strain to the
cific mechanical
Name properties.
Maximum Force (N) Maximum Stress (N/mm2 ) Maximum Distance (mm)
The sample SN-PO-01
Q-GL-01 70.38 showed the0.16496
best results (Figure 9). 2.84
This sample was pr
in a FS-GL-19
similar manner as the ones made with
38.39 cardboard. Sheets of
0.08997 4.52newspaper were
in water
SN-PO-01for 24 h, which
276.95 were then torn into small pieces by 3.65
0.64909 hand. These sampl
formed five times better
SC-PO-02 89.49
than the average values for beech sawdust.
0.20973 4.23
SC-PO-02, the s
containing
S-FS-PO-03
shredded29.18 cardboard, showed lower results than the1.64
0.06840
ones containing shr
newspaper,
S-FS-PO-04
yet it performed
46.05
better compared
0.10792
to the sawdust composites.
2.74
The card
and newspaper pieces were approximately 15 mm long.
The last two pieces containing sawdust and sand in different ratios, S-FS-PO-
S-FS-PO-04, showed no unexpected results. Adding sand did not improve f
strength.

Table 5. Three-point flexural test of various samples.

 Biomimetics 2022, 7, 51 11 of 16

The sample SN-PO-01 showed the best results (Figure 9). This sample was prepared
in a similar manner as the ones made with cardboard. Sheets of newspaper were soaked
in water for 24 h, which were then torn into small pieces by hand. These samples per-
formed five times better than the average values for beech sawdust. SC-PO-02, the sample
Biomimetics 2022, 7, x FOR PEER REVIEW 11 o
containing shredded cardboard, showed lower results than the ones containing shredded
newspaper, yet it performed better compared to the sawdust composites. The cardboard
and newspaper pieces were approximately 15 mm long.

9. Three-point
Figure 9.
Figure Three-point flexural testtest
flexural of various samples.
of various samples.
The last two pieces containing sawdust and sand in different ratios, S-FS-PO-03 and
Samples
S-FS-PO-04, CS1-PO-06
showed and CS1-PO-07
no unexpected consisted
results. Adding sand did entirely of bleached
not improve flexural cellulose
strength. pulp a
exhibited
Samplesexcellent
CS1-PO-06mechanical properties
and CS1-PO-07 consisted when compared
entirely of bleached tocellulose
the other pulp tested
and samp
Another
exhibited technique was explored
excellent mechanical by when
properties varying the ratios
compared to theofother
the tested
two basic components,
samples. An-
other technique was explored by varying the ratios of the two basic
lulose pulp and beech sawdust. The first, CS1-FS-PO-01, was made up of 30% cellulcomponents, cellulose
pulp and beech sawdust. The first, CS1-FS-PO-01, was made up of 30% cellulose and 70%
and 70% sawdust, while the second, CS1-FS-PO-02, was made up of 70% cellulose a
sawdust, while the second, CS1-FS-PO-02, was made up of 70% cellulose and 30% sawdust.
30% sawdust. These were used to investigate if adding another organic component
These were used to investigate if adding another organic component improved or degraded
proved or degraded
the qualities that were the qualities
being that
measured. were
Even being
though themeasured.
sample withEven morethough
cellulosethe sample w
pulp
more cellulose
performed better,pulp performed
the 30% sawdust better, the 30%
in the sample sawdust
reduced in the
its total sample reduced
performance when its to
compared to the
performance whensamples that solely
compared contained
to the samples cellulose.
that solely Adding 30% cellulose
contained to the
cellulose. Adding 3
sample consisting
cellulose mainly of
to the sample sawdust did
consisting not drastically
mainly of sawdust changedidthe notresult when compared
drastically change the res
to the average of the sawdust samples (Table 6, Figure 10).
when compared to the average of the sawdust samples (Table 6, Figure 10).
Table 6. Three-point flexural test of bleached cellulose pulp compared to blend of bleached cellulose
Table 6. Three-point
pulp with flexural test of bleached cellulose pulp compared to blend of bleached cellu
beech sawdust.
pulp with beech sawdust.
Name Maximum Force (N) Maximum Stress (N/mm2 ) Maximum Distance (mm)
Name Maximum
CS1-PO-06
Force (N)147.95 Maximum Stress
0.34675
(N/mm2) Maximum
3.83
Distance (mm)
CS1-PO-06 147.95
CS1-PO-07 151.95
0.34675
0.35614 3.92
3.83
CS1-PO-07 151.95
average 149.95 0.35614
0.35145 3.88 3.92
average 149.95
CS1-FS-PO-01 44.87 0.35145
0.10516 2.09 3.88
CS1-FS-PO-01 44.87
CS1-FS-PO-02 75.17 0.10516
0.17617 3.92 2.09
CS1-FS-PO-02 75.17
average 60.02 0.17617
0.14067 3.01 3.92
average 60.02 0.14067 3.01
 CS1-PO-07 151.95 0.35614 3.92
average 149.95 0.35145 3.88
CS1-FS-PO-01 44.87 0.10516 2.09
CS1-FS-PO-02 75.17 0.17617 3.92
Biomimetics 2022, 7, 51 12 of 16
average 60.02 0.14067 3.01

Figure 10. Three-point flexural test of bleached cellulose pulp compared to blend of bleached cellulose
Figure 10. Three-point flexural test of bleached cellulose pulp compared to blend of bleached ce
pulp with beech sawdust.
lose pulp with beech sawdust.
3.5. Capillary Water Absorption
The testing included several steps—firstly, the samples were completely dried out, and
their surface was sealed with ethylene–vinyl acetate, which was applied using a standard
glue gun. Once the dry weight of the samples was determined (M0), the container was
filled with distilled water until the samples were immersed by 5 cm. The samples were
placed in the filled container on a slant to ensure there was no air trapped beneath them,
because of their uneven surfaces. The samples were weighed in a defined time range—after
10, 30, 60, 90 min, 2, 4, 8, and 24 h (Table 7). Five samples were examined, all of which had
the same dimensions of 10 × 10 × 10 cm. Three of them were identical, consisting of beech
sawdust inoculated with Pleurotus ostreatus, another with the same substrate but inoculated
with Ganoderma lucidum, and an oak sawdust inoculated with Ganoderma lucidum.

Table 7. Weight after certain periods of time. A water absorption coefficient greater than 2 is classified
as strongly absorbent, while less than 2 is classified as water resistant, less than 0.5 is classified as
water repellent, and less than 0.001 is classified as waterproof.

M0: M1: M2: M3: M4: M5: M6: M7: M8:

Weight Weight Weight Weight Weight Weight Weight Weight Weight C (kg/(m2 ×
Name M8-M1 M4-M1
Initial after after after after after 2 h after 4 h after 8 h after min05 ))
(g) 10 min (g) 30 min (g) 60 min (g) 90 min (g) (g) (g) (g) 24 h (g)
FS-PO-17 285 291 299 313 325 338 393 472 565 274 34 0.0034
FS-PO-18 280 286 292 302 308 311 327 254 423 137 22 0.0022
FS-PO-19 273 281 289 304 311 318 346 399 505 224 30 0.0030
Q-GL-02 240 241 244 247 248 249 258 290 577 336 7 0.0007
FS-GL-20 233 233 234 237 239 244 272 355 546 313 6 0.0006

For calculating the coefficient of water absorption, the following formula applies:
 
C = 0.1( M4 − M1)kg/ m2 × min0.5 .

The last column of Table 7 shows a distinct dissociation of values; the samples inocu-
lated with Pleurotus ostreatus and Ganoderma lucidum, with the latter showing a lower water
absorption than the first sample group. These findings suggest that Ganoderma lucidum
increases water repellence in samples inoculated with sawdust. Pleurotus ostreatus samples
show water-repellent qualities, and Ganoderma lucidum samples show waterproof qualities.
Biomimetics 2022, 7, 51 13 of 16

4. Discussion
The substrates were chosen for their performance, but availability was also an im-
portant selection criterion. Out of the four initial materials that were evaluated, beech
sawdust and cellulose pulp were considered to have potential for further research. Both
exhibited adequate stiffness, growth density and a satisfactory quality impression. If com-
pressed during inoculation, straw can also be considered a viable substrate, as it creates
very lightweight and porous composites.
Density was measured on samples with initial dimensions of 10 × 10 × 10 cm, which
was naturally decreased by mycelium digesting the substrate, compacting it and also by
the drying process. In addition to these parameters, manually filling and compressing the
moulds may have also influenced the density values of each sample.
The compression strength of beech sawdust composites had an average value of
2.49 MPa, with an interesting differentiation of samples whose growth lasted three days
longer and exhibited higher values. Beech sawdust inoculated with Ganoderma lucidum did
not perform as well as samples inoculated with Pleurotus ostreatus.
Three-point flexural tests were carried out on six beech sawdust samples with an
average value of 0.11 N/mm2 . Two samples inoculated with Ganoderma lucidum were
compared, as the sample inoculated on oak sawdust showed better results. Shredded
newspaper performed well as a substrate, with a value of 0.649 N/mm2 .
Cardboard and newspaper are the materials worth considering for future experiments.
Both exhibit excellent compression strength values and the highest density from purely
organic samples after mycelial growth. They are also usually discarded and can be recycled
in this manner. However, additional research on these two materials is necessary, since
they were tested on individual samples. Cellulose pulp exhibited excellent mechanical
properties, but its dimensions after drying are not as predictable due to its high shrinkage.
Using sand as an additive has shown stable results while documenting shrinkage, yet it is
to be considered an improper additive, since it does not enhance the mechanical properties.
Adding clay to the organic substrate was beneficial for the plasticity of the samples, and it
will be researched further.
Finally, capillary water absorption was tested with two mycelium strains, Pleurotus
ostreatus and Ganoderma lucidum. The samples inoculated with the former strain are water
repellent, while the ones inoculated with the latter exhibit waterproof qualities. There was
no difference between oak and beech sawdust in terms of water absorption.
The goal of this research was to evaluate various mycelium and lignocellulosic sub-
strate combinations. This is important if mycelium-based composites are to be introduced
into the building industry—consequently, the material samples were standardised and
tested to be comparable with conventional building materials in terms of potential future
applications. This series of tests was used to characterise and assess their properties. The
data of this research will be used by the authors in order to further develop methods
of evaluating properties of mycelium-based composites for specific applications, i.e., for
researching heterogeneities in mycelium-based composites.

5. Conclusions
Mycelium-based composites exhibit structural properties that open up the possibility
of their implementation in the building industry. Their applications include compression-
only structures, temporary assemblies, art installations [23] and materials for acoustic
and thermal insulation [24]. These have already been implemented as case studies and
products developed in several companies. However, their application as a widely accepted
alternative to some building components and commercialisation is yet to be seen. Moulds
are a viable solution for shaping the material mixtures, yet the necessity of a sterile working
environment, as well as the time mycelial growth takes, are somewhat limiting factors.
Within this research, it was possible to develop a fabrication process for mycelium-
based composites on a scale of architectural elements similar to masonry units. Moreover,
a sterile work environment was established, and a productive shaping method developed.
 Biomimetics 2022, 7, 51 14 of 16

The aim of this work was to gather data on several material properties and, as a result, to
select the ones most suitable for composite materials depending on their application.
Another aspect that will be investigated is the correlation between growth time and
the mechanical properties of mycelium-based composites, as seen in the results for different
compression strength values of samples with a longer growth period. A series of samples
will be made in which growth is interrupted in different samples and on numerous occa-
sions, with a few days between each interruption. Another aspect that is planned to be
looked into is the correlation between growth time, mechanical properties and weight loss
of mycelium-based composites, as it has already been investigated on the decay of wood
by brown-rot fungi [25]. This is important for defining the optimal growth advancements
in the composite while retaining its maximal mechanical capacities.
In addition to the fine-tuning of the composite by enhancing the desired material
properties, the results presented here will be used for making heterogeneous material
mixtures in complex geometries, including varying mechanical requirements. The major
Biomimetics 2022, 7, x FOR PEER REVIEW 14
potential that is yet to be explored is controlled material distribution within a specific
element. An initial experiment to test this hypothesis was conducted by creating a series
of trusses consisting of different cellulose types, each exhibiting different mechanical
properties
properties (Figure
(Figure 11). After
11). After thoroughly
thoroughly analysinganalysing theofproperties
the properties of mixtures,
homogeneous homogeneous
atures, a heterogeneous
heterogeneous material distribution
material distribution will be implemented.
will be implemented.

Figure11.11.
Figure Truss
Truss structure
structure mademade from cellulose
from cellulose pulp. pulp.

In addition to exploring natural coatings to prevent the degradation of organic com-

In addition to exploring natural coatings to prevent the degradation of organic
posites caused by moisture, the waterproof qualities of Ganoderma lucidum can be beneficial
posites
for caused
its future use inby moisture, thecomposite
mycelium-based waterproof qualities
materials. An of Ganodermamethodology
experimental lucidum can be be
cial for its future use in mycelium-based composite materials. An
has recently been developed in one study on the subject of biodegradability of mycelium- experimental met
ologycomposites
based has recently been
based developed
on soil burial testsin[26].
oneThese
study on the subject
experimental methodsof biodegradability
are crucial if of
mycelium-based composites will be used in exterior applications. Using a
celium-based composites based on soil burial tests [26]. These experimental method certain mycelium
strain
crucialthat
if is more resistant to composites
mycelium-based the water uptakewill is
bepromising but still applications.
used in exterior highly dependent Using a ce
on the type of substrate used [10]. Yet, their rapid biodegradability is one of their quali-
mycelium strain that is more resistant to the water uptake is promising but still hi
ties that make the material appealing in terms of sustainability and waste management.
dependent on
Consequently, thethe type
focus of substrate
of application used [10]. Yet,composites
of mycelium-based their rapid stillbiodegradability
remains in dry is on
their qualities
interior locations. that make the material appealing in terms of sustainability and waste m
agement. Consequently, the focus of application of mycelium-based composites sti
Supplementary
mains in dryMaterials: The following supporting information can be downloaded at: https:
interior locations.
//www.mdpi.com/article/10.3390/biomimetics7020051/s1, Table S1: Measurements during the
drying process and average water content calculation.
Supplementary Materials: The following supporting information can be downloade
www.mdpi.com/xxx/s1, Table S1: Measurements during the drying process and average water
tent calculation.

Author Contributions: Conceptualization, H.V., L.G. and J.J.; Writing—Original Draft Prepara
H.V.; Writing—Review and Editing, H.V., L.G. and J.J.; Experiment—Conduction, J.J.; Visualiza
Biomimetics 2022, 7, 51 15 of 16

Author Contributions: Conceptualization, H.V., L.G. and J.J.; Writing—Original Draft Preparation,
H.V.; Writing—Review and Editing, H.V., L.G. and J.J.; Experiment—Conduction, J.J.; Visualization,
L.G.; Supervision, M.S. All authors have read and agreed to the published version of the manuscript.
Funding: This work was funded by the Austrian Science Fund (FWF) project F77 (SFB “Advanced
Computational Design”).
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: Not applicable.
Acknowledgments: We are most grateful for the technical support of the Institute of Technology
and Testing of Building Materials, Graz University of Technology and Anita Klaus, Department for
Industrial Microbiology, University of Belgrade.
Conflicts of Interest: The authors declare no conflict of interest.

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