Tiziano Derme

Growth Based Fabrication Daniela Mitterberger

Independent researcher/MäID

Techniques for Bacterial Cellulose Umberto Di Tanna

Independent researcher

Three-Dimensional Grown Membranes and Scaffolding Design

for Biological Polymers

ABSTRACT
Self-assembling manufacturing for natural polymers is still in its infancy, despite the urgent need 1 Bacterial cellulose differential
growing. Physical model of the
for alternatives to fuel-based products. Non-fuel based products, specifically bio-polymers, possess
membrane and representation of
exceptional mechanical properties and biodegradability. Bacterial cellulose has proven to be a remark- the funicular system.
ably versatile bio-polymer, gaining attention in a wide variety of applied scientific applications such as
electronics, biomedical devices, and tissue-engineering. In order to introduce bacterial cellulose as a
building material, it is important to develop bio-fabrication methodologies linked to material-informed
computational modeling and material science. This paper emphasizes the development of three-di-
mensionally grown bacterial cellulose (BC) membranes for large-scale applications, and introduces new
manufacturing technologies that combine the fields of bio-materials science, digital fabrication, and
material-informed computational modeling. This paper demonstrates a novel method for bacterial cellu-
lose bio-synthesis as well as in-situ self-assembly fabrication and scaffolding techniques that are able
to control three-dimensional shapes and material behavior of BC. Furthermore, it clarifies the factors
affecting the bio-synthetic pathway of bacterial cellulose—such as bacteria, environmental conditions,
nutrients, and growth medium—by altering the mechanical properties, tensile strength, and thickness
of bacterial cellulose. The transformation of the bio-synthesis of bacterial cellulose into BC-based
bio-composite leads to the creation of new materials with additional functionality and properties.
Potential applications range from small architectural components to large structures, thus linking forma-
tion and materialization, and achieving a material with specified ranges and gradient conditions, such as
hydrophobic or hydrophilic capacity, graded mechanical properties over time, material responsiveness,
and biodegradability.

488
INTRODUCTION of architecture suggests a bottom-up, material-driven design
Contemporary digital fabrication tools are able to produce with specified ranges and gradient conditions (Soldevila 2015).
geometrically complex objects and structures, yet most of This paper emphasizes the development of new manufacturing
the constructs are not generally sustainable nor energy effi- technologies for the production of 3D-grown bacterial cellulose
cient (Oxman et al. 2012). In contrast, organic self-assembling (BC) membranes.
processes produce little to no waste, using small amounts of
energy to produce multi-functional and adaptable systems In this paper, this claim is substantiated and a solution is offered,
(Vincent 2012). Despite the recognized capabilities of natural making the following contributions:
processes to generate complex structures of organic and inor-
ganic multi-functional composites (shells, corals, teeth, wood, 1. Investigation of bio-inspired fabrication methodologies and virtual
silk, horn, collagen, and muscle fibers), the use of bio-materials and physical prototyping. The factors affecting cellulose bio-syn-
for large scale architectural and engineering applications is still thesis are demonstrated—mainly growth medium, environmental
underdeveloped (Benyus 1997; Vincent 2012). Their structural conditions, and the formation of derivatives (§1.1 and §2.1). The
and functional diversity, especially that of bio-polymers, under- design of the culture medium is a key influence for the growth
lines their capacity to replace existing synthetic polymers and to of microorganisms, and therefore in stimulating the formation of
provide new methods of bio-fabrication as well as new applica- three-dimensional membranes.
tions and structures.
2. The transformation of the bio-synthesis of bacterial cellulose
Cellulose, one of these bio-polymers, is one of the most abun- into BC based bio-composite is shown, leading to the creation
dant biodegradable materials in nature, and has been the topic of new materials with additional functionalities and properties.
of wide investigations in macromolecular chemistry (Mohite et al. Furthermore, we clarify the factors that affect the bio-synthetic
2014). The high water content of bio-synthetic cellulose (99%) pathway of bacterial cellulose, such as bacteria, nutrients, and
and its mechanical properties make it a versatile material that can medium culture properties. Particular focus is given to the
be manufactured in various sizes and shapes. Bacterial cellulose creation of a natural polymer which could grow to any thickness,
has many unique properties, including high purity, high water shape, and robust structure (§1.2 and §2.3, 2.4).
retention, and a hydrophilic nature, tensile strength, thermal
stability, and biodegradability. 3. In-situ self-assembly fabrication techniques and scaffolding tech-
niques for bacteria cellulose are presented and show the creation
Because of these unique properties, it is an attractive candidate of a bio-composite via the fermentation of bacteria strain A.
for a wide range of applications, including within architecture and Xylinum along the surface of natural fibers (§1.3 and § 2.2, 2.5).
engineering (i.e.: water retaining structures, architectural compo-
nents, etc.), but due to the lack of suitable fabrication methods 1.1 Bio-Inspired Fabrication Methodologies and Virtual and
and digital design tools, cellulose is still disregarded as a building Physical Prototyping
material. Although recent developments within biochemistry and Through the combination of non-fuel-based materials with
microelectronic engineering have improved knowledge of biological
materials, it is still not possible to produce bacterial cellulose on an
industrial scale and control the three-dimensional (3D) outcomes
through standard manufacturing and digital techniques (Fernandez
et al. 2013). Current approaches towards virtual and physical
prototyping with non-fuel-based materials also lack the capacity to
model and fabricate with continuously varying material properties
(Oxman 2011). In order to introduce cellulose as a building material,
it is therefore important to develop bio-fabrication methodologies
linked to materially informed computational modeling.

In nature, morphogenesis is a biological process describing the

formation of a shape of an organism, inseparably linking forma-
tion and materialization (Menges 2007). By contrast, architecture
is characterized by prioritizing form-generation over inherent 2 Consolidation through drying. Bacterial cellulose three-dimensional morphology,
with variable thickness.
material logic. The integration of morphogenesis within the field

MATERIAL FRONTIERS 489

 3 Bio-synthesis of bacterial cellu-
lose: strikers for preparation of
Acetobacter xylinum culture.

4 Bio-synthesis of bacterial cellulose:

Microfluidics system to provide
continuous nutrient to the culture.

5 Bio-synthesis of bacterial cellulose:

bacterial cellulose growing after 5
days.

4 5

advances in biological sciences, genetics, and bio-engineering of composites by generating novel morphogenetic mechanisms
bacteria, a new set of possible bio-fabrication technologies can linked to bacteria and different states of matter. This process is
be developed. clearly described within §2.3, focusing on the different levels
of oxygen within the water medium. As a result, 3D growth of
Current design practice is mostly characterized by the domi- cellulose can be directly manipulated.
nation of shape over matter, consequently prioritizing virtual
shape-defining parameters over physical material and fabrication 1.2 Bio-Nanocomposite and Bacteria Engineering
constraints, leading to a geometric-centric design phase (Menges Engineering the bio-synthesis of bacterial cellulose (BC) into
2007; Oxman 2011). Nevertheless, some recent developments BC-based nanocomposites leads to better mechanical and
in direct digital manufacturing enable a shift towards a mate- thermal properties, or additional functionalities that are useful
rial-centric design practice, such as water-based fabrication in many applications and fields. They could be categorized, for
techniques (Oxman 2011). Additive manufacture (AM) tech- instance, as high-strength materials, plant-mimicking materials,
nologies for rapid prototyping employ virtual, computer-aided electrically-conductive materials, catalytic materials, antimicrobial
designed models, and translate them into thin horizontal succes- materials, thermo-responsive materials, and many others. More
sive cross-sections to define three-dimensional physical objects explicitly, the process describes a structure with growth-induced
(Sachs et al. 1993). material properties reacting to external stimuli and resulting in
hierarchically structured forms (Soldevila 2015). Bacterial cellu-
AM technologies have become an efficient and common means lose production depends heavily on several factors, including
to deliver geometrically precise functional prototypes in relatively culture medium and environmental conditions. The culture
short periods of time (Oxman 2012). At the same time, there is medium (Figure 3) contains a carbon source, nitrogen source,
a need to expand manufacturing processes towards bio-fabri- and other nutrients required for the bacteria to grow. In normal
cation-based approaches, borrowing techniques from biological static and aerobic conditions, the bacteria will form a pellicle
science and tissue engineering. In contrast to AM technologies, (flake) (Figure 5) at the surface of the culture medium. This
which relate to a specific controlled output, bio-fabrication pellicle will grow in thickness slightly, but the thickness is limited
techniques in a water medium aim to create dynamic feedback by the supply of oxygen and nutrients to the bacteria. Recent
or reciprocity within a specific context. This process certainly studies found it possible to optimize the thickness and strength
escapes the pitfalls of bio-mimicry in favor of bio-synthesis, as of BC by creating continuous systems of sub-ministration of
a substance forms through interaction with a living organism. nutrients (Figure 4). This creates a potential condition where the
As a result, this will proliferate methods to fabricate synthetic natural polymer could grow to any thickness and shape to form a

490 Growth-based fabrication techniques for bacterial cellulose

Derme, Mitterberger, Di Tanna
structure (Gateholm et al. 2012). The variation of culture medium
components can alter the metabolism of the bacteria (Figures 13
and 14, and §2.3) and consequently the mechanical properties
and bio-synthesis pathways. Such 3D bacterial cellulose-based
structures and membranes can be prepared with a target topo-
logical condition, thickness, and strength. In this approach, the
distribution of material properties is informed by structural and
environmental performance criteria contributing to the internal
6 7
physical makeup of the membrane. It thus requires a set of
virtual and physical prototyping tools and methods to support
variable fabrication approaches and modeling (Oxman 2011).

It is precisely this complex and dynamic exchange between

organism, environment, and functionality that makes synthetic
life valuable for architecture.

1.3 Self-Assembly Fabrication Techniques

Self-assembly fabrication of 3D structures and membranes can
be described as a multi-step fermentation process in the pres-
ence of natural fibers or other polymers. This can furthermore
lead to the formation of BC-based hybrids or nanocomposites
(Qiu and Netravali 2014). Ultimately, advances in self-assembly
fabrication techniques will lead to different manufacturing
methods, implying a condition of growing and morphogenesis.
Taking as reference the biosynthetic process of bacterial cellulose,
these methods are able to control three-dimensional shapes and
8
material behavior. Self–assembly fabrication can be achieved by
providing a scaffold to support and guide the process of growing
to define precise target geometry (Figures 6–9 and §2.2).

Cellulose is a polymer which forms the cell wall of eukaryotic

plants and algae and can also be found as the major constit-
uent of the cell wall of fungi. Despite this, a few bacteria can
also secrete cellulose. A notable example is Acetobacter xylinum,
which is known to secrete cellulose as part of its metabolism
of glucose and other carbohydrates. The bacteria strains of A.
Xylinum have been found to grow preferentially on the surface of
natural fibers (Figures 17 and 18) or certain polymer molecules,
rather than freely in the medium. Therefore, starch, soy resin, or
polyvinyl alcohol (Figures 6 and 7) provide ideal substrates for
the bacteria to grow on and can therefore lead to the formation
9
of BC-based hybrids or bio-composites. Bio-composites are
composite materials reinforced via natural plant-based fibers or 6 BC-induced growth over permanent scaffold. PLA scaffolding and PVA
immersion.
certain polymers (Qiu and Netravali, 2014). Biosynthesizing and
engineering the original BC into-BC based composites could 7 BC-induced growth over permanent scaffold. Immersion of scaffold into culture
potentially find several applications in the fields of architec- medium.

ture, engineering, and product design, opening up a substantial 8 BC-induced growth over permanent scaffold. Image of the membrane during the
change in how products and buildings are form-found, designed, drying process.

and fabricated (Oxman 2011). A variety of methods, digital and 9 BC-induced growth over permanent scaffold. Detail of the resulting membrane.
analog, have been utilized for achieving this purpose. Polyvinyl

MATERIAL FRONTIERS 491

 10

11 12

alcohol hardening (Figures 6–9 and §2.2) and sisal fiber deposi- Preparation of the culture medium (Figure 4): 5% sugar and 0.5%
tion (Figures 17 and 18 and §2.5) can be used as excellent in-situ of ammonium sulphate were added to mature coconut water,
bio-fabrication techniques. and the mixture was brought to boil for 5 minutes. 1% acetic
acid (99.85%) was added and mixed. A micro-fluid system (Figure
A key point for potential future development is the augmen- 5) was applied to provide a continuous flow of nutrients in order
tation of the mechanical properties of the polymer, including to favor an unlimited growth of BC (Gateholm et al. 2012).
its strength and stiffness. In all cases, consolidation is usually
achieved by drying, sintering, or solidification techniques. BC in 2.2 In Situ Self-Assembling (Figures 6–9, 11, 12)
particular has the capacity to be calcified (Figure 10). This solid- We prepared different scaffolding methods to induce BC growth
ification method is currently used in biomedical applications for over a predetermined shape. Two strategies of scaffolding
tissue and bone engineering using hydroxyapatite, chitosan, algi- were introduced: permanent and bio-degradable. Figures 6–9
nate, and agarose. In this case, it may be possible to generate a and 12 show the creation of shells and membranes using BC
material with properties similar to those of bones if we introduce as a binding agent. The scaffold remains embedded within the
an agent in its mineral phase into the bacteria culture medium structure. Bio-degradability (dissolving after the bio-synthesis
during the formation of BC. process) of a scaffold is shown in figure 11, using a sodium-algi-
nate scaffold.
METHODS
This process is intended to show novel BC fermentation methods The culture medium was prepared in a glass beaker (2 liters) and
and scaffolding techniques for controlling shape, thickness, and 10% of AXy culture was added. The flask was left at 25 C° for
material-driven computing experiments. 7 days without disturbance. Oxygen was supplied during the
photosynthetic period to direct the growth on the scaffolding.
2.1 Preparation of Acetobacter xylinum (A Xy) Culture
(Figures 3–5) 2.3 Adhesion Growing (Figures 13 and 14)
Preparation of the AXy culture (Figure 3): The bacteria was This methodology showed a potential to grow three-dimensional
grown in broth and subsequently streaked on agar with an incu- morphology with a target geometry. Static and anaerobic
bation period of 24 hours. A single colony of AXy was inoculated conditions caused the inversion of the metabolism of the
in a flask containing mature coconut water, which was sterilized bacteria, producing a growing which was no longer superficial,
at 121 C° for 15 minutes. The flask was left at 25 C° or room but instead adhered to the morphology of the containing
temperature for 2–3 days. element.

492 Growth-based fabrication techniques for bacterial cellulose

Derme, Mitterberger, Di Tanna
 10 Images of cellulose-based scaffolds
a) pure cellulose and composite
scaffolds of c) cellulose agarose,
and e) chitosan–alginate.

11 Sodium alginate medium led to

the creation of result of a bio-film
soluble scaffolding.

12 BC-induced growth over permanent

PLA scaffold led to the creation of a
growth shell structure.

13 Three-dimensional growth
morphology under static conditions:
resulting geometry.

14 Three-dimensional growth
morphology under static condi-
tions,removal operations from the
containing boundary.

13 14

The culture medium was prepared in a latex flask (1.5 liter) and
10% of AXy culture was added. The flask was left at 25 C° for 10
days without disturbance.

2.4 Bacterial Cellulose with Differential Growing Patterns

(Figures 1, 15, 16)
We prepared BC membranes with different growing patterns
and different thicknesses, and used a particle-spring system as
a “tension-only” hypothetical funicular model to simulate the
behavior of the membrane (Figure 16).

15
The culture medium was prepared in a glass beaker (2 liter) and
10% of AXy culture was added. The flask was left at 25 C° for 4
days without shaking and nutrients were continuously added.
The flask was then left at 25 C° for 5 days. Drying method:
freezing (4 hours) and drying (4 days at 10 C°) .

2.5 Scaffolding Technique Using Sisal Fiber (Figures 17, 18)

We prepared a scaffolding technique with BC-modified sisal
fibers (produced in an incubating shaker), using 60 cm long
sterilized sisal fibers. These were added to the culture media
and sterilized. AXy was inoculated into the culture media and
BC-modified sisal fibers were extracted after 3 days culture.
Preliminary studies were done by simulating the material
behavior according to the pattern deposition of the fibers.
16

RESULTS 15 BC differential growing. Physical model of the membrane and representation of

the funicular system.
This approach demonstrated the first steps in the design of a
complementary technology able to expand the potential of 16 Particle spring simulation, “tension only” funicular system of the membrane.

MATERIAL FRONTIERS 493

 prototyping processes for bio-synthesis. Specifically, experiments
2.1 and 2.4 showed that BC is moldable in cultivation. By metic-
ulous control of the addition of fermentation media through the
use of a microfluidic system, BC can grow to a potentially unlim-
ited thickness and shape. Moreover, other experiments confirmed
the hypothesis that BC grows preferentially on natural fibres such
as sisal. It has been noted that the creation of BC-based bio-com-
posites increase the mechanical properties of the bio-polymer,
offering the potential to create structural and graded proper-
ties. Finally it has been successfully proven that it is possible
to grow BC around a predefined shape; the addition of oxygen
during the fermentation drove the cellulose to follow and adhere
to specific surface conditions. Despite the results, all the exper-
iments are still far away from a real application on a large scale
and the accuracy of material-driven computing methodologies,
relative to prototypes as grown, needs to be further explored.

CONCLUSIONS
This paper outlined novel bio-fabrication and scaffolding
17
techniques for the control of 3D membranes and morpholo-
gies of bacterial cellulose, positioning the results in the field of
bio-materials science, digital fabrication, and material-informed
computational modeling. Future developments of the present
research include:

• Developing a range of structural BC bio-composites using

hydroxyapatite, chitosan, or lining as agents to calcify the 3D
membranes.
• Exploring the conditions for target-geometry-based growing.
Define the role of the scaffold compared to the growing
pattern of BC.
• Investigating BC as a matrix for growth and cultivation of
photosynthetic microorganisms such as algae and cyano-
bacteria in order to generate a material able to perform
photosynthesis and react to photo stimuli.

ACKNOWLEDGEMENTS
We thank our colleagues from University of Tokyo and
Yusuke Obuchi and Jutamat Klinsoda from the Department of
Applied Microbiology Institute of Food Research and Product
Development, Kasetsart University, Thailand who provided
insight and expertise that greatly assisted the research.

18
REFERENCES
17 Result of BC bio-composite using sisal fibers. Bacterial cellulose as binding Benyus, Janine M., 1997. Biomimicry: Innovation Inspired by Nature. New
material has been observed to grow preferably around natural fibers.
York: Quill.
18 Recursive system for sisal fiber deposition and scaffolding creation. The aggrega-
tion of the fibers relative to a hypothetical growing ratio of BC.

494 Growth-based fabrication techniques for bacterial cellulose

Derme, Mitterberger, Di Tanna
Vincent, Julian. 2012. Structural Biomaterials. Princeton, NJ: Princeton Tiziano Derme is teaching at University of Melbourne a design and
University Press. research studio. Tiziano received his Architectural Masters degree with
honors at the University of Rome “La Sapienza ”. He worked several years
Fernandez, Javier G. and Donald E. Ingber. 2013. “Bioinspired Chitinous
for “New-Territories” with Francois Roche and in 2015 he was researcher
Material Solutions for Environmental Sustainability and Medicine.”
and tutor assistant at University of Tokyo at T(ADS) Advanced Design
Advanced Functional Materials 23 (36): 4454–4466.
Studies. His body of work ranges between Sustainable Prototyping,
Mohite, Bhavna V. and Satish V. Patil. 2014. “A Novel Bio Material: Design Computing and synthetic biology. Since 2016 Tiziano is also
Bacterial Cellulose and its New Era Applications” Biotechnology and director and co-founder of multidisciplinary design practice MäID
Applied Biochemistry 61 (2): 101–110. FutureRetrospectiveNarrative.

Oxman, Neri, Elizabeth Tsai, and Michal Firstenberg. 2012. “Digital

Anisotropy: A Variable Elasticity Rapid Prototyping Platform.” Virtual and Daniela Mitterberger is currently teaching at University of Melbourne
Physical Prototyping 7 (4): 261–274. a design and research studio. Daniela received her Architectural
Masters degree with honors at the Academy of Fine Arts Vienna and
Oxman, Neri. 2011. “Variable Property Rapid Prototyping.” Virtual and was nominated for the big price of the Academy of Fine Arts and the
Physical Prototyping 6 (1): 3–31. Hunter Douglas Award 2015. Her work ranges between human body as
Qiu, Kaiyan and Anil N. Netravali. 2014. “Fabrication and Applications architecture and the definition of technology and tools within the field of
of Bacterial Cellulose Based Nanocomposites” Polymer Reviews 54 (4): architecture. In 2015 Daniela worked for “New-Territories” with Francois
598–626. Roche and since 2016 Daniela is director and co-founder of the multidis-
ciplinary design practice MäID FutureRetrospectiveNarrative.
Sachs, Emanuel, Michael Cima, James Cornie, David Brancazio, Jim
Bredt, Alain Curodeau, Tailin Fan, Satbir Khanuja, Alan Lauder, John Lee,
Umberto Di Tanna received his architectural Master degree at the
and Steve Michaels. 1993. “Three-Dimensional Printing: The Physics
University of Rome “La Sapienza”. In 2012 he was researcher for CRR(
and Implications of Additive Manufacturing.” CIRP Annals: Manufacturing
Rome Research Centre) on a spin-off regarding open source manufac-
Technology 42 (1): 257–260
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Menges, Achim. 2007. “Computational Morphogenesis: Integral Form prototyping, biomaterials and scenography. In 2013 Umberto worked for
Generation and Materialization Processes.” In Proceedings of the 3rd SHSH Architecture and scenography and in 2015 he was co-founder of
International ASCAAD Conference on Embodying Virtual Architecture, edited Quyala_synthetic Environments, an independent research cluster focused
by Ahmad Okeil, Aghlab Al-Attili, and Zaki Mallasi. Alexandria, Egypt: on biologically inspired fabrication, based in Bangkok.
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Gateholm, Paul, Henrik Backdahl, Theodore Jon Tzavaras, Rafael V.

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US Patent 0190078A, filed September 28, 2010, and issued July 26,
2012.

Soldevila, Laia Mogas. 2015. “Water-based Digital Design and Fabrication”

MS Thesis, Massachusetts Institute of Technology.

IMAGE CREDITS
Figures 1–9, 11–18: Derme, Mitterberger, Di Tanna, 2015
Figure 10: Jang et al., 2012

MATERIAL FRONTIERS 495