Colab Biomaterials

Description

This workshop focuses on biomaterials, bio-inspired, and biological materials. We attempt to mix biology and material science with design thinking and rapid prototyping. The aim of this workshop is to connect local researchers, artists and designers to explore new applications of biomaterials. We encourage participants to explore alternative ways of conducting scientific research in an interdisciplinary setting.

1. NatureInspired #collaboration #Biomimicry #RapidPrototyping
2. research
   

The workshop consisted of 2 day and hosted at EPFL and Hackarium in Lausanne. Participants worked on an interdisciplinary project around bioinspired and biomaterials. The programme includes 2D printing with Bacteria, alginate encapsulation, kombucha processing, prototyping, and series of participatory lectures.

The three main take away of the workshop was:
Understand physical + biological property of biomaterials
Being playful with materials
Consider how the objects you design interacts with local environments

more info here colab biomaterials, Open Science School https://docs.google.com/document/d/1HmrTohpbVGg-HXVKxy01vH_xnnN_fm7cKQlgShhzzWc/edit#heading=h.yo78etrsp471

Content of the workshop

Structure of workshop

The 2 day workshop consisted practicals, lectures, idea generation activities and prototyping with activities that bridges together artists, scientists and designers to brainstorm and work on an interdisciplinary project around synthetic biology and life engineering. The first day of the workshop was hosted in Hackarium and the second day of the workshop was hosted at EPFL.

Schedule (insert the image of the schedule)

SECTION 1: Consider how the objects you design interacts with local environments

Debate + Ice Breaker: Anthropocene

The debate was opened by Vanessa Lorenzo and Gabriella Sanchez.

Peer-to-Peer teaching: Biology of Materials

One of the valuable aspects of an interdisciplinary team is the asymmetry of knowledge and skills, which becomes very impactful if managed well among the members. However, when trying to go together deeper on one specific topic, it can be very hard to keep the motivation and the learning curve for everyone. Biologists and people more familiar with life sciences would be very bored if they have to attend a Biology 101 lesson about bioremediation. Biology 101 presented some foundational basis of biology, biochemistry, and bioengineering needed to understand bioremediation and how life produces, stores, and makes energy.

We had a collaborative lecture divided on three different topics: the chemical basis of life and the type of biomolecules; the type of energy metabolisms and diversity of life; and the use of synthetic biology and genetic engineering to change life. The format of the lecture was specifically designed to involve all biologists in the room in the learning and teaching process. We divided all participants in groups that were composed of a mixture of people already knowing about the subject and people not so familiar with life sciences. We did a few cycles of lecture in the following manner: first, the lecturers were presenting a few topics about biology; secondly, the participants were discussing about this and explaining to each other the main ideas; at the end, we debriefed of the content that we were supposed to learn or follow. SECTION 2 : Understand physical + biological property of biomaterials Showroom & debate: Innovative materials and biomaterials.

Innovative Biomaterials Table was installed to showcase over the whole day a range of biomaterials and their usage in fashion, construction and design. Here are some of the materials that were exposed: Biocement - bacterially produced cement, EPFL Laboratory of Soil MEchanics Structural color surfaces inspired by butterfly wings, Morphotonix Self cleaning surface, by prof. Yves Leterrier, LPAC lan EPFL Mushroom materials, from Hackuarium Edible bittles, http://www.davidideas.com/details/wikicells Kombucha materials Biogranules, http://biogranulats.com/ Cellulose textiles - biodegradable packaging and bags - BioApply Straw materials - http://artdepaille.com/ Natural untreated materials: Bamboo Moss Paine cones Bird feathers

The session was based on 2 presentations and discussion sessions:

Made by Nature-Mechanical properties of Natural Materials, by Darja Dubravcic The presentation questioned how biological organisms use materials to obtained properties interesting for us humans, like: protective packaging, permeable surfaces, color, solar cells, homes, lightweight structures…. Nature design the same thing we humans do, but with the minimum use of energy, maximum use of materials, no waste and it does it all with resource abundant materials that at biodegradable almost by definition. The presentation focused on 3 main ways that nature achieves that: 1) creating different functions by modifying the form, as opposed to us humans who for every new function use new material, 2) lightweight construction algorithms that give rise to strong and light constructions that minimize the use of energy and materials, and 3) multifuncional design where nature always combines several function within a given product - materials that are light, flexible, stiff, self-clean, sense, repair...

Innovative use of Natural Materials in fashion, design and construction, by Juliette Lenouvelle.

Over more than dozens of industrial cases Juliette showed us how natural materials are used today to create new construction materials, electricity, textiles.... She finally focused on the need for global life cycle assessment for every materials used and every purpose because this is where the true sustainability decisions come to place Practical #1: Printing with pigment-producing bacteria on petri dishes using a hacked 3D printer. (Vanessa Lorenzo)

Microbiology of pigment-producing bacteria Open Science School’s Co-lab Biomaterial EPFL & Hackuarium December 2016. EPFL & Hackuarium, Lausanne

Content: Growth conditions of microorganisms. Physical (temperature, light), chemical (pH, osmolarity), toxicity (antibiotic, heavy metal), or nutrient requirement (auxotrophy, bioavailability) conditions. Pigments and physical nature of color.

Activities: Observe the different microorganisms Play with the bacteria printer Experiment different supports for their growth.

Introduction

Bacterial growth is affected by (1) temperature, (2) nutrient availability, (3) water supply, (4) oxygen supply, and (5) acidity of the medium.

Temperature: Theoretically, bacteria can grow at all temperatures between the freezing point of water and the temperature at which protein or protoplasm coagulates. Somewhere between these maximum and minimum points lies the optimum temperature at which the bacteria grow best. Temperatures below the minimum stop bacterial growth but do not kill the organism. However, if the temperature is raised above the maximum, bacteria are soon killed. Most cells die after exposure to heat treatments in the order of 70°C for 15 seconds, although spore-forming organisms require more severe heat treatment, e.g. live steam at 120°C for 30 minutes. Bacteria can be classified according to temperature preference: Psychrophilic bacteria grow at temperatures below 16°C, mesophilic bacteria grow best at temperatures between 16 and 40°C, and thermophilic bacteria grow best at temperatures above 40°C.

Nutrients: Bacteria need nutrients for their growth and some need more nutrients than others. Lactobacilli live in milk and have lost their ability to synthesise many compounds, while Pseudomonas can synthesise nutrients from very basic ingredients. Bacteria normally feed on organic matter; as well as material for cell formation organic matter also contains the necessary energy. Such matter must be soluble in water and of low molecular weight to be able to pass through the cell membrane. Bacteria therefore need water to transport nutrients into the cell. If the nutrient material is not sufficiently broken down, the microorganism can produce exoenzymes which split the nutrients into smaller, simpler components so they can enter the cell. Inside the cell the nutrients are broken down further by other enzymes, releasing energy which is used by the cell.

Water: Bacteria cannot grow without water. Many bacteria are quickly killed by dry conditions whereas others can tolerate dry conditions for months; bacterial spores can survive dry conditions for years. Water activity (AW) is used as an indicator of the availability of water for bacterial growth. Distilled water has an AW of 1. Addition of solute, e.g. salt, reduces the availability of water to the cell and the AW drops; at AW less than 0.8 cell growth is reduced. Cells that can grow at low AW are called osmophiles.

Oxygen: Animals require oxygen to survive but bacteria differ in their requirements for, and in their ability to utilise, oxygen. Bacteria that need oxygen for growth are called aerobic. Oxygen is toxic to some bacteria and these are called anaerobic. Anaerobic organisms are responsible for both beneficial reactions, such as methane production in biogas plants, and spoilage in canned foods and cheeses. Some bacteria can live either with or without oxygen and are known as facultative anaerobic bacteria.

Acidity: The acidity of a nutrient substrate is most simply expressed as its pH value. Sensitivity to pH varies from one species of bacteria to another. The terms pH optimum and pH maximum are used. Most bacteria prefer a growth environment with a pH of about 7, i.e. neutrality. Bacteria that can tolerate low pH are called aciduric. Lactic acid bacteria in milk produce acid and continue to do so until the pH of the milk falls to below 4.6, at which point they gradually die off. In canning citrus fruits, mild heat treatments are sufficient because the low pH of the fruit inhibits the growth of most bacteria. (adapted from International Livestock Research Institute)

In this experiment, we will manipulate and grow different pigment-producing bacteria and observe their growth. Pigmentation is a characteristic that is common to many species of Bacteria. Pigments are light-absorbing compounds that are responsible for the colors that organisms display. Diverse groups of pigments are produced by organisms of the Bacteria domain, and they play important roles in the survival of the organisms which produce them. For example, the pigment xanthomonadin protects the Bacteria Xanthomonas oryzae from damage due to light, also called photodamage (extracted from MicrobeWiki).

We will use 3 different strains in our experiment:

Micrococcus luteus:Gram-positive, Coccus, saprotrophic bacterium that belongs to the family Micrococcaceae. An obligate aerobe, M. luteus is found in soil, dust, water and air, and as part of the normal flora of the mammalian skin. The bacterium also colonizes the human mouth, mucosae, oropharynx and upper respiratory tract.

Chemical structure of xanthophyll, a yellow pigment found in Micrococcus luteus.

Micrococcus roseus: Gram positive bacterial cell that grows in the tetrad arrangement. The normal habitat for this Micrococcus species is skin, soil, and water. It derives its name from the pink carotenoid pigment that it secretes.

Chemical structure of canthaxanthin, a pink pigment found in Micrococcus roseus.

Janthinobacterium lividum: aerobic, gram-negative, soil-dwelling bacterium that has a distinctive dark-violet (almost black) color due to violacein production. Its anti-fungal properties are of particular interest since J. lividum is found on the skin of certain amphibians, including the red-backed salamander (Plethodon cinereus), where it prevents infection by the devastating chytrid fungus (Batrachochytrium dendrobatidis).

Chemical structure of violacein, a violet pigment found in Janthinobacterium lividum.

Materials

Nutrient agar plates Luria Broth plates Luria Broth plates with Ampicillin (100 mg/ml), IPTG (1 mM) and tryptophan (2 g/l) Lb liquide Lb liquide + 1%Glycerol

Protocol: make your own experiment

Now you are free to draw colorful painting on your petri dishes, or experiment and use the bacteria on different conditions to check their growth. Please note the results and anticipate the results beforehand as an exercise. What is so special about natural pigments? Research ideas.

Pigments are normally produced by bacteria, plants, or even animals as a reaction to environmental stress or defense.

J. lividum For example, the pigment violacein from J. lividum inhibits the toxic effect and growth of a fungus known as Batrachochytrium. This fungus causes a disease known as Chytridiomycosis in amphibians The disease devastated amphibian populations around the world, in a global decline towards multiple extinctions, part of the Holocene extinction. Because of this, understanding the uses of this bacteria has been of major interest. A study conducted in 2009 explored the effects of Bd and the use of J. lividium in the lab for survival. They used three experimental treatments: frogs infected with Bd, frogs given the bacteria J. lividium and frogs with the given bacteria and then exposed to Bd. Nearly all of the frogs exposed to Bd experienced mortality while none of the other treatments had any deaths. This effectively introduced the use of J. lividium as a possible method for Bd prevention in the lab setting.

Care: As you see, this bacteria can help you fight some diseases on amphibians (salamanders, frogs, etc). Lb agar plate, and a cozy 25 - 30 °C Temperature would make them happy. Purple babies will colonize your plate after 4 - 7 days. Wanna ink? The best is to transport them with a sterile spatula to a Lb (liquid) or Lb +1% glycerol (liquid) bottle, shake it constantly at around 270 rpm and a cozy 25 - 30 °C Temperature to make them happy.

Micrococcus Luteus Yellow mates that can be found almost everywhere: in soil, dust, water, air, in our skin… they are strong and they can live (almost) forever. Does a time machine of 30.000 years tell you something? Also, they block UV radiation… let’s go to the beach!

Care: Lb agar plate, and a warm up of 37°C Temperature for about 3 - 4 days would make them happy. Wanna ink? The best is to transport them with a sterile spatula to a Lb (liquid) shake it constantly at around 270 rpm at 37°C to have their shiniest bright yellow.

Micrococcus Roseus Siblings of Luteus and living on mammalian skin, so, your skin as well! Uv protector and beautiful color pigment micro - factories. Are you thinking about a new tattoo?

Care: Lb agar plate, and a warm up of 25 - 37°C (they are chill and relaxed about the temperature) for about 3 - 4 days would make them happy. Wanna ink? The best is to transport them with a sterile spatula to a Lb (liquid). Shake them constantly at around 270 rpm at 37°C to have their beautiful and elegant pink-salmon babies.

Experiment Kit (10 Kits)

Step one: 1 Mixed bacteria plate 1 Spatula 1 Lb agar plate Spot and isolate your colorful babies!

Step two: 3 ink tonner 1 Paper 1 Brush Paint!

Step three: 1 Squared petri 1 Bio-printing session Make your digital design come alive!

General Material: Gas bottle (clean air circulation) Ethanol (96%) Gloves Kitchen paper

Experiment Description Overall explanation of 1, 2, 3 steps. In groups of 3 people, choose 2 steps and 10 min experiment for 1st and 2nd step, 5min for 3rd step.

1st step, clean your spatula,open the mixed color petri, take a sample of one of the colors at a time. Scatter them into one area of the petri dish.

2nd step, soak your brush in the bacterial ink and paint your paper.

3rd step, design your path and print with it.

Biosafety All the samples will stay at the lab and destroyed after 5 days (to see and document development), only the prototypes sprayed with ethanol or blitch will be kept.

Practical #2: Adopt a kombucha! Kombucha SCOBY processing and inoculation.

Production and processing of Kombucha sheets Open Science School’s Co-lab Biomaterial EPFL December 2016. EPFL, Lausanne

Content: Molecular properties (molecule type, structure, functional groups) and macroscopic properties of a biomaterial (elasticity, color, stiffness). Role of material processing and adaptation. Modification of the properties of Kombucha, make hypotheses on how to do it, experiment with different methods for the treatment of Kombucha, and draw conclusions upon results. Biofilm growth and properties

Activities: Brainstorm about the applications of Kombucha. Inoculate sugary tea with Kombucha for the week after. Treat the pre-made Kombucha sheets to modify their properties.

Introduction

Kombucha is a symbiotic living material made of a variable composition of different species of bacteria and yeast. Among the most common ones participating in the symbiosis we can find: Bacteria: Acetobacter xylinum, A. xylinoides, A. aceti, A. pasteurianus, Bacterium gluconicum. Yeasts: Schizosaccharomyces pombe, Kloeckera apiculata, Saccharomycodes ludwigii, Saccharomyces cerevisiae, Zygosaccharomyces bailii, Brettanomyces bruxellensis, B. lambicus, B. custersii and Pichia. Kombucha tea is a traditional health-promoting fermented beverage that exists since several thousand years. It is produced by the fermentation of sugared tea with a symbiotic colony of bacteria and yeast. Kombucha is a traditional beverage drunk for its wide range of health benefits. The cellulose matrix formed in the culture medium can be used as a bio-cellulose tissue to create clothes, or bio-paper. But it can also be used as a cosmetology for its anti-inflammatory, antioxidant and anticancer properties.

Content: What is a biofilm?

In fact, the kombucha sheet forming on top of the culture is a type of biofilm. Biofilms are the most common way in which microorganisms exist in nature. They are very resistant and have very interesting physical properties.

A biofilm is any group of microorganisms in which cells stick to each other and often these cells adhere to a surface. These adherent cells are frequently embedded within a self-produced matrix of extracellular polymeric substance (EPS). Biofilm extracellular polymeric substance, which is also referred to as slime (although not everything described as slime is a biofilm), is a polymeric conglomeration generally composed of extracellular DNA, proteins, and polysaccharides (like the bacterial cellulose we find in kombucha). Biofilms may form on living or non-living surfaces and can be prevalent in natural, industrial and hospital settings

Biofilms are usually found on solid substrates submerged in or exposed to an aqueous solution, although they can form as floating mats on liquid surfaces and also on the surface of leaves, particularly in high humidity climates. Given sufficient resources for growth, a biofilm will quickly grow to be macroscopic (visible to the naked eye). Biofilms can contain many different types of microorganism, e.g. bacteria, archaea, protozoa, fungi and algae; each group performs specialized metabolic functions. However, some organisms will form single-species films under certain conditions. The social structure (cooperation/competition) within a biofilm depends highly on the different species present.

Biofilms can be found in many different places and conditions: Biofilms can be found on rocks and pebbles at the bottom of most streams or rivers and often form on the surface of stagnant pools of water. In fact, biofilms are important components of food chains in rivers and streams and are grazed by the aquatic invertebrates upon which many fish feed. Biofilms can grow in the most extreme environments: from, for example, the extremely hot, briny waters of hot springs ranging from very acidic to very alkaline, to frozen glaciers. In the human environment, biofilms can grow in showers very easily since they provide a moist and warm environment for the biofilm to thrive. Biofilms can form inside water and sewage pipes and cause clogging and corrosion. Biofilms on floors and counters can make sanitation difficult in food preparation areas. Biofilms are present on the teeth of most animals as dental plaque, where they may cause tooth decay and gum disease. Biofilms are used in microbial fuel cells (MFCs) to generate electricity from a variety of starting materials, including complex organic waste and renewable biomass.

Picture: Electron microscopy of biofilm of Staphylococcus aureus biofilm on an indwelling catheter. Public Domain, https://commons.wikimedia.org/w/index.php?curid=2740748

Materials

Kombucha sheets (about one or two per person) Grean tea, vinegar, sugar Ink and organic colorants Dryers and ovens Press Knives Blender Bleach and hydrogen peroxide NaOH 1M Water and paper press Needles

Protocol: how start a culture of bacterial cellulose?

Instructions: Boil 1L of distilled water and add 20g tea in bags. Let infuse during 5 minutes and add 113 g of glucose. Let cool at room temperature and add 100mL of a previous batch (liquid medium and a piece of the cellulosic matrix). Add 100ml of cider vinegar. Seal the bottle and let ferment and grow the culture at room temperature, between 5 and 14 days.

The culture doesn’t need too much oxygen. Yeasts hydrolyze sucrose into glucose and fructose, producing ethanol and carbon dioxide, as metabolites. Acetic acid bacteria converts glucose into gluconic acid and fructose into acetic acid. The final pH reading should be between 2.5 and 3.2, to prevent the symbiotic culture from becoming contaminated by undesirable microorganisms.

Picture: Dr Peter Musk, a scientist catalyst at The Edge Southbank, with some of the vegan leather being produced from kombucha. Photo credits: Queensland University of Technology (Queensland, Australia).

Protocol: how to use and modify bacterial cellulose?

Before starting the protocol, we need to inoculate sugary tea with Kombucha for the week after. To do so, prepare the medium and the inoculum as described above and let it at room temperature in the soom, covered. After this, you can experiment with the Kombucha you got and treat the pre-made Kombucha sheets to modify their properties. Some options include: coloring / decoloring, blending, and desiccation. The information you’ll see below is mostly adapted from the iGEM Team Imperial College 2014 (you can check more on their website: http://http://2014.igem.org/Team:Imperial).

The team from iGEM Imperial 2014 did an intensive production and prototyping during some months and can serve as an example of what we would be able to do in a short term project around biomaterials. The goal of their iGEM project was to optimize the production of bacterial cellulose by engineering Gluconacetobacter xylinus. They also explored the processing of the biomaterial, producing and testing water filters, and functionalizing them with binding proteins to trap specific contaminants.

Bacterial cellulose (BC) exhibits a multitude of different properties depending on the processing, growth conditions, functionalization and strain used for production of the material. Acquiring large quantities of cellulose produced would allow testing of a broad variety of cellulose processing methods and functionalization steps. By mass producing cellulose this enables a better understanding of what material properties can be realistically produced during the short duration of iGEM. More importantly, it improves the likelihood of finding suitable processing candidates for the project’s aim of making a customisable ultrafiltration membrane, at the same time as allowing room for creativity and exploration of the remarkable properties of cellulose.

Kombucha processed sheets from iGEM Imperial College 2014. From left to right: thin kombucha, thick NaOH processed kombucha, and blue-dyed kombucha. Minimum requirements

Treatment of BC requires killing the cells, particularly if the cells are genetically engineered, which is the aim for putting the customisable in ultrafiltration membranes. Based on brainstorming with Central Saint Martins student Zuzana, removing the colour of BC is required as it looks displeasing to the eye otherwise, and seems counter-intuitive to filter clean water with cellulose coloured like turbid water. Removal of the smell of BC has also been raised as a requirement, particularly by producers who work in close contact with the processing facilities. Mass Production Methods

Setting up the mass production of cellulose was done according to the Kombucha media protocol , which involved setting up 61 trays with media and G. xylinus and yeast co-culture. The trays were left to grow up over 7 days, after which diminishing pellicle growth was detected. Upon harvesting, the pellicles were sorted according to granular pellicles and even pellicles (see pictures above). All pellicles were kept in distilled water in large plastic buckets or containers. Below shows the general workflow (adapted from IGEM Imperial 2014) employed to mass produce the cellulose and illustrates the process of manufacturing biomaterials with significantly different properties despite originating from the same BC source.

Inoculate sugary tea and incubate for 7 days at room temperature Harvest pellicules and assess quality of the BC Blend the bacterial cellulose to a paste Wash in water Treat chemically Dyson ocean blue stain in 900ml of distilled water and 100ml of olive oil Air drying without treatment Blend 200g with 250ml of water. Incubate at 0.1 M NaOH at 80°C for 3 hours Oven drying with press after NaOH treatment for 120 minutes Incubation in 1M NaHCO3 for 60 min at 120°C Put into a shape and dry Press dry Produces the most white cellulose if the film that covers the bottom of the pellicle is removed before treatment. BC capable of immersion into water, then it could be reshaped and re dried Produces poor results: brown cellulose Blue flexible cellulose, thin layer, not fragile Brownish cellulose with high moisture content left, material is flexible

Appendices: methods tested at iGEM Imperial 2014 (as stated in their wiki page)

0.1M NaOH for 60 min at 120C: Produces the most white cellulose but the process has been shown to produce some yellow/brownish cellulose if the film that covers the bottom of the pellicle was not removed before treatment. 1M NaHCO3 for 60 min at 120C: Produces quite poor results, 3 samples have been tested and even after 4 hours of treatment at 120C the samples were less white than similar samples treated in distilled water for the same duration. Heat treatment in distilled water at 120C: 3 samples still contained some

brownish tint after 4 hours of incubation, but the samples produced were considerably whiter than those treated with baking soda solution

Air drying without treatment first: Produces brownish cellulose with high moisture content left, material is flexible 120C Oven drying without press, without treatment for 180 min: Produces brittle paper like cellulose, it is fragile, brownish and prone to tears. Oven drying with press (1l Duran bottle on top of two tiles) after NaOH treatment for 120 min: Produces flexible more plastic cellulose capable of being shaped into a cone that filters water through. The cellulose was capable of immersion into water, which produced wet cellulose that could be reshaped and re dried. NaOH for 20 min at 120 C, followed by blending: produces cellulose that seems like it is much less ductile. Disadvantage: the functionalization will be blended just like the cellulose, so the proteins may be broken down mechanically. Distilled water treatment over 48 hours: Produced more white cellulose than what was harvested. The distilled water turned yellow giving evidence that the surface of cellulose actually did dissolve some of the medium’s colour 60 C incubation in tightly wrapped autoclave tape: New tape was applied every 3-8 hours during a 36 hour period. The pressure allowed water to escape and create a compact material of high hardness. Quite a promising result for hard cellulose.

Kombucha Textile : Cellulose Baterial Textile & “Vegetable” Leather

@ Suzanne Lee creation made from Kombucha

Here tuto super cool :) : Cuir végétale-Kombucha https://www.youtube.com/watch?v=vXrAxGx8dTY&feature=share

Protocol: Kombucha Textile

You Need : 200 cl water 20 cl cider vinegar 200 cl sugar 20 g of green tea (2 tea bag) 1 elastic 1piece of gaze fabric 1 strain of Kombucha SCOBY The “starter” : The liquid where the strain has growth (if you have not this liquid, don’t worry*) Glass Jar

Boil water (stop to heat the water) Add tea and let it infuse 15 min Put sugar in the pot and mix with a whisk Wait that the water cool down below 30°C Put vinegar + Water-Sugar-tea liquid + Starter into the Glass jar Put the Kombucha strain on the surface (white face in the top) Put the gaze fabric to close the jarr Leave the Jarr in a room without sun,

• If you don’t have starter, put your strain of Kombucha on a plate. Put 5 tablespoons of white vinegar (pur, distilled, boiled : to avoid competition with other organisms)

Kombucha : What we can do with it & Future (To put on the presentation google slide):

“Now a team from Imperial College London have developed a set of DNA tools to control and engineer a strain of bacteria - normally found in a fermented green tea drink called kombucha tea - to produce modified bacterial cellulose on command. This technique also enables the team to “weave” proteins and other biomolecules into the fabric of the bacterial cellulose as it grows.” http://www3.imperial.ac.uk/newsandeventspggrp/imperialcollege/newssummary/news_27-5-2016-14-46-16

Practical #3: Learn how to make 3D biogels! Alginate encapsulation using sodium alginate and calcium chloride.

Alginate encapsulation of Chlorella spp. Open Science School’s Co-lab Biomaterial EPFL December 2016. EPFL, Lausanne

Content: What is alginate, where do we find it, why do we use it? Encapsulation at an industrial level. Biopolymers.

Activities: Prepare sodium alginate, calcium chloride solution and encapsulate algae. Change the protocol to make an alternative formula. Experiment with the deposition flow to make a 3D gel.

Introduction

Alginate is a biopolymer coming from the cell walls of kelp among other algae. Alginate can create mesh-like structures composed of polysaccharides that become insoluble in contact with calcium. They can keep living bacteria, algae, or even enzymes in the inside. They can be immobilized while remaining biologically active or alive.

Researchers have found many applications of these biogels. Among them: scaffold for human tissue, structures that replicate 3D bacterial communities, or bioreactors that keep microorganisms inside while flowing water constantly.

Chlorella is a genus of single-cell green algae belonging to the phylum Chlorophyta. It is spherical in shape, about 2 to 10 μm in diameter, and is without flagella. The cultivation of microalgae in photobioreactors by continuous culture has been used worldwide to generate large amounts of biomass. This type of culture systems is well established and applied to the production of microbial biomass in fermenters (bacteria and fungi), for example, the alcohol industry.

Content: Bioencapsulation

Encapsulation by a permeable membrane could allow for sustained growth without contamination. Porous membrane allows passage of nutrients, but not cells. This can be useful in different settings in biotechnology, like when we are co-culturing two different species together or when we have to isolate biomass from the medium or simply take a soluble molecule and no biomass. The process of separating biomass and medium can be very costly or time-consuming. Aggregating the microorganisms in beads eases this process.

Microencapsulation and alginate, collagen, or cellulose scaffolds for cells have become quite popular recently and have grabbed a lot of attention from the media. Here we want to present a very simple procedure of encapsulation and 3D printing that has already been used for some time in science, and even in kitchen (“cuisine moléculaire”). It was also used to print layer of bacteria in a biofilm-like confirmation by the iGEM team TU Delft in 2015.

The protocol that follows uses encapsulation by calcium chloride. Calcium alginate is a water-insoluble, gelatinous, cream-coloured substance that can be created through the addition of aqueous calcium chloride to aqueous sodium alginate.

Picture: Encapsulated Chlorella spp. in alginate beads. By Juliette Lenouvel, from Open Science School. Licensed CC BY-SA-NC 3.0.

Picture: Encapsulated Chlorella spp. in alginate beads. By Paloma Portela, from Open Science School. Licensed CC BY-SA-NC 3.0. Experiment: Encapsulating microalgae

Materials Chlorella or Spirulina powder, or a living culture. Sodium alginate, Calcium chloride Plastic pasteur pipettes Beakers or plastic cups (not provided) 100 ml of deionized or distilled water (not provided)

Protocol

Prepare the following solutions: 150 ml of 4% Sodium alginate solution in water (6 g per 150 ml). 200 ml of 4% Calcium chloride solution in water (8 g per 200 ml). Chlorella spp. or Spirulina culture (either a concentrated living culture or a solution of 1g per 50 ml of water). Mix the Sodium alginate and the algae solution 1:1. Pour the Calcium chloride solution into a cup or a beaker. To create the encapsulated balls or fibers, you just need to mix some concentrated algae solution with the the alginate solution. Drop by drop, pour the alginate-algae mix into the Calcium chloride solution. You can try alternative ways of encapsulating, printing, making wires, or changing the proportions of algae/alginate/calcium.

Troubleshooting

The consistency and stiffness of the resulting objects will depend on you extrusion method as well as the final concentrations of alginate and calcium chloride. The solutions will almost immediately become solid when they are in contact at that concentration (20 grams per litre, or 2% each). If the beads are too soft, you might want to increase the concentration of each of the components or leave the beads longer in the calcium chloride media. If the beads are too hard, try to add more deionised/distilled water to the alginate-algae solution. Be careful not to put calcium chloride into the sodium alginate beaker or bottle, because it will polymerize the entire bottle and decrease the efficiency of your experiment.

Further research ideas

Encapsulation is a biocompatible (you can put with living things) gel → you can use this for structure for bacteria to live in, to put enzyme in so that the enzymes are living things because you can control it. Algae is an option to grow things - algae right now is good at producing lipid molecules, vitamins and biofuels because they are energy efficient. Their media is not easily contaminated - the only contaminant is algae. SECTION 2: Being playful with materials

GameJam

Mourdjen Bari, a game teacher from Center of Research and Interdisciplinarity (CRI), ran a 2 hour session on GameJam and gamification. GameJam is an activity frequently ran by the GameLab in the CRI called Game. We asked the participants to gamify what they learnt in the peer-2-peer teaching: Biology of Materials.

Design: Idea Generation! Kickstart your project & explore project ideas with your group.

Project Development:

We produced a card game to help construct narratives of the project based on the materials provided at the workshop.

The rules of the workshop are the following:

Project Development: Storyboard Development Example: Petin’ Sound:

y

Prototyping:

Due to limited material availability, participants got creative with the material available. We focused on low-tech prototyping material and focused on the visualisation of the ideas rather than the functionality of the material.

Petin’ sound - inspired by the practical “encapsulating alginate”

� Outline of the projects

Project #1: Art in the Dark

Team members: Yan Su (Master of arts student), Saziye Yorulmaz (PhD in Biology), Elisa Sia (Master of arts student) and Saziye Yorulmaz (PhD in Biology) Abstract:

The main objective is to bring a whole new experience to our color perception in art works and during expositions. The group aims to enable blind people feel colors maximizing the use of bio materials. The prototype aims to express how to incorporate bio-inspiration and nature to the arts.

Colors in the paintings will be expressed through the temperature and texture of the surface. Active electrical heating - creating a network of resistors, heated via electric current to create “temperature” pixels. Heating and cooling could also be controlled using water, air or some other gas/liquid.

Music and sounds of nature will contribute to create a richer experience. The painting will be printed with a 3D printer (plastic + various concentration of metal) for different conductivity and consequently various touch feeling.

Another proposition is to create a manual mosaic out of various materials to create textures. Patterns could be expressed using bio-compatible material and placing different kinds of plants, that are able to grow in the dark (moss, mint, fungi, etc.)

Motivation, purpose, and science or design references

Plants, that can grow in the dark: http://www.apartmenttherapy.com/5-interesting-plants-that-can-119497

• Maidenhair Ferns are a great option because they have frilly fun leaves that vary from the usual thick leaves of indoor plants. Most Ferns do well inside with low light (and ferns look great in terrariums) so check out others like Silver Lace Fern with variegated leaves. • Begonias: These plants offer a wide range of leaf colors and shapes and if you get a Rex Begonia, it will live quite happily without any direct light. Just make sure you don't overwater it. Soak it and let it dry out, soak and dry. • Mint: Mint will normally grow in a bog, so as long as you keep it moist and it gets a little bit of light, you should be able to harvest mint for tea, for fruit salads and it has the added advantage of giving off a nice scent indoors. • Swedish Ivy: This plant has an old fashioned look that sort of reminds us of gramma, but consider a new way to grow it, like as a part of a vertical garden. • A Moss Terrarium: If you seriously have very little light, consider creating a terrarium of moss. It just needs moisture and glances of light, position it near a window where it will get bounced light and it should thrive. If you don't know where to start, consider a kit to get you going. References Temperature pixels Material mosaic Plant patterns

Project #2: Bio-inspired adventure Tent Team members: Eléonore Wild, Adrien Vergès, Hugo Moreno, Jessica Girard and Nicolas Zooler Abstract:

WHY a tent ? We wanted to show that we could also build object that is bio-inspired in fields like sports and hobbies. We wanted to offer a practical and innovative tent that would help people get closer to nature. We also wanted it to be accessible to everyone. We decided to add some functionalities to our tent. As water is always vital, we designed a water collecting tent inspired on the namibian beetle shell. We also wanted that tent to be a small cosy place to be, and therefore decided to work on sound insulation and modulation is possible if coupled with an other one. Finally, we wanted it easy to carry, light and compact, so we thought to use memory-shape materials technology. Motivation, purpose, and science or design references:

SOLUTION : SOUND INSULATION TEXTILE Inspired on mushroom mycelium. The textile will be made out of porous material and the structure will be sound absorbent.

OUTSIDE LAYER OF TEXTILE is designed to collect water - Inspired on Namibian beetle shell. This takes water from moisture (humidity). The combination of hydrophilic and hydrophobic structures

SOLUTION : The frame of the tent has a memory-shape frame. It is easily put away and minimising space occupied for transportation. Memory-shape metallic alloy: Demo Metal alloy: for example nickel-titanium Titanium : excellent strength /weight ratio Resists corrosion

         Goal → Reducing frame size to have more space in your backpack !

Detailed Description References Bug repellent: https://asknature.org/resource/superhydrophobicity-the-lotus-effect/#.WE1YTvnhBE https://upload.wikimedia.org/wikipedia/commons/thumb/1/13/Lotus3.jpg/800px-Lotus3.jpg Shape-memory alloy: Kula D and Ternaux E. Materiology: the creative industry’s guide to materials and technologies. Basel. Birkhäuser. p.230 and p.242

Namibian Beetle Shell: https://www.pinterest.com/pin/444449056957240027/ http://news.nationalgeographic.com/news/2001/11/1101_TVdesertbeetle.html https://ninithi.com/2015/07/20/how-a-desert-beetle-make-self-filling-water-bottles-possible/ Ternaux E and all. Industry of Nature: another approach to ecology. Amsterdam. Frame Publishers. p239

Mycelium sound insulation: https://radicalmycologypdx.wordpress.com/2011/11/23/mycelium/ www.paroc.bc Patnaik, A., Mvubu, M., Muniyasamy, S., Botha, A., & Anandjiwala, R. D. (2015). Thermal and sound insulation materials from waste wool and recycled polyester fibers and their biodegradation studies. Energy and Buildings, 92, 161-169. Project 3: EchoVision

Team members: Marina Buckel (Chief designer), Imane Baïz (Design Advisor), Daiana Mirzarafie-Ahi (Play-Doh Designer), Antoine Vigouroux (Biologist) and Hubert Taïeb (Material engineer)

Abstract:

Video: https://www.youtube.com/watch?v=2WDNn_RkwHU

Motivation, purpose, and science or design references: EchoVision is a project that aims at guiding blind people thanks to a jacket that gives the direction to take via a device that comprises an emitter and receptors. The device detects obstacles (bio-inspired by the bat which emits waves to detect its prey) and passes on this information to smart fibres covering the inside of the jacket. These fibres can expand and contract (bio-inspired by the Mimosa pudica) so as to guide the person to where he or she is going. Our motivation is to offer a new way to guide blind people using technology and biomaterials, in addition to the cane and audio-devices that already exist, in order to vary the senses that are stimulated.

References:

Caress of the Gaze (https://vimeo.com/138697237): “What if our outfit could recognize and respond to the gaze of the other? This is an interactive 3D printed wearable which can detect other people’s gaze and respond accordingly with life-like behavior”, created by Behnaz Farahi http://www.dailymail.co.uk/sciencetech/article-3252668/The-anti-ogling-shirt-3D-printed-outfit-changes-shape-men-stare-it.html#ixzz4ZWOkpVIV

Caress of the Gaze (https://vimeo.com/138697237): “What if our outfit could recognize and respond to the gaze of the other? This is an interactive 3D printed wearable which can detect other people’s gaze and respond accordingly with life-like behavior”, created by Behnaz Farahi.

Project 4: Book of Infinity

Team members: Celine Tchao, Vivien Russel and Ariana Abstract:

The world is facing increasing shortages of resources including food and shelter. Myths about this go as far back as Jesus, as seen in the biblical story of the Feeding of the 5000 where he feeds many just on five loaves of bread and two fish. We have taken this story and attempted to turn it into reality through the means of a book, the book of infinity. This book is embedded with high tech seeds to create a new foundation of life, community and materials such as plant seeds that are edible, kombucha samples for filtering that can be shared, and a plastic degrading fungus species. Motivation, purpose, and science or design references:

The Book of the Infinity

References https://www.ncbi.nlm.nih.gov/pubmed/10904168

Project #5: Petin’ Sound Team members: Oceane Patiny, Juanma Garcia, Hasnaa Benlafkih, Adrien Rigobello and Vanessa Lorenzo. Abstract:

Petin’ sound is about growing microorganisms and caring them so they can encapsulate us into a more intimate atmosphere within our noisy urban and public spaces. Alginating tiny microcosms containing algae, nutrients and other organisms, we create self sufficient and portable ecosystems that we can take with us and wear. After a busy day, we can keep them like precious bio gems in our domestic aquarium so they can have a good rest. Motivation, purpose, and science or design references:

The motivation of the project is to create a symbiotic soundscape to inhabit when a break from the world around us is needed. The purpose is to allow a human-nature interaction through a multi species ecosystem.

Scientific reference:

http://sandv.com/downloads/1007croc.pdf

Design Reference:

Burtonnita. Algae Culture, 2010. Self-sufficient and symbiotic ecosystem that would allow the user to nurture from algae and feed them back with exhalating CO2. http://www.burtonnitta.co.uk/Algaculture.html

Story Board:

Prototype: Headphones & Earplugs

Detailed Description: Peting Sound is a self sufficient and portable soundscape and ecosystem that we can take with us and wear. By mixing sodium alginate and calcium chloride we create spherical capsules that can contain algae and other organisms in water. The biomaterial encapsulating these ecosystem will serve as both bioreactor and biopolymeric earplugs. The combination with new porous and sound absorbing materials will keep the ecosystem slightly isolated from the exterior noise. The contact with the (human) user’s body will provide the right temperature to the whole ecosystem while the user will benefit from the noise “filtering effect” of the earplug. There would be two versions: headphones and earplugs.

References: http://sandv.com/downloads/1007croc.pdf http://www.burtonnitta.co.uk/Algaculture.html

� Conclusion Learning outcomes and good practices from organizers

Experiment:

Considering the limited resources available we did best to organise a workshop.

There were suggestions from the organisers that our programme was very rushed. This will forever remains as an issue for Co-lab workshop series.

The presentations were designed differently compared to the past and was focused more to make them inspiring for the participants. On the other hand, the practical seemed to be simplified from the scientific side. Some participants and organisers found the scientific content uninspiring due to the long lectures and lack of hands on activity.

Design/Idea Generation

The ideas were of high quality, and we believe the success comes from the good time management of the programme, flow of the whole workshop and the success of the card game designed with the help of Pablo Garcia. It was inspirational to adopt a 'business' point of view - like engineers. Considering this series being the shorted workshop, the quality of the ideas were high. The storyboard activity ran very smoothly and deserve to be further developed in future workshops.

It was a shame that we could not take full advantage of the discussion which occurred in the Anthropocene debate. We not only failed to connect the content created in the discussion, but failed to prove the link between anthropocene and

Prototype

This workshop was a realisation that we will encounter times where we will have limited resources to create prototypes. Dealing with such circumstance was an opportunity to realise the necessity to come up with a methodology where we can hold workshops in such situations. We have always depended on a workshop which allows sophisticated prototyping, but regardless of the limited resources, the participants of this workshop did very well.

It was amazing to see participants to getting hands on by drawings or creating visual material during the prototyping, but we should have had a time to mix the drawings.

We received some comments on how the aim of the prototype was confusing. Some participants found it challenging to understand that during the prototype we do not expect anything concrete - we just provide a place for free exploration of creation of ideas.

General flow of the workshop

The Anthropocene, powerful but a bit disconnected from the rest. Maybe with more disruptive examples of applications that takes in account design role on the climate change while designing an object or application? This comment also applies to the GameJam.

We were in the rush. Especially the last day when they were prototyping. We should had incite them more to go the lab and prototype, like Peti’n Sound group. But this is also because there was not a lot equipement in the lab.

Overall, we were able to create a good atmosphere and group dynamic, however, there are problems that we could not look over. The lack of budget was a challenging obstacle and we could not provide lunches for organiers and participants both days. Possible Suggestions

The debate worked so well. We should be able to introduce a more personalised twist to each workshop if we tailored the content of the workshop based on the debate between the participants.

Organisation has always been an issue with Co-lab workshops. We suggest a concrete documentation method, clearly allocated roles amongst organisers. These things include things such as, allocating a photographer who is in charge of documenting high quality photos of the prototypes created by the participants.

We propose to introduce a pitch for when designing the programme for future workshops. There may be a necessity to organise a pre-workshop for workshop organisers. This could involve 30 min planned activities to give each organiser to try out their plan with other organisers.

For future Co-lab workshops, documentation and debriefing will be done on the last 2 days. The organisers should organise an extra workshop for themselves to finish all of the documentation right after the workshop. This involves producing the report, editing the videos and uploading them on social media and organising the photographs taken during the workshop.

It would be nice if there was a platform or choices/support for the continuation of some of the projects. That would give solid arguments to the workshop. We should think of how to provide opportunities for the ideas to be exposed to local institutions and communities for a chance for further development.