INTRODUCTION
The reticular assemblies formed by the mycelium, which is fungi’s root-like vegetative structure, consist of thread-like hyphae. They develop until they are heated to block any further change in shape. They have been increasingly investigated as the replacement for some types of plastic materials, in particular polystyrene foam (Abhijith et al. 2018). The substrate on which mycelium develops is formed by lignocellulosic biomass, often waste or by-product of other industrial sectors. In this way, the matrix is composed and bound together by the mycelium growth. The process for the creation of the composite includes the collection, grinding, and sterilization of the biomass. This is followed by inoculation, incubation at temperatures between 23 and 28 °C and at a humidity content of 50 to 70%, and finally drying at temperatures in the order of 70 °C, which stops development and gives the final geometry to the composite (Elsacker et al. 2019).
The described procedure makes it possible to establish a new class of composite materials, which is prevalent, yet not exclusive, and of interest in the construction industry (Javadian et al. 2020). Additionally, there have been suggestions for various applications of these bio-blocks, which would depend on their growth characteristics after inoculation and to the temperature (around 70 to 90 °C) and time of the “baking” process that ends the mycelial growth process (Joshi et al.2020). Of course, amongst the many factors that control the self-growing of the material, is also the selection of the substrate and that of the fungal species to allow the maximum effectiveness of the production (Islam et al. 2017). A number of factors do influence the mechanical properties of the obtained materials: first the nature of the substrate used for growth, then the structure of the mycelium matrix obtained, including architecture of hyphal network, cell wall structure, and growth kinetics and the substrate-matrix adhesion characteristics (Jones et al. 2017).
One main question is whether it is feasible to dispose of general and mixed lignocellulosic waste by employing it as the substrate for mycelium growth. However, when trying a comparison with polystyrene foams, the density obtained by mycelium foams does not come down to less than 50 kg/m3, with compression strength not exceeding 20 kPa and an even lower flexural strength (Angelova et al. 2021). The above properties suggest that, given the difficulty to further reduce the density, the need to achieve higher compression properties raises interest in other industries, such as footwear. With this aim, the insertion of a natural fiber (e.g., jute) fabric has been proposed (Silverman et al.2020).
However, there are a number of possible issues, or else factors, that may lead to potential success or failure of the process (Fig. 1). These have been integrated with further information, drawing from what was suggested by Sydor et al. (2022). In general terms, to improve the properties of the obtained foam, it appears important to be as specific as possible about the kind of biomass used in producing myco-composites.
The Role of Hemp in Myco-composites
Hemp (Cannabis sativa) is at the core of an important productive system, whose products span from food (oil, seeds, flour etc.) to non-food (textiles, mats, ropes, bricks, bio-fuel, phytoremediation, etc.) (Rheay et al. 2021). However, to improve the competitiveness of the industry based on this crop, it needs to be inserted into a circular economy philosophy. Attention must be paid to the use of waste biomass generated in the various productions and thereby potentially reducing the carbon footprint of this production (Scrucca et al. 2020). The development of myco-composites based on hemp by-products can be an opportunity in this sense.
Based on the above evidence, this review concentrates on the use of hemp-derived biomass (hurds, fibers, mats) on the development of myco-composites, and the range of properties obtained in this case. Figure 2 reports the different types of hemp-derived materials used in the literature to serve as the mycelium matrix bonding. These can all be waste or by-products of the industrial system based on this plant. Hemp has attracted considerable interest in the field of composites and is therefore widely treated in the literature, including recent reviews, such as that of Müssig et al. (2020). Utilization of the waste from hemp production system is also of interest, which includes the possible re-use of hemp hurds in the field of composites, combining it with pottery clay (Scardecchia et al. (2020), or more in general with concrete, considering that hurds constitute 70% of the whole material weight obtained from hemp system (Barbhuiya and Das 2022). It needs also to be noticed that hemp by-products are particularly adapted to binding into structures aimed for the construction industry, such as hempcrete, which is constituted by hemp shives and binder. This is another reference for myco-composites, other than the usual one with polystyrene foams (Demir and Doğan 2020).
Another possible by-product from hemp, which is usually discarded in industrial processing, is hemp pith. This relatively small part of the stem has appeared promising for acoustic properties and was used as a substrate for mycelium growth by Pelletier et al. (2013). Various combinations including among other materials, sorghum straw, flax shive, kenaf pith, cotton, were considered.
The lack of definition of the amount of mycelial growth obtained appears to have limited interest in such work, though the comparison indicated that, exposed to an A-weighted road noise, a 100% substrate in hemp pith offered an attenuation of around 7 dB with respect to a ceiling tile.
A further work exploiting hemp pith together with cotton ginning waste with different fiber-fungal strain combinations was performed by adding fiber fabric on the surface, therefore not pertaining, in the stricter sense, to pure myco-composites (Ziegler et al. 2016). This offered a maximum value of 1.06 GPa for compression strength, which was therefore superior to Styrofoam, though at the expense of around 350% water absorption. It has been suggested that hemp by-products would be among the possible candidates to serve as semi-structural myco-composite boards.
In light of the versatility of their properties, they were included in the database recently provided with this aim by Shen et al. (2023). This is also demonstrated by other investigations that suggest the use of hemp hurds in myco-composites as a wood replacement material. In the case of Bajwa et al. (2017), the enhancement of the resistance to termite attack was offered in hemp pith grown mycelium inoculated by Trametes versicolor by coating the board using guayule resin.
Hemp Hurds and Other Byproducts for Manufacturing of Myco-composites
The substrate, which is bonded with the mycelial material, would better undergo chemical characterization, so that at least the relevant lignin, cellulose, and hemicellulose content is measured. A large number of substrates have been applied over the last 15 years for mycelium growth, and this section is aimed at comparing their performance over the production of composites. To attempt a classification of the different substrates, they can be divided among prevalently ligneous (wood-like ones), ligno-cellulosic (based on those plant fibers that contain also some lignin), and cellulosic (e.g., from paper or straw) substrates. In some studies, a blend of different biomass materials has been used as the substrate. Specifically, hemp hurds have been reported as amongst the first waste sources, namely coming from the textile industry, that have been researched for mycelial growth using a Pleurotus ostreatus strain. Hemp hurds were compared with a number of other substrates, coming from other industrial sectors, such as sawdust of deciduous trees, wheat, and rye straw. The combination of hemp hurds and wheat straw offered the best results in terms of yield, reaching 69 g of Pleurotus ostreatus over 100 g of substrate (Sobieralski et al. 2011). A more organic study on eight fungi species and a similar variety of waste indicated that Flammulina velutipes offered the highest mycelial growth when coupled with a bare hemp hurds substrate, yet this did not lead to further investigations on the production of myco-composites, as from the explored literature (Siwuulski et al. 2010).
As from the data reported by Sydor et al. (2022), the most popular substrates in the production of mycocomposites are pine wood or undefined wood, cotton fibers, wheat bran or straw, or cellulose fibers. Hemp fibers, hurds, or mats do pertain to ligno-cellulosic biomass. Despite some variability that can be encountered among the three by-products, they always contain some amount of lignin (no more than 10%), though their main component is polysaccharidic (cellulose, hemicellulose).
Myco-composites with hemp shives substrate achieved a compression strength of 235 kPa, fast biodegradability in 12 weeks, as compatible with composting standards, yet water absorption peaked at 1000% (Loris et al. 2022). Another study incorporating hemp shives substrate in a Trametes versicolormycelium resulted in a mycelial growth of 0.8:1, a compression strength of 190 to 200 kPa, yet an increase in stability and comparatively reduced water absorption, between 600 and 880% (Irbe et al. 2022). These findings, which demonstrate a very high water absorption tendency, have suggested a need for more specific studies on myco-composites having a substrate composed of biomass formed by hemp hurds and a species growing as an infesting plant in a lake environment, Lagarosiphon major, which is a typical infesting plant from internal waters (Pittau et al. 2022). Hemp myco-composites absorbed up to 145 g/m2of water for tests at relative humidity between 50 and 80%, and a water absorption of 48 kg/m2 was measured, in both cases after 12 hours. In both cases, this was around 4 to 5 times higher than the lake plant, which was attributed to the presence of a much more diffuse and effective capillary network in hemp hurds.
Biodegradation after 12 weeks was more precisely assessed in hemp hurds myco-composites by Zimele et al. (2020), again with Trametes versicolor. It was clarified that over 70% of the mass had been lost at that moment and the compression strength obtained reached 360 ± 50 kPa, with 160 ± 20 kPa flexural strength and a density of 134 kg/m3. Compostability was not considered as the only option for the disposal of mycocomposites grown on hemp hurds, though. A recent study on packaging materials based on this substrate suggested that at end-of-life it might serve as a precursor for a jet fuel using a one-pot ionic liquid technology (Choi et al. 2023).
Though hurds represent the most popular substrate for mycelial growth among hemp by-products, other options were also explored, also with comparative studies. The previously mentioned study by Elsacker et al. (2019), again with Trametes versicolor, suggested the compared use of hemp waste in different formats, namely hurds, compressed or not, in the dimension range of 5 to 25 mm, and chopped fibers, smaller than 5 mm. In no case did the dry density exceed 100 kg/m3and the compression Young’s modulus was revealed to be in the region of 1 MPa. Water absorption in these composites was in the range of 2 to 3.5 kg/m2. Around the same interval was also observed the competing flax or straw biomass as a substrate.
Moreover, the relation between the types of fungi for mycelium growth and the relevant substrate needs to be investigated. Trametes versicolor appears, as from the literature, to be among the most suitable for the use of hemp biomass. However, a further recent study did investigate the application of edible fungi, namely Pleurotus ostreatus, on hemp hurds for the production of myco-composites with a relatively low incubation time of 16 days, though the mechanical properties of the obtained boards were not reported (Reiss II 2022). It is also noteworthy that the same species of edible fungi has been used in a solid-state fermentation process of hemp biomass to enhance its nutritional profile to provide a highly proteinaceous food for animals, which suggests a high level of compatibility between hemp cellulosic products and Pleurotus ostreatus (Eliopoulos et al. 2022).
Another possibility was to combine hemp with other cellulosic materials, to serve as a hybrid substrate for mycelium. In particular, a study performed by Elsacker et al. (2021) blended 5 to 25 mm hemp hurds with bacterial cellulose in a mycelium (Trametes versicolor) matrix. The boards were densified at 70 and then at 200 °C. Mechanical testing was performed after a curing process of 4 weeks at 21 °C and 65% relative humidity. In particular, tensile (largely varying values, with maximum at 76.4 MPa), flexural (average value 2.94 MPa), and internal bond testing (average value at 54 Pa) was carried out. A different type of hybridization using hemp hurds as the support for Pleurotus ostreatusgrowth in myco-composites was performed by the addition of laterite by Etinosa et al. (2023). This proved effective up to 70 wt.%, to try to offer a higher compression strength, up to 800 kPa, though losing in the ductility of the material.
The comparison of different forms of hemp by-products (hurds, loose fibers, non-woven mats) together with two fungi species, Coriolus versicolor and Pleurotus ostreatus, for mycelium growth was developed by Lelivelt et al. (2015). The most promising results were obtained using hemp mat with Coriolus versicolor,which resulted in the highest level of densification of the foam during testing, achieving a level of compressive strength between 24 and 93 kPa with a density between 170 and 250 kg/m3 (Lelivelt 2015).
Despite the aforementioned difficulties in using mixed agrowaste for the production of mycocomposites, on the other side, hybrid substrates would also be worthy of consideration in view of the maximization of profits and yields as regards the biorefinery concept, using widely available agrowaste, such as straw (Jones et al. 2020).
An example of a hybrid substrate that has been attempted in combination with hemp hurds is wheat bran (Sisti et al. 2021). Hemp shives presented dimension ranges of 10 to 15 mm length, 3 to 6 mm width, and 1 to 1.5 mm thickness, whilst wheat bran was a waste from pasta production, in the form of flakes with an approximate volume of 1 mm3. In the above study, for the first time, superhydrophobic properties of mycelium-based materials have been investigated, with a contact angle of 131° for a 20 wt% wheat bran content. However, mechanical properties started decreasing from the addition to hemp of more than 10 wt% wheat bran, though the values were significantly high, 820 MPa compression strength for 75% deformation, and 100 MPa flexural strength for 70% deformation. The introduction of wheat bran in combination with hemp hurds does appear promising, though the advantage might be variable, due to the wide dispersion of the particles’ aspect ratio (Rahman et al. 2021).
To try to develop usable products, for example acoustic boards based on myco-composites, sometimes a generic approach is used, mixing the various by-products previously indicated of the hemp system. This was the case for pollutant absorbing panels, where hemp-based myco-composites (204 mg/kg) proved superior with respect to rice straw (169 mg/kg) and lacquer tree wood chip (59 mg/kg) ones (Lee and Choi 2021). These results are promising as a part of a trend in trying to introduce myco-composites grown from hemp hurds in the actual building paneling as complement for plywood rather than a self-supported structure, such as reported by Almpani-Lekka et al. (2021), on a hemp hurds myco-composite grown on Fomes fomentarius. A more structural application has been proposed in the creation of hemp hurds myco-composites based on wood veneer with an inoculation of Ganoderma lucidum, and prepared in comparison with the bare myco-composite not including wood veneer molded to a density of 145 kg/m2 (Őzdemir et al. 2022). Tensile, flexural, pull-out, and compression tests have been performed. In particular, the coverage with the low-density veneer brought some improvement in the flexural strength from 170 to 190 kPa, while it offered 30.6 MPa of tensile strength, and 360 kPa of pull-out strength. In contrast, compression strength was in the order of 1.2 MPa.
CONCLUSIONS
The so-called myco-composites, where lignocellulosic material adhere together by the insertion of an adapted networking material provided by the mycelium hyphae, has shown increasing success, as demonstrated by the exponentially growing number of publications over the last years. This can be attributed to the large availability of both the matrix and the filler material, particularly enhanced by the need to dispose of agrowaste, which in some cases presents both a scarce adaptability to integration into compost and is even less suitable to be applied as energy recovery by carbonization. A production system that offers the possibility of a variety of ligno-cellulosic waste is hemp. In particular, hurds or shives, as well as fibre waste and more generally hemp textile scraps have been used in a number of occasions to serve as the support for the production of myco-composites with obvious limitations due to their high water absorption and capillarity. However, they are able to offer successful combinations with other biomass, such as wheat straw, and have proved successful with inoculations of different fungi, such as Trametes versicolor and Pleurotus ostreatus. Future challenges are in the control of the biomass properties in terms of absence of contamination and of limited variability of the properties, which might also reduce the required incubation times, considering that hemp yielding as a substrate appeared generally among the highest for biomass used in myco-composites production.
Data Availability Statement
Data available on request from the authors.
Declaration of Conflicting Interests
The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
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