IL305612B2

IL305612B2 - Edible scaffolds, their preparation and uses

Info

Publication number: IL305612B2
Application number: IL305612A
Authority: IL
Original assignee: Redefine Meat Ltd
Priority date: 2023-08-31
Filing date: 2023-08-31
Publication date: 2025-02-01
Also published as: IL305612A; IL305612B1; WO2025046585A1

Description

0294472181- EDIBLE SCAFFOLDS, THEIR PREPARATION AND USES TECHNOLOGICAL FIELD The present disclosure relates to the food industry and more specifically to the food alternative industry.

BACKGROUND ART References considered to be relevant as background to the presently disclosed subject matter are listed below: - International patent application publication No. WO2022147357 - European patent application publication No. EP41442 - European patent application publication No. EP41660 - International patent application publication No. WO2022079717 Acknowledgement of the above references herein is not to be inferred as meaning that these are in any way relevant to the patentability of the presently disclosed subject matter.

BACKGROUND Plant based food products require the mimicking of organoleptic properties in order to meet consumers' needs. When dealing with whole cut meat alternatives, a specific challenge resides in the mimicking of connective tissue.

International patent application publication No. WO2022147357 describes a method of preparing a connective tissue analog and meat analog comprising the same; the method comprises combining ingredients comprising a hydrocolloid base and a dietary fiber additive to form a substantially homogenous mixture; hydrating the substantially homogenous mixture to form a hydrated gel; and at least partially dehydrating the gel to obtain an at least partially dehydrated gel comprising a non-covalently cross-linked polymer network, thereby forming the connective tissue analog. 0294472181- European patent application publication No. EP4144225 describes a scaffold including a plant protein and in-vitro meat produced by using the scaffold.

European patent application publication No. EP4166005 describes a method for obtaining a sterilizable macroporous three-dimensional (3D) tissue engineering scaffold which comprising a network of at least an inter-crosslinked biocompatible polymer. The method comprises preparing a dissolution of the at least a biocompatible polymer, pouring out the solution into molds and freeze.

International patent application publication No. WO2022147357 describes an edible meat analogue and a method of producing the same, the meat analogue comprising a plurality of protein strands and inter- strand sheaths material.

GENERAL DESCRIPTION The present disclosure provides, in accordance with a first of its aspects, a method of producing an edible 3D scaffold, the method comprising - introducing into a mold an edible gel forming formulation, the mold comprising a boundary and an internal pattern, the internal pattern constituting a negative of a 3-dimensional scaffold structure, - providing conditions that cause formation of a gel in said internal pattern; and - releasing the gel from the mold, said gel having a 3D scaffold structure; the 3D scaffold is defined as an arrangement of elongated channels interconnected by channel walls, said channel walls having a thickness that is less than half a channel's cross-sectional dimension.

Also provided, according to a second aspect of the presently disclosed subject matter, is a scaffold comprising a three-dimensional gel structure (referred to herein, at times, by the term "3D scaffold") including an arrangement of elongated channels interconnected by channel walls; wherein said channel walls comprise an edible gel and said channel walls have a thickness that is less than half a channel's cross-sectional diameter. 0294472181- In accordance with a third of its aspects, the present disclosure provides a method of producing an alternative whole muscle meat, the method comprises introducing edible protein mass into an edible 3D scaffold gel defined as an arrangement of elongated channels interconnected by channel walls.

BRIEF DESCRIPTION OF THE DRAWINGS In order to better understand the subject matter that is disclosed herein and to exemplify how it may be carried out in practice, embodiments will now be described, by way of non-limiting example only, with reference to the accompanying drawings, in which: Fig. 1 is an image of a silicon mold used for forming a connective tissue scaffold according to some examples of the presently disclosed subject matter.

Fig. 2 is an image of a plastic mold used for forming the silicon mold of Fig. 1.

Fig. 3 is an image of a 3D scaffold obtained using the silicon mold of Fig. 1.

Fig. 4A-4B are images of 3D scaffold having a Honeycomb structure with 0.wall thickness after being glued to T shape jig and put in between Llyod grippers for tensile test (Fig. 4A), and showing the specimen stretched while being held between two gripper (Fig. 4B) Fig. 5A-5F are graphs showing the tensile stress-strain curves and elastic moduli calculations of the evaluated specimens with or without post treatment and with the different wall thickness, as defined in Table 2.

Fig. 6 is an image of an alternative meat product including the connective tissue scaffold and protein mass.

Fig. 7A-7B are schematic illustration of a 3D scaffold having elongated parallel channels, with intersecting channels, showing specifically a cross sectional view (Fig. 7A) and top isometric view (Fig. 7B).

DETAILED DESCRIPTION The present disclosure is based on the development of edible scaffolds having a predesigned arrangement of thin-walled elongated channels into which edible protein 0294472181- mass is or can be introduced. The combination of scaffold, filled with the protein mass result in a product having organoleptic properties that resemble those of alternative whole muscle cuts.

The edible scaffolds and the formation of the thin-walled elongated channels is achieved using a mold designed as the negative of the desired arrangement of the thin- walled elongated channels.

The formation of the edible scaffold involves casting of gel forming material in the specifically designed mold and thereafter the release of the molded 3D structure from the mold.

Specifically, and in accordance with a first aspect of the presently disclosed subject matter, there is provided a method for producing an edible 3D scaffold, the method comprising: - introducing into a mold an edible gel forming formulation, the mold comprising a boundary and an internal pattern, the internal pattern constituting a negative of a 3-dimensional scaffold structure, - providing conditions that cause formation of a gel in said internal pattern; and - releasing the gel from the mold, said gel having said 3D scaffold structure; the 3D scaffold being defined as an arrangement of elongated channels interconnected by channel walls, said channel walls have a thickness that is less than half a channel's cross-sectional diameter.

In accordance with a second of its aspects, the presently disclosed subject matter provides a scaffold comprising a three-dimensional gel structure including an arrangement of elongated channels interconnected by channel walls; wherein the channel walls comprise an edible gel and wherein said channel walls have a thickness that is less than half a channel's cross-sectional diameter.

For the sake of simplicity, the following description including the different examples and definitions are applicable to both the presently disclosed method and the presently disclosed 3D scaffold, with the necessary changes according to the specific context. Thus, for example, when referring to the elongated channels, it is to be 30 0294472181- understood to refer to those of the mold, forming part of the first aspect of the presently disclosed subject matter, as well as to the 3D scaffold of the second aspect of the presently disclosed subject matter.

In the context of the presently disclosed subject matter, when referring to a "mold", it is to be understood to refer to a tangible structure having distinct boundaries (i.e. a defined physical perimeter) within which resides the internal pattern. This internal pattern embodies the negative impression or, in other words, the exact reverse configuration of the 3D scaffold to be produced therewith.

More specifically, the internal pattern provides at least the negative of the arrangement of the elongated channels of the 3D scaffold.

In the context of the presently disclosed subject matter, when referring to elongated channels it is to be understood to mean essentially linear, extended passages suitable to receive and accommodate protein mass, the channels having a length (longitudinal dimension) greater than their width (cross sectional dimension).

In some examples of the presently disclosed subject matter, the elongated channels have a length of at least 5mm. Accordingly, the internal pattern of the mold is constructed so as to provide channels with such length of at least 5mm.

In some examples of the presently disclosed subject matter, the elongated channels have an inner cross-sectional dimension of between about 1 mm and about mm. This is to be understood to mean also that the mold is constructed with an internal pattern is configured to provide elongated channels with an inner cross-sectional dimension of between about 1 mm and about 5 mm.

In the context of the presently disclosed subject matter, the term "inner cross-sectional dimension" is to be understood to refer to the largest dimension perpendicular to the channel's longitudinal axis, and without accounting to the thickness of the channel's wall at the cross section (i.e. net cross section).

The inner cross-sectional dimension is greater than the wall's thickness.

In some examples of the presently disclosed subject matter, the walls of the channel have a thickness that is less than half the channel's cross-sectional dimension. 0294472181- In some examples of the presently disclosed subject matter, the walls of the channel have a thickness that is less than a third the channel's cross-sectional dimension.

In some examples of the presently disclosed subject matter, the walls of the channel have a thickness that is less than quarter the channel's cross-sectional dimension.

In some examples of the presently disclosed subject matter, the walls of the channels have a thickness that is less than 0.4mm; at times, less than 0.3mm, when the 3D scaffold is in wet form; or less than 0.2mm; at time, less than 0.1mm when the 3D scaffold is in dried form (irrespective of the cross-sectional dimension of the channel).

When referring to dried or dry form, in the context of the presently disclosed subject matter, it is to be understood to encompass the 3D scaffold including up to about 20wt% water; at times, up to about 15wt% water; or even, at times, up to 10wt% water. Drying can be achieved by any method known in the art, as further described herein below.

When referring to wet form, in the context of the presently disclosed subject matter, it is to be understood to encompass the 3D scaffold including at least 20wt% water; at times, at least 30wt% water; at times, at least 40wt%; at times, at least 50wt%; at time, at least 60wt%; at times, at last 70wt%; at times, at least 80wt%; at times, at least 90wt%; at times, at least 95wt%; at least 98wt% or even up to about 98wt%.

In some examples of the presently disclosed subject matter, when the cross-section dimension is 1 mm, the wall's thickness is less than 0.5mm, at times, less than 0.3mm; at times, less than 0.25mm.

In some examples of the presently disclosed subject matter, when the cross-section dimension of the channel is 5mm, the wall's thickness will be less than 2.5mm; at times, less than 0.825mm; at times, less than 0.625mm.

In some examples of the presently disclosed subject matter, the channel walls have a thickness that is less than 0.2mm, when the 3D scaffold is in dried form.

In some examples of the presently disclosed subject matter, the channel walls have a thickness that is less than 0.1mm; when the 3D scaffold is in dried form. 0294472181- In some examples of the presently disclosed subject matter, the channel walls have a thickness that is in between about 0.01mm and about 0.2mm; when the 3D scaffold is in dried form.

In some examples of the presently disclosed subject matter, the channel walls have a thickness that is in between about 0.05mm and about 0.15mm; when the 3D scaffold is in dried form.

In some examples of the presently disclosed subject matter, the elongated channels within the 3D scaffold have essentially the same cross-sectional dimensions and/or a same average wall thickness.

In some examples of the presently disclosed subject matter, the elongated channels within the 3D scaffold may vary in their cross-sectional dimensions and/or in their wall thickness.

In some examples of the presently disclosed subject matter, the mold (i.e. the internal pattern) is constructed to provide channels with an inner cross-sectional area of between about 1mm and about 25mm. As a result, the channels within the 3D scaffold have inner cross-sectional areas of between about 1mm and about 25mm.

In some examples of the presently disclosed subject matter, the channels within the 3D scaffold have inner cross-sectional areas of between about 2mm and about 20mm; at times, between about 2mm and about 15mm; at times, between about 2mm and about 10mm. At times, about 3mm.

In the context of the presently disclosed subject matter, the term "inner cross-sectional area" is to be understood to refer to the two-dimensional surface spanned within the internal boundaries of the channel's wall, when viewed perpendicular to the channel's longitudinal axis, and without accounting to the thickness of the channel's wall at the cross section (i.e. net cross section).

The internal pattern is further configured to provide channels that not all channels are parallel. As such, at least a portion of the channels intersect, so that an organized collection of interconnected channels is formed. Yet, the internal pattern is configured such that at least 50% of the elongated channels have the same nominal direction, when viewed in a segment of the 3D scaffold having dimensions of about 10mm* 20mm* 30 0294472181- 20mm (4000mm); the short, 10mm dimension being the dimension aligned with the direction of the channels.

For the sake of simplicity, in the above and below description and in the appended claims, when referring to a 3D scaffold or to a sample of the 3D scaffold having dimensions of 10mm* 20mm* 20mm it is to be understood to have the short dimension (10mm) in the direction of the longitudinal axis of the elongated channels. Since the other two dimensions in the sample are greater than the longitudinal dimension of the elongated channels, the short dimension can be regarded as the thickness of the sample.

At times, at least 60% of the elongated channels have a same nominal direction, when viewed in a segment of the 3D scaffold having dimensions of about 10mm* 20mm* 20mm; at times, at least 70% of the elongated channels have a same nominal direction, when viewed in a segment of the 3D scaffold having dimensions of about 10mm* 20mm* 20mm; at times, at least 80% of the elongated channels have a same nominal direction, when viewed in a segment of the 3D scaffold having dimensions of about 10mm* 20mm* 20mm at least 90% of the elongated channels have a same nominal direction, when viewed in a segment of the 3D scaffold having dimensions of about 10mm* 20mm* 20mm.

The term " nominal direction " as used herein refers to a direction where significantly more than 50% of the channels (or the negative impression of the channels, in the mold) have a direction of up-to ±45 degrees from that nominal direction, when the channels (or their negative impression in the mold) are viewed from any direction perpendicular to the channel's direction. The term " nominal direction" may also refer to the average of the channels' direction as found using high magnification imaging as described herein. The nominal direction is a solid angle, where its projection on each of the 2 views, is the average direction found at this view.

In some examples of the presently disclosed subject matter, the channels (or their negative impression in the mold) are essentially parallel or essentially aligned to have a nominal direction.

In the context of the present disclosure, the term "essentially" is used to denote some level of deviation, such as 1%, 2%, 3%, 10%, or even up to 20%, from a defined parameter. 0294472181- In this context, when referring to "essentially parallel channels" or "parallelly oriented channels" or "essentially aligned channels" it is to be understood to refer to the orientation of at least 80% of the channels, at times, at least 85%; at times, at least 90%; at times, at least 95% of the channels one with respect to the other when viewed within a specimen, to be generally parallel. In this context, the essential alignment is within a specimen having a dimension of at least 1cm*2cm*2cm.

The term "essentially parallel" or "generally parallel" should be understood to encompass the nominal direction of the longitudinal axis to be at most ± 10º, at times, at most ± 3º, at most ±1º.

In some examples of the presently disclosed subject matter, the internal pattern is configured such that at least part of the elongated channels are intersected by one other channel, when viewed in a segment of the 3D scaffold having dimensions of about 10mm* 20mm*20mm. The dimensions of the intersecting channels need not to be the same as those of the elongated channels.

In some examples of the presently disclosed subject matter, the intersecting channels form an angle with the longitudinal axis of the elongated channel.

In some examples of the presently disclosed subject matter, the intersecting channels are essentially perpendicular to the longitudinal axis of the elongated channel.

A schematic illustration of elongated channels being intersected by other channels (or holes) is provided in Figs 7A-7B.

Without being bound thereto, it is believed that there is an advantage of having intersecting holes (also referred to as intersecting channels) between the columns is that during the material filling inside the cell, the mass will pass through the holes between the columns and create a connection between the phase of the filled mass. Such interactions may affect adhesion of the whole structure and better interlocking of the structure.

Thus, in the context of the present disclosure, when referring to intersecting channels, it also encompasses holes, and these are not necessarily elongated channels. 0294472181- In some examples of the presently disclosed subject matter, at least a portion of the elongated channels traverses the entire 3D scaffold, spanning from one end point to the opposite end point extend from one end to another end of the 3D scaffold.

In some examples of the presently disclosed subject matter, elongated channels are open at least at one end thereof.

In some examples of the presently disclosed subject matter, elongated channels are open from both ends thereof.

The cross section of the channel may have different shapes.

In some examples of the presently disclosed subject matter, at least a portion of the channels have a polygonal contour, e.g. have a polygonal cross section.

In some examples of the presently disclosed subject matter, the polygonal cross section includes polygonal shapes with at least 3 sides, this includes, without being limited thereto, quadrilaterals (4 sides), pentagons (5 sides), hexagons (6 sides), and heptagon (7 sides).

In some examples of the presently disclosed subject matter, the internal pattern is configured to provide channels with a hexagon cross section; and accordingly, the 3D scaffold comprises channels with a hexagon cross section.

In some examples of the presently disclosed subject matter, the internal pattern is configured to provide essentially aligned channels with a hexagon cross section. This may be regarded as an arrangement of essentially aligned pillars having a hexagonal cross section. According to this non-limiting example, the 3D scaffold thus comprises essentially aligned channels with a hexagon cross section.

In some examples of the presently disclosed subject matter, the internal pattern comprises the negative impression of a 3D honeycomb scaffold structure; and accordingly, the 3D scaffold comprises a 3D honeycomb scaffold structure. Figure 1 provides an image of an exemplary honeycomb mold and Figure 2 provides an image of an exemplary 3D scaffold having an internal pattern of honeycomb.

In some examples of the presently disclosed subject matter, the scaffold walls pattern is defined by applying the Voronoi function, i.e. the channel walls have a cross section structure defined by Voronoi function, and accordingly, the 3D scaffold 30 0294472181- comprises a Voronoi scaffold structure. The inputs of the Voronoi function are selected in such a manner that the channels dimensions and cross-sectional area fit the previously defined dimensions for the channel wall, channel cross section and other disclosed dimensions of the 3D scaffold.

In the context of the presently disclosed subject matter, when referring to a Voronoi scaffold or cross section it is to be understood to refer to a three dimensional architectural or structural framework created using the Voronoi tessellation concept. In this connection, it is to be appreciated that applying Voronoi principles is only an example of possible tessellation and other techniques can be used to construct the 3D pattern, such as mathematical or algorithmic methods or image processing of livestock meat or even manual design of a pattern, e.g. by industrial designer. The design of the 3D scaffold pattern can be, for example, inspired by existing and accessible patterns of meat fibers structures, determined via photography, microscopy and/or histology, bearing in mind that the channels structure and pattern is aimed, inter alia, to imitate the transverse section of muscle fascicles.

In some examples of the presently disclosed subject matter, at least a portion of the channels are designed to have or have a round or curved contour, e.g. have a round or curved cross section.

In some examples of the presently disclosed subject matter, at least a portion of the channels are designed to have or have a hybrid cross section, i.e. at least part of the cross section is defined by straight lines and angles therebetween and at least some other parts of the cross section are curved lines.

The method comprises introducing into the mold an edible gel forming formulation.

In the context of the present disclosure, the term "introducing" in connection with preparing the 3D scaffold, is to be understood to encompass encompasses any action that results in the essentially full embedment of edible gel (formed from the edible gel forming formulation) within the internal pattern of the mold.

In some examples of the presently disclosed subject matter, the gel forming formulation is in the form of semi solid or solid particles, and the introducing of the gel forming formulation into the mold (and eventually the formation of a gel within the 0294472181- internal pattern of the mold) comprises placing the solid or semi-solid edible gel forming particles over the mold and heating the mold to a temperature sufficient to cause melting of the solid or semi solid edible gel forming particles into a gel forming molten and to flow of gel forming molten into the internal pattern.

In some examples of the presently disclosed subject matter, the introducing of the gel forming formulation into the mold comprises placing molten gel forming formulation onto the mold.

In some examples of the presently disclosed subject matter, the introducing of the gel forming formulation into the mold comprises injection molding of molten gel forming formulation into the internal pattern of said mold.

Heating of the mold, when necessary, can be prior to, concomitant with or after placing the solid or semi-solid edible gel forming particles thereon.

In some examples of the presently disclosed subject matter, the heating of the mold is prior to placing the solid or semi-solid edible gel forming particles thereon.

In some examples of the presently disclosed subject matter, the mold is heated to a temperature sufficient to maintain the gel forming formulation in molten state, and to allow flow of the gel forming melt into the spaces formed by the internal pattern.

In some examples of the presently disclosed subject matter, the method also comprises cooling of the mold and the gel forming molten therein. Thus, according to the presently disclosed method, some of the conditions for the formation of the 3D scaffold using the defined mold involve heating of the mold and after the gel forming formulation is embedded within the internal pattern (the internal spaces) of the mold, the conditions comprise cooling the mold and the gel material therein. Without being limited thereto, the cooling causes the hardening of the gel material and thus of the 3D scaffold, and facilitates the extraction of the 3D scaffold from the mold.

Extraction of the 3D scaffold from the mold can be performed using techniques known in the art. For example, and without being limited thereto, the 3D scaffold can be removed using mechanical ejection tools, such as draft angles, mold extraction devices, lifting surfaces; air ejection techniques, such as compressed air; vacuum assisted ejection, spray release agents (suitable for the food industry) etc. 30 0294472181- In some examples of the presently disclosed subject matter, the method also comprises drying the 3D scaffold gel, after being extracted from the mold.

When referring to drying, in the context of the presently disclosed subject matter, it is to be understood to encompass any technique of water removal. The drying does not need to be to complete dryness. In some examples of the presently disclosed subject matter, when referring, with respect to the 3D scaffold, to dry or drying or dried, it is to be understood to define that the 3D scaffold has up to about 20wt% water; at times, up to about 15wt% water; at times, up to about 10wt% water; at times, up to about 5wt%; at times, up to about 3wt%; at times, up to about 1wt%. Water content in the scaffold gel can be determined using, for example, Differential Scanning Calorimetry (DSC), or by moisture analyzer as known in the art.

In some examples of the presently disclosed subject matter, the drying is by any one or combination of freeze drying, heating and air dry of the 3D scaffold gel.

In some examples of the presently disclosed subject matter, the drying comprises at least freeze drying of the 3D scaffold gel.

Accordingly, in some examples of the presently disclosed subject matter, when drying is applied, the 3D scaffold in accordance with the presently disclosed subject matter is in dried or dry form.

In some examples of the presently disclosed subject matter, the method also comprises applying onto surfaces of the internal pattern an edible protein binding agent.

In the context of the presently disclosed subject matter, the term "edible protein binding agent" denotes any edible agent that facilitates or promotes the binding of the protein mass to be introduced into the elongated channels, to the channel walls.

Non-limiting examples of edible protein binding agent include gluten, glucomannan, locus bean gum, mastic resin, casein, guar gum, gum Arabic, xanthan gum, a psyllium, a chitin, an inulin, a pectin, a dextrin, a starch, a cellulose, a hemicellulose, a lignin, a citrus fiber extract, dietary enzyme, a transglutaminase analogs or derivatives thereof, or any combination thereof. Preferably, the protein binding agent is a non-animal derived/vegan protein (e.g. artificially produced casein).

In some examples of the presently disclosed subject matter, the edible protein binding agent comprises gluten. 0294472181- In some examples of the presently disclosed subject matter, the edible protein binding agent comprises gluten powder.

The mold per se can be made from materials known in the art of molding, such as, silicon, polyurethane, latex rubber, metal, aluminum, fiberglass-reinforced plastics (FRP), wood and others.

Further, the mold per se can be made by a variety of techniques known in the art for the fabrication of rigid molds, including, for example, binder jetting, material jetting, direct energy deposition (DED), powder bed fusion (PBF), fused deposition molding (FDM), mechanical milling, laser machining and others. For example, the mold of Fig. 1 was prepared using a 3D printing Stereolithography (SLA) using "Smooth SIL 940" silicon model, with Shore 40 food grade. Other techniques may include CNC.

The gel forming formulation can comprises a variety of edible gels.

In some examples of the presently disclosed subject matter, the gel forming formulation comprises material that forms hydrogel. Thus, in such cases, the gel forming formulation is a hydrogel forming formulation.

In some examples of the presently disclosed subject matter, the gel forming formulation comprises material that forms a hydrocolloid. Thus, in such cases, the gel forming formulation is a hydrocolloid forming formulation.

In some examples of the presently disclosed subject matter, the hydrocolloid forming formulation comprises water and at least one hydrocolloid forming agent.

In some examples of the presently disclosed subject matter, the gel (or at times, hydrocolloid) forming agent is selected from the group consisting of Carrageenan, Agar, Gelatin, Sodium Alginate, Gellan Gum, Pectin, Xanthan Gum, Guar Gum, Methylcellulose, Starch, Konjac, Locust Bean Gum, Tara Gum, Curdlan and Chitosan.

In some examples of the presently disclosed subject matter, the hydrocolloid forming agent is Carrageenan (CAR).

In some examples of the presently disclosed subject matter, the hydrocolloid forming agent is Kappa Carrageenan (κ-CAR). 0294472181- In some examples of the presently disclosed subject matter, the gel forming formulation comprises at least 5% CAR (out of total wet weight), preferably at least 5% κ-CAR.

In some examples of the presently disclosed subject matter, the hydrocolloid forming formulation comprises a combination of two or more hydrocolloid forming agents, at least one being selected from the above non-limiting list of hydrocolloid forming agents.

Non-limiting Examples of gel forming materials that can be used in the context of the presently disclosed subject matter and their physical properties is provided in Table below, each formulation constituting an independent example of the presently disclosed subject matter.

Table 1: physical properties of exemplary gel forming material Formulation Average Load (N) Average Strain Average Stress (MPa)5% CAR 3.46 0.089 0.65% CAR+5%coconut oil 2.38 0.494 0.0 % CAR+10%coconut oil 2.25 0.443 0.0 % CAR+5%glycerol 2.83 0.624 0.15% CAR+10% glycerol 3.46 0.687 0.15% CAR+5%pectine 1.94 0.449 0.05% CAR+5%gellan 6.08 1.009 0.15% CAR+5% +Methyl D-lactate 1.45 0.452 0.0 % CAR+5% glycerol +1%gellan 3.38 0.829 0.1 Thus, without being limited to these examples, it is clear that by variations of the gel forming formulation can allow the variability in the physical properties. Further, the 15 0294472181- above examples show an advantage of combining gellan and/or glycerol with CAR as well as an advantage of adding to the gel forming formulation gellan.

In some examples of the presently disclosed subject matter, the gel forming formulation comprises other additives, not necessarily gel forming agents.

In some examples of the presently disclosed subject matter, the gel forming formulation comprises water-soluble polyol(s). In some examples, the polyol is selected from the group consisting of glycerol, Maltitol, sorbitol, xylitol, erythritol, and isomalt and combinations of same.

In some examples, the polyol is or comprises at least glycerol (also known as glycerin).

In some examples of the presently disclosed subject matter, the formulation comprises at least 5% CAR with glycerol.

In some examples of the presently disclosed subject matter, the amount of glycerol in the gel forming formulation is designed to be in the range of about 1%w/w and 10%w/w out of the total wet weight.

In some examples of the presently disclosed subject matter, the formulation comprises Gellan and at least 5% CAR out of the total wet weight.

In some examples of the presently disclosed subject matter, the amount of Gellan in the gel forming formulation, if present, is designed to be in the range of up to 3%w/w out of the total wet weight.

In some examples of the presently disclosed subject matter, the formulation comprises glycerin and Gellan with at least 5% CAR out of the total wet weight.

The amount of water mixed with the dry components of the gel forming formulation may range based on the selected components.

In some examples of the presently disclosed subject matter, the gel forming formulation can comprise animal free fat or oil, e.g. plant-based oil(s).

Without being limited thereto, the gel forming material can comprise any one or combination of the following oils: olive oil, coconut oil, sunflower oil, soybean oil, canola oil, avocado oil, almond oil, grapeseed oil, flaxseed oil, palm oil, hemp seed oil, 0294472181- Without being limited thereto, the gel forming material can comprise any one or combination of cacao butter, shea butter, coconut fat.

Without being limited thereto, the gel forming material can comprise a combination of at least one animal free fat (such as those listed above) and at least one animal free oil (such as those listed above).

For example, and without being limited thereto, the plant-based oil can include coconut oil and the formulation can include between about 1%w/w and 15%w/w plant-based oil.

The presently disclosed subject matter also provides, in accordance with a further aspect, a method of producing an alternative whole muscle meat, the method comprises introducing edible protein mass into an edible 3D scaffold gel defined as an arrangement of elongated channels interconnected by channel walls (the walls comprising the edible gel). In this context, there is also provided the alternative whole muscle meat/cut obtained using the disclosed 3D scaffold and the methods of using it.

Without being bound by theory, the combination of the presently disclosed 3D gel scaffold and the protein mass introduced therein provides improved properties in terms of texture and other organoleptic properties desired for alternative, plant based whole muscle meat. It is envisaged that the 3D gel structure imitates the functionality of the connective tissue surrounding bundles of muscle fibers also known as fascicles, while the protein mass provides the functionality of the muscle fibers.

Thus, for the sake of simplicity, it is to be understood that the above description and different definitions of features and alternative examples provided in connection with the presently disclosed method of producing the edible 3D scaffold and the presently disclosed edible 3D scaffold per se, are also applicable and define different examples and alternatives to the method of producing the alternative whole muscle cut.

The protein mass introduced into the edible 3D scaffold is an edible protein mass.

The term "protein mass" as used herein, denotes a composition comprising one or more edible proteins. In some examples, the protein mass is pliable, with a sufficient degree of flowability (under pressure) to allow its introduction into the elongated channels of the 3D scaffold, without causing any damage to the scaffold structure. 30 0294472181- In some examples of the presently disclosed subject matter, the protein mass is in fluid form.

In some examples of the presently disclosed subject matter, the protein mass has a physical state that permits its pouring or injection into the channels.

In some examples of the presently disclosed subject matter, the protein mass comprises the one or more edible proteins, in combination with other substances, such as hydrogels, fats, coagulation agents etc.

In some examples of the presently disclosed subject matter, the protein mass comprises at least 10%w/w, at times, at least 20%w/w, at times, at least 30%w/w protein(s).

In some examples of the presently disclosed subject matter, the protein mass comprises between 10%w/w and 30%w/w protein(s).

In some examples of the presently disclosed subject matter, the protein mass comprises between 15%w/w and 25%w/w protein(s).

The protein mass being introduced into the 3D scaffold is to be considered as a wet composition. In some examples, the wet protein mass comprises at least 50%w/w water.

In some examples of the presently disclosed subject matter, the protein mass comprises between 55% and 80% water; at times, between 60% and 75% water; at times between 65% and 70% water.

In some examples of the presently disclosed subject matter, the protein mass comprises plant derived (e.g. isolate or concentrate) proteins (including peptides and amino acids). Without being limited thereto, the plant source for the protein mass can be any one or combination of soy, wheat, legume (e.g. pea, chickpea, beans), rapeseed and corn as well as many other plant based protein sources as known in the food industry.

In some examples of the presently disclosed subject matter, the protein mass can comprise protein derived from sources other than plants, such as algae, fungi (e.g. yeast), bacteria and microorganisms in general.

In some examples of the presently disclosed subject matter, the protein mass can comprise texturized vegetable protein (TVP). TVP is known in the art to be used as a 30 0294472181- meat extender or vegetarian meat and is usually created by extruding protein isolates or concentrates using high shear, pressure and heat, from vegetable sources such as wheat, pea and others. TVP is commercially available in different sizes from large chunks to small flaks.

In some examples of the presently disclosed subject matter, the protein mass comprises TVP. In some examples of the presently disclosed subject matter, the minimal amount of TVP within the protein mass is at least 10%w/w of dry weight TVP, out of the total protein in the protein mass. At times, the minimal amount is at least 15%w/w of dry TVP, at times at least 20%w/w of dry TVP; at times at least 25%w/w of dry TVP; at times at least 30%w/w of dry TVP; at times at least 35%w/w of dry TVP; at times at least 40%w/w of dry TVP; at times at least 45%w/w of dry TVP; at times at least 50%w/w of dry TVP; at times at least 55%w/w of dry TVP; at times at least 60%w/w of dry TVP.

In some examples of the presently disclosed subject matter, the minimal amount of TVP within the protein mass (out of the total protein), as determined before being introduced into the scaffold, is at least 20%w/w of wet TVP out of the total protein; at times, at least 30%w/w of wet TVP out of the total protein ; at least 35%w/w of wet TVP out of the total protein. At times, the minimal amount is at least 40%w/w of wet TVP; at times, at least 45%w/w wet TVP; at times, at least 50% w/w TVP; at times, at least 55% w/w TVP; at times, at least 60% w/w TVP; at times, at least 65% w/w TVP; at times, at least 70% w/w TVP; at times, at least 75% w/w TVP.

The protein mass does not include only TVP. In some examples, the protein containing material contains at least 30% non-TVP protein matter.

In some examples of the presently disclosed subject matter, the protein mass also comprises fibrous material, i.e. natural fibers.

The protein mass can be introduced into the elongated channels of the 3D scaffold by any techniques known in the art that is suitable for such protein mass deposition.

Without being limited thereto, the introduction of the protein mass into the 3D scaffold's elongated channels, can be by the use of fluid filling techniques, such as, piston filling machines (e.g. SGP-250 Liquid Piston Filler Machine), peristaltic filling machines, gravity filling machines, vacuum filling machines, overflow filling machines, time- 30 0294472181- pressure filling machines, net weight filling machines, electrical caulking gun (e.g. Markita DCGI80) equipped with a nozzle (in the mm size, e.g. 4 mm).

To facilitate the introduction of the protein mass without causing unnecessary pressure on the channel walls, and further to facilitate flow of the protein mass within the channel, the dispensing nozzle is selected to have a cross sectional dimension that is smaller than that of the cross-sectional dimension of the elongated channel.

In some examples of the presently disclosed subject matter, the introduction of the protein mass is into a single 3D scaffold.

In some examples of the presently disclosed subject matter, the introduction of the protein mass is into two or more 3D scaffold, stacked one on top of the other.

In some examples of the presently disclosed subject matter, the method of producing the alternative whole cut comprises stacking one on top of another 3D scaffolds, as disclosed herein, already filled with protein mass.

In some examples of the presently disclosed subject matter, when the method comprises stacking one on top of another 3D scaffolds already filled with protein mass, the protein mass of the different scaffold layers may be the same or different.

In some examples of the presently disclosed subject matter, the method of producing the alternative whole cut comprise intruding edible fat into at least part of the elongated channels.

In some examples of the presently disclosed subject matter, the method of producing the alternative whole cut comprise intruding edible fat in between two stacked 3D scaffolds holding the protein mass.

DESCRIPTION OF NON-LIMITING EXAMPLESThe following non-limiting Example illustrates formation of a matrix-like connective tissue structure into which edible, non-animal proteins are introduced creating a whole meat alternative muscle cut structure.

EXAMPLE 1 - Preparation of a Connective Tissue Scaffold For forming the edible scaffold that forms the connective tissue of the whole meat alternative muscle cut structure, a silicon mold was constructed to function as a negative 0294472181- of the desired cellular matrix shape and porosity of the connective tissue to be produced. In this non-limiting Example, the mold, shown in Figure 1,had the shape suitable to provide a scaffold with an internal configuration of a honeycomb, defined by hexagon open cells.

Specifically, the silicon mold was created to provide hexagon cells having a circumcircle diameter of about 2.5mm and a hexagonal cell wall thickness of about 0.2mm.

Preparation of a mold The mold was prepared from food grade Silicon, with 40 Shore A hardness that was poured into a plastic rigid mold as shown in Figure 2 , having hexagonal cells with a wall thickness of about 0.2mm. In fact, the plastic mold of Figure 2, is essentially a mirror structure of the scaffold to be obtained.

Preparation of connective tissue using mold The connective tissue (CT) forming material was prepared from a mixture comprising 5% Carrageenan, 5% Glycerol, 1% Gellan, 89% Water, which was heated to 90°C and steered in thermomixer until a homogeneous CT forming material was obtained.

Process A: The homogeneous CT forming material was cooled to room temperature and then broken into pieces and placed over the Silicon mold until completely covered. The mold with the CT pieces were placed into an oven, pre-heated to 90°C for 20 minutes until all CT forming material melted and penetrated into the Silicon mold leaving a thin layer of CT forming material over the mold. Onto the thin layer of CT forming material, an additional portion of molten CT forming material is poured and put back into the oven set to a temperature of 115°C, for an additional 10 minutes, allowing any trapped air bubbles to be released from the molded CT forming material.

Process B: Molten CT forming material was poured onto the oven heated mold and then spread and pressed against the mold's open cells. The mold with the pressed CT forming material was then placed back in the open, set at 95°C, for 10 minutes, after which it was allowed to cool outside at room temperature. 30 0294472181- The mold with the scaffold made of the CT forming material was cooled at 4°C for 15 minutes and after removing excess CT forming material (e.g. the layers on top of the mold), the CT scaffold was extracted from the silicon mold.

The CT scaffold is shown in Figure 3 , which has a honeycomb internal structure.

Post-formation Processing of the CT Scaffold Freeze-drying For preserving geometric structure of the CT scaffold, the CT scaffold is subject to freeze-drying so as to remove water from the scaffold's walls. Freeze drying provides improved long term structural stability as compared to CT scaffold prepared by the same procedure, yet, without the freeze drying.

The conditions of freeze drying are provided in Table 2 including temperature, duration, vacuum.

Table 2: Freeze-Drying Conditions Shelf Temperature Ramp (min) Hold (min) Vac(microbar) Freezing-10 0 30 92 -25 30 0 93 -25 0 120 9 Sublimation-First drying-25 0 30 75 -25 0 90 66 -20 10 0 67 -10 0 30 68 0 0 120 6 Desorption- Secondary Drying10 0 360 610 15 0 60 511 15 0 360 412 20 0 240 200 0294472181- Shelf Temperature Ramp (min) Hold (min) Vac(microbar)20 0 240 Surface treatment To increase adhesion of the protein mass introduced into the open cells of the CT scaffold, the cells surface is scattered with edible gluten powder and placed in an oven heated to 50°C for 24 hours.

EXAMPLE 2 - Connective Tissue Scaffold Characteristics The physical properties of the 3D scaffold samples were determined. Specifically, honeycomb scaffolds were prepared from a formulation comprising 5wt% CAR, 5wt% glycerol and 1wt% gellan F, to provide honeycomb specimens (90~110*40~50*10mm) with 20 micron, 50 micron, 200 micron or 400 micron wall. Some of the specimens were subjected to post treatment by either freeze-drying according to Table 2 above, or by immersing within ionic bath (anti-solvent including 18 g KCl dissolved in 250ml filtered water, into which 250 ml ethanol is added). Table 3 provides the different formulations and post treatments of the characterized specimens.

Table 3 – Specimens for characterization Specimen No. Wall thickness (micron) Freeze drying Ionic bath 1 400 - - 2 200 - - 3 50 √ - 4 20 √ - 400 - √ 6 200 - √ The tensile strength of combined specimens was analyzed. To this end, the specimens were glued with cyanoacrylate glue at their edges (top and down) to a T shape accessory which are needed for gripping behavior. The specimens were gripped by Llyod 0294472181- gripper for tensile tests (see Figure 4A). Then, at room temperature (0C±0C), each of the specimens was stretched using LLOYD TPA instrument equipped with a 100N load cell at a speed of 5mm/s. Figure 4B provides an image of a tensile strength measurement of the same 3D scaffold having a Honeycomb structure with 0.4mm wall thickness, showing the specimen stretched while being held between two gripper.

The results are presented in Table 4 . Presenting the max load [N], tensile strength [MPa] and strain at break (mm/mm).

Table 4: mechanical properties of tested specimens Table 4 shows that the different specimens can withstand a max load of at least 3N. Further, it is clear from these results that post treatment, e.g. freeze drying or ionic bath immersion significantly improved the mechanical properties of the specimens.

As appreciated, the tensile strength graphs of a tensile test displays stress (σ) on the y-axis and strain (ε) on the x-axis. Stress is the force applied per unit area, while strain is the proportional change in length of the material.

In the honeycomb structures of Table 3, the true cross-sectional area was determined by addition of the total walls thickness in the cross-section area, excluding the voids in between.

Figs. 5A-5F are the graphs showing the tensile stress-strain curves and elastic moduli calculations of the evaluated specimens with or without post treatment and with the different wall thickness, defined in Table 3 (Fig. 5A – specimen 1, Figure 5B- specimen 2 etc).

The tested specimens exhibited properties of hyperplastic materials.

Specimen No. 1 2 3 4 5 6 Max Load [N] 7.4 3.8 44.5 27.8 63.7 26.

Max Stress (MPa)0.06 0.05 2.86 2.58 0.74 0.

Max Strain (mm/mm)0.63 0.49 0.49 0.94 1.09 1.23 0294472181- Generally, tensile test graph of a hyperelastic material differs from that of conventional linear materials because hyperelastic materials display nonlinear stress-strain behavior, often associated with significant deformations. Hyperelastic materials are a class of materials that can undergo large elastic deformations while still returning to their original shape upon unloading. They are commonly used to model rubber-like materials, soft tissues, and elastomers.

Further, Generally, a hyperelastic material graph of a tensile test is divided into segments: Initial Behavior: In the initial phase of the test, as stress is applied, the hyperelastic material behaves elastically. However, unlike linear materials, the stress-strain curve is nonlinear from the beginning. The stress response increases more rapidly than the strain due to the material's highly deformable nature.

Nonlinear Elastic Region: As the material continues to deform under increasing stress, the stress-strain curve maintains its nonlinear trend. Hyperelastic materials often exhibit strain-stress relationships described by various mathematical models, such as the Neo-Hookean, Mooney-Rivlin, or Ogden models. These models capture the nonlinear behavior of the material under deformation.

High Deformation Range: Hyperelastic materials can withstand significant deformations without undergoing plastic deformation or permanent damage. This behavior allows them to be stretched or compressed to large strains and still return to their original shape when the load is removed.

Ultimate Stretch Limit: The stress-strain curve will eventually reach a point where the material's deformation capabilities are exhausted. This could be represented by the point of maximum stress, often referred to as the ultimate stretch limit. Beyond this point, the material might begin to show signs of failure or exhibit more pronounced nonlinear behavior. 30 0294472181- Failure or Necking (If Applicable): Depending on the specific behavior of the hyperelastic material, it might experience necking or localized deformation before ultimate failure. In some cases, the material might rupture or tear, leading to a rapid decrease in stress.

EXAMPLE 3 - Preparation of a Whole Muscle Cut using the Connective Tissue Scaffold To obtain the alternative whole muscle cut, the open cells of the CT scaffold of Example 1 were filled with protein mass using a piston filling machine (Makita equipped with a 3mm nozzle). The protein composition included 20% Soy protein isolate, 10% Gluten, 10% Canola oil and 60% Water.

The resulting alternative whole muscle cut contained a connective tissue scaffold with protein mass occupying the scaffold’s open cells, as clearly shown in Figure 6.

Claims

- 27 - 30 56 12 / 02944721178- CLAIMS:

1. A method of producing an edible 3D scaffold, the method comprising - introducing into a mold an edible gel forming formulation, the mold comprising a boundary and an internal pattern, the internal pattern constituting a negative of a 3-dimensional scaffold structure, - providing conditions that cause formation of a gel in said internal pattern; and - releasing the gel from the mold, said gel having a 3D scaffold structure; the 3D scaffold is defined as an arrangement of elongated channels interconnected by channel walls, said channel walls having a thickness that is less than half a channel's cross-sectional dimension; and said elongated channels are free for filling with edible protein mass.

2. The method of claim 1, wherein said internal pattern is configured to provide channels with a length of at least 5mm.

3. The method of claim 1 or 2, wherein said internal pattern is configured to provide any one of the following: - channels with an inner cross-sectional dimension of between 1 mm and mm; channels with an inner cross-sectional area of between 1mm and 25mm and/or channel walls with a thickness of less than 0.2mm, when said 3D scaffold is in dried form; and/or - channel walls with a thickness of less than 0.4mm, when said 3D scaffold is in wet form.

4. The method of any one of claims 1 to 3, wherein said internal pattern is configured such that at least 50% of the elongated channels (i) have a same nominal direction, when viewed in a segment of the 3D scaffold having dimensions of 10mm*20mm*20mm, and/or intersect at least one other elongated channel, when viewed in a segment of the 3D scaffold having dimensions of 10mm*20mm*20mm.

5. The method of any one of claims 1 to 4, wherein said internal pattern is configured to provide channels with a hexagon cross section. - 28 - 30 56 12 / 02944721178-

6. The method of any one of claims 1 to 5, wherein said internal pattern is configured to provide a 3D honeycomb scaffold structure.

7. The method of any one of claims 1 to 6, wherein said internal pattern is configured to provide channels with a Voronoi cross section.

8. The method of any one of claims 1 to 7, wherein said gel forming formulation comprises solid or semi-solid edible gel forming particles and said introducing of the gel forming formulation into said mold comprises placing the gel forming formulation including the solid or semi-solid edible gel forming particles over said mold and heating the mold to a temperature sufficient to cause melting of the solid or semi solid edible gel forming particles into a gel forming molten and flow of said gel forming molten into said internal pattern.

9. The method of claim 8, comprising heating the mold prior to placing the solid or semi-solid edible gel forming particles thereon.

10. The method of any one of claims 1 to 8, wherein said introducing of the gel forming formulation into said mold comprises placing molten gel forming formulation onto the mold.

11. The method of claim 10, comprising heating the mold prior to placing said molten gel forming formulation.

12. The method of any one of claims 9 to 11, comprising heating the mold to a temperature sufficient to maintain said gel forming formulation in molten state, and allowing flow of said gel forming molten into said internal pattern.

13. The method of any one of claims 1 to 12, wherein said introducing of the gel forming formulation into said mold comprises injection molding of molten gel forming formulation into the internal pattern of said mold.

14. The method of any one of claims 1 to 13, wherein the conditions that cause formulation of the gel in said internal pattern comprises cooling of said mold and said gel forming molten.

15. The method of any one of claims 1 to 14, comprising drying said 3D scaffold gel by any one or combination of freeze drying, heating, air dry of the 3D scaffold gel. - 29 - 30 56 12 / 02944721178-

16. The method of claim 15, wherein said freeze drying is of the 3D scaffold gel after being released from the mold.

17. The method of any one of claims 1 to 16, comprising applying onto surfaces of the internal pattern an edible protein binding agent.

18. The method of claim 17, wherein said edible protein binding agent comprises gluten.

19. The method of any one of claims 1 to 18, wherein said gel forming formulation is a hydrogel forming formulation.

20. The method of any one of claims 1 to 19, wherein gel forming formulation comprises water and at least one hydrocolloid forming agent selected from the group consisting of Carrageenan, Agar, Gelatin, Sodium Alginate, Gellan Gum, Pectin, Xanthan Gum, Guar Gum, Methylcellulose, Starch, Konjac, Locust Bean Gum, Tara Gum, Curdlan and Chitosan and combinations of same.

21. The method of claim 20, wherein said gel forming formulation comprises at least Carrageenan.

22. A scaffold comprising a three-dimensional gel structure including an arrangement of elongated channels interconnected by channel walls; wherein said channel walls comprise an edible gel and said channel walls have a thickness that is less than half a channel's cross-sectional diameter; and said elongated channels are free for filling with edible protein mass.

23. The 3D scaffold of claim 22, wherein said elongated channels have a length of at least 5mm.

24. The 3D scaffold of claim 22 or 23, wherein said elongated channels are characterized by at least one of the following: - said elongated channels have an inner cross-sectional dimension of between 1 mm and 5 mm; - said elongated channels have an inner cross-sectional area of between 1mm and 25mm; - said elongated channels have a wall a thickness of less than 0.2mm. - 30 - 30 56 12 / 02944721178-

25. The 3D scaffold of any one of claims 22 to 24, wherein at least 50% of said elongated channels have a same nominal direction, when viewed in a segment of the 3D scaffold having dimensions of 10mm(width)*20mm*20mm.

26. The 3D scaffold of any one of claims 22 to 25, wherein said elongated channels have a hexagon cross section.

27. The 3D scaffold of any one of claims 22 to 26, wherein said elongated channels provide a 3D honeycomb scaffold structure.

28. The 3D scaffold of any one of claims 22 to 27, wherein said elongated channels have a Voronoi cross section.

29. The 3D scaffold of any one of claims 22 to 28, in wet form or in dried form.

30. The 3D scaffold of claim 38, being in freeze-dried.

31. The 3D scaffold of any one of claims 22 to 30, wherein said edible gel is or comprises a hydrogel.

32. The 3D scaffold of any one of claims 22 to 31, wherein said edible gel is or comprises a hydrocolloid.

33. The 3D scaffold of any one of claims 22 to 32, wherein said edible gel is or comprises water and at least one hydrocolloid forming agent selected from the group consisting of Carrageenan, Agar, Gelatin, Sodium Alginate, Gellan Gum, Pectin, Xanthan Gum, Guar Gum, Methylcellulose, Starch, Konjac, Locust Bean Gum, Tara Gum, Curdlan and Chitosan and combinations of same.

34. The 3D scaffold of any one of claims 22 to 33, wherein said edible gel is or comprises at least Carrageenan.

35. The 3D scaffold of any one of claims 22 to 34, wherein said edible gel is or comprises at least 5%w/w Carrageenan.

36. The 3D scaffold of any one of claims 22 to 35, wherein said edible gel is or comprises at least Carrageenan and Gellan.

37. The 3D scaffold of any one of claims 22 to 36, wherein said edible gel is or comprises at least Carrageenan and glycerin. - 31 - 30 56 12 / 02944721178-

38. The 3D scaffold of any one of claims 22 to 37, wherein said edible gel is or comprises at least Carrageenan, glycerin and Gellan.

39. A method of producing an alternative whole muscle meat, the method comprises introducing edible protein mass into an edible 3D scaffold gel defined as an arrangement of elongated channels interconnected by channel walls; wherein the elongated channels are free for filling with said edible protein mass.

40. The method of claim 39, wherein at least one of the following conditions is fulfilled: said channel walls have a thickness that is less than half a channel's cross-sectional diameter; said channel walls have a thickness that is less than 0.2mm.

41. The method of claim 39 or 40, wherein said elongated channels are characterized by at least one of the following: - said elongated channels have a length of at least 5m; - said elongated channels have an inner cross-sectional diameter of between mm and 5 mm; - said elongated channels have an inner cross-sectional area of between 1mm and 25mm.

42. The method of any one of claims 39 to 41, wherein said channel walls have a thickness of less than 0.2mm, when said 3D scaffold is in dried form, or less than 0.4mm when said 3D is in wet form.

43. The method of any one of claims 39 to 42, wherein at least 50% of the elongated channels have a same nominal direction when viewed in a segment of the 3D scaffold having dimensions of 10mm*20mm*20mm.

44. The method of any one of claims 39 to 43, wherein at least 50% of the elongated channels intersect at least one other elongated channel when viewed in a segment of the 3D scaffold having dimensions of 10mm*20mm*20mm.

45. The method of any one of claims 39 to 44, wherein said 3D scaffold gel has channels have with a hexagon cross section. - 32 - 30 56 12 / 02944721178-

46. The method of any one of claims 39 to 45, wherein said 3D scaffold gel has 3D honeycomb scaffold structure.

47. The method of any one of claims 39 to 46, wherein said 3D scaffold gel has channels with a Voronoi cross section.

48. The method of any one of claims 39 to 47, wherein said gel is a hydrogel.

49. The method of any one of claims 39 to 48, wherein said gel is a hydrocolloid.

50. The method of any one of claims 39 to 49, wherein said gel comprises at least one hydrocolloid forming agent selected from the group consisting of Carrageenan, Agar, Gelatin, Sodium Alginate, Gellan Gum, Pectin, Xanthan Gum, Guar Gum, Methylcellulose, Starch, Konjac, Locust Bean Gum, Tara Gum, Curdlan and Chitosan and combinations of same.