Decellularised cell wall structures from plants and fungus and use thereof as scaffold materials
Inventors
Pelling, Andrew Edward • Cuerrier, Charles Michel • Modulevsky, Daniel J. • Hickey, Ryan Joseph
Assignees
Publication Number
US-11045582-B2
Publication Date
2021-06-29
Expiration Date
2037-02-10
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Abstract
Provided herein are scaffold biomaterials comprising a decellularised plant or fungal tissue from which cellular materials and nucleic acids of the tissue are removed, the decellularised plant or fungal tissue comprising a cellulose- or chitin-based 3-dimensional porous structure. Methods for preparing such scaffold biomaterials, as well as uses thereof as an implantable scaffold for supporting animal cell growth, for promoting tissue regeneration, for promoting angiogenesis, for a tissue replacement procedure, and/or as a structural implant for cosmetic surgery are also provided. Therapeutic treatment and/or cosmetic methods employing such scaffolds are additionally described.
Core Innovation
The invention provides scaffold biomaterials comprising decellularised plant or fungal tissue from which cellular materials and nucleic acids have been removed. These decellularised tissues contain cellulose- or chitin-based three-dimensional porous structures that serve as scaffolds capable of supporting animal cell growth, promoting tissue regeneration and angiogenesis, and functioning as structural implants for various surgical and cosmetic applications.
The problem being solved addresses the limitations of current biomaterials, which often involve complex and costly production methods, and that are frequently of human or animal origin. Such materials can cause immune rejection, adverse responses, disease transmission risks, environmental impact, and ethical concerns. Additionally, many existing biomaterials lose shape over time, compromising tissue repair or replacement. There is a need for alternative biomaterials that are biocompatible, maintain structure, are cost-effective, and have minimal environmental impact.
This invention addresses these issues by utilizing decellularised plant or fungal tissues as scaffold biomaterials which are non-resorbable, highly porous, and maintain their shape over time. The biomaterials can be produced using relatively simple and efficient methods such as detergent treatment with removal of residual detergents, and may be further functionalized chemically or structurally to tailor properties for specific applications. The plant- or fungus-derived scaffolds demonstrate biocompatibility, promote vascularization and tissue repair, and offer a versatile and sustainable source material with low environmental footprint.
Claims Coverage
The patent contains one independent claim directed to a method for preparing a decellularised plant or fungal tissue scaffold including detergent treatment and subsequent removal of residual detergent.
Method for preparing decellularised scaffold with detergent treatment and residual removal
The method involves providing plant or fungal tissue of predetermined size and shape, decellularising it through treatment with sodium dodecyl sulphate (SDS), and removing residual SDS using an aqueous divalent salt solution that precipitates SDS micelles from the tissue.
Removal of divalent salt residue and SDS micelles
Following precipitation, the residual salt solution, salt residue, and/or SDS micelles are removed by treatments such as deionized water, acetic acid, dimethylsulfoxide (DMSO), sonication, or combinations thereof.
Use of specific divalent salts in residual detergent removal
The divalent salt used in the aqueous solution to remove residual SDS comprises magnesium chloride (MgCl2) or calcium chloride (CaCl2).
Specific detergent concentration and washing protocol
Decellularisation uses an SDS solution ranging from about 0.1% to about 1% in water, with residual SDS removed post-treatment using approximately 100 mM CaCl2 aqueous solution followed by incubation in deionized water.
Post-decellularisation processing and functionalization
The method can include processing the decellularised tissue to introduce microarchitecture features and/or functionalizing free hydroxyl groups via acylation, alkylation, or other covalent modifications.
Functionalization agents for scaffold modification
Functionalization of the decellularised tissue hydroxyl groups may include collagen, factors promoting cell specificity, cell growth factors, or pharmaceutical agents.
Cell seeding on the scaffold structure
The method can comprise introducing living animal cells onto the cellulose- or chitin-based porous scaffold and allowing their adhesion.
Animal cell types for scaffold seeding
The living animal cells used on the scaffold may be mammalian cells.
Human cell seeding on scaffold
The living animal cells seeded onto the scaffold may be human cells.
Overall, the claims cover a method of producing decellularised plant or fungal tissue scaffolds through SDS treatment and an inventive residual detergent removal step using divalent salt solutions, along with potential functionalization and cell seeding steps to tailor scaffold properties for biomedical applications.
Stated Advantages
The biomaterial is relatively low-cost and can be produced using efficient and time-condensed procedures.
It maintains its shape over long periods and is resistant to deformation post-implantation due to its non-resorbable nature.
The biomaterial exhibits high biocompatibility with minimal or almost non-existent immunogenic response.
It induces rapid vascularization (angiogenesis) promoting integration and tissue regeneration.
The scaffold has a minimal footprint, making it nearly invisible before and after cell invasion and angiogenesis.
Derived from plants or fungi, the biomaterial has a relatively low environmental impact and can be produced from food waste, offering sustainable and ethical sourcing.
Documented Applications
Use as implantable scaffolds for supporting animal cell growth and promoting tissue regeneration.
Use as scaffolds for promoting angiogenesis (blood vessel formation) within tissues.
Application as structural implants for tissue replacement surgeries including skin grafts, bone replacement and filling, spinal cord repair, heart, muscle, nerve, and blood vessel regeneration.
Use as structural implants for cosmetic surgery.
Use as vitreous humour replacement in hydrogel form.
Application as artificial bursae, with the scaffold forming sac-like structures containing scaffold hydrogel.
Use in repair or regeneration following spinal cord injury by implantation at the spinal cord to promote repair and functional recovery.
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