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Decellularized Biomaterial from Non-Mammalian Tissue

a biomaterial and non-mammalian technology, applied in the direction of drugs, peptide/protein ingredients, prosthesis, etc., can solve the problems of life-threatening, the use of conventional tissue scaffold products is not without drawbacks, and the biomechanical function of damaged tissue is restored. a true challenge, the effect of preserving structural and functional integrity

Inactive Publication Date: 2019-08-22
ISE PROFESSIONAL TESTING & CONSULTING SERVICES INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes a process for creating a biological scaffold biomaterial from urodele tissue, which can be used for wound healing and tissue regeneration purposes. The process involves decellularizing the tissue sample to remove cellular components and reduce antigenicity. The resulting biomaterial has a structure and function that promotes skin regeneration and reduces inflammation. The invention also provides a method for preparing the biomaterial and a tissue graft that can be used for wound healing. The technical effects of this patent are an improved understanding of the immunologically privileged state of neonates and the use of biological scaffolds to promote skin regeneration and reduce inflammation during wound healing.

Problems solved by technology

Tissue engineering efforts are ongoing to produce methods and materials for replacing biological functions, typically repairing or replacing whole tissues or portions thereof In this regard, wound treatment and skin repair are areas of predominant focus, as the loss of skin integrity due to illness or injury can lead to chronic, life threatening complications.
The ability to restore biomechanical function of damaged tissue presents a true challenge.
The use of conventional tissue scaffold products is not without drawbacks, however.
Tissue harvesting from human donors can produce undesirable consequences such as donor site morbidity or infection associated with removal of skin for donation.
Disease transmission risk and intersample variation are additional drawbacks associated with biological scaffold products.
In addition, it may be difficult to obtain sufficient tissue components necessary to cover large areas of damaged tissue.
Furthermore, conventional biological and synthetic materials can be costly, not effective in many instances, and limited in availability.

Method used

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  • Decellularized Biomaterial from Non-Mammalian Tissue
  • Decellularized Biomaterial from Non-Mammalian Tissue
  • Decellularized Biomaterial from Non-Mammalian Tissue

Examples

Experimental program
Comparison scheme
Effect test

example 1

Processing Axolotl Dermis

[0098]Axolotl dermis samples can be decellularized by preparing excised samples from healthy or healing axolotl dermal tissue and then subjecting the samples to hypo / hyperosmotic soaks for cell lysis, solvent dehydration, and oven drying. Specific processing of these grafts includes storage in 15-26% NaCl, multiple hypo / hyperosmotic soaks (utilizing NaCl solutions and water), and then solvent dehydration using ethanol, and then evaporation of the solvent either with air drying or oven drying at 37° C.

[0099]Histological examination of native axolotl dermal tissue was performed to identify the presence of the notable ECM elements, such as the basement membrane. See FIG. 1 and FIG. 4. Comparative histological and immunohistochemical analysis of native axolotl dermal tissue and human amniotic membrane was performed to compare the ECM structure and constituents, and to assess relative concentration and distribution of critical constituents. See FIG. 2, FIG. 3, an...

example 2

Splitting and Lamination of Acellular Dehydrated Axolotl Dermis

[0100]Decellularized dehydrated axolotl dermis can be split, via a mechanical splitter, to isolate heterogeneous matrix into homogenous sections. Isolated sections of desired thickness then can be rehydrated and lyophilized to obtain multilayered laminate structures of desired orientation with facial surface features. More specifically, dual-sided basement membrane structure, with interior open porous matrix obtained from the reticular dermis region of the dermal matrix, can be constructed to obtain desired facial surface properties. Alternatively, isolated native section can be used in native form for desired clinical outcome. For example, open porous homogenous matrix of the reticular dermis can be used to obtain augmentation of soft tissue structures.

[0101]A laminated custom construct with sulfated gags on both facial surface and collagen IV and laminin could be obtained for desirable dual-surface, anti-adhesion and a...

example 3

Preparation of Solubilized Acellular Dehydrated Axolotl Dermis, Pericardium, Fascia Lata, Periosteum, Peritoneum, or Dura Mater

[0102]Decellularized dehydrated axolotl native or isolated section of acellular urodele connectivue tissye matrix can be prepared by sectioning decellularized soft tissue structures into 1 cm2 sections and homogenizing the sections in a Warring blender (˜100 grams of tissue) in aqueous 1M glacier acetic acid for 30-60 seconds. Preparation of sponge can be obtained by the addition of varying volumes of water followed then neutralization and lypoholization of the slurry in a mold of desired geometric shape. The resultant porosity will correlate to the volume of water added to the matrix. Additionally, a selected range of bioactive extracts can be added to the slurry prior to neutralization, including particulated or small protein constituents extracted from digested human or urodele mineralized and nonmineralized connective tissues, such as demineralized bone ...

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Abstract

The growth factor profile, connective tissue matrix constituents, and immunoprivileged status of urodele extracellular matrix (ECM) and accompanying cutaneous tissue, plus the presence of antimicrobial peptides there, render urodele-derived tissue an ideal source for biological scaffolds for xenotransplantation. In particular, a biological scaffold biomaterial can be obtained by a process that entails (A) obtaining a tissue sample from a urodele, where the tissue comprises ECM, inclusive of the basement membrane, and (B) subjecting the tissue sample to a decellularization process that maintains the structural and functional integrity of the extracellular matrix, by virtue of retaining its fibrous and non-fibrous proteins, glycoaminoglycans (GAGs) and proteoglycans, while removing sufficient cellular components of the sample to reduce or eliminate antigenicity and immunogenicity for xenograft purposes. The resultant urodele-derived biomaterial can be used to enhance restoration of skin homeostasis, to reduce the severity, duration and associated damage caused by post-surgical inflammation, and to promote progression of natural healing and regeneration processes. In addition, the biomaterial promotes the formation of remodeled tissue that is comparable in quality, function, and compliance to undamaged human tissue.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims the benefit under 35 USC § 119(e) to U.S. provisional application 61 / 750,555, filed Jan. 9, 2013, the entire contents of which are incorporated herein by reference in their entirety.BACKGROUND OF THE INVENTION[0002]Tissue engineering efforts are ongoing to produce methods and materials for replacing biological functions, typically repairing or replacing whole tissues or portions thereof In this regard, wound treatment and skin repair are areas of predominant focus, as the loss of skin integrity due to illness or injury can lead to chronic, life threatening complications.[0003]Wound healing involves complex interactions between cells, growth factors, and extracellular matrix (ECM) components to reconstitute tissue following injury. The wound healing process in adult mammalian tissue has been well characterized and can be broken down into three stages—inflammation, proliferation, and remodeling.[0004]Typically, in re...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61L27/36A61L27/38A61L27/52C12N5/071A61K9/70A61L27/60A61L27/54A61K35/65A61L27/58A61K35/12A61K38/17
CPCA61L27/3687A61L27/3604A61L27/3633A61L27/38A61L27/52C12N5/0625A61K9/70A61L27/60A61L27/54A61L27/362A61K35/65A61L27/3625A61L27/58A61K35/12A61K38/1703A61L27/3691A61L2300/604A61L2300/412C12N2533/92C12N2533/90A61L2430/40A61L2300/606A61L2300/252A61L2300/64A61L2430/34A61P17/02A61P41/00A61P43/00
Inventor EARLY, RYANNE
Owner ISE PROFESSIONAL TESTING & CONSULTING SERVICES INC
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