Hypothermic and cryogenic preservation of tissue‐engineered human bone

Tam, Edmund; McGrath, Madison; Sladkova, Martina; AlManaie, Athbah; Alostaad, Anaam; Peppo, Giuseppe Maria de

doi:10.1111/nyas.14264

Cited by 11 publications

(9 citation statements)

References 41 publications

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“…Recently, Tam et al compared the effects of two potential preservation methods on the survival, quality and function of human tissue-engineered bone grafts from iPSCs. They found that storage at − 80 °C resulted in cell death and structural alteration of the extracellular matrix, whilst hypothermic storage at 4 °C did not significantly affect tissue viability and integrity [ 171 ].…”

Section: Current Applications Of Cryopreservationmentioning

confidence: 99%

Winter is coming: the future of cryopreservation

et al. 2021

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The preservative effects of low temperature on biological materials have been long recognised, and cryopreservation is now widely used in biomedicine, including in organ transplantation, regenerative medicine and drug discovery. The lack of organs for transplantation constitutes a major medical challenge, stemming largely from the inability to preserve donated organs until a suitable recipient is found. Here, we review the latest cryopreservation methods and applications. We describe the main challenges—scaling up to large volumes and complex tissues, preventing ice formation and mitigating cryoprotectant toxicity—discuss advantages and disadvantages of current methods and outline prospects for the future of the field.

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Section: Current Applications Of Cryopreservationmentioning

confidence: 99%

Winter is coming: the future of cryopreservation

et al. 2021

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“…Tissue engineered bone maintained viability and extracellular matrix integrity following refrigeration but not freezing. 11 Synthetic epidermis remained viable for up to 9 days of refrigeration. 8 Articular cartilage maintained viability, glycosaminoglycan content and collagen content for 28 days at both room temperature and 4℃ (at least partly due to the low metabolic demands of chondrocytes native to avascular tissue beds in vivo).…”

Section: Discussionmentioning

confidence: 98%

“…These include hypothermic and cryogenic storage of tissue engineered bone (48 h at 4 or −80℃), synthetic human epidermis (up to 13 days at 4℃), articular cartilage (up to 56 days at room temperature, 4 or 37℃), human adipose stem cell sheets (3 or 7 days at 4℃), and aligned SC columns in a collagen matrix (2 days at 4℃ or 1 day at −80℃ followed by 2 days in liquid nitrogen). 6,8,11,33,34 These studies exclusively employ in vitro assays to evaluate viability and potency. Tissue engineered bone maintained viability and extracellular matrix integrity following refrigeration but not freezing.…”

Section: Discussionmentioning

confidence: 99%

“…2 Among these, hypothermic storage has been most widely adopted for clinical use, as most protocols do not require the use of toxic cryoprotective agents or specialized equipment beyond standard refrigerators. To date, hypothermic preservation strategies have been implemented to extend shelf lives of cells, tissues and organs such as red blood cells, 3 hepatocytes, 4 mesenchymal stromal cells, 5 adipose stem cells, 6 cornea, 7 native and engineered skin, 8,9 nerve allografts, 10 engineered bone, 11 and solid organs. 12 Hypothermic storage exploits the temperature dependence of nearly all biochemical reaction rates to effectively slow biological time.…”

Section: Introductionmentioning

confidence: 99%

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Biopreservation of living tissue engineered nerve grafts

et al. 2021

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Tissue engineered nerve grafts (TENGs) built from living neurons and aligned axon tracts offer a revolutionary new approach as “living scaffolds” to bridge major peripheral nerve defects. Clinical application, however, necessitates sufficient shelf-life to allow for shipping from manufacturing facility to clinic as well as storage until use. Here, hypothermic storage in commercially available hibernation media is explored as a potential biopreservation strategy for TENGs. After up to 28 days of refrigeration at 4℃, TENGs maintain viability and structure in vitro. Following transplantation into 1 cm rat sciatic defects, biopreserved TENGs routinely survive and persist for at least 2 weeks and recapitulate pro-regenerative mechanisms of fresh TENGs, including the ability to recruit regenerating host tissue into the graft and extend neurites beyond the margins of the graft. The protocols and timelines established here serve as important foundational work for the manufacturing, storage, and translation of other neuron-based tissue engineered therapeutics.

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“…The presence of collagen fibers in the matrix and deposition of bone glycoproteins were studied on demineralized samples following pullout testing. Samples were washed in DPBS, fixed in 4% (v/v) PFA in PBS at 4 °C for 24 h, and then decalcified in Immunocal (Decal Chemical Corp.) for 48 h at RT as previously reported 66 . After decalcification, the samples were dehydrated through graded concentrations of ethanol prior to paraffin embedding.…”

Section: Scaffoldsmentioning

confidence: 99%

A biomimetic engineered bone platform for advanced testing of prosthetic implants

Sladkova‐Faure

Pujari-Palmer

Öhman‐Mägi

et al. 2020

Sci Rep

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Existing methods for testing prosthetic implants suffer from critical limitations, creating an urgent need for new strategies that facilitate research and development of implants with enhanced osseointegration potential. Herein, we describe a novel, biomimetic, human bone platform for advanced testing of implants in vitro, and demonstrate the scientific validity and predictive value of this approach using an assortment of complementary evaluation methods. We anchored titanium (Ti) and stainless steel (SS) implants into biomimetic scaffolds, seeded with human induced mesenchymal stem cells, to recapitulate the osseointegration process in vitro. We show distinct patterns of gene expression, matrix deposition, and mineralization in response to the two materials, with Ti implants ultimately resulting in stronger integration strength, as seen in other preclinical and clinical studies. Interestingly, RNAseq analysis reveals that the TGF-beta and the FGF2 pathways are overexpressed in response to Ti implants, while the Wnt, BMP, and IGF pathways are overexpressed in response to SS implants. High-resolution imaging shows significantly increased tissue mineralization and calcium deposition at the tissue-implant interface in response to Ti implants, contributing to a twofold increase in pullout strength compared to SS implants. Our technology creates unprecedented research opportunities towards the design of implants and biomaterials that can be personalized, and exhibit enhanced osseointegration potential, with reduced need for animal testing.

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Hypothermic and cryogenic preservation of tissue‐engineered human bone

Cited by 11 publications

References 41 publications

Winter is coming: the future of cryopreservation

Winter is coming: the future of cryopreservation

Biopreservation of living tissue engineered nerve grafts

A biomimetic engineered bone platform for advanced testing of prosthetic implants

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