Functional Imaging of Matrix Structure of Cryopreserved Engineered Tissues Using Back-Directional Gated Mesoscopic Imaging

Author(s):  
Young L. Kim ◽  
Zhengbin Xu ◽  
Altug Ozcelikkale ◽  
Bumsoo Han

Successful cryopreservation of engineered tissues (ETs) can greatly advance the access and availability of cell/tissue engineering products for clinical use. One of the key challenges in cryopreserving ETs is that the functionality of ETs should be maintained throughout the preservation process. Many of the functionalities are associated with the extracellular matrix (ECM) microstructure, which in turn can be a crucial marker for the post-thaw functionality. Recent studies also reported that the ECM microstructure can be affected by freezing-induced cell-fluid-matrix interactions.1–3 Thus, it is critical to assess three-dimensional (3-D) matrix structure of cryopreserved ETs in a non-destructive, non-invasive, and rapid manner.

Author(s):  
Ka Yaw Teo ◽  
J. Craig Dutton ◽  
Frederick Grinnell ◽  
Bumsoo Han

Long-term cryopreservation of functional engineered tissues (ETs) is a key enabling technology for tissue engineering and regenerative medicine. However, a limited understanding of tissue-level biophysical phenomena during freeze/thaw (F/T) and their effects on cells and ECM microstructure poses significant challenges for i) preserving tissue functionality, and ii) controlling highly tissue-type dependent cryopreservation outcomes.


2021 ◽  
Vol 22 (17) ◽  
pp. 9367
Author(s):  
Yeonggwon Jo ◽  
Seung Hyeon Hwang ◽  
Jinah Jang

Tissues and organs are not composed of solely cellular components; instead, they converge with an extracellular matrix (ECM). The composition and function of the ECM differ depending on tissue types. The ECM provides a microenvironment that is essential for cellular functionality and regulation. However, during aging, the ECM undergoes significant changes along with the cellular components. The ECM constituents are over- or down-expressed, degraded, and deformed in senescence cells. ECM aging contributes to tissue dysfunction and failure of stem cell maintenance. Aging is the primary risk factor for prevalent diseases, and ECM aging is directly or indirectly correlated to it. Hence, rejuvenation strategies are necessitated to treat various age-associated symptoms. Recent rejuvenation strategies focus on the ECM as the basic biomaterial for regenerative therapies, such as tissue engineering. Modified and decellularized ECMs can be used to substitute aged ECMs and cell niches for culturing engineered tissues. Various tissue engineering approaches, including three-dimensional bioprinting, enable cell delivery and the fabrication of transplantable engineered tissues by employing ECM-based biomaterials.


2021 ◽  
Author(s):  
Mattia Saggioro ◽  
Stefania D'Agostino ◽  
Anna Gallo ◽  
Sara Crotti ◽  
Sara D'Aronco ◽  
...  

Three-dimensional (3D) culture systems are progressively getting attention given their potential in overcoming limitations of the classical 2D in vitro systems. Among different supports for 3D cell culture, hydrogels (HGs)...


Polymers ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 1444 ◽  
Author(s):  
Sun Hee Cho ◽  
Jeong In Kim ◽  
Cheol Sang Kim ◽  
Chan Hee Park ◽  
In Gi Kim

To date, many researchers have studied a considerable number of three-dimensional (3D) cotton-like electrospun scaffolds for tissue engineering, including the generation of bone, cartilage, and skin tissue. Although numerous 3D electrospun fibrous matrixes have been successfully developed, additional research is needed to produce 3D patterned and sophisticated structures. The development of 3D fibrous matrixes with patterned and sophisticated structures (FM-PSS) capable of mimicking the extracellular matrix (ECM) is important for advancing tissue engineering. Because modulating nano to microscale features of the 3D fibrous scaffold to control the ambient microenvironment of target tissue cells can play a pivotal role in inducing tissue morphogenesis after transplantation in a living system. To achieve this objective, the 3D FM-PSSs were successfully generated by the electrospinning using a directional change of the sharply inclined array collector. The 3D FM-PSSs overcome the current limitations of conventional electrospun cotton-type 3D matrixes of random fibers.


Materials ◽  
2020 ◽  
Vol 13 (16) ◽  
pp. 3522
Author(s):  
Su Jeong Lee ◽  
Jun Hee Lee ◽  
Jisun Park ◽  
Wan Doo Kim ◽  
Su A Park

Recently, many research groups have investigated three-dimensional (3D) bioprinting techniques for tissue engineering and regenerative medicine. The bio-ink used in 3D bioprinting is typically a combination of synthetic and natural materials. In this study, we prepared bio-ink containing porcine skin powder (PSP) to determine rheological properties, biocompatibility, and extracellular matrix (ECM) formation in cells in PSP-ink after 3D printing. PSP was extracted without cells by mechanical, enzymatic, and chemical treatments of porcine dermis tissue. Our developed PSP-containing bio-ink showed enhanced printability and biocompatibility. To identify whether the bio-ink was printable, the viscosity of bio-ink and alginate hydrogel was analyzed with different concentration of PSP. As the PSP concentration increased, viscosity also increased. To assess the biocompatibility of the PSP-containing bio-ink, cells mixed with bio-ink printed structures were measured using a live/dead assay and WST-1 assay. Nearly no dead cells were observed in the structure containing 10 mg/mL PSP-ink, indicating that the amounts of PSP-ink used were nontoxic. In conclusion, the proposed skin dermis decellularized bio-ink is a candidate for 3D bioprinting.


Author(s):  
Soham Ghosh ◽  
J. Craig Dutton ◽  
Bumsoo Han

Freezing of biomaterials is emerging as one of the key biotechnologies in cell/tissue engineering, medicine and biology. Its applications include — 1) preservation of cell/tissue engineering products, 2) quality control of biospecimens cryopreserved in tissue banks and repositories, and 3) synthesis procedures of biomaterials such as decellularization of native tissues to create acellular (i.e., cell-free) complex three-dimensional extracellular matrices (ECMs). Traditionally, research efforts have focused on determining optimal freeze/thaw (F/T) protocols with chemical additives, so called cryoprotective agents, for a given cell/tissue-type by comparing the outcomes of F/T protocols, which are mainly gauged by cell viability. Although cell viability is the major constituent, it has recently been recognized that other features beyond viability are also critical to the functionality of biomaterials, including the microstructure of the ECM, the status of cell-matrix adhesion, and the cytoskeletal structure and organization [1, 2, 3].


2019 ◽  
Vol 13 (1) ◽  
Author(s):  
Sonia Fathi Karkan ◽  
Soodabeh Davaran ◽  
Reza Rahbarghazi ◽  
Roya Salehi ◽  
Abolfazl Akbarzadeh

Abstract Attention has recently increased in the application of electrospun fibers because of their putative capability to create nanoscale platforms toward tissue engineering. To some extent, electrospun fibers are applicable to the extracellular matrix by providing a three-dimensional microenvironment in which cells could easily acquire definite functional shape and maintain the cell-to-cell connection. It is noteworthy to declare that placement in different electrospun substrates with appropriate physicochemical properties enables cells to promote their bioactivities, dynamics growth and differentiation, leading to suitable restorative effects. This review paper aims to highlight the application of biomaterials in engineered vascular grafts by using electrospun nanofibers to promote angiogenesis and neovascularization


Author(s):  
Angela Seawright ◽  
Altug Ozcelikkale ◽  
J. Craig Dutton ◽  
Bumsoo Han

Cryopreservation can provide long-term storage of various biological tissues, which has significant impact on tissue engineering and regenerative medicine. For successful cryopreservation of tissues, tissue functionality must be maintained including physical properties such as mechanical, optical, and transport properties, as well as cellular viability. Such properties are associated with the extracellular matrix (ECM) microstructure. Thus, the preservation of the ECM microstructure may lead to successful cryopreservation [1,2]. Yet, there is still very little known about changes in the ECM microstructure during freezing/thawing.


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