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Advanced Cell Therapy for Asherman's Syndrome
Published in Carlos Simón, Carmen Rubio, Handbook of Genetic Diagnostic Technologies in Reproductive Medicine, 2022
Jordi Ventura, Xavier Santamaria
Another interesting approach in tissue engineering is the use of decellularized scaffolds with high biocompatibility and low rejection effects compared with organ transplantation (80). The use of decellularized uterus and posterior regeneration by autologous cells has already been successfully used for uterus regeneration in different rat (81–83) and pig models (84). However, this approach has its limitations because of the limited availability of uterus from human donors for decellularization and the presence of some cell residues after decellularization (85).
Stem Cells and Nanotechnology
Published in Stavros Hatzopoulos, Andrea Ciorba, Mark Krumm, Advances in Audiology and Hearing Science, 2020
The first studies by tissue engineering on design, fabrication, and characterization of scaffolds for ossiculoplasty concerned a partial ossicular replacement prosthesis (PORP). This prosthesis was cultured in vitro with human MSC and osteoinductive factors (Danti et al., 2009, 2010; D’Alessandro et al., 2012). This 3D porous scaffold of the PORP, made of poly(propylene fumarate)/poly(propylene fumarate)-diacrylate (PPF/PPF-DA), a rigid polymer biodegradable in the long term, was designed by a photo-crosslinking particulate-leaching technique with pore size and porosity suitable to support human MSC differentiation into osteoblasts. The PORP scaffolds were cultured with osteoinduced human MSC to generate in vitro bone extracellular matrix (ECM) within the scaffold porosity. After 12 days, an early bone matrix was detected, consisting of collagen type I fibers and calcium phosphate nodules (Danti et al., 2010). After decellularization, these scaffolds were used for a short term in vitro cultures of undifferentiated human MSC. The results of this process showed cellular viability, distribution, and quality of extracellular collagen type I and high mineralization (Danti et al., 2009; D’Alessandro et al., 2012). These concepts are summarized in Figure 9.2.
Biological reactions to reconstructive materials
Published in Steven J. Kronowitz, John R. Benson, Maurizio B. Nava, Oncoplastic and Reconstructive Management of the Breast, 2020
Steven J. Kronowitz, John R. Benson, Maurizio B. Nava
The decellularization process involves a balance between the competing aims of complete removal of antigenic cellular material (nucleic acids, lipids, and certain cellular proteins) and preserving the structure of ECM (collagen, elastin, GAGs, and growth factors), such that it provides an optimal substrate for regeneration of functional host tissues. In contrast to thinner tissues such as intestinal submucosa, the thickness and complexity of dermis requires more intensive mechanical and chemical methods to achieve adequate decellularization.8 These harsher processing methods have been shown to result in variable degradation of extracellular matrix structure as well as reduction in GAG and growth factor content. However, failure of a decellularization method to remove major histocompatibility complex (MHC) antigens to below a critical level will result in a failure of matrix remodeling and incorporation.7 A variety of decellularization methods have been described, including mechanical, chemical, enzymatic, and detergent-based protocols, and often a combination of these.1 The optimal decellularization protocol depends on tissue type, thickness, shape, cell density, and matrix density and thus will vary by tissue of origin and reconstructive purposes.1 While decellarization protocols of particular commercially available ADMs are proprietary, they presumably involve some combination of the above-reported methods.
Use of Decellularized SMILE (Small-Incision Lenticule Extraction) Lenticules for Engineering the Corneal Endothelial Layer: A Proof-of-Concept
Published in Current Eye Research, 2023
Swatilekha Hazra, Jacquelyn Akepogu, Supriya Krishna, SriRavali Pulipaka, Bhupesh Bagga, Charanya Ramachandran
Any tissue that is used for engineering must have limited residual native DNA content to reduce the chances of rejection following transplantation. Several methods have been employed to decellularize the cornea. Since complete decellularization is difficult to achieve without compromising the tissue integrity, Crapo et al.30 came up with the following recommendation to consider a tissue “sufficiently decellularized” and safe for transplantation: (a) the residual cellular components especially the DNA should be less than 50 ng/mg of the tissue, (b) DNA fragments should be <200 bp, and (c) absence of visible nuclear material within the ECM confirmed using DAPI or H&E staining. Decellularization of SMILE lenticules with serum and Opti-MEM, as shown in this paper, comes very close to meeting the aforementioned criteria. The residual DNA was reduced to ∼4 and 6.3% of the fresh lenticule when treated with Opti-MEM and serum, respectively. This is very similar to the values reported by Yam et al.,18 with the use of 0.1% SDS for decellularization with extensive washing and agitation.
Surgical Models to Explore Acellular Liver Scaffold Transplantation: Step-by-Step
Published in Organogenesis, 2020
Marlon L. Dias, Cíntia M. P. Batista, Victor J. K. Secomandi, Alexandre C. Silva, Victoria R. S. Monteiro, Lanuza A. Faccioli, Regina C. S. Goldenberg
Chronic liver diseases affect more than 500 million people worldwide.1 Currently, transplantation is the only treatment available for liver failure. However, problems such as organ availability, graft viability and immune rejection create a long waiting list. Tissue engineering appears as a promising alternative with the production of acellular organs and tissues from the extracellular matrix (ECM) of potential use in Regenerative Medicine. ECM is a complex and dynamic environment characterized by biophysical, mechanical and biochemical properties specific to each organ. Acellular scaffolds can be produced by decellularization techniques.2 The decellularization process removes all cell content of an organ or tissue, leaving only the components of the extracellular matrix specific to the organ or tissue such as collagen, fibronectin, laminin and others.3 The technique of decellularization has already been used for several organs such as heart, lung, kidney, placenta and liver, as well as tissues such as skin, intestinal mucosa, heart valve, among others.4-11 Some studies have shown that these acellular organs can be transplanted in animals.12, 13 Unfortunately, regarding acellular liver transplantation several aspects have not yet been reported, such as endogenous ECM potency to cell recruitment in vivo, acellular liver graft long-term function and contribution to recipient rat liver functions post-transplantation.14
Development of Bioengineered Organ Using Biological Acellular Rat Liver Scaffold and Hepatocytes
Published in Organogenesis, 2020
Tanya Debnath, Chandra Shekar Mallarpu, Lakshmi Kiran Chelluri
There are a variety of approaches such as chemical, enzymatic, and physical methods for effective decellularization of whole organs.9 However, other bioactive ligands such as integrins, cadherins, are also important for cell–cell interaction, cell–ECM interaction, and new ECM formation.10 The tissue ultra-structure, matrix density, and architecture vary among different types of organs and hence play a major role in the utilization of decellularization techniques. In case of a complex and dense organ such as liver, decellularization is usually achieved through the use of the hepatic vascular network and dynamic culture systems which can support the delicate vasculature of the organ during processing.11 Completely decellularized organs or tissues can eradicate antagonistic immune response elicited by cell membrane epitopes, allogeneic or xenogeneic DNA, and damage-associated molecular pattern molecules.12