scaffold matrix
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BACKGROUND: A biological scaffold from extracellular matrix can be produced by a variety of decellularization methods whose caveat consists in efficiently eliminating cells from the treated tissue. This scaffold can be used in diverse applications for tissue engineering and organ regeneration. Preservation of the extracellular matrix ultrastructure is highly desirable because of its unique architecture, contained growth factors and decreased immunological response. All of these properties provide attachment sites and adequate environment for cells colonizing this scaffold, reconstituting the decellularized organ. This review briefly describes chemical decellularization methods, evaluation of these protocols and the role of ECM in tissue engineering. CONCLUSION: Chemical decellularization is an often used method for scaffold preparation and makes possible a well-preserved three dimensional structure of extracellular matrix.
The objective of our study was to compare the cellular and extracellular matrix (ECM) structure and the biomechanical properties of human pericardium (HP) with the normal human aortic heart valve (NAV). HP tissues (from 12 patients) and NAV samples (from 5 patients) were harvested during heart surgery. The main cells in HP were pericardial interstitial cells, which are fibroblast-like cells of mesenchymal origin similar to the valvular interstitial cells in NAV tissue. The ECM of HP had a statistically significantly (p < 0.001) higher collagen I content, a lower collagen III and elastin content, and a similar glycosaminoglycans (GAGs) content, in comparison with the NAV, as measured by ECM integrated density. However, the relative thickness of the main load-bearing structures of the two tissues, the dense part of fibrous HP (49 ± 2%) and the lamina fibrosa of NAV (47 ± 4%), was similar. In both tissues, the secant elastic modulus (Es) was significantly lower in the transversal direction (p < 0.05) than in the longitudinal direction. This proved that both tissues were anisotropic. No statistically significant differences in UTS (ultimate tensile strength) values and in calculated bending stiffness values in the longitudinal or transversal direction were found between HP and NAV. Our study confirms that HP has an advantageous ECM biopolymeric structure and has the biomechanical properties required for a tissue from which an autologous heart valve replacement may be constructed.
- MeSH
- aorta * MeSH
- biomechanika MeSH
- biopolymery chemie MeSH
- extracelulární matrix metabolismus MeSH
- lidé MeSH
- mechanické jevy * MeSH
- perikard cytologie MeSH
- pevnost v tahu MeSH
- srdeční chlopně cytologie MeSH
- testování materiálů MeSH
- tkáňové inženýrství * MeSH
- tkáňové podpůrné struktury chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Trypanosoma brucei has a complex life cycle during which its single mitochondrion is subjected to major metabolic and morphological changes. While the procyclic stage (PS) of the insect vector contains a large and reticulated mitochondrion, its counterpart in the bloodstream stage (BS) parasitizing mammals is highly reduced and seems to be devoid of most functions. We show here that key Fe-S cluster assembly proteins are still present and active in this organelle and that produced clusters are incorporated into overexpressed enzymes. Importantly, the cysteine desulfurase Nfs, equipped with the nuclear localization signal, was detected in the nucleolus of both T. brucei life stages. The scaffold protein Isu, an interacting partner of Nfs, was also found to have a dual localization in the mitochondrion and the nucleolus, while frataxin and both ferredoxins are confined to the mitochondrion. Moreover, upon depletion of Isu, cytosolic tRNA thiolation dropped in the PS but not BS parasites.
- MeSH
- aktivní transport - buněčné jádro MeSH
- buněčné jádro metabolismus MeSH
- ferredoxiny metabolismus MeSH
- jaderné lokalizační signály MeSH
- lyasy štěpící vazby C-S chemie genetika metabolismus MeSH
- mitochondriální proteiny metabolismus MeSH
- mitochondrie metabolismus MeSH
- molekulární sekvence - údaje MeSH
- multimerizace proteinu MeSH
- proteiny asociované s jadernou matrix chemie genetika metabolismus MeSH
- proteiny vázající železo metabolismus MeSH
- protozoální proteiny chemie genetika metabolismus MeSH
- sekvence aminokyselin MeSH
- Trypanosoma brucei brucei enzymologie genetika metabolismus MeSH
- vazba proteinů MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
ECM is composed of different collagenous and non-collagenous proteins. Collagen nanofibers play a dominant role in maintaining the biological and structural integrity of various tissues and organs, including bone, skin, tendon, blood vessels, and cartilage. Artificial collagen nanofibers are increasingly significant in numerous tissue engineering applications and seem to be ideal scaffolds for cell growth and proliferation. The modern tissue engineering task is to develop three-dimensional scaffolds of appropriate biological and biomechanical properties, at the same time mimicking the natural extracellular matrix and promoting tissue regeneration. Furthermore, it should be biodegradable, bioresorbable and non-inflammatory, should provide sufficient nutrient supply and have appropriate viscoelasticity and strength. Attributed to collagen features mentioned above, collagen fibers represent an obvious appropriate material for tissue engineering scaffolds. The aim of this minireview is, besides encapsulation of the basic biochemical and biophysical properties of collagen, to summarize the most promising modern methods and technologies for production of collagen nanofibers and scaffolds for artificial tissue development.
- MeSH
- cytoskelet chemie MeSH
- extracelulární matrix - proteiny chemie MeSH
- extracelulární matrix chemie MeSH
- kolagen chemie MeSH
- lidé MeSH
- mechanický stres MeSH
- nanostruktury * MeSH
- pružnost MeSH
- testování materiálů MeSH
- tkáňové inženýrství * MeSH
- tkáňové podpůrné struktury * MeSH
- viskozita MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Lately, the need for three-dimensional (3D) cell culture has been recognized in order to closely mimic the organization of native tissues. Thus, 3D scaffolds started to be employed to facilitate the 3D cell organization and enable the artificial tissue formation for the emerging tissue engineering applications. 3D scaffolds can be prepared by various techniques, each with certain advantages and disadvantages. Decellularization is an easy method based on removal of cells from native tissue sample, yielding extracellular matrix (ECM) scaffold with preserved architecture and bioactivity. This chapter provides a detailed protocol for decellularization of pig lung and also some basic assays for evaluation of its effectivity, such as determination of DNA content and histological verification of the selected ECM components. Such decellularized scaffold can subsequently be used for various tissue engineering applications, for example, for recellularization with cells of interest, for natural ECM hydrogel preparation, or as a bioink for 3D bioprinting.
- MeSH
- extracelulární matrix MeSH
- hydrogely MeSH
- plíce * MeSH
- prasata MeSH
- tkáňové inženýrství * metody MeSH
- tkáňové podpůrné struktury * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Seriously compromised function of some organs can only be restored by transplantation. Due to the shortage of human donors, the need to find another source of organs is of primary importance. Decellularized scaffolds of non-human origin are being studied as highly potential biomaterials for tissue engineering. Their biological nature and thus the ability to provide a naturally-derived environment for human cells to adhere and grow highlights their great advantage in comparison to synthetic scaffolds. Nevertheless, since every biomaterial implanted in the body generates immune reaction, studying the interaction of the scaffold with the surrounding tissues is necessary. This review aims to summarize current knowledge on the immunogenicity of semi-xenografts involved in transplantation. Moreover, positive aspects of the interaction between xenogeneic scaffold and human cells are discussed, focusing on specific roles of proteins associated with extracellular matrix in cell adhesion and signalling.
Introduction: The aim of this study was to assess the long-term impact and potential effectiveness of our specialized acellular dermal matrix (ADM) in a two-stage breast reconstruction process. Objective: Opinions regarding the use of ADMs are currently divided. While their positive contribution to reconstructive breast surgery is evident, the results of studies vary depending on specific procedures, patient selection, and techniques employed. Material and methods: In a retrospective study conducted between January 2015 and October 2023, it was examined a cohort of patients who underwent delayed two-stage breast reconstruction with the addition of ADM prepared by Central Tissue Bank (CTB) the Burn and Reconstructive Surgery Department University Hospital Ružinov. Our primary focus was on the occurrence of significant postoperative complications during both the initial and subsequent reconstruction periods, taking into account patients’ medical history, comorbidities, and adjuvant therapy. Results: We examined a total of 46 patients (49 breasts) who underwent two-stage breast reconstruction. The average age of the patients was 46 and the average BMI was 23.1. The average length of outpatient follow-up for female patients was 32 months. We observed a total of 4 cases of capsular contracture, ranging from grade I to grade III, with 2 cases requiring surgical revision through capsulotomy and implant exchange. Postoperative complications, such as infection and dehiscence leading to expander/implant loss, occurred in one case. The occurrence of seroma was noted in 3 cases. Complications were more frequently observed in the group of patients with post-radiation chest changes and comorbidities such as diabetes or hypertension, and in patients with a lower BMI than the group’s average (23.1). In the group of patients who were smokers, we did not observe an increased rate of complications, with the exception of wound dehiscence in cases where there was no expander exposure. Conclusion: In experienced hands, ADM prepared by CTB and used in delayed two-stage breast reconstruction, can be beneficial as an adjunct to prosthetic breast reconstruction while also reducing costs.
A number of implantable biomaterials derived from animal tissues are now used in modern surgery. Xe-Derma is a dry, sterile, acellular porcine dermis. It has a remarkable healing effect on burns and other wounds. Our hypothesis was that the natural biological structure of Xe-Derma plays an important role in keratinocyte proliferation and formation of epidermal architecture in vitro as well as in vivo. The bioactivity of Xe-Derma was studied by a cell culture assay. We analyzed growth and differentiation of human keratinocytes cultured in vitro on Xe-Derma, and we compared the results with formation of neoepidermis in the deep dermal wounds treated with Xe-Derma. Keratinocytes cultured on Xe-Derma submerged in the culture medium achieved confluence in 7-10 days. After lifting the cultures to the air-liquid interface, the keratinocytes were stratified and differentiated within one week, forming an epidermis with basal, spinous, granular, and stratum corneum layers. Immunohistochemical detection of high-molecular weight cytokeratins (HMW CKs), CD29, p63, and involucrin confirmed the similarity of organization and differentiation of the cultured epidermal cells to the normal epidermis. The results suggest that the firm natural structure of Xe-Derma stimulates proliferation and differentiation of human primary keratinocytes and by this way improves wound healing.
- MeSH
- extracelulární matrix metabolismus MeSH
- fibroblasty cytologie fyziologie MeSH
- hojení ran fyziologie MeSH
- keratinocyty cytologie fyziologie MeSH
- kultivované buňky MeSH
- lidé MeSH
- proliferace buněk MeSH
- řízená tkáňová regenerace přístrojové vybavení metody MeSH
- tkáňové inženýrství přístrojové vybavení metody MeSH
- tkáňové podpůrné struktury MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, N.I.H., Extramural MeSH
Východiska: Primární lidské B buňky chronické lymfocytární leukemie (CLL) podléhají při kultivaci in vitro buněčné smrti, nicméně jejich přežití lze signifikantně prodloužit kontaktem se stromálními buňkami nebo přítomností specifických solubilních faktorů. Pro účely výzkumu chování CLL buněk jsme vytvořili 3D in vitro model, ve kterém bylo simulováno vhodné mikroprostředí pro CLL buňky umožňující studium mechanizmu jejich přežívání v dlouhodobé kultivaci. Materiál a metody: Naším cílem bylo, aby struktura scaffoldu byla geometricky podobná 3D morfologii kostní dřeně, která vyplňuje trabekulární kost, aby měl 3D scaffold dostatečně velký povrch pro zachycení buněk a zároveň velkou pórovitost pro buněčnou migraci a transport živin. Dalším požadavkem byla také alespoň částečná transparentnost potřebná pro pozorování buněčného modelu pomocí optických metod. Připravili jsme 3D scaffoldy z porózního hydrogelu poly (2-hydroxyetyl metakrylát) (pHEMA), poly (2-hydroxyetyl metakrylát-co-2-aminoetyl metakrylát) p (HEMA-co-AEMA) a p (HEMA-co-AEMA) modifikovaný s často používaným adhezním peptidem Arg-Gly-Asp (RGD). Všechny hydrogelové scaffoldy byly vyrobeny ve čtyřech velikostech pórů (125, 200, 300 a 350–450 μm). Scaffoldy byly testovány pomocí HS-5 buněčné linie odvozené z lidských stromálních buněk kostní dřeně a HEK293 buněčné linie odvozené z lidských embryonálních buněk ledvin. Výsledky: Hydrogelový scaffold p (HEMA-co-AEMA) modifikovaný adhezním peptidem Arg-Gly-Asp (RGD) s velikostí pórů 350–450 μm prokázal, že je vhodným systémem pro 3D kultivace buněk, neboť podporuje interakce mezi buňkami navzájem a také mezi buňkami a materiálem. Tento scaffold byl použit pro nasazení kultivace složené z HS-5 buněk a CLL buněk, které byly stimulovány pomocí ligandu CD40 a cytokinu IL-4. Viabilita CLL buněk byla vyšší v přítomnosti obou stimulátorů zároveň než v případě každého zvlášť. Závěr: Ukázali jsme, že technologie 3D scaffoldů je velmi dobře využitelná pro modelování mikrosystémů, kde se nádorové buňky chovají jako ve svém přirozeném mikroprostředí. Klíčová slova: hematoonkologie – leukemie – hydrogel – stromální buňky
Background: Primary human B cells chronic lymphocytic leukemia undergoes apoptosis, from which they can be rescued by contact with stromal cells or by the addition of specific soluble factor, when cultured in vitro. For research purposes of the behavior of CLL cells we created 3D in vitro model in which we simulated appropriate microenvironment for CLL cells to allow study the mechanism of survival of these cells in long-term cultivation. Material and Methods: Our aim was the scaffold structure to be geometrically similar to the 3D morphology of supporting bone marrow tissue in a trabecular bone; the 3D scaffold was also designed to conform to biocompatibility, sufficiently large surface area for cell attachment, high porosity for cell migration, proliferation and transport of nutrients. Another requirement was a partial transparency for inspection of cell model with optical techniques. We prepared 3D scaffolds from porous hydrogel poly (2-hydroxyethyl methacrylate) (pHEMA), poly (2-hydroxyethyl methacrylate-co-2-aminoethyl methacrylate) p (HEMA-co-AEMA) and p (HEMA-co-AEMA) modified with frequently used cell adhesion peptide Arg-Gly-Asp (RGD). All hydrogel scaffolds were manufactured in four pore diameters (125, 200, 300 and 350–450 μm). Scaffolds were tested with human bone marrow stromal cell line HS-5 and human embryonic kidney cell line HEK293. Results: Hydrogel scaffold p (HEMA-co-AEMA) modified with adhesion peptide Arg-Gly-Asp (RGD) with pore diameter of 350–450 μm demonstrated that it is a convenient system for 3D cell cultivation, since it promotes interaction between the cells and also between the cells and the material. This scaffold was used for seeding of co-cultivation system of HS-5 cells with CLL-cells, which were stimulated through the CD40L signaling pathway as well as via the IL-4 pathway. Viability of B-CLL cells was higher in the presence of both stimulators than with each alone. Conclusions: We have shown that 3D scaffold technology is very useful for modeling of microsystems where the cancer cells behave like in their natural microenvironment. Key words: hematooncology – leukemia – hydrogel – stromal cells This work was supported by grant COST CZ LD15144 “Cellular and acellular grounds for regeneration of bones and teeth” awarded by the Ministry of Education, Youth and Sport of the Czech Republic. The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 6. 3. 2017 Accepted: 26. 3. 2017
- MeSH
- biokompatibilní materiály MeSH
- biologické modely MeSH
- chronická lymfatická leukemie patologie MeSH
- hydrogely * MeSH
- kultivační techniky metody MeSH
- mezenchymální kmenové buňky MeSH
- nádorové buňky kultivované * MeSH
- nádorové mikroprostředí MeSH
- techniky in vitro MeSH
- tkáňové podpůrné struktury * MeSH
- Publikační typ
- práce podpořená grantem MeSH
In this work, we report on the preparation of a novel biodegradable textile scaffold made of palmitoyl-hyaluronan (palHA). Monofilament fibres of palHA with a diameter of 120μm were prepared by wet spinning. The wet-spun fibres were subsequently processed into a warp-knitted textile. To find a compromise between swelling in water and degradability of the final textile scaffold, a series of palHA derivatives with different degrees of substitution of the palmitoyl chain was synthesized. Freeze-drying not only provided shape fixation, but also speeded up scaffold degradation in vitro. Fibronectin, fibrinogen, laminin and collagen IV were physically adsorbed on the textile surface to enhance cell adhesion on the material. The highest amount of adsorbed cell-adhesive proteins was achieved with fibronectin (89%), followed by fibrinogen (81%). Finally, textiles modified with fibronectin or fibrinogen both supported the adhesion and proliferation of normal human fibroblasts in vitro, proving to be a useful cellular scaffold for tissue engineering.
- MeSH
- biokompatibilní materiály chemie metabolismus farmakologie MeSH
- buněčná adheze účinky léků MeSH
- fibroblasty cytologie účinky léků MeSH
- hydrofobní a hydrofilní interakce * MeSH
- kyselina hyaluronová chemie metabolismus farmakologie MeSH
- lidé MeSH
- povrchové vlastnosti MeSH
- textilie * MeSH
- tkáňové inženýrství MeSH
- tkáňové podpůrné struktury chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
