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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Biochemistry".

Deadline for manuscript submissions: closed (31 December 2020) | Viewed by 34512

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Prof. Dr. Marek J. Łos

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Guest Editor

Centre of Biotechnology, Silesian University of Technology, 44-100 Gliwice, Poland
Interests: cancer immunology; cell death execution pathways; cancer stem cell biology; epigenetic reprogramming (production of iPS-cells); transdifferentiation technologies; other aspects of regenerative pharmacology
Special Issues, Collections and Topics in MDPI journals

Dr. Artur Cieślar-Pobuda

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Stem Cell Group, Nordic EMBL Partnership Center for Molecular Medicine Norway, University of Oslo, P.O. box 1137, Blindern, 0318 Oslo, Norway

Special Issue Information

Dear Colleagues,

The 21^st century is witnessing a rapid progress in biotechnology that will likely revolutionize medical interventions not only in the area of transplantology but also in that of drug delivery, with the identification of the most suitable cell subpopulations as primary therapeutic targets in oncology and in tissue regeneration. For example, in 2006 Yamanaka and colleagues developed a technology to produce isogenic (embryonic-like) induced pluripotent stem cells (iPSCs) from somatic cells by the process of reprogramming. iPSCs could then be differentiated into virtually any cell type in the body. In the meantime, an alternative technology of customized cell production from somatic cells through the process of transdifferentiation has progressed. Furthermore, our understanding of the biology of cancer allows for pinpointing cell populations (cancer stem cells, cancer initiating cells, cancer propagating cells) that are responsible for therapeutic failure. On the other hand, in the material science area, new composite materials (biodegradable and non-biodegradable) are being developed that may replace specific components of the connective tissue in different specialized organs. The proposed Special Issue aims to present diverse advances in biotechnology (i) at the level of cell and tissue engineering, (ii) at the level of advanced cancer therapy development, and (iii) at the level of biomaterial and general biotechnology science. Hence, it aims to bridge the gap between the above-mentioned research areas, while presenting specific aspects in an understandable manner to researchers from distinct research fields.

Prof. Dr. Marek J. Łos
Dr. Artur Cieślar-Pobuda
Guest Editors

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Keywords

stem cells
biomaterials
tissue engineering
reprogramming
transdifferentiation
electrospinning
3D printing

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Published Papers (8 papers)

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Research

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19 pages, 3028 KiB

Open AccessArticle

Comparison of Physicochemical, Mechanical, and (Micro-)Biological Properties of Sintered Scaffolds Based on Natural- and Synthetic Hydroxyapatite Supplemented with Selected Dopants

by Andrzej Hudecki, Dorota Łyko-Morawska, Anna Kasprzycka, Alicja Kazek-Kęsik, Wirginia Likus, Jolanta Hybiak, Kornelia Jankowska, Aleksandra Kolano-Burian, Patryk Włodarczyk, Weronika Wolany, Jarosław Markowski, Wojciech Maziarz, Iwona Niedzielska, Wojciech Pakieła, Mariusz Nowak and Marek J. Łos

Int. J. Mol. Sci. 2022, 23(9), 4692; https://doi.org/10.3390/ijms23094692 - 23 Apr 2022

Cited by 4 | Viewed by 2447

Abstract

The specific combinations of materials and dopants presented in this work have not been previously described. The main goal of the presented work was to prepare and compare the different properties of newly developed composite materials manufactured by sintering. The synthetic- (SHAP) or natural- (NHAP) hydroxyapatite serves as a matrix and was doped with: (i) organic: multiwalled carbon nanotubes (MWCNT), fullerenes C60, (ii) inorganic: Cu nanowires. Research undertaken was aimed at seeking novel candidates for bone replacement biomaterials based on hydroxyapatite—the main inorganic component of bone, because bone reconstructive surgery is currently mostly carried out with the use of autografts; titanium or other non-hydroxyapatite -based materials. The physicomechanical properties of the developed biomaterials were tested by Scanning Electron Microscopy (SEM), Dielectric Spectroscopy (BSD), Nuclear Magnetic Resonance (NMR), and Differential Scanning Calorimetry (DSC), as well as microhardness using Vickers method. The results showed that despite obtaining porous sinters. The highest microhardness was achieved for composite materials based on NHAP. Based on NMR spectroscopy, residue organic substances could be observed in NHAP composites, probably due to the organic structures that make up the tooth. Microbiology investigations showed that the selected samples exhibit bacteriostatic properties against Gram-positive reference bacterial strain S. epidermidis (ATCC 12228); however, the property was much less pronounced against Gram-negative reference strain E. coli (ATCC 25922). Both NHAP and SHAP, as well as their doped derivates, displayed in good general compatibility, with the exception of Cu-nanowire doped derivates. Full article

(This article belongs to the Special Issue Recent Advances in Biotechnology)

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19 pages, 28137 KiB

Open AccessArticle

Micellar Carriers of Active Substances Based on Amphiphilic PEG/PDMS Heterograft Copolymers: Synthesis and Biological Evaluation of Safe Use on Skin

by Justyna Odrobińska, Magdalena Skonieczna and Dorota Neugebauer

Int. J. Mol. Sci. 2021, 22(3), 1202; https://doi.org/10.3390/ijms22031202 - 26 Jan 2021

Cited by 3 | Viewed by 3100

Abstract

Amphiphilic copolymers containing polydimethylsiloxane (PDMS) and polyethylene glycol methyl ether (MPEG) were obtained via an azide-alkyne cycloaddition reaction between alkyne-functionalized copolymer of MPEG methacrylate and azide-functionalized PDMS. “Click” reactions were carried out with an efficiency of 33–47% increasing grafting degrees. The grafted copolymers were able to carry out the micellization and encapsulation of active substances, such as vitamin C (VitC), ferulic acid (FA) and arginine (ARG) with drug loading content (DLC) in the range of 2–68% (VitC), and 51–89% (FA or ARG). In vitro release studies (phosphate buffer saline, PBS; pH = 7.4 or 5.5) demonstrated that the maximum release of active substances was mainly after 1–2 h. The permeability of released active substances through membrane mimicking skin evaluated by transdermal tests in Franz diffusion cells indicated slight diffusion into the solution (2–16%) and their remaining in the membrane. Studies on the selected carrier with FA showed no negative effect on cell viability, proliferation capacity or senescence, as well as cell apoptosis/necrosis differences or cell cycle interruption in comparison with control cells. These results indicated that the presented micellar systems are good candidates for carriers of cosmetic substances according to physicochemical characterization and biological studies. Full article

(This article belongs to the Special Issue Recent Advances in Biotechnology)

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12 pages, 4338 KiB

Open AccessArticle

Clonal Expansion of Tumor-Infiltrating T Cells and Analysis of the Tumor Microenvironment within Esophageal Squamous Cell Carcinoma Relapsed after Definitive Chemoradiation Therapy

by Takahiro Mori, Kenichi Kumagai, Keisuke Nasu, Takamasa Yoshizawa, Koji Kuwano, Yoshiki Hamada, Hideki Kanazawa and Ryuji Suzuki

Int. J. Mol. Sci. 2021, 22(3), 1098; https://doi.org/10.3390/ijms22031098 - 22 Jan 2021

Cited by 6 | Viewed by 3195

Abstract

(1) Background: Comparable prognoses after definitive chemoradiation therapy (CRT) to surgery alone for esophageal squamous cell carcinoma (ESCC) have been previously reported; however, no robust prognostic markers have been established. The clonality of tumor-infiltrating lymphocytes (TILs) and tumor microenvironments (TMEs) in ESCC relapsed after CRT were examined to explore prognostic markers. (2) Methods: Clonality of TIL and TME were examined in ESCC with and without preceding CRT, as well as oral squamous cell carcinoma (OSCC) and healthy volunteers as controls. The clonality of TIL was assessed by T-cell receptor (TCR) α and β repertoire analyses and evaluated by diversity indices. The TME was assessed by quantitative polymerase chain reaction evaluating PD-L1 and CD8B. (3) Results: The clonal expansion of TIL was significantly induced within ESCCs and OSCCs, when compared to healthy volunteers, and was mostly induced within ESCCs after definitive CRT. Diversity indices of TIL were not associated with the prognosis, but the ratio of PD-L1 mRNA to CD8B mRNA in TME was significantly associated with a poor prognosis after salvage surgery (p = 0.007). (4) Conclusions: The clonal expansion of TIL is induced after definitive CRT for ESCC, and the ratio of PD-L1 mRNA to CD8B mRNA within tumor tissues is a prognostic marker candidate for salvage esophagectomy after CRT. Full article

(This article belongs to the Special Issue Recent Advances in Biotechnology)

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24 pages, 5271 KiB

Open AccessArticle

Bovine Organospecific Microvascular Endothelial Cell Lines as New and Relevant In Vitro Models to Study Viral Infections

by Anne-Claire Lagrée, Fabienne Fasani, Clotilde Rouxel, Marine Pivet, Marie Pourcelot, Aurore Fablet, Aurore Romey, Grégory Caignard, Damien Vitour, Sandra Blaise-Boisseau, Claudine Kieda, Henri-Jean Boulouis, Nadia Haddad and Catherine Grillon

Int. J. Mol. Sci. 2020, 21(15), 5249; https://doi.org/10.3390/ijms21155249 - 24 Jul 2020

Cited by 8 | Viewed by 3741

Abstract

Microvascular endothelial cells constitute potential targets for exogenous microorganisms, in particular for vector-borne pathogens. Their phenotypic and functional variations according to the organs they are coming from provide an explanation of the organ selectivity expressed in vivo by pathogens. In order to make available relevant tools for in vitro studies of infection mechanisms, our aim was to immortalize bovine organospecific endothelial cells but also to assess their permissivity to viral infection. Using transfection with SV40 large T antigen, six bovine microvascular endothelial cell lines from various organs and one macrovascular cell line from an umbilical cord were established. They display their own panel of endothelial progenitor/mature markers, as assessed by flow cytometry and RT-qPCR, as well as the typical angiogenesis capacity. Using both Bluetongue and foot-and-mouth disease viruses, we demonstrate that some cell lines are preferentially infected. In addition, they can be transfected and are able to express viral proteins such as BTV8-NS3. Such microvascular endothelial cell lines bring innovative tools for in vitro studies of infection by viruses or bacteria, allowing for the study of host-pathogen interaction mechanisms with the actual in vivo target cells. They are also suitable for applications linked to microvascularization, such as anti-angiogenic and anti-tumor research, growing fields in veterinary medicine. Full article

(This article belongs to the Special Issue Recent Advances in Biotechnology)

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24 pages, 3517 KiB

Open AccessArticle

Investigating Programmed Cell Death and Tumor Invasion in a Three-Dimensional (3D) Microfluidic Model of Glioblastoma

by Ehsan Samiei, Amir Seyfoori, Brian Toyota, Saeid Ghavami and Mohsen Akbari

Int. J. Mol. Sci. 2020, 21(9), 3162; https://doi.org/10.3390/ijms21093162 - 30 Apr 2020

Cited by 36 | Viewed by 6312

Abstract

Glioblastoma multiforme (GBM) is a rapidly progressive and deadly form of brain tumor with a median survival rate of ~15 months. GBMs are hard to treat and significantly affect the patient’s physical and cognitive abilities and quality of life. Temozolomide (TMZ)—an alkylating agent that causes DNA damage—is the only chemotherapy choice for the treatment of GBM. However, TMZ also induces autophagy and causes tumor cell resistance and thus fails to improve the survival rate among patients. Here, we studied the drug-induced programmed cell death and invasion inhibition capacity of TMZ and a mevalonate cascade inhibitor, simvastatin (Simva), in a three-dimensional (3D) microfluidic model of GBM. We elucidate the role of autophagy in apoptotic cell death by comparing apoptosis in autophagy knockdown cells (Atg7 KD) against their scrambled counterparts. Our results show that the cells were significantly less sensitive to drugs in the 3D model as compared to monolayer culture systems. An immunofluorescence analysis confirmed that apoptosis is the mechanism of cell death in TMZ- and Simva-treated glioma cells. However, the induction of apoptosis in the 3D model is significantly lower than in monolayer cultures. We have also shown that autophagy inhibition (Atg7 KD) did not change TMZ and Simva-induced apoptosis in the 3D microfluidic model. Overall, for the first time in this study we have established the simultaneous detection of drug induced apoptosis and autophagy in a 3D microfluidic model of GBM. Our study presents a potential ex vivo platform for developing novel therapeutic strategies tailored toward disrupting key molecular pathways involved in programmed cell death and tumor invasion in glioblastoma. Full article

(This article belongs to the Special Issue Recent Advances in Biotechnology)

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16 pages, 7156 KiB

Open AccessArticle

Alteration of the Dopamine Receptors’ Expression in the Cerebellum of the Lysosomal Acid Phosphatase 2 Mutant (Naked–Ataxia (NAX)) Mouse

by Mehdi Mehdizadeh, Niloufar Ashtari, Xiaodan Jiao, Maryam Rahimi Balaei, Asghar Marzban, Farshid Qiyami-Hour, Jiming Kong, Saeid Ghavami and Hassan Marzban

Int. J. Mol. Sci. 2020, 21(8), 2914; https://doi.org/10.3390/ijms21082914 - 21 Apr 2020

Cited by 8 | Viewed by 3302

Abstract

A spontaneous mutation in the lysosomal acid phosphatase (Acp2) enzyme (nax: naked–ataxia) in experimental mice results in delayed hair appearance and severe cytoarchitectural impairments of the cerebellum, such as a Purkinje cell (PC) migration defect. In our previous investigation, our team showed that Acp2 expression plans a significant role in cerebellar development. On the other hand, the dopaminergic system is also a player in central nervous system (CNS) development, including cerebellar structure and function. In the current investigation, we have explored how Acp2 can be involved in the regulation of the dopaminergic pathway in the cerebellum via the regulation of dopamine receptor expression and patterning. We provided evidence about the distribution of different dopamine receptors in the developing cerebellum by comparing the expression of dopamine receptors on postnatal days (P) 5 and 17 between nax mice and wild–type (wt) littermates. To this aim, immunohistochemistry and Western blot analysis were conducted using five antibodies against dopamine receptors (DRD1, –2, –3, –4, and –5) accompanied by RNAseq data. Our results revealed that DRD1, –3, and –4 gene expressions significantly increased in nax cerebella but not in wt, while gene expressions of all 5 receptors were evident in PCs of both wt and nax cerebella. DRD3 was strongly expressed in the PCs’ somata and cerebellar nuclei neurons at P17 in nax mice, which was comparable to the expression levels in the cerebella of wt littermates. In addition, DRD3 was expressed in scattered cells in a granular layer reminiscent of Golgi cells and was observed in the wt cerebella but not in nax mice. DRD4 was expressed in a subset of PCs and appeared to align with the unique parasagittal stripes pattern. This study contributes to our understanding of alterations in the expression pattern of DRDs in the cerebellum of nax mice in comparison to their wt littermates, and it highlights the role of Acp2 in regulating the dopaminergic system. Full article

(This article belongs to the Special Issue Recent Advances in Biotechnology)

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16 pages, 3863 KiB

Open AccessArticle

HSP70/IL-2 Treated NK Cells Effectively Cross the Blood Brain Barrier and Target Tumor Cells in a Rat Model of Induced Glioblastoma Multiforme (GBM)

by Farzaneh Sharifzad, Soura Mardpour, Saeid Mardpour, Esmaeil Fakharian, Adeleh Taghikhani, Amirhossein Sharifzad, Sahar Kiani, Yasaman Heydarian, Marek J. Łos, Zahra Azizi, Saeid Ghavami, Amir Ali Hamidieh and Marzieh Ebrahimi

Int. J. Mol. Sci. 2020, 21(7), 2263; https://doi.org/10.3390/ijms21072263 - 25 Mar 2020

Cited by 30 | Viewed by 6290

Abstract

Natural killer (NK) cell therapy is one of the most promising treatments for Glioblastoma Multiforme (GBM). However, this emerging technology is limited by the availability of sufficient numbers of fully functional cells. Here, we investigated the efficacy of NK cells that were expanded and treated by interleukin-2 (IL-2) and heat shock protein 70 (HSP70), both in vitro and in vivo. Proliferation and cytotoxicity assays were used to assess the functionality of NK cells in vitro, after which treated and naïve NK cells were administrated intracranially and systemically to compare the potential antitumor activities in our in vivo rat GBM models. In vitro assays provided strong evidence of NK cell efficacy against C6 tumor cells. In vivo tracking of NK cells showed efficient homing around and within the tumor site. Furthermore, significant amelioration of the tumor in rats treated with HSP70/Il-2-treated NK cells as compared to those subjected to nontreated NK cells, as confirmed by MRI, proved the efficacy of adoptive NK cell therapy. Moreover, results obtained with systemic injection confirmed migration of activated NK cells over the blood brain barrier and subsequent targeting of GBM tumor cells. Our data suggest that administration of HSP70/Il-2-treated NK cells may be a promising therapeutic approach to be considered in the treatment of GBM. Full article

(This article belongs to the Special Issue Recent Advances in Biotechnology)

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Review

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16 pages, 44507 KiB

Open AccessReview

Nanotechnology Facilitated Cultured Neuronal Network and Its Applications

by Satnam Singh, Sachin Mishra, Song Juha, Manojit Pramanik, Parasuraman Padmanabhan and Balázs Gulyás

Int. J. Mol. Sci. 2021, 22(11), 5552; https://doi.org/10.3390/ijms22115552 - 24 May 2021

Cited by 4 | Viewed by 3717

Abstract

The development of a biomimetic neuronal network from neural cells is a big challenge for researchers. Recent advances in nanotechnology, on the other hand, have enabled unprecedented tools and techniques for guiding and directing neural stem cell proliferation and differentiation in vitro to construct an in vivo-like neuronal network. Nanotechnology allows control over neural stem cells by means of scaffolds that guide neurons to reform synaptic networks in suitable directions in 3D architecture, surface modification/nanopatterning to decide cell fate and stimulate/record signals from neurons to find out the relationships between neuronal circuit connectivity and their pathophysiological functions. Overall, nanotechnology-mediated methods facilitate precise physiochemical controls essential to develop tools appropriate for applications in neuroscience. This review emphasizes the newest applications of nanotechnology for examining central nervous system (CNS) roles and, therefore, provides an insight into how these technologies can be tested in vitro before being used in preclinical and clinical research and their potential role in regenerative medicine and tissue engineering. Full article

(This article belongs to the Special Issue Recent Advances in Biotechnology)

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