Biomedicines

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18 pages, 8562 KiB

Open AccessArticle

Comparison between Platelet Lysate, Platelet Lysate Serum, and Fetal Bovine Serum as Supplements for Cell Culture, Expansion, and Cryopreservation

by Juan Manuel Duarte Rojas, Luz Marina Restrepo Múnera and Sergio Estrada Mira

Biomedicines 2024, 12(1), 140; https://doi.org/10.3390/biomedicines12010140 - 09 Jan 2024

Cited by 2 | Viewed by 1268

Abstract

As cell culture supplements, human platelet lysate (PL) and human platelet lysate serum (PLS) are alternatives to fetal bovine serum (FBS) due to FBS-related issues such as ethical concerns, variability between batches, and the possible introduction of xenogenic contaminants. This study compared the [...] Read more.

As cell culture supplements, human platelet lysate (PL) and human platelet lysate serum (PLS) are alternatives to fetal bovine serum (FBS) due to FBS-related issues such as ethical concerns, variability between batches, and the possible introduction of xenogenic contaminants. This study compared the composition and efficacy of PL, PLS, and FBS as supplements in the culture and cryopreservation of human dermal fibroblasts, Wharton’s jelly-derived mesenchymal stem cells (WJ-MCS), and adipose tissue (AdMSC). Biochemical components, some growth factors, and cytokines present in each of them were analyzed; in addition, the cells were cultured in media supplemented with 5% PL, 5% PLS, and 10% FBS and exposed to different freezing and thawing solutions with the supplements under study. Biochemical parameters were found to be similar in PL and PLS compared to FBS, with some differences in fibrinogen and calcium concentration. Growth factors and cytokines were higher in PL and PLS compared to FBS. Cell proliferation and morphology showed no significant differences between the three culture media. Regarding the cryopreservation and thawing of cells, better results were obtained with PLS and FBS. In conclusion, PL and PLS are an excellent choice to replace the standard supplement of animal origin (FBS) in the media used for the culture and cryopreservation of fibroblasts, WJ-MSC, and AdMSC. Full article

(This article belongs to the Special Issue Advanced Research in Tissue Engineering and Cellular Culture and Their Applications)

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19 pages, 3532 KiB

Open AccessArticle

Combining Electrostimulation with Impedance Sensing to Promote and Track Osteogenesis within a Titanium Implant

by Nadja Engel, Michael Dau, Vivien Engel, Denise Franz, Fabian Klemmstein, Christiane Thanisch, Jürgen F. Kolb, Marcus Frank, Armin Springer, Rüdiger Köhling, Rainer Bader, Bernhard Frerich, Nadine Wiesmann, Diana Heimes and Peer W. Kämmerer

Biomedicines 2023, 11(3), 697; https://doi.org/10.3390/biomedicines11030697 - 24 Feb 2023

Cited by 2 | Viewed by 1540

Abstract

(1) Background: Electrical stimulation is a promising alternative to promote bone fracture healing but with the limitation of tracking the osteogenesis progress in vivo. To overcome this issue, we present an opportunity to combine the electrical stimulation of a commercial titanium implant, which [...] Read more.

(1) Background: Electrical stimulation is a promising alternative to promote bone fracture healing but with the limitation of tracking the osteogenesis progress in vivo. To overcome this issue, we present an opportunity to combine the electrical stimulation of a commercial titanium implant, which promotes osteogenesis within the fracture, with a real-time readout of the osteogenic progress by impedance sensing. This makes it possible to adjust the electrical stimulation modalities to the individual patient’s fracture healing process. (2) Methods: In detail, osteogenic differentiation of several cell types was monitored under continuous or pulsatile electrical stimulation at 0.7 V AC/20 Hz for at least seven days on a titanium implant by electric cell-substrate impedance sensing (ECIS). For control, chemical induction of osteogenic differentiation was induced. (3) Results: The most significant challenge was to discriminate impedance changes caused by proliferation events from those initiated by osteogenic differentiation. This discrimination was achieved by remodeling the impedance parameter Alpha (α), which increases over time for pulsatile electrically stimulated stem cells. Boosted α-values were accompanied by an increased formation of actin stress fibers and a reduced expression of the focal adhesion kinase in the cell periphery; morphological alterations known to occur during osteogenesis. (4) Conclusions: This work provided the basis for developing an effective fracture therapy device, which can induce osteogenesis on the one hand, and would allow us to monitor the induction process on the other hand. Full article

(This article belongs to the Special Issue Advanced Research in Tissue Engineering and Cellular Culture and Their Applications)

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9 pages, 1031 KiB

Open AccessArticle

In Vitro Evaluation of the Cytotoxic Potential of Thiosemicarbazide Coordinating Compounds in Hepatocyte Cell Culture

by Valeriana Pantea, Vitalie Cobzac, Olga Tagadiuc, Victor Palarie and Valentin Gudumac

Biomedicines 2023, 11(2), 366; https://doi.org/10.3390/biomedicines11020366 - 26 Jan 2023

Cited by 2 | Viewed by 1212

Abstract

Cancer is a global medical problem and, despite research efforts in the field of tumor treatment, there is currently a shortage of specific anticancer drugs. Most anticancer drugs show significant side effects. The liver is the organ that has central functions in drug [...] Read more.

Cancer is a global medical problem and, despite research efforts in the field of tumor treatment, there is currently a shortage of specific anticancer drugs. Most anticancer drugs show significant side effects. The liver is the organ that has central functions in drug metabolism, being a major target of the harmful action of anticancer compounds. In this context, it is essential to evaluate the cytotoxic effects of potential anticancer substances. Therefore, hepatotoxicity and hepatocyte viability were determined in vitro to evaluate the action of seven new local thiosemicarbazide coordination compounds (CCT) on normal liver cells. Doxorubicin was used as a reference substance. The control group consisted of hepatocytes not exposed to CCT action. The cell viability of hepatocytes treated with CCT decreased significantly by 5–12% compared to the control, but was statistically significantly higher by 5–14% compared to doxorubicin, except after CMD-8 and CMT-67 influence, when it does not change. Thus, new local CCT had a selective effect on hepatocytes in vitro and were less hepatotoxic compared to doxorubicin, which may be the basis for further study of its potential in anticancer drugs. Full article

(This article belongs to the Special Issue Advanced Research in Tissue Engineering and Cellular Culture and Their Applications)

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15 pages, 2279 KiB

Open AccessArticle

Local and Systemic Injections of Human Cord Blood Myeloid-Derived Suppressor Cells to Prevent Graft Rejection in Corneal Transplantation

by Jae-young Lee, Hyun-Jung Sohn, Chang-Hyun Kim, Tai-Gyu Kim and Hyun Soo Lee

Biomedicines 2022, 10(12), 3223; https://doi.org/10.3390/biomedicines10123223 - 12 Dec 2022

Cited by 4 | Viewed by 1392

Abstract

Myeloid-derived suppressor cells (MDSCs) are therapeutic agents to prevent graft rejection in organ transplants by modulating inflammation. Herein, the immunosuppressive effect of human cord blood MDSCs on corneal allograft models was confirmed. CB-MDSCs were locally (subconjuctival, 5 × 10⁵) or systemically [...] Read more.

Myeloid-derived suppressor cells (MDSCs) are therapeutic agents to prevent graft rejection in organ transplants by modulating inflammation. Herein, the immunosuppressive effect of human cord blood MDSCs on corneal allograft models was confirmed. CB-MDSCs were locally (subconjuctival, 5 × 10⁵) or systemically (intravenous, 1 × 10⁶) injected twice on days 0 and 7. A corneal transplantation model was established using C57BL/6 and BALB/c mice, and corneal graft opacity was measured to evaluate graft rejection up to 6 weeks. Results showed that graft survival in the MDSCs groups increased compared to vehicle groups after 42 days. Systemic and local MDSC administration inhibited the maturation (MHC-II^hi CD11c+) of dendritic cells (DCs) and the differentiation of interferon γ+ CD4+ Th1 in draining lymph nodes (LNs). However, vehicle groups increased the infiltration of CD3+ T cells and F4/80+ macrophages and produced prominent neovascular and lymphatic vessels into the graft site with increased mRNA expression of VEGF-A/C and VEGFR-1/R-3. Local MDSCs administration showed prominent anti-angiogenic/anti-lymphangiogenic effects even at lower MDSCs doses. Thus, CB-MDSCs could relatively suppress the infiltration of pathological T cells/macrophages into the corneas and the migration of mature DCs into draining LNs Therefore, ocular and systemic MDSCs administration showed therapeutic potential for preventing corneal allograft rejection. Full article

(This article belongs to the Special Issue Advanced Research in Tissue Engineering and Cellular Culture and Their Applications)

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19 pages, 4847 KiB

Open AccessArticle

A Strategy for Magnetic and Electric Stimulation to Enhance Proliferation and Differentiation of NPCs Seeded over PLA Electrospun Membranes

by Irene Cuenca-Ortolá, Beatriz Martínez-Rojas, Victoria Moreno-Manzano, Marcos García Castelló, Manuel Monleón Pradas, Cristina Martínez-Ramos and Jorge Más Estellés

Biomedicines 2022, 10(11), 2736; https://doi.org/10.3390/biomedicines10112736 - 28 Oct 2022

Cited by 3 | Viewed by 1598

Abstract

Neural progenitor cells (NPCs) have been shown to serve as an efficient therapeutic strategy in different cell therapy approaches, including spinal cord injury treatment. Despite the reported beneficial effects of NPC transplantation, the low survival and differentiation rates constrain important limitations. Herein, a [...] Read more.

Neural progenitor cells (NPCs) have been shown to serve as an efficient therapeutic strategy in different cell therapy approaches, including spinal cord injury treatment. Despite the reported beneficial effects of NPC transplantation, the low survival and differentiation rates constrain important limitations. Herein, a new methodology has been developed to overcome both limitations by applying a combination of wireless electrical and magnetic stimulation to NPCs seeded on aligned poly(lactic acid) nanofibrous scaffolds for in vitro cell conditioning prior transplantation. Two stimulation patterns were tested and compared, continuous (long stimulus applied once a day) and intermittent (short stimulus applied three times a day). The results show that applied continuous stimulation promotes NPC proliferation and preferential differentiation into oligodendrocytic and neuronal lineages. A neural-like phenotypic induction was observed when compared to unstimulated NPCs. In contrast, intermittent stimulation patterns did not affect NPC proliferation and differentiation to oligodendrocytes or astrocytes morphology with a detrimental effect on neuronal differentiation. This study provides a new approach of using a combination of electric and magnetic stimulation to induce proliferation and further neuronal differentiation, which would improve therapy outcomes in disorders such as spinal cord injury. Full article

(This article belongs to the Special Issue Advanced Research in Tissue Engineering and Cellular Culture and Their Applications)

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Review

Jump to: Research

23 pages, 1121 KiB

Open AccessReview

Three-Dimensional Bioprinting of Naturally Derived Hydrogels for the Production of Biomimetic Living Tissues: Benefits and Challenges

by Elena Merotto, Piero G. Pavan and Martina Piccoli

Biomedicines 2023, 11(6), 1742; https://doi.org/10.3390/biomedicines11061742 - 17 Jun 2023

Cited by 2 | Viewed by 1460

Abstract

Three-dimensional bioprinting is the process of manipulating cell-laden bioinks to fabricate living structures. Three-dimensional bioprinting techniques have brought considerable innovation in biomedicine, especially in the field of tissue engineering, allowing the production of 3D organ and tissue models for in vivo transplantation purposes [...] Read more.

Three-dimensional bioprinting is the process of manipulating cell-laden bioinks to fabricate living structures. Three-dimensional bioprinting techniques have brought considerable innovation in biomedicine, especially in the field of tissue engineering, allowing the production of 3D organ and tissue models for in vivo transplantation purposes or for in-depth and precise in vitro analyses. Naturally derived hydrogels, especially those obtained from the decellularization of biological tissues, are promising bioinks for 3D printing purposes, as they present the best biocompatibility characteristics. Despite this, many natural hydrogels do not possess the necessary mechanical properties to allow a simple and immediate application in the 3D printing process. In this review, we focus on the bioactive and mechanical characteristics that natural hydrogels may possess to allow efficient production of organs and tissues for biomedical applications, emphasizing the reinforcement techniques to improve their biomechanical properties. Full article

(This article belongs to the Special Issue Advanced Research in Tissue Engineering and Cellular Culture and Their Applications)

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17 pages, 2692 KiB

Open AccessReview

Key Challenges in Diamond Coating of Titanium Implants: Current Status and Future Prospects

by Maria Letizia Terranova

Biomedicines 2022, 10(12), 3149; https://doi.org/10.3390/biomedicines10123149 - 06 Dec 2022

Cited by 1 | Viewed by 1307

Abstract

Over past years, the fabrication of Ti-based permanent implants for fracture fixation, joint replacement and bone or tooth substitution, has become a routine task. However, it has been found that some degradation phenomena occurring on the Ti surface limits the life or the [...] Read more.

Over past years, the fabrication of Ti-based permanent implants for fracture fixation, joint replacement and bone or tooth substitution, has become a routine task. However, it has been found that some degradation phenomena occurring on the Ti surface limits the life or the efficiency of the artificial constructs. The task of avoiding such adverse effects, to prevent microbial colonization and to accelerate osteointegration, is being faced by a variety of approaches in order to adapt Ti surfaces to the needs of osseous tissues. Among the large set of biocompatible materials proposed as an interface between Ti and the hosting tissue, diamond has been proven to offer bioactive and mechanical properties able to match the specific requirements of osteoblasts. Advances in material science and implant engineering are now enabling us to produce micro- or nano-crystalline diamond coatings on a variety of differently shaped Ti constructs. The aim of this paper is to provide an overview of the research currently ongoing in the field of diamond-coated orthopedic Ti implants and to examine the evolution of the concepts that are accelerating the full transition of such technology from the laboratory to clinical applications. Full article

(This article belongs to the Special Issue Advanced Research in Tissue Engineering and Cellular Culture and Their Applications)

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27 pages, 3044 KiB

Open AccessReview

Modifying Orthobiological PRP Therapies Are Imperative for the Advancement of Treatment Outcomes in Musculoskeletal Pathologies

by Peter A. Everts, Timothy Mazzola, Kenneth Mautner, Pietro S. Randelli and Luga Podesta

Biomedicines 2022, 10(11), 2933; https://doi.org/10.3390/biomedicines10112933 - 15 Nov 2022

Cited by 3 | Viewed by 2713

Abstract

Autologous biological cellular preparations have materialized as a growing area of medical advancement in interventional (orthopedic) practices and surgical interventions to provide an optimal tissue healing environment, particularly in tissues where standard healing is disrupted and repair and ultimately restoration of function is [...] Read more.

Autologous biological cellular preparations have materialized as a growing area of medical advancement in interventional (orthopedic) practices and surgical interventions to provide an optimal tissue healing environment, particularly in tissues where standard healing is disrupted and repair and ultimately restoration of function is at risk. These cellular therapies are often referred to as orthobiologics and are derived from patient’s own tissues to prepare point of care platelet-rich plasma (PRP), bone marrow concentrate (BMC), and adipose tissue concentrate (ATC). Orthobiological preparations are biological materials comprised of a wide variety of cell populations, cytokines, growth factors, molecules, and signaling cells. They can modulate and influence many other resident cells after they have been administered in specific diseased microenvironments. Jointly, the various orthobiological cell preparations are proficient to counteract persistent inflammation, respond to catabolic reactions, and reinstate tissue homeostasis. Ultimately, precisely delivered orthobiologics with a proper dose and bioformulation will contribute to tissue repair. Progress has been made in understanding orthobiological technologies where the safety and relatively easy manipulation of orthobiological treatment tools has been demonstrated in clinical applications. Although more positive than negative patient outcome results have been registered in the literature, definitive and accepted standards to prepare specific cellular orthobiologics are still lacking. To promote significant and consistent clinical outcomes, we will present a review of methods for implementing dosing strategies, using bioformulations tailored to the pathoanatomic process of the tissue, and adopting variable preparation and injection volume policies. By optimizing the dose and specificity of orthobiologics, local cellular synergistic behavior will increase, potentially leading to better pain killing effects, effective immunomodulation, control of inflammation, and (neo) angiogenesis, ultimately contributing to functionally restored body movement patterns. Full article

(This article belongs to the Special Issue Advanced Research in Tissue Engineering and Cellular Culture and Their Applications)

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