scholarly journals A microfluidic-based cell encapsulation platform to achieve high long-term cell viability in photopolymerized PEGNB hydrogel microspheres

2017 ◽  
Vol 5 (1) ◽  
pp. 173-180 ◽  
Author(s):  
Zhongliang Jiang ◽  
Bingzhao Xia ◽  
Ralph McBride ◽  
John Oakey
Keyword(s):  
Tissue Engineering ◽  
Polyethylene Glycol ◽  
Regenerative Medicine ◽  
Cell Viability ◽  
Cell Encapsulation ◽  
Cell Delivery ◽  
Cellular Behavior ◽  
Hydrogel Scaffolds

Cell encapsulation within photopolymerized polyethylene glycol (PEG)-based hydrogel scaffolds has been demonstrated as a robust strategy for cell delivery, tissue engineering, regenerative medicine, and developing in vitro platforms to study cellular behavior and fate.

Abstract 9: Temperature-responsive Cell Delivery Biopolymers for Cardiac Tissue Engineering

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Brisa Pena ◽  
Valentina Martinelli ◽  
Susanna Bosi ◽  
Carmen Sucharov ◽  
Mark Jeong ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Body Temperature ◽  
Cell Viability ◽  
Cultured Cells ◽  
Ventricular Myocytes ◽  
3D Culture ◽  
Polymeric Matrix ◽  
Neonatal Rat ◽  
Cell Delivery

Background: Advances in cell therapy and material science have made tissue engineering a promising strategy for heart regeneration. We developed an injectable biomimetic reverse thermal gel (RTG) that is liquid at room temperature but gel-like at body temperature, with the ultimate goal of being able to serve as a vehicle for cell-based delivery (liquid) to targeted tissue areas (gel-phase at 37°C). In this study we tested the suitability of this biomimetic RTG on cell viability. Methods and results: We tested different biomimetic RTG systems with and without the chemical incorporation of lysine. In vitro 3D culture experiments were performed with neonatal rat ventricular myocytes (NRVM) by mixing 3x104 cells with 50 μl of polymeric solution and allowing gel formation at 37°C. The cultured cells were incubated for 21 days. For controls we used NRVMs plated on 2D traditional gelatin coated dishes. We found that the 3D polymeric matrix induces rapid coordinated contraction with improved functionality when compared with standard 2D-cultured NRVM. By immunostaining for the morphology of the sarcomere (alpha-actinin) and DAPI, we also observed that the 3D polymeric matrix stimulates cells to spread and form 3D syncytia. Conclusion: These proof-of-concept results demonstrate long-term cell viability in this unique biomimetic system and therefore provide feasibility of a polymeric cell delivery system that permits reversible liquid-to-gel transition at body temperature. These results offer potential for a tissue engineering approach to cardiac regeneration.

scholarly journals A Perfusion Bioreactor for Longitudinal Monitoring of Bioengineered Liver Constructs

Nanomaterials ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 275
Author(s):  
Lisa Sassi ◽  
Omolola Ajayi ◽  
Sara Campinoti ◽  
Dipa Natarajan ◽  
Claire McQuitty ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Liver Disease ◽  
Cell Viability ◽  
3D Culture ◽  
Culture Conditions ◽  
Disease Models ◽  
Non Invasive ◽  
And Function

In the field of in vitro liver disease models, decellularised organ scaffolds maintain the original biomechanical and biological properties of the extracellular matrix and are established supports for in vitro cell culture. However, tissue engineering approaches based on whole organ decellularized scaffolds are hampered by the scarcity of appropriate bioreactors that provide controlled 3D culture conditions. Novel specific bioreactors are needed to support long-term culture of bioengineered constructs allowing non-invasive longitudinal monitoring. Here, we designed and validated a specific bioreactor for long-term 3D culture of whole liver constructs. Whole liver scaffolds were generated by perfusion decellularisation of rat livers. Scaffolds were seeded with Luc+HepG2 and primary human hepatocytes and cultured in static or dynamic conditions using the custom-made bioreactor. The bioreactor included a syringe pump, for continuous unidirectional flow, and a circuit built to allow non-invasive monitoring of culture parameters and media sampling. The bioreactor allowed non-invasive analysis of cell viability, distribution, and function of Luc+HepG2-bioengineered livers cultured for up to 11 days. Constructs cultured in dynamic conditions in the bioreactor showed significantly higher cell viability, measured with bioluminescence, distribution, and functionality (determined by albumin production and expression of CYP enzymes) in comparison to static culture conditions. Finally, our bioreactor supports primary human hepatocyte viability and function for up to 30 days, when seeded in the whole liver scaffolds. Overall, our novel bioreactor is capable of supporting cell survival and metabolism and is suitable for liver tissue engineering for the development of 3D liver disease models.

scholarly journals In Vitro Biological Performance of Alginate Hydrogel Capsules for Stem Cell Delivery

2021 ◽  
Vol 9 ◽  
Author(s):  
Jaqueline Brandão de Souza ◽  
Gustavo dos Santos Rosa ◽  
Mariana Correa Rossi ◽  
Fernanda de Castro Stievani ◽  
João Pedro Hübbe Pfeifer ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cell ◽  
Cell Viability ◽  
Sodium Alginate ◽  
Cell Encapsulation ◽  
Alginate Hydrogel ◽  
Stem Cell Delivery ◽  
Biological Performance

Encapsulation of biological components in hydrogels is a well described method for controlled drug delivery of proteins, tissue engineering and intestinal colonization with beneficial bacteria. Given the potential of tissue engineering in clinical practice, this study aimed to evaluate the feasibility of encapsulation of adipose tissue-derived mesenchymal stem cells (MSCs) of mules in sodium alginate. We evaluated capsule morphology and cell viability, immunophenotype and release after encapsulation. Circular and irregular pores were observed on the hydrogel surface, in which MSCs were present and alive. Capsules demonstrated good capacity of absorption of liquid and cell viability was consistently high through the time points, indicating proper nutrient diffusion. Flow cytometry showed stability of stem cell surface markers, whereas immunohistochemistry revealed the expression of CD44 and absence of MHC-II through 7 days of culture. Stem cell encapsulation in sodium alginate hydrogel is a feasible technique that does not compromise cell viability and preserves their undifferentiated status, becoming a relevant option to further studies of tridimensional culture systems and in vivo bioactive agents delivery.

scholarly journals Extracellular Matrix-Based Biomaterials for Cardiovascular Tissue Engineering

2021 ◽  
Vol 8 (11) ◽  
pp. 137
Author(s):  
Astha Khanna ◽  
Maedeh Zamani ◽  
Ngan F. Huang
Keyword(s):  
Tissue Engineering ◽  
Extracellular Matrix ◽  
Regenerative Medicine ◽  
Three Dimensional ◽  
Vascular Differentiation ◽  
Cardiovascular Tissue ◽  
Cellular Behavior ◽  
Cardiovascular Tissue Engineering ◽  
And Function

Regenerative medicine and tissue engineering strategies have made remarkable progress in remodeling, replacing, and regenerating damaged cardiovascular tissues. The design of three-dimensional (3D) scaffolds with appropriate biochemical and mechanical characteristics is critical for engineering tissue-engineered replacements. The extracellular matrix (ECM) is a dynamic scaffolding structure characterized by tissue-specific biochemical, biophysical, and mechanical properties that modulates cellular behavior and activates highly regulated signaling pathways. In light of technological advancements, biomaterial-based scaffolds have been developed that better mimic physiological ECM properties, provide signaling cues that modulate cellular behavior, and form functional tissues and organs. In this review, we summarize the in vitro, pre-clinical, and clinical research models that have been employed in the design of ECM-based biomaterials for cardiovascular regenerative medicine. We highlight the research advancements in the incorporation of ECM components into biomaterial-based scaffolds, the engineering of increasingly complex structures using biofabrication and spatial patterning techniques, the regulation of ECMs on vascular differentiation and function, and the translation of ECM-based scaffolds for vascular graft applications. Finally, we discuss the challenges, future perspectives, and directions in the design of next-generation ECM-based biomaterials for cardiovascular tissue engineering and clinical translation.

scholarly journals Colloidal quantum dots suitability for long term cell imaging

2018 ◽  
Author(s):  
Patricia M. A. Farias ◽  
André Galembeck ◽  
Raquel Milani ◽  
Wilson S. Mendonca ◽  
Andreas Stingl
Keyword(s):  
Quantum Dots ◽  
Polyethylene Glycol ◽  
Cell Viability ◽  
Glial Cells ◽  
Aqueous Suspension ◽  
High Rate ◽  
Term Monitoring ◽  
Observation Period

AbstractFluorescent semiconductor nanoparticles in tree-dimensional quantum confinement, quantum dots (QDs), synthesized in aqueous medium, and functionalized with polyethylene glycol, were used as probes for the long-term imaging of glial cells. In vitro living healthy as well as cancer glial cells were labelled by direct insertion of a small volume of QDs contained in aqueous suspension into the culture wells. A long-term monitoring (over 7 days) of the cells was performed and no evidence of cell fixation and/or damage was observed. Two control groups, healthy and cancer glial cells, were used to compare cell viability. During the observation period, labelled and non labelled cells presented the same dynamics and no difference was observed regarding cell viability. To our knowledge, this is the first report of the viability of hydrophilic prepared quantum dots without any further surface treatment than the polyethylene-glycol coverage for the long-term imaging of living cells. Further, the study also permitted the observation of two distinct interaction mechanisms between cells and QDs. Healthy glial cells were mainly labelled at their surface, while non-healthy glial cells have shown a high rate in the uptake of QDs.

scholarly journals Long-Term Cryostorage of Mesenchymal Stem Cell-Containing Hybrid Hydrogel Scaffolds Based on Fibrin and Collagen

Gels ◽  
2020 ◽  
Vol 6 (4) ◽  
pp. 44
Author(s):  
Marfa N. Egorikhina ◽  
Yulia P. Rubtsova ◽  
Diana Ya. Aleynik
Keyword(s):  
Tissue Engineering ◽  
Proliferative Activity ◽  
Experimental Protocol ◽  
Hydrogel Scaffold ◽  
Hybrid Hydrogel ◽  
Hydrogel Scaffolds ◽  
High Viability ◽  
Scaffold Structure ◽  
Difficult Issue

The most difficult issue when using tissue engineering products is enabling the ability to store them without losing their restorative capacity. The numbers and viability of mesenchymal stem cells encapsulated in a hydrogel scaffold after cryostorage at −80 °C (by using, individually, two kinds of cryoprotectors—Bambanker and 10% DMSO (Dimethyl sulfoxide) solution) for 3, 6, 9, and 12 months were determined, with subsequent assessment of cell proliferation after 96 h. The analysis of the cellular component was performed using fluorescence microscopy and the two fluorochromes—Hoechst 3334 and NucGreenTM Dead 488. The experimental protocol ensured the preservation of cells in the scaffold structure, retaining both high viability and proliferative activity during storage for 3 months. Longer storage of scaffolds led to their significant changes. Therefore, after 6 months, the proliferative activity of cells decreased. Cryostorage of scaffolds for 9 months led to a decrease in cells’ viability and proliferative activity. As a result of cryostorage of scaffolds for 12 months, a decrease in viability and proliferative activity of cells was observed, as well as pronounced changes in the structure of the hydrogel. The described scaffold cryostorage protocol could become the basis for the development of storage protocols for such tissue engineering products, and for helping to extend the possibilities of their clinical use while accelerating their commercialization.

scholarly journals Placenta Derived Mesenchymal Stem Cells Hosted on RKKP Glass-Ceramic: A Tissue Engineering Strategy for Bone Regenerative Medicine Applications

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Mario Ledda ◽  
Marco Fosca ◽  
Angela De Bonis ◽  
Mariangela Curcio ◽  
Roberto Teghil ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Regenerative Medicine ◽  
Glass Ceramic ◽  
Phase Distribution ◽  
Sol Gel ◽  
Pcr Analysis ◽  
Cycle Phase ◽  
Cell Delivery ◽  
Limiting Factor

In tissue engineering protocols, the survival of transplanted stem cells is a limiting factor that could be overcome using a cell delivery matrix able to support cell proliferation and differentiation. With this aim, we studied the cell-friendly and biocompatible behavior of RKKP glass-ceramic coated Titanium (Ti) surface seeded with human amniotic mesenchymal stromal cells (hAMSCs) from placenta. The sol-gel synthesis procedure was used to prepare the RKKP glass-ceramic material, which was then deposited onto the Ti surface by Pulsed Laser Deposition method. The cell metabolic activity and proliferation rate, the cytoskeletal actin organization, and the cell cycle phase distribution in hAMSCs seeded on the RKKP coated Ti surface revealed no significant differences when compared to the cells grown on the treated plastic Petri dish. The health of of hAMSCs was also analysed studying the mRNA expressions of MSC key genes and the osteogenic commitment capability using qRT-PCR analysis which resulted in being unchanged in both substrates. In this study, the combination of the hAMSCs’ properties together with the bioactive characteristics of RKKP glass-ceramics was investigated and the results obtained indicate its possible use as a new and interesting cell delivery system for bone tissue engineering and regenerative medicine applications.

scholarly journals Tuning the Properties of PNIPAm-Based Hydrogel Scaffolds for Cartilage Tissue Engineering

Polymers ◽  
2021 ◽  
Vol 13 (18) ◽  
pp. 3154
Author(s):  
Md Mohosin Rana ◽  
Hector De la Hoz Siegler
Keyword(s):  
Tissue Engineering ◽  
Physical Properties ◽  
Tissue Regeneration ◽  
Structural Integrity ◽  
Cartilage Tissue ◽  
Design Parameters ◽  
Comprehensive Overview ◽  
Hydrogel Scaffolds ◽  
Design Variables

Poly(N-isopropylacrylamide) (PNIPAm) is a three-dimensional (3D) crosslinked polymer that can interact with human cells and play an important role in the development of tissue morphogenesis in both in vitro and in vivo conditions. PNIPAm-based scaffolds possess many desirable structural and physical properties required for tissue regeneration, but insufficient mechanical strength, biocompatibility, and biomimicry for tissue development remain obstacles for their application in tissue engineering. The structural integrity and physical properties of the hydrogels depend on the crosslinks formed between polymer chains during synthesis. A variety of design variables including crosslinker content, the combination of natural and synthetic polymers, and solvent type have been explored over the past decade to develop PNIPAm-based scaffolds with optimized properties suitable for tissue engineering applications. These design parameters have been implemented to provide hydrogel scaffolds with dynamic and spatially patterned cues that mimic the biological environment and guide the required cellular functions for cartilage tissue regeneration. The current advances on tuning the properties of PNIPAm-based scaffolds were searched for on Google Scholar, PubMed, and Web of Science. This review provides a comprehensive overview of the scaffolding properties of PNIPAm-based hydrogels and the effects of synthesis-solvent and crosslinking density on tuning these properties. Finally, the challenges and perspectives of considering these two design variables for developing PNIPAm-based scaffolds are outlined.

scholarly journals Natural Architectures for Tissue Engineering and Regenerative Medicine

2020 ◽  
Vol 11 (3) ◽  
pp. 47
Author(s):  
Floris Honig ◽  
Steven Vermeulen ◽  
Amir A. Zadpoor ◽  
Jan de Boer ◽  
Lidy E. Fratila-Apachitei
Keyword(s):  
Tissue Engineering ◽  
Regenerative Medicine ◽  
Surface Properties ◽  
State Of The Art ◽  
Cell Behaviour ◽  
Cell Responses ◽  
Underlying Mechanisms ◽  
Functional Biomaterials

The ability to control the interactions between functional biomaterials and biological systems is of great importance for tissue engineering and regenerative medicine. However, the underlying mechanisms defining the interplay between biomaterial properties and the human body are complex. Therefore, a key challenge is to design biomaterials that mimic the in vivo microenvironment. Over millions of years, nature has produced a wide variety of biological materials optimised for distinct functions, ranging from the extracellular matrix (ECM) for structural and biochemical support of cells to the holy lotus with special wettability for self-cleaning effects. Many of these systems found in biology possess unique surface properties recognised to regulate cell behaviour. Integration of such natural surface properties in biomaterials can bring about novel cell responses in vitro and provide greater insights into the processes occurring at the cell-biomaterial interface. Using natural surfaces as templates for bioinspired design can stimulate progress in the field of regenerative medicine, tissue engineering and biomaterials science. This literature review aims to combine the state-of-the-art knowledge in natural and nature-inspired surfaces, with an emphasis on material properties known to affect cell behaviour.

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