scholarly journals Making microenvironments: A look into incorporating macromolecular crowding into in vitro experiments, to generate biomimetic microenvironments which are capable of directing cell function for tissue engineering applications

2017 ◽  
Vol 8 ◽  
pp. 204173141773046 ◽  
Author(s):  
Paula Benny ◽  
Michael Raghunath
Keyword(s):  
Tissue Engineering ◽  
Cell Culture ◽  
Cell Function ◽  
Macromolecular Crowding ◽  
Matrix Composition ◽  
In Vitro Experiments ◽  
Current Cell ◽  
In Vitro Systems

Biomimetic microenvironments are key components to successful cell culture and tissue engineering in vitro. One of the most accurate biomimetic microenvironments is that made by the cells themselves. Cell-made microenvironments are most similar to the in vivo state as they are cell-specific and produced by the actual cells which reside in that specific microenvironment. However, cell-made microenvironments have been challenging to re-create in vitro due to the lack of extracellular matrix composition, volume and complexity which are required. By applying macromolecular crowding to current cell culture protocols, cell-made microenvironments, or cell-derived matrices, can be generated at significant rates in vitro. In this review, we will examine the causes and effects of macromolecular crowding and how it has been applied in several in vitro systems including tissue engineering.

Nitrite and Red Cell Function in Carp: Control Factors for Nitrite Entry, Membrane Potassium Ion Permeation, Oxygen Affinity and Methaemoglobin Formation

1990 ◽  
Vol 152 (1) ◽  
pp. 149-166 ◽  
Author(s):  
FRANK B. JENSEN
Keyword(s):  
Cell Function ◽  
Oxygen Affinity ◽  
Red Cells ◽  
Red Cell ◽  
In Vitro Experiments ◽  
K Uptake ◽  
The Mean ◽  
K Release

Red cell function was studied in carp by a combination of in vivo and in vitro experiments with nitrite as the perturbing agent. In vivo accumulation of nitrite caused a marked increase in the red cell methaemoglobin content, and reduced the mean cellular volume. The oxygen affinity of unoxidized haemoglobin was strongly decreased, partly as result of the elevated concentration of cellular nucleoside triphosphates and haemoglobin associated with red cell shrinkage. Red cell pH was unchanged compared to controls, but reduced when referred to constant extracellular pH and O2 saturation. The mean cellular K+ content decreased, reflecting a K+ loss from the red cells during their shrinkage. This K+ loss contributed significantly to the large plasma hyperkalaemia during nitrite exposure. In vitro experiments revealed that nitrite influx into deoxygenated red cells was much larger than into oxygenated red cells. Nitrite permeation of the red cell membrane was not inhibited by DIDS and did not change extracellular pH. Methaemoglobin (MetHb) formation was more pronounced in deoxygenated blood than in oxygenated blood, but quasi-steady states were reached, reflecting a balance between nitrite-induced MetHb formation and the action of MetHb reductase. Red cells incubated in the oxygenated state released K+, whereas a net K+ uptake occurred in deoxygenated cells. Nitrite did not change the K+ loss from oxygenated cells, but shifted the K+ uptake in deoxygenated cells to a pronounced K+ release by the time high MetHb levels were reached. Both types of red cell K+ release were inhibited by DIDS and appeared to occur via a route involving Band 3. The data are consistent with the hypothesis that a significant DIDS-sensitive K+ efflux from the red cells occurs whenever a large fraction of the haemoglobin molecules assumes an R-like quaternary structure.

scholarly journals A Multimodal Stimulation Cell Culture Bioreactor for Tissue Engineering: A Numerical Modelling Approach

Polymers ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 940
Author(s):  
João Meneses ◽  
João C. Silva ◽  
Sofia R. Fernandes ◽  
Abhishek Datta ◽  
Frederico Castelo Ferreira ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Cell Culture ◽  
Numerical Modelling ◽  
3D Design ◽  
Digital Twins ◽  
Numerical Stimulation ◽  
Modelling Methods ◽  
Selection Of

The use of digital twins in tissue engineering (TE) applications is of paramount importance to reduce the number of in vitro and in vivo tests. To pursue this aim, a novel multimodal bioreactor is developed, combining 3D design with numerical stimulation. This approach will facilitate the reproducibility between studies and the platforms optimisation (physical and digital) to enhance TE. The new bioreactor was specifically designed to be additive manufactured, which could not be reproduced with conventional techniques. Specifically, the design suggested allows the application of dual stimulation (electrical and mechanical) of a scaffold cell culture. For the selection of the most appropriate material for bioreactor manufacturing several materials were assessed for their cytotoxicity. Numerical modelling methods were then applied to the new bioreactor using one of the most appropriate material (Polyethylene Terephthalate Glycol-modified (PETG)) to find the optimal stimulation input parameters for bone TE based on two reported in vitro studies.

Poly(glycerol sebacate) – a revolutionary biopolymer

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Israd H. Jaafar ◽  
Sabrina S. Jedlicka ◽  
John P. Coulter
Keyword(s):  
Mechanical Properties ◽  
Tissue Engineering ◽  
Cell Culture ◽  
Surface Modification ◽  
Mammalian Cell Culture ◽  
Chemical Properties ◽  
Tunable Mechanical Properties ◽  
And Chemical Properties

Abstract Novel materials possessing physical, mechanical, and chemical properties similar to those found in vivo provide a potential platform for building artificial microenvironments for tissue engineering applications. Poly(glycerol sebacate) is one such material. It has tunable mechanical properties within the range of common tissue, and favorable cell response without surface modification with adhesive ligands, and biodegradability. In this chapter, an overview of the material is presented, focusing on synthesis, characterization, microfabrication, use as a substrate in in vitro mammalian cell culture, and degradation characteristics.

Developments in three-dimensional cell culture technology aimed at improving the accuracy of in vitro analyses

2010 ◽  
Vol 38 (4) ◽  
pp. 1072-1075 ◽  
Author(s):  
Daniel J. Maltman ◽  
Stefan A. Przyborski
Keyword(s):  
Cell Culture ◽  
Cell Growth ◽  
Cultured Cells ◽  
Ex Vivo ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Proliferation And Differentiation ◽  
In Vitro Systems

Drug discovery programmes require accurate in vitro systems for drug screening and testing. Traditional cell culture makes use of 2D (two-dimensional) surfaces for ex vivo cell growth. In such environments, cells are forced to adopt unnatural characteristics, including aberrant flattened morphologies. Therefore there is a strong demand for new cell culture platforms which allow cells to grow and respond to their environment in a more realistic manner. The development of 3D (three-dimensional) alternative substrates for in vitro cell growth has received much attention, and it is widely acknowledged that 3D cell growth is likely to more accurately reflect the in vivo tissue environments from which cultured cells are derived. 3D cell growth techniques promise numerous advantages over 2D culture, including enhanced proliferation and differentiation of stem cells. The present review focuses on the development of scaffold technologies for 3D cell culture.

scholarly journals Cell Microenvironment Engineering and Monitoring for Tissue Engineering and Regenerative Medicine: The Recent Advances

2014 ◽  
Vol 2014 ◽  
pp. 1-18 ◽  
Author(s):  
Julien Barthes ◽  
Hayriye Özçelik ◽  
Mathilde Hindié ◽  
Albana Ndreu-Halili ◽  
Anwarul Hasan ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Regenerative Medicine ◽  
Control Cell ◽  
Matrix Composition ◽  
Tissue Engineering Scaffolds ◽  
Cell Microenvironment ◽  
Engineered Tissues ◽  
Cellular Behaviour

In tissue engineering and regenerative medicine, the conditions in the immediate vicinity of the cells have a direct effect on cells’ behaviour and subsequently on clinical outcomes. Physical, chemical, and biological control of cell microenvironment are of crucial importance for the ability to direct and control cell behaviour in 3-dimensional tissue engineering scaffolds spatially and temporally. In this review, we will focus on the different aspects of cell microenvironment such as surface micro-, nanotopography, extracellular matrix composition and distribution, controlled release of soluble factors, and mechanical stress/strain conditions and how these aspects and their interactions can be used to achieve a higher degree of control over cellular activities. The effect of these parameters on the cellular behaviour within tissue engineering context is discussed and how these parameters are used to develop engineered tissues is elaborated. Also, recent techniques developed for the monitoring of the cell microenvironmentin vitroandin vivoare reviewed, together with recent tissue engineering applications where the control of cell microenvironment has been exploited. Cell microenvironment engineering and monitoring are crucial parts of tissue engineering efforts and systems which utilize different components of the cell microenvironment simultaneously can provide more functional engineered tissues in the near future.

Hollow Shells Development and Characterization for Cells Carrying Purpose

2016 ◽  
Vol 696 ◽  
pp. 238-242
Author(s):  
Thomas Miramond ◽  
Pascal Borget ◽  
Françoise Moreau ◽  
Borhane Fellah ◽  
G. Daculsi
Keyword(s):  
Tissue Engineering ◽  
Cell Adhesion ◽  
Bone Tissue Engineering ◽  
Surface Area ◽  
In Vitro Experiments ◽  
Hollow Shell ◽  
Physico Chemical ◽  
Set Up

Bioceramics draw attention in bone tissue engineering field since their biomimetic properties regarding bone attribute. In this context, a concept of smart bioceramics granules made of Hydroxyapatite have been set up, enhancing surface area available to body fluids containing proteins and cell adhesion for bone forming respectively thanks to microporosities and macropore concavities. New “hollow shell” granules were developed and assessed by physico-chemical characterizations, in-vitro experiments and in-vivo implantation in comparison with classical round granules. This new original galenic formulation showed promising potential in cell carrying and osteoconduction matter.

scholarly journals Comparison of chondrogenesis-related biological behaviors between human urine-derived stem cells and human bone marrow mesenchymal stem cells from the same individual

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jiachen Sun ◽  
Fei Xing ◽  
Min Zou ◽  
Min Gong ◽  
Lang Li ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Bone Marrow ◽  
Cartilage Repair ◽  
Cartilage Defects ◽  
In Vitro Experiments ◽  
Rabbit Knee ◽  
Marrow Mesenchymal Stem Cells

Abstract Background Stem cells are the main choice for seed cells in tissue engineering, but using most traditional stem cells requires invasive and complicated procedures. Human urine-derived stem cells (hUSCs) are an alternative stem cell source with the advantages of being isolated noninvasively and repetitively from the same individual. The aim of this study was to compare chondrogenesis-related biological behaviors between hUSCs and human bone marrow mesenchymal stem cells (hBMSCs) from the same individual. Methods hUSCs and hBMSCs were isolated from six patients who underwent iliac bone grafting. Cell morphology, proliferation, colony-forming, migration, and multidifferentiation analyses were performed in vitro. Then, acellular cartilage extracellular matrix (ACM) scaffolds were fabricated for in vivo implantation. The comparisons of cell viability, morphology, proliferation, and chondrogenesis between hUSCs and hBMSCs cultured on scaffolds were performed before implantation. The scaffolds loaded with hUSCs or hBMSCs were implanted into a rabbit knee model to repair cartilage defects. Magnetic resonance imaging (MRI) and micro-computed tomography (μCT) Analyses, inflammation and toxicity assays, gross observation, and histological evaluation were performed to evaluate the cartilage repair effects. Results In in vitro experiments, hUSCs had better capacity for proliferation, colony-forming, and migration compared to hBMSCs in the same passage, while hBMSCs had greater osteogenic, adipogenic, and chondrogenic abilities compared to hUSCs in the same passage. Both hUSCs and hBMSCs at passage 3 had the strongest potential for proliferation, colony-forming, and multilineage differentiation compared to cells in other passages. The ACM scaffolds loaded with hUSCs or hBMSCs both significantly promoted the repair of cartilage defects in the rabbit knee model at 12 weeks’ postimplantation, and the new tissue was mainly hyaline cartilage. However, there was no significant difference in cartilage repair effects between hUSCs and hBMSCs. Conclusions In in vitro experiments, hUSCs presented better capacity for proliferation, while hBMSCs had greater chondrogenic ability. However, hUSCs and hBMSCs had similar cartilage repair effects in vivo. Results indicated that hUSCs can be a stem cell alternative for cartilage regeneration and provide a powerful platform for cartilage tissue engineering and clinical transformation. Graphical abstract

Decellularized bone extracellular matrix in skeletal tissue engineering

2020 ◽  
Vol 48 (3) ◽  
pp. 755-764
Author(s):  
Benjamin B. Rothrauff ◽  
Rocky S. Tuan
Keyword(s):  
Tissue Engineering ◽  
Bone Regeneration ◽  
Tissue Regeneration ◽  
Animal Studies ◽  
Tumor Resection ◽  
Regenerative Capacity ◽  
Skeletal Tissue ◽  
Large Bone

Bone possesses an intrinsic regenerative capacity, which can be compromised by aging, disease, trauma, and iatrogenesis (e.g. tumor resection, pharmacological). At present, autografts and allografts are the principal biological treatments available to replace large bone segments, but both entail several limitations that reduce wider use and consistent success. The use of decellularized extracellular matrices (ECM), often derived from xenogeneic sources, has been shown to favorably influence the immune response to injury and promote site-appropriate tissue regeneration. Decellularized bone ECM (dbECM), utilized in several forms — whole organ, particles, hydrogels — has shown promise in both in vitro and in vivo animal studies to promote osteogenic differentiation of stem/progenitor cells and enhance bone regeneration. However, dbECM has yet to be investigated in clinical studies, which are needed to determine the relative efficacy of this emerging biomaterial as compared with established treatments. This mini-review highlights the recent exploration of dbECM as a biomaterial for skeletal tissue engineering and considers modifications on its future use to more consistently promote bone regeneration.

In vitro- / in vivo- Untersuchungen zur Biokompatibilität und Bioresorption amorpher Kieselgelfaser für das Tissue engineering humaner Knorpelgewebe

2004 ◽  
Vol 83 (02) ◽  
Author(s):  
A Haisch ◽  
A Evers ◽  
K Jöhrens-Leder ◽  
S Jovanovic ◽  
B Sedlmaier ◽  
...  
Keyword(s):  
Tissue Engineering
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