Macromolecular Crowding: The Next Frontier in Tissue Engineering

2014 ◽  
Vol 96 ◽  
pp. 1-8 ◽  
Author(s):  
Pramod Kumar ◽  
Abhigyan Satyam ◽  
Diana Gaspar ◽  
Daniela Cigognini ◽  
Clara Sanz-Nogués ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Self Assembly ◽  
Macromolecular Crowding ◽  
Fold Increase ◽  
Collagen Type I ◽  
Dermal Fibroblasts ◽  
Cell Phenotype ◽  
Type I ◽  
Matrix Deposition

Tissue engineering by self-assembly hypothesises that optimal repair and regeneration can be achieved best by using the cells’ inherent ability to create organs with proficiency still unmatched by currently available scaffold fabrication technologies. However, the prolonged culture time required to develop an implantable device jeopardises clinical translation and commercialisation of such techniques. Herein, we report that macromolecular crowding, a biophysical in vitro microenvironment modulator, dramatically accelerates extracellular matrix deposition in cultured human corneal, lung and dermal fibroblasts and human bone marrow mesenchymal stem cells. In fact, an almost 5 to 30 fold increase in collagen type I deposition was recorded as early as 48 hours in culture, without any negative effect in cell phenotype and function.

scholarly journals Collagen-Based Electrospun Materials for Tissue Engineering: A Systematic Review

2021 ◽  
Vol 8 (3) ◽  
pp. 39
Author(s):  
Britani N. Blackstone ◽  
Summer C. Gallentine ◽  
Heather M. Powell
Keyword(s):  
Systematic Review ◽  
Tissue Engineering ◽  
Collagen Type I ◽  
The Body ◽  
Pool Data ◽  
Type I ◽  
High Concentrations ◽  
Increased Strength

Collagen is a key component of the extracellular matrix (ECM) in organs and tissues throughout the body and is used for many tissue engineering applications. Electrospinning of collagen can produce scaffolds in a wide variety of shapes, fiber diameters and porosities to match that of the native ECM. This systematic review aims to pool data from available manuscripts on electrospun collagen and tissue engineering to provide insight into the connection between source material, solvent, crosslinking method and functional outcomes. D-banding was most often observed in electrospun collagen formed using collagen type I isolated from calfskin, often isolated within the laboratory, with short solution solubilization times. All physical and chemical methods of crosslinking utilized imparted resistance to degradation and increased strength. Cytotoxicity was observed at high concentrations of crosslinking agents and when abbreviated rinsing protocols were utilized. Collagen and collagen-based scaffolds were capable of forming engineered tissues in vitro and in vivo with high similarity to the native structures.

Inhibition Of Collagen And ADP-Induced Platelet Aggregation By Plasma Fibronectin

1981 ◽  
Author(s):  
D G Moon ◽  
J E Kaplan
Keyword(s):  
Gradient Centrifugation ◽  
Fold Increase ◽  
Lag Time ◽  
Collagen Type I ◽  
Type I ◽  
Plasma Fibronectin ◽  
Aggregation Rate ◽  
Tail Tendon ◽  
Induced Aggregation

Platelets contain fibronectin a glycoprotein with an established affinity for collagen. This observation has led other investigators to postulate that fibronectin is the platelet collagen receptor. The much greater concentration of fibronectin in the plasma surrounding platelets, however, has led us to suggest that plasma fibronectin may bind to collagen and competitively inhibit the platelet- collagen interaction. Rat platelets were isolated by Stractan density gradient centrifugation and aggregated with acid-solubilized rat tail tendon collagen (Type I) in a Payton 300B Aggregometer. Fibronectin was twice purified by affinity chromatography with gelatin linked to CNBr- activated Sepharose 4B. Simultaneous addition of 50 μg fibronectin and 25 μg collagen to platelets suspended in Tyrodes solution at 37°C resulted in a 2-fold increase in lag time and a 30% decrease in aggregation rate as compared to control values. When collagen was preincubated in Tyrodes solution for 12 minutes at 26°C without platelets to allow for prior fibrillogenesis, the addition of 50 μg fibronectin with the platelets resulted in <20% increase in lag time and a 20-30% decrease in aggregation rate. In a separate series of experiments, fibronectin was also found to inhibit ADP-induced aggregation. In this case, the initial rate of aggregation was comparable with and without fibronectin, but this maximal rate was maintained for a shorter period in the presence of fibronectin. Thus, fibronectin reduced the in vitro aggregation response to two different physiological stimuli. Our data supports previous studies which indicate that fibronectin reduces the reactivity of platelets with collagen and provides evidence of a role for fibronectin in modulating platelet responses in the absence of collagen.

scholarly journals Padina pavonica Extract Promotes In Vitro Differentiation and Functionality of Human Primary Osteoblasts

Marine Drugs ◽  
2019 ◽  
Vol 17 (8) ◽  
pp. 473
Author(s):  
Mariagiulia Minetti ◽  
Giulia Bernardini ◽  
Manuele Biazzo ◽  
Gilles Gutierrez ◽  
Michela Geminiani ◽  
...  
Keyword(s):  
Healthy Lifestyle ◽  
Fold Increase ◽  
Nuclear Factor Κb ◽  
Collagen Type I ◽  
Bone Diseases ◽  
Type I ◽  
Bone Homeostasis ◽  
Safe Product ◽  
Differentiation Stage

Marine algae have gained much importance in the development of nutraceutical products due to their high content of bioactive compounds. In this work, we investigated the activity of Padina pavonica with the aim to demonstrate the pro-osteogenic ability of its extract on human primary osteoblast (HOb). Our data indicated that the acetonic extract of P. pavonica (EPP) is a safe product as it did not show any effect on osteoblast viability. At the same time, EPP showed to possess a beneficial effect on HOb functionality, triggering their differentiation and mineralization abilities. In particular EPP enhanced the expression of the earlier differentiation stage markers: a 5.4-fold increase in collagen type I alpha 1 chain (COL1A1), and a 2.3-fold increase in alkaline phosphatase (ALPL), as well as those involved in the late differentiation stage: a 3.7-fold increase in osteocalcin (BGLAP) expression and a 2.8-fold in osteoprotegerin (TNFRSF11B). These findings were corroborated by the enhancement in ALPL enzymatic activity (1.7-fold increase) and by the reduction of receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) ratio (0.6-fold decrease). Moreover, EPP demonstrated the capacity to enhance the bone nodules formation by 3.2-fold in 4 weeks treated HOb. Therefore, EPP showed a significant capability of promoting osteoblast phenotype. Given its positive effect on bone homeostasis, EPP could be used as a useful nutraceutical product that, in addition to a healthy lifestyle and diet, can be able to contrast and prevent bone diseases, especially those connected with ageing, such as osteoporosis (OP).

In Vitro Osteogenic, Angiogenic, and Inflammatory Effects of Copper in β-Tricalcium Phosphate

MRS Advances ◽  
2019 ◽  
Vol 4 (21) ◽  
pp. 1253-1259
Author(s):  
Weiguo Han ◽  
Haley Cummings ◽  
Murali Krishna Duvuuru ◽  
Sarah Fleck ◽  
Sahar Vahabzadeh ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Tricalcium Phosphate ◽  
Physical And Mechanical Properties ◽  
Early Time ◽  
Collagen Type I ◽  
Type I ◽  
Dependent Manner ◽  
Dental Tissue ◽  
Engineering Applications

AbstractTricalcium phosphate (TCP) is a promising candidate in bone and dental tissue engineering applications. Though osteoconductive, its low osteoinductivity is a major concern. Trace elements addition at low concentrations are known for their impact on not only the osteoinductivity, but also physical and mechanical properties of TCP. Copper (Cu) is known for its role in vascularization and angiogenesis in biological systems. Here, we studied the effects of Cu addition on phase composition, porosity, microstructure and in vitro interaction with osteoblast (OB) cells. Our results showed that Cu stabilized the TCP structure, while no significant effect of microstructure and porosity was found. Cu at concentrations less than 1 wt.% did not have any cytotoxic effect while decreased proliferation of OBs were observed at 1 wt.% Cu doped TCP. Addition of Cu upregulated collagen type I and vascular endothelial growth factor expression in a dose dependent manner at early time-point. Furthermore, Cu reduced inflammatory gene expression by human osteoblasts. These findings show that addition of Cu to TCP may provide a therapeutic strategy that can be applied in bone tissue engineering applications.

scholarly journals 3D PCL/Gelatin/Genipin Nanofiber Sponge as Scaffold for Regenerative Medicine

Materials ◽  
2021 ◽  
Vol 14 (8) ◽  
pp. 2006
Author(s):  
Markus Merk ◽  
Orlando Chirikian ◽  
Christian Adlhart
Keyword(s):  
Tissue Engineering ◽  
Self Assembly ◽  
Ex Vivo ◽  
Three Dimensional ◽  
Dermal Fibroblasts ◽  
Engineering Applications ◽  
Sem Images ◽  
Biologically Relevant ◽  
Naturally Occurring

Recent advancements in tissue engineering and material science have radically improved in vitro culturing platforms to more accurately replicate human tissue. However, the transition to clinical relevance has been slow in part due to the lack of biologically compatible/relevant materials. In the present study, we marry the commonly used two-dimensional (2D) technique of electrospinning and a self-assembly process to construct easily reproducible, highly porous, three-dimensional (3D) nanofiber scaffolds for various tissue engineering applications. Specimens from biologically relevant polymers polycaprolactone (PCL) and gelatin were chemically cross-linked using the naturally occurring cross-linker genipin. Potential cytotoxic effects of the scaffolds were analyzed by culturing human dermal fibroblasts (HDF) up to 23 days. The 3D PCL/gelatin/genipin scaffolds produced here resemble the complex nanofibrous architecture found in naturally occurring extracellular matrix (ECM) and exhibit physiologically relevant mechanical properties as well as excellent cell cytocompatibility. Samples cross-linked with 0.5% genipin demonstrated the highest metabolic activity and proliferation rates for HDF. Scanning electron microscopy (SEM) images indicated excellent cell adhesion and the characteristic morphological features of fibroblasts in all tested samples. The three-dimensional (3D) PCL/gelatin/genipin scaffolds produced here show great potential for various 3D tissue-engineering applications such as ex vivo cell culturing platforms, wound healing, or tissue replacement.

scholarly journals Adapting the Scar-in-a-Jar to Skin Fibrosis and Screening Traditional and Contemporary Anti-Fibrotic Therapies

2021 ◽  
Vol 9 ◽  
Author(s):  
João Q. Coentro ◽  
Ulrike May ◽  
Stuart Prince ◽  
John Zwaagstra ◽  
Olli Ritvos ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Histone Deacetylases ◽  
Transforming Growth Factor ◽  
Dermal Fibroblast ◽  
Trichostatin A ◽  
Macromolecular Crowding ◽  
Skin Fibrosis ◽  
Type I ◽  
Matrix Deposition

Skin fibrosis still constitutes an unmet clinical need. Although pharmacological strategies are at the forefront of scientific and technological research and innovation, their clinical translation is hindered by the poor predictive capacity of the currently available in vitro fibrosis models. Indeed, customarily utilised in vitro scarring models are conducted in a low extracellular matrix milieu, which constitutes an oxymoron for the in-hand pathophysiology. Herein, we coupled macromolecular crowding (enhances and accelerates extracellular matrix deposition) with transforming growth factor β1 (TGFβ1; induces trans-differentiation of fibroblasts to myofibroblasts) in human dermal fibroblast cultures to develop a skin fibrosis in vitro model and to screen a range of anti-fibrotic families (corticosteroids, inhibitors of histone deacetylases, inhibitors of collagen crosslinking, inhibitors of TGFβ1 and pleiotropic inhibitors of fibrotic activation). Data obtained demonstrated that macromolecular crowding combined with TGFβ1 significantly enhanced collagen deposition and myofibroblast transformation. Among the anti-fibrotic compounds assessed, trichostatin A (inhibitors of histone deacetylases); serelaxin and pirfenidone (pleiotropic inhibitors of fibrotic activation); and soluble TGFβ receptor trap (inhibitor of TGFβ signalling) resulted in the highest decrease of collagen type I deposition (even higher than triamcinolone acetonide, the gold standard in clinical practice). This study further advocates the potential of macromolecular crowding in the development of in vitro pathophysiology models.

scholarly journals Dose-Response Tendon-Specific Markers Induction by Growth Differentiation Factor-5 in human bone marrow and umbilical cord Mesenchymal Stem Cells

2020 ◽  
Author(s):  
Maria Camilla Ciardulli ◽  
Luigi Marino ◽  
Erwin P. Lamparelli ◽  
Maurizio Guida ◽  
Nicholas R. Forsyth ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Cell Types ◽  
Collagen Type I ◽  
Width Ratio ◽  
Type I ◽  
Anti Inflammatory

Mesenchymal Stem Cells derived from bone marrow (hBM-MSCs) are utilized in tendon tissue‐engineering protocols while extra-embryonic cord-derived, including from Wharton&rsquo;s Jelly (hWJ-MSC), are emerging as useful alternatives. To explore the tenogenic responsiveness of hBM-MSCs and hWJ-MSCs to hGDF-5 we supplemented each at doses of 1, 10, and 100 ng/mL and determined proliferation, morphology and time-dependent expression of tenogenic markers. We evaluated expression of Collagen types 1 (COL1A1) and 3 (COL3A1), Decorin (DCN), Scleraxis A (SCX-A), Tenascin-C (TNC) and Tenomodulin (TNMD) noting the earliest and largest increase with 100 ng/mL. With 100 ng/mL, hBM-MSCs showed upregulation of SCX-A (1.7-fold) at day 1, TNC (1.3-fold) and TNMD (12-fold) at Day 8. hWJ-MSCs, at the same dose, showed up-regulation of COL1A1 (3-fold), DCN (2.7-fold), SCX (3.8-fold) and TNC (2.3-fold) after 3 days of culture. hWJ-MSCs also showed larger proliferation rate and marked aggregation into a tubular shaped system at Day 7 (with 100 ng/mL of hGDF-5). Simultaneous to this we explored expression of pro-inflammatory (IL-6, TNF, IL-12A, IL-1&beta;) and anti-inflammatory (IL-10, TGF-&beta;1) cytokines across for both cell types. hBM-MSCs exhibited a better balance of pro-inflammatory and anti-inflammatory cytokines upregulating IL-1&beta; (11-fold) and IL-10 (10-fold) at Day 8; hWJ-MSCs, had a slight expression of IL-12A (1.5-fold) but a greater up-regulation of IL-10 (2.5-fold). Collagen type I and tenomodulin proteins, detected by immunofluorescence, confirming the greater protein expression when 100 ng/mL were supplemented. In the same conditions, both cell types showed specific alignment and shape modification (fibroblast-like) with a Lenght/Width ratio increase at value higher than 1, suggesting their response in activating tenogenic commitment events, and they both potential use in 3D in vitro tissue engineering protocols.

Needleless electrospinning of PVP/PGS fibrous scaffolds for skin tissue engineering applications

2018 ◽  
Author(s):  
Antonios Keirouz ◽  
Giuseppino Fortunato ◽  
Anthony Callanan ◽  
Norbert Radacsi
Keyword(s):  
Tissue Engineering ◽  
Tensile Modulus ◽  
Dermal Fibroblasts ◽  
Skin Tissue ◽  
Skin Substitute ◽  
Electrospinning Technique ◽  
Skin Tissue Engineering ◽  
Needleless Electrospinning ◽  
High Concentrations

Scaffolds and implants used for tissue engineering need to be adapted for their mechanical properties with respect to their environment within the human body. Therefore, a novel composite for skin tissue engineering is presented by use of blends of Poly(vinylpyrrolidone) (PVP) and Poly(glycerol sebacate) (PGS) were fabricated via the needleless electrospinning technique. The formed PGS/PVP blends were morphologically, thermochemically and mechanically characterized. The morphology of the developed fibers related to the concentration of PGS, with high concentrations of PGS merging the fibers together plasticizing the scaffold. The tensile modulus appeared to be affected by the concentration of PGS within the blends, with an apparent decrease in the elastic modulus of the electrospun mats and an exponential increase of the elongation at break. Ultraviolet (UV) crosslinking of PGS/PVP significantly decreased and stabilized the wettability of the formed fiber mats, as indicated by contact angle measurements. In vitro examination showed good viability and proliferation of human dermal fibroblasts over the period of a week. The present findings provide important insights for tuning the elastic properties of electrospun material by incorporating this unique elastomer, as a promising future candidate for skin substitute constructs.

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