Tissue Engineering Using Plant-Derived Cellulose Nanofibrils (CNF) as Scaffold Material

Author(s):  
Kristin Syverud
Author(s):  
Melanie Krüger ◽  
Bart Spee ◽  
Andreas Walther ◽  
Laura De Laporte ◽  
Linda M. Kock

Abstract Nanofibrillar cellulose as a naturally biocompatible scaffold material is very promising for tissue engineering. It is shear thinning but has the downside of not being degradable in animals, it can only be degraded by cellulase enzymes. In this study, a newly developed bioreactor was used to culture fibroblast spheroids under flow conditions inside nanocellulose hydrogels with and without the presence of cellulase. The aim was to control the tissue size and ideally find a match between degradation and tissue formation within this promising material. Both the concentration of cellulase and the flow rate were varied and their influence on the activity and growth of fibroblast clusters was assessed. Cluster diameters, degradation, metabolic activity, and tissue production increase with higher cellulase concentration, although concentrations above 1 g/l does not have an additional benefit. Flow leads to more viable cells, more proliferation and migration, leading to overall larger tissue constructs compared to static conditions. This is most likely due to the shear thinning effect of flow on cellulose nanofibrils (CNFs) in addition to the increased nutrient supply through perfusion. At a constant cellulase concentration of 1 g/l, a flow of 2 ml/min proved to be optimal for tissue production. Therefore, degradation in combination with flow leads to more effective tissue production in CNF hydrogels, which is a very potent scaffold material for tissue engineering.


2009 ◽  
Vol 21 (1) ◽  
pp. 237 ◽  
Author(s):  
D. Kim ◽  
A. J. Maki ◽  
H.-J. Kong ◽  
E. Monaco ◽  
M. Bionaz ◽  
...  

Adipose tissue presents an appealing alternative to bone marrow as a source of mesenchymal stem cells (MSC). However, in order to enhance cell proliferation and differentiation, 3-dimensional (3-D) culture may be required. A 3-D culture has benefits due to its more in vivo-like environment. Further, to form a functional tissue, a scaffold material is required to ensure proper shape and allow for efficient delivery of nutrients and growth factors. Alginate, a resorbable hydrogel, is a potential injectable scaffold for fat and bone tissue engineering due to its high biocompatibility, gelation with calcium and slow dissolution in a physiologic environment. In the present study, we examined the viability, gene expression and morphology of MSC, isolated from porcine adipose (ADSC) and bone marrow (BMSC), during osteogenic and adipogenic differentiation in a 3D alginate hydrogel environment for 0, 7 and 14 days (d). ADSC and BMSC were infused into alginate hydrogels, which polymerized upon the addition of Ca+2 ions. Both stem cell types were differentiated into osteoblasts using 0.1 μm dexamethasone, 10 mm beta glycerophosphate and 50 μm ascorbic acid, whereas adipocytes were differentiated using 10 μm insulin, 1 μm dexamethasone, and 0.5 mm IBMX. Osteogenic differentiation was confirmed using alkaline phosphatase, Von Kossa, and alizarin red S staining and adipogenic differentiation was confirmed using Oil Red O. Cell viability and proliferation was quantified using the MTT assay. Gene expression was measured using qPCR. The morphology of ADSC and BMSC differentiated toward osteogenic lineages changed with both cell types forming osteogenic nodules over time. The nodules formed by ADSC were larger in diameter than those formed by BMSC. Unlike the osteogenic cells that formed nodules, the ADSC and BMSC differentiated into adipogenic cells showed no significant changes in cell size or aggregation. Gene expression results indicated increased PPARG expression in BMSC with time whereas ADSC showed a peak of expression on day 7 and then decreased. ADSC showed increased (14-fold) PPRG expression when compared with BMSC. ADSC had 160-fold less expression of ALP than BMSC. BMSC showed a 16-fold higher expression level of BGLAP than ADSC. ADSC showed a 15.8% higher expression than BMSC for COL1a1. Both ADSC and BMSC showed similar trends SPARC expression, but BMSC had a 12-fold higher expression of SPP1 than ADSC. In summary, both types of mesenchymal stem cells successfully differentiated into both lineages and maintained viability in the hydrogel over time. In conclusion, alginate is a viable scaffold material for the differentiation of mesenchymal stem cells for tissue engineering applications. These results allow for future studies using the pig as an in vivo fat and bone tissue engineering model. This research was supported by the Illinois Regenerative Medicine Institute.


Author(s):  
Ana L. F. Soares ◽  
Maria Stekelenburg ◽  
Frank Baaijens

Small-diameter vessels tissue engineering (TE) seeks to provide viable replacements to the native ones. The approach involves seeding autologous cells onto a biodegradable scaffold material and delivering the suitable environmental factors (biochemical and biomechanical stimuli) during culture via a conditioning protocol. Still, these replacements appear to lack the native mechanical integrity. TE protocols should then be improved and optimized.


Author(s):  
Andrew P. Breidenbach ◽  
Nathaniel A. Dyment ◽  
Yinhui Lu ◽  
Jason T. Shearn ◽  
David W. Rowe ◽  
...  

Tendon and ligament injuries account for one-third of all musculoskeletal injuries [1]. Collagen fibrils in these mechanosensitive tissues transmit forces to mobilize and stabilize joint movement. Donor tissues used to repair these tissues often lack the mechanical properties of the tissue they are replacing. One promising alternative using tissue engineering combines stem/progenitor cells in three-dimensional tissue engineered constructs (TECs).


MRS Advances ◽  
2017 ◽  
Vol 2 (24) ◽  
pp. 1309-1314 ◽  
Author(s):  
Pengfei Duan ◽  
Nehir Kandemir ◽  
Jiajun Wang ◽  
Jinju Chen

ABSTRACTHydrogels have been widely used in many applications from tissue engineering to drug delivery systems. For both tissue engineering and drug delivery, the mechanical properties are important because they would affect cell-materials interactions and injectability of drugs encapsulated in hydrogel carriers. Therefore, it is important to study the mechanical properties of these hydrogels, particularly at physiological temperature (37°C). This study adopted strain sweep and frequency sweep rotational rheological tests to investigate the rheological characteristics of various tissue engineering relevant hydrogels with different concentrations at 37°C. These hydrogels include alginate, RGD-alginate, and copolymerized collagen/alginate/fibrin. It has revealed that the addition of RGD has negligible effect on the elastic modulus and viscosity of alginate. Alginate gels have demonstrated shear thinning behavior which indicates that they are suitable candidates as carriers for cells or drug delivery. The addition of collagen and fibrin would reinforce the mechanical properties of alginate which makes it a strong scaffold material.


2013 ◽  
Vol 16 (4) ◽  
pp. 843-848 ◽  
Author(s):  
O. Drewnowska ◽  
B. Turek ◽  
B. Carstanjen ◽  
Z. Gajewski

Abstract Biomaterials originate from natural substances and are widely used in medicine. Although they have to satisfy many conditions to be useful for treatment, more and more research is carried out with new types of biomaterials that can help replace various tissues such as tendons and bones. Chitosan is a very promising material, revealing unique features, which makes it useful for veterinary medicine - antimicrobial activity, biocompatibility, biodegradability. It is also known as good scaffold material, especially when combined with other polymers. This article describes chitosan as a biomaterial and tissue engineering scaffold with possible applications in veterinary medicine


RSC Advances ◽  
2015 ◽  
Vol 5 (31) ◽  
pp. 23999-24008 ◽  
Author(s):  
Navdeep Manhas ◽  
K. Balasubramanian ◽  
P. Prajith ◽  
Prashant Rule ◽  
Sunil Nimje

The process of extraction of cellulose nanofibrils by steam explosion followed by electrospinning with biodegradable polymers to yield PCL/PVA nanoencapsulated cellulosic reinforcing fillers for tissue engineering applications.


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