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admin | Natural Testosterone Replacement | 02.10.2014
Hyaluronic acid (HA) is a biological polymer found ubiquitously in the extracellular matrix (ECM) of several tissues. The ability for a hydrogel to bind proteins (such as growth factors or cytokines) is an important design parameter to consider for protein-mediated tissue remodeling.
To address this limitation of HA, recent research in the Burdick Lab at the University of Pennsylvania focuses on designing HA polymers with sulfate groups to create robust hydrogels capable of protein binding and release. Researchers took advantage of carboxylic acid residues on HA to generate modified HA polymers with hydroxyethyl methacrylate (HEMA-HA).
The binding affinities of HEMA-HA and HEMA-SHA were characterized using protein-binding assays of stromal cell-derived factor-1? (SDF-1?).
After characterizing the binding efficiency of the polymers, HEMA-SHA and HEMA-HA were used to fabricate hydrogels. This work describes a novel method to easily incorporate sulfate groups into HA hydrogels for enhanced binding and prolonged release of protein. To keep up-to-date with all the latest research, sign-up to our RSS feed or Table of contents alert. This study used specific biomaterials that draw inspiration from nature.  Specifically, the researchers used elastin, a commonly found biomaterial, as a template, and created a biomaterial with a similar protein composition.
This work is an elegant demonstration of how biomaterials and cells can be combined to produce a viable tissue-engineered construct that may one day be used as corneal replacement therapy.
The ability for a stem cell to differentiate into a specific cell type depends on the cell’s surrounding microenvironment. Researchers at Tohoku University in Japan have fabricated “honeycomb” topological substrates to regulate the differentiation of mesenchymal stem cells. After obtaining four different honeycomb films with varying pore size and strut width, human mesenchymal stem cells (hMSCs) were seeded on the surfaces to evaluate cell differentiation into osteo-specific (bone) and myo-specific (muscle) cells. Interestingly, when both the pore size and strut width were smaller than hMSC size, cells showed globular morphologies and increased expression of osteopotin (consistent with osteo-specific differentiation). This study demonstrates that geometries of honeycomb topographical substrates may have a direct influence on stem cell differentiation.
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HA has an abundance of carboxylic acid groups along the polymer chain, making HA amenable to a variety of chemical modifications – including the addition of sulfate groups.
Interestingly, SDF-1? binding to sulfated HA was comparable to heparin controls, and virtually no SDF-1? was bound to non-sulfated HA.  The enhanced binding to HEMA-SHA is likely due to the increased negative charge from the sulfate groups on the HEMA-SHA (confirmed with zeta potential measurements). Using a 90%-10% blend of HEMA-HA and HEMA-SHA and a 100% solution HEMA-HA as a control, the polymer solutions were cross-linked into hydrogels using a redox initiator system for free radical cross-linking.
Candidate and NSF Fellow in the Biomedical Engineering department at Northwestern University. To further test these materials, the biomaterial sheets were then immersed in a saline solution for 4 weeks to study non-enzyme based degradation, and the crosslinked sheets again resisted degradation better than the non-crosslinked sheets.
The cells were able to survive on the biomaterial and grew well on the micropatterned biomaterial sheets.
Variables such as substrate stiffness, presence of certain growth factors, fluid flow, and extracellular matrix composition in the cell culture environment all play a role in the cell’s ability to differentiate.
The substrates are manufactured using a chloroform solution of polystyrene and an amphiphilic co-polymer, poly(N-dodecyl acrylamide-co-6-acrylamidehexanoic acid), or dioleoylphosphatidylethanolamine (DOPE). To determine the cell differentiation state, cells were stained with osteoblast and myoblast markers, osteopontin and MyoD1, respectively. On the other hand, when the pore size was the same size as hMSCs, the cells expressed MyoD1 and had elongated morphologies (consistent with myo-specific differentiation).
Surface topology of biomaterial surfaces must be taken into account for future designs of cell culture environments for stem cell differentiation studies.
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The ester groups present in the cross-links also make the gel susceptible to hydrolytic degradation, which is an important feature for temporal release of bound protein from the hydrogel.


Rheometry data show that the presence of sulfate groups in the hydrogel does not alter gel cross-linking kinetics and gel stiffness. The crosslinked sheets were also able to transmit light, which is crucial for their application as corneal replacements.
The cells also followed the alignment of the micropatterns on the surface of the biomaterials. Additionally, there is great interest in studying the effects of surface topology on stem cell differentiation in vitro to more accurately mimic the in vivo microenvironment. The solutions were cast on a glass slide and exposed to a flow of humid air to generate polystyrene films that resemble honeycomb from a beehive. Given the promising morphological and staining results, further studies are required to fully characterize the molecular mechanisms of the differentiation process. This is available in different flavors smooth coconut, chocolate, caramel cheesecake, vanilla, cookies and cream, banana, chocolate mint and strawberry. ZMA increase pro-anabolic hormones at the same time peptide bonded glutamine enhance skeletal protein synthesis. This support maximizes lean muscle development and recovers muscles after intense training. Diet Matrix is a unique supplement that is powerful protein powder which enables you to develop and maintain muscle tone. Additionally, HEMA-HA polymers were allowed to react with SO3 to form sulfated HEMA-HA (HEMA-SHA).
Additionally, the sulfated HA hydrogel showed prolonged release of SDF-1? over time compared to the non-sulfated hydrogel control.
The researchers then placed multiple layers of the micropatterned scaffolds together such that each successive patterned layer was perpendicular to the previous layer, and ‘glued’ the scaffolds together using droplets of collagen.  Cells were then seeded onto each layer using a syringe. Using various micro-fabrication technologies, 2D and 3D substrates with varying topographic features including grooves, pits, pores, and posts can be fabricated and tested with cells to evaluate cell response. The honeycomb substrates had varying pore size and strut width depending on the chloroform evaporation time.
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