Heat denaturation of fibrinogen to develop a biomedical matrix

Marx, Gerard; Mou, Xiaode; Hotovely-Salomon, Anna; Levdansky, Lilia; Gaberman, Elena; Belenky, Dimitry; Gorodetsky, Raphael

doi:10.1002/jbm.b.30842

Cited by 35 publications

(26 citation statements)

References 45 publications

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“…We suspected the large difference in the amount of protein recovered was due to precipitation of fibrinogen in the plasma samples. Indeed, removal of the fibrinogen by pre-incubation at 56°C for 3 min (33,34) reduced the levels of extraneous protein in the acetate precipitated samples to levels comparable to those obtained for the serum samples with essentially no loss of exosomes. As discussed above, extraneous proteins in the exosome preparation should not pose any problems for transcriptomic analysis.…”

Section: Discussionmentioning

confidence: 77%

A novel “salting-out” procedure for the isolation of tumor-derived exosomes

Brownlee

Lynn

Thorpe

et al. 2014

Journal of Immunological Methods

105

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The last decade has seen an exponential growth in the number of exosome-related publications. Although many of these studies have used exosomes from biological fluids (blood, and ascites or pleural effusions) the vast majority employed vesicles isolated from large volumes of tissue culture supernatants. While several techniques are available for their isolation, all require a significant reduction in volume to obtain sufficient concentrations for study. One approach is to concentrate the medium before proceeding with their isolation, however, these procedures are very time consuming and require specialized laboratory equipment. Here we provide a new and effective method for the isolation of tumor-derived exosomes based on “charge neutralization” with acetate. We show that titration of tissue culture supernatants with 0.1M acetate to pH 4.75 results in immediate precipitation of virtually all the exosomes. The precipitated exosomes can be washed to remove residual media and are readily “resolubilized” upon resuspension in acetate-free buffer at neutral pH. This simple cost effective method significantly increases the yield of exosomes from an unlimited quantity of culture supernatants. Exosomes isolated by this technique are indistinguishable from exosomes recovered by direct ultracentrifugation.

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Section: Discussionmentioning

confidence: 77%

A novel “salting-out” procedure for the isolation of tumor-derived exosomes

Brownlee

Lynn

Thorpe

et al. 2014

Journal of Immunological Methods

105

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“…Little denaturation by heat occurs during a fabrication step, but prepared microspheres still maintain its major functions. 26 By controlling stirring speed and heat, we could prepare the spherical microspheres with highly porous surface structure, offering cells sufficient space for attachment. 27 However, the differences between bare-Fbg microspheres and DOX-linker-Fbg microspheres were found on their surface density.…”

Section: Discussionmentioning

confidence: 99%

Biocompatible and biodegradable fibrinogen microspheres for tumor-targeted doxorubicin delivery

Joo¹,

Park²,

An³

2015

IJN

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In the development of effective drug delivery carriers, many researchers have focused on the usage of nontoxic and biocompatible materials and surface modification with targeting molecules for tumor-specific drug delivery. Fibrinogen (Fbg), an abundant glycoprotein in plasma, could be a potential candidate for developing drug carriers because of its biocompatibility and tumor-targeting property via arginine–glycine–aspartate (RGD) peptide sequences. Doxorubicin (DOX), a chemotherapeutic agent, was covalently conjugated to Fbg, and the microspheres were prepared. Acid-labile and non-cleavable linkers were used for the conjugation of DOX to Fbg, resulting in an acid-triggered drug release under a mild acidic condition and a slow-controlled drug release, respectively. In vitro cytotoxicity tests confirmed low cytotoxicity in normal cells and high antitumor effect toward cancer cells. In addition, it was discovered that a longer linker could make the binding of cells to Fbg drug carriers easier. Therefore, DOX–linker–Fbg microspheres could be a suitable drug carrier for safer and effective drug delivery.

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“…Due to the conformational transformations of fibrinogen at these high temperatures, FMBs are highly haptotactic to various cell types, including osteogenic bone marrow-derived progenitors. 99,100 For bone tissue engineering studies, the obtained FMBs were incubated for 2 days in a suspension containing whole 97,98,101 Next, authors cultured FMB-MSCs constructs in osteogenic medium and obtained a bone-like structure which was implanted into a mouse skull defect. After 2 months, results showed that defects filled with bone-like tissue had a Ca/P ratio similar to that of the native bone while no repair was seen in the control animals without implants or when the defect was filled with FMBs alone.…”

Section: Different Forms Of Fibrin Applications In Bone Tissue Enginementioning

confidence: 99%

A review of fibrin and fibrin composites for bone tissue engineering

Noori

Ashrafi

Vaez-Ghaemi

et al. 2017

IJN

377

256

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Tissue engineering has emerged as a new treatment approach for bone repair and regeneration seeking to address limitations associated with current therapies, such as autologous bone grafting. While many bone tissue engineering approaches have traditionally focused on synthetic materials (such as polymers or hydrogels), there has been a lot of excitement surrounding the use of natural materials due to their biologically inspired properties. Fibrin is a natural scaffold formed following tissue injury that initiates hemostasis and provides the initial matrix useful for cell adhesion, migration, proliferation, and differentiation. Fibrin has captured the interest of bone tissue engineers due to its excellent biocompatibility, controllable biodegradability, and ability to deliver cells and biomolecules. Fibrin is particularly appealing because its precursors, fibrinogen, and thrombin, which can be derived from the patient’s own blood, enable the fabrication of completely autologous scaffolds. In this article, we highlight the unique properties of fibrin as a scaffolding material to treat bone defects. Moreover, we emphasize its role in bone tissue engineering nanocomposites where approaches further emulate the natural nanostructured features of bone when using fibrin and other nanomaterials. We also review the preparation methods of fibrin glue and then discuss a wide range of fibrin applications in bone tissue engineering. These include the delivery of cells and/or biomolecules to a defect site, distributing cells, and/or growth factors throughout other pre-formed scaffolds and enhancing the physical as well as biological properties of other biomaterials. Thoughts on the future direction of fibrin research for bone tissue engineering are also presented. In the future, the development of fibrin precursors as recombinant proteins will solve problems associated with using multiple or single-donor fibrin glue, and the combination of nanomaterials that allow for the incorporation of biomolecules with fibrin will significantly improve the efficacy of fibrin for numerous bone tissue engineering applications.

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Heat denaturation of fibrinogen to develop a biomedical matrix

Cited by 35 publications

References 45 publications

A novel “salting-out” procedure for the isolation of tumor-derived exosomes

A novel “salting-out” procedure for the isolation of tumor-derived exosomes

Biocompatible and biodegradable fibrinogen microspheres for tumor-targeted doxorubicin delivery

A review of fibrin and fibrin composites for bone tissue engineering

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