Silk Materials Functionalized via Genetic Engineering for Biomedical Applications

Deptuch, Tomasz; Dams‐Kozlowska, Hanna

doi:10.3390/ma10121417

Cited by 29 publications

(34 citation statements)

References 115 publications

(207 reference statements)

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“…The materials are often functionalized to better address the needs of the desired application. As mentioned above, bioengineered spider silks are encoded by synthetic genes that are constructed at the DNA level, which provides the opportunity to introduce a DNA sequence that encodes a functional peptide, domain or other protein [14][15][16]. Hybrid (chimeric) silk materials functionalized with binding domains have been reported to efficiently bind nucleic acids, small drugs or proteins [14].…”

Section: Introductionmentioning

confidence: 99%

“…As mentioned above, bioengineered spider silks are encoded by synthetic genes that are constructed at the DNA level, which provides the opportunity to introduce a DNA sequence that encodes a functional peptide, domain or other protein [14][15][16]. Hybrid (chimeric) silk materials functionalized with binding domains have been reported to efficiently bind nucleic acids, small drugs or proteins [14]. Furthermore, silk polymers have also been successfully modified with motifs possessing targeting properties towards specific cells [17][18][19].…”

Section: Introductionmentioning

confidence: 99%

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Drug affinity and targeted delivery: double functionalization of silk spheres for controlled doxorubicin delivery into Her2-positive cancer cells

et al. 2020

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Background: The optimal drug delivery system should be biocompatible, biodegradable, and allow the sustained release of the drug only after it reaches the target cells. Silk, as a natural polymer, is a great candidate for building drug carriers. Genetically engineered silks offer the possibility of functionalization. Previously, we characterized bioengineered silk spheres that were functionalized with H2.1 peptide that selectively delivered a drug to Her2-positive cancer cells. However, drug leakage from the silk spheres showed the need for improved control. Results: To control the drug loading and release, we designed and produced functional silk (DOXMS2) that contains a DOX peptide with an affinity for doxorubicin. The DOXMS2 spheres showed the decreased release of doxorubicin compared with MS2 particles. Next, the DOXMS2 silk was blended with the H2.1MS1 polymer to improve the control of doxorubicin binding and release into Her2-positive cancer cells. The H2.1MS1:DOXMS2 particles showed the highest doxorubicin-loading capacity and binding per cell, which resulted in the highest cytotoxic effect compared with that of other sphere variants. Since drug release at a pH of 7.4 from the blended H2.1MS1:DOXMS2 particles was significantly lower than from blended spheres without DOXMS2 silk, this indicated that such particles could control the release of the drug into the circulatory system before the carrier reached the tumor site. Conclusions: This strategy, which is based on the blending of silks, allows for the generation of particles that deliver drugs in a controlled manner.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Drug affinity and targeted delivery: double functionalization of silk spheres for controlled doxorubicin delivery into Her2-positive cancer cells

et al. 2020

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“…8,9 Moreover, bioengineered spider silk can be functionalized via genetic engineering. 10,11 Nanoparticles made of bioengineered silk that are functionalized with tumor-homing peptides have shown enhanced cellular uptake. [12][13][14][15][16] Previously, we designed a system for targeted cancer therapy that was composed of blended bioengineered silks functionalized with a Her2-binding peptide, ie, H2.1.…”

Section: Introductionmentioning

confidence: 99%

<p>Cellular uptake, intracellular distribution and degradation of Her2-targeting silk nanospheres</p>

Florczak¹,

Maćkiewicz²,

Dams‐Kozlowska³

2019

IJN

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Background: The development of nanocarrier technology has attracted great interest in the last decade. Biodegradable spheres made of functionalized silk have considerable potential to be used as drug delivery systems for cancer treatment. A targeting ligand displayed at the surface of a carrier, with a specific affinity towards a particular receptor, can further enhance the accumulation and uptake of nanoparticles at the site of a tumor. Materials and methods: The hybrid constructs were obtained by adding a Her2-binding peptide (H2.1) to MS1 and MS2 bioengineered silks based on the MaSp1 and MaSp2 proteins from N. clavipes, respectively. The H2.1MS1 and H2.1MS2 proteins were blended at a weight ratio of 8:2. Stable silk particles were formed by mixing a soluble protein with potassium phosphate using a micromixing technique. We used specific inhibitors of endocytosis to determine the cellular uptake pathway of the silk nanoparticles in human Her2positive breast cancer cells. The subcellular distribution of silk particles was investigated by evaluating the signal colocalization with organelle-specific tracker. Moreover, lysosomal and exosomal inhibitors were implemented to evaluate their impact on the silk spheres behavior and degradation. Results: The functionalized spheres were specifically taken up by Her2-positive cancer cells. Silk particles facilitated the entry into cells through both the clathrin-and caveoladependent pathways of endocytosis. Upon entering the cells, the particles accumulated in the lysosomes, where intracellular degradation occurred. Conclusions: The present study demonstrated directly that the lysosomal function was essential for silk-based carrier elimination. The degradation of the carrier is of great importance to develop an optimal drug delivery system.

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“…Recombinant spider silk proteins represent very promising candidates due to their non‐cytotoxic and customizable properties as well as their simple processability into various morphologies, such as scaffolds, hydrogels, films, microcapsules, and spheres . In addition, the genetic engineering enables the functionalization of recombinant spider silk proteins, for example, by adding the sequences that encode ligands targeting tumor cells to increase the efficiency of cancer therapy …”

Section: Introductionmentioning

confidence: 99%

“…9,10 In addition, the genetic engineering enables the functionalization of recombinant spider silk proteins, for example, by adding the sequences that encode ligands targeting tumor cells to increase the efficiency of cancer therapy. [11][12][13][14] The potential application of spider silk particles as drug carriers has been already successfully evaluated in in vitro studies. 12,[15][16][17][18][19] However, the optimization of particle preparation is needed to obtain spheres with a defined, nanometric size in a controlled and reproducible manner.…”

Section: Introductionmentioning

confidence: 99%

Optimization of spider silk sphere formation processing conditions to obtain carriers with controlled characteristics

Florczak

Jastrzębska

Białas

et al. 2018

J Biomedical Materials Res

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Bioengineered spider silk is a focus of research due to its biocompatibility, biodegradability, and excellent mechanical properties. Functionalized silk can be processed into spheres and employed as selective drug delivery vehicles in targeted cancer treatment. Efficient, repeatable and controllable processing conditions are essential to drug carrier development. This study aimed to optimize the processing conditions of silk spheres formation, scale‐up, and automation of the silk spheres production process. The automated micromixing system provided substantial amounts of silk spheres under repetitive production conditions. Micromixing resulted in smaller sphere sizes and narrower sphere size distributions than mixing with a pipette. Furthermore, the particle size and size distribution of silk spheres could be tailored by varying mixing process parameters, such as protein concentration, silk and salting out buffer ratio, mixing speed, and the size of the tubes and mixing zone. In addition, the implementation of ultrafiltration techniques provided a fast and efficient concentration of spheres in water. Furthermore, the shear forces introduced by micromixing did not impede the properties of the Her2 binding peptide (H2.1) since the functionalized H2.1MS1:H2.1MS2 silk spheres selectively were internalized by Her2‐positive cancer cells. This study indicated that micromixing in combination with ultrafiltration enabled scale‐up of the sphere production process under controllable and repeatable conditions. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 3211–3221, 2018.

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Silk Materials Functionalized via Genetic Engineering for Biomedical Applications

Cited by 29 publications

References 115 publications

Drug affinity and targeted delivery: double functionalization of silk spheres for controlled doxorubicin delivery into Her2-positive cancer cells

Drug affinity and targeted delivery: double functionalization of silk spheres for controlled doxorubicin delivery into Her2-positive cancer cells

<p>Cellular uptake, intracellular distribution and degradation of Her2-targeting silk nanospheres</p>

Optimization of spider silk sphere formation processing conditions to obtain carriers with controlled characteristics

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