A20/Tnfaip3, an early NF-κB response gene and key negative regulator of NF-κB signaling, suppresses proinflammatory responses. Its ubiquitinase and deubiquitinase activities mediate proteasomal degradation within the NF-κB pathway. This study investigated the involvement of A20 signaling alterations in podocytes in the development of kidney injury. The phenotypes of A20Δpodocyte (podocyte-specific knockout of A20) mice were compared with those of control mice at 6 months of age to identify spontaneous changes in kidney function. A20Δpodocyte mice presented elevated serum urea nitrogen and creatinine levels, along with increased accumulation of inflammatory cells-neutrophils and macrophages-within the glomeruli. Additionally, A20Δpodocyte mice displayed significant podocyte loss. Ultrastructural analysis of A20 podocyte-knockout mouse glomeruli revealed hypocellularity of the glomerular tuft, expansion of the extracellular matrix, podocytopenia associated with foot process effacement, karyopyknosis, micronuclei, and podocyte detachment. In addition to podocyte death, we also observed damage to intracapillary endothelial cells with vacuolation of the cytoplasm and condensation of nuclear chromatin. A20 expression downregulation and CRISPR-Cas9 genome editing targeting A20 in a podocyte cell line confirmed these findings in vitro, highlighting the significant contribution of A20 activity in podocytes to glomerular injury pathogenesis. Finally, we analyzed TNFAIP3 transcription levels alongside genes involved in apoptosis, anoikis, NF-κB regulation, and cell attachment in glomerular and tubular compartments of kidney biopsies of patients with various renal diseases.
- MeSH
- Cytoskeleton * metabolism MeSH
- Glomerulonephritis * pathology metabolism genetics MeSH
- Kidney Glomerulus pathology metabolism MeSH
- Humans MeSH
- Mice, Inbred C57BL MeSH
- Mice, Knockout * MeSH
- Mice MeSH
- NF-kappa B metabolism MeSH
- Podocytes * metabolism pathology MeSH
- Signal Transduction MeSH
- Tumor Necrosis Factor alpha-Induced Protein 3 * metabolism genetics MeSH
- Inflammation * pathology genetics MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Microbial colonization on the titanium condenser material (TCM) used in the cooling system leads to biofouling and corrosion and influences the water supply. The primary investigation of the titanium condenser was infrequently studied on characterizing biofilm-forming bacterial communities. Different treatment methods like electropotential charge, ultrasonication, and copper coating of titanium condenser material may influence the microbial population over the surface of the titanium condensers. The present study aimed to catalog the primary colonizers and the effect of different treatment methods on the microbial community. CFU (1.7 × 109 CFU/mL) and ATP count (< 5000 × 10-7 relative luminescence units) showed a minimal microbial population in copper-coated surface biofilm as compared with the other treatments. Live and dead cell result also showed consistency with colony count. The biofilm sample on the copper-coated surface showed an increased dead cell count and decreased live cells. In the metagenomic approach, the microbiome coverage was 10.06 Mb in samples derived from copper-coated TCM than in other treated samples (electropotential charge-17.94 Mb; ultrasonication-20.01 Mb), including control (10.18 Mb). Firmicutes preponderate the communities in the biofilm samples, and Proteobacteria stand next in the population in all the treated condenser materials. At the genus level, Lactobacillaceae and Azospirillaceae dominated the biofilm community. The metagenome data suggested that the attached community is different from those biofilm samples based on the environment that influences the bacterial community. The outcome of the present study depicts that copper coating was effective against biofouling and corrosion resistance of titanium condenser material for designing long-term durability.
The adherence of bladder uroepithelial cells, subsequent expression, and regulation of type 1 fimbrial genes (key mediator of attachment) in clinical multidrug-resistant uropathogenic Escherichia coli (MDR-UPECs) isolated from individuals with asymptomatic bacteriuria (ABU) remain unexplored till date. Therefore, this study aimed to investigate the underlying molecular mechanisms associated with the adherence of clinical MDR-ABU-UPECs to human a uroepithelial cell line (HTB-4), both in the absence and presence of D-Mannose. These investigations focused on phase variation, expression, and regulation of type 1 fimbriae and were compared to a prototype ABU-strain (E. coli 83972) and symptomatic MDR-UPECs. Discordant to the ABU prototype strain, MDR-ABU-UPECs exhibited remarkable adhesive capacity that was significantly reduced after D-mannose exposure, fairly like the MDR symptomatic UPECs. The type 1 fimbrial phase variation, determined by the fim switch analysis, asserted the statistically significant incidence of "both OFF and ON" orientation among the adherent MDR-ABU-UPECs with a significant reduction in phase-ON colonies post-D-mannose exposure, akin to the symptomatic ones. This was indicative of an operative and alternating type 1 fimbrial phase switch. The q-PCR assay revealed a coordinated action of the regulatory factors; H-NS, IHF, and Lrp on the expression of FimB and FimE recombinases, which further controlled the function of fimH and fimA genes in ABU-UPECs, similar to symptomatic strains. Therefore, this study is the first of its kind to provide an insight into the regulatory crosstalk of different cellular factors guiding the adhesion of ABU-UPECs to the host. Additionally, it also advocated for the need to accurately characterize ABU-UPECs.
- MeSH
- Adhesins, Escherichia coli genetics metabolism MeSH
- Bacterial Adhesion * MeSH
- Fimbriae, Bacterial * genetics metabolism MeSH
- Bacteriuria microbiology MeSH
- Cell Line MeSH
- Epithelial Cells * microbiology MeSH
- Escherichia coli Infections * microbiology MeSH
- Humans MeSH
- Mannose metabolism pharmacology MeSH
- Drug Resistance, Multiple, Bacterial * genetics MeSH
- Fimbriae Proteins * genetics metabolism MeSH
- Escherichia coli Proteins genetics metabolism MeSH
- Gene Expression Regulation, Bacterial MeSH
- Uropathogenic Escherichia coli * genetics drug effects isolation & purification MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Prostate cancer (PCa) is the second most common cancer. In this paper, the isolation and properties of exosomes as potential novel liquid biopsy markers for early PCa liquid biopsy diagnosis are investigated using two prostate human cell lines, i.e., benign (control) cell line RWPE1 and carcinoma cell line 22Rv1. Exosomes produced by both cell lines are characterised by various methods including nanoparticle-tracking analysis, dynamic light scattering, scanning electron microscopy and atomic force microscopy. In addition, surface plasmon resonance (SPR) is used to study three different receptors on the exosomal surface (CD63, CD81 and prostate-specific membrane antigen-PMSA), implementing monoclonal antibodies and identifying the type of glycans present on the surface of exosomes using lectins (glycan-recognising proteins). Electrochemical analysis is used to understand the interfacial properties of exosomes. The results indicate that cancerous exosomes are smaller, are produced at higher concentrations, and exhibit more nega tive zeta potential than the control exosomes. The SPR experiments confirm that negatively charged α-2,3- and α-2,6-sialic acid-containing glycans are found in greater abundance on carcinoma exosomes, whereas bisecting and branched glycans are more abundant in the control exosomes. The SPR results also show that a sandwich antibody/exosomes/lectins configuration could be constructed for effective glycoprofiling of exosomes as a novel liquid biopsy marker.
- MeSH
- Exosomes * chemistry MeSH
- Carcinoma * metabolism pathology MeSH
- Lectins analysis metabolism MeSH
- Humans MeSH
- Polysaccharides analysis metabolism MeSH
- Liquid Biopsy MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
Konjugáty protilátka-léčivo (ADC) jsou velmi aktuální terapeutickou skupinou kancerostatik. Tvoří je monoklonální protilátka, nejčastěji typu IgG, na kterou je přes vhodný linker (kovalentní řetězec) konjugována jedna nebo více molekul cytotoxického léčiva. Konjugovaná forma léčiva má výrazně nižší toxicitu než forma volná, kterou v terapii nelze užívat samostatně. Databáze SÚKL registruje zatím 9 ADC přípravků pro použití v onkologii. Vzhledem k tomu, že možných kombinací párů protilátka-léčivo je obrovské množství, do roku 2025 se očekává „boom“ této oblasti a až čtyřnásobný nárůst prodejů. V krevní cirkulaci směřuje ADC přípravek specificky k antigenu přítomnému na povrchu nádorové buňky. Linker mezi protilátkou a léčivem musí být stabilní v prostředí krevní cirkulace a teprve až po internalizaci konjugátu být v cílové nádorové buňce zcela degradován nebo zůstat vázaný na léčivo po degradaci proteinové části konjugátu (tzv. štěpitelné nebo neštěpitelné linkery). Následně léčivo způsobí různými mechanismy apoptózu nádorové buňky. Léčiva použitá v ADC přípravcích 2. generace byla až 1000krát toxičtější než chemoterapeutikum doxorubicin a jednalo se především o deriváty auristatinů a maytansinů. V současné 3. generaci vyvíjených konjugátů se zkouší i méně toxická léčiva na bázi kamptothecinů, amanitinů aj. Léčivo s linkerem je k mAb připojeno pomocí různých biokonjugačních metod. Zde se uplatňuje celá škála technik syntetické chemie, přičemž biokonjugace může být buď nespecifická nebo specifická. Konjugovatelné jsou především periferní aminokyseliny protilátky – cystein, lysin, histidin, tyrosin, glutamin a redukované disulfidové můstky mezi dvěma těžkými nebo mezi těžkým a lehkým řetězcem. Pro specifickou konjugaci byly vyvinuty např. techniky glykoinženýrství, založené na N-glykosylaci protilátky na asparaginu (N297). Konjugační techniky, ale i syntéza „nahé“ humánní mAb jsou předmětem utajovaného „know-how“ řady progresivních firem a laboratoří, které v oblasti ADC přípravků působí.
Antibody-drug conjugates are a promising therapeutic class of cytostatic agents. They consist of a monoclonal antibody, usually of the IgG type, to which one or more molecules of a cytotoxic drug are conjugated via a suitable linker (covalent chain). The conjugated form of the drug has significantly lower toxicity than the free form, which cannot be used alone in therapy. The Czech State Institute for Drug Control database has so far registered 9 antibody-drug conjugates for use in oncology. Given the vast number of possible combinations of antibody-drug pairs, aboom in sales is expected by 2025. In the blood circulation, the antibody-drug conjugate specifically targets the antigen expressed on the surface of the tumor cell. The linker between the antibody and the drug must be stable in the blood circulation environment and only be completely degraded in the target tumor cell after internalization of the conjugate or remain bound to the drug after degradation of the protein part of the conjugate (the so-called cleavable or non-cleavable linkers). Subsequently, the drug causes apoptosis of the tumor cell by various mechanisms. The drugs used in the 2nd generation antibody-drug conjugates were up to 1,000 times more toxic than the chemotherapeutic drug doxorubicin and were mainly auristatin and maytansine derivatives. Less toxic drugs based on camptothecins, amanitins, etc. are also being tested in the current 3rd generation of conjugates under development. The linker drug is attached to the antibody by various bioconjugation methods. Here, a range of synthetic chemistry techniques are applied, and bioconjugation can be either non-specific or specific. In particular, the peripheral amino acids of the antibody – cysteine, lysine, histidine, tyrosine, glutamine and reduced disulfide bridges between the two heavy chains or between the heavy and light chains – are conjugated. For a specific conjugation, e.g. glycoengineering techniques based on N-glycosylation of antibody on asparagine (N297) have been developed. Conjugation techniques, as well as the synthesis of "naked" human antibodies are the subject of classified "know-how" of a number of commercial progressive companies and laboratories.Full text English translation is available in the on-line version.
Konjugáty protilátka-léčivo (ADC) jsou velmi aktuální terapeutickou skupinou kancerostatik. Tvoří je monoklonální protilátka, nejčastěji typu IgG, na kterou je přes vhodný linker (kovalentní řetězec) konjugována jedna nebo více molekul cytotoxického léčiva. Konjugovaná forma léčiva má výrazně nižší toxicitu než forma volná, kterou v terapii nelze užívat samostatně. Databáze SÚKL registruje zatím 9 ADC přípravků pro použití v onkologii. Vzhledem k tomu, že možných kombinací párů protilátka-léčivo je obrovské množství, do roku 2025 se očekává „boom“ této oblasti a až čtyřnásobný nárůst prodejů. V krevní cirkulaci směřuje ADC přípravek specificky k antigenu přítomnému na povrchu nádorové buňky. Linker mezi protilátkou a léčivem musí být stabilní v prostředí krevní cirkulace a teprve až po internalizaci konjugátu být v cílové nádorové buňce zcela degradován nebo zůstat vázaný na léčivo po degradaci proteinové části konjugátu (tzv. štěpitelné nebo neštěpitelné linkery). Následně léčivo způsobí různými mechanismy apoptózu nádorové buňky. Léčiva použitá v ADC přípravcích 2. generace byla až 1000krát toxičtější než chemoterapeutikum doxorubicin a jednalo se především o deriváty auristatinů a maytansinů. V současné 3. generaci vyvíjených konjugátů se zkouší i méně toxická léčiva na bázi kamptothecinů, amanitinů aj. Léčivo s linkerem je k mAb připojeno pomocí různých biokonjugačních metod. Zde se uplatňuje celá škála technik syntetické chemie, přičemž biokonjugace může být buď nespecifická nebo specifická. Konjugovatelné jsou především periferní aminokyseliny protilátky – cystein, lysin, histidin, tyrosin, glutamin a redukované disulfidové můstky mezi dvěma těžkými nebo mezi těžkým a lehkým řetězcem. Pro specifickou konjugaci byly vyvinuty např. techniky glykoinženýrství, založené na N-glykosylaci protilátky na asparaginu (N297). Konjugační techniky, ale i syntéza „nahé“ humánní mAb jsou předmětem utajovaného „know-how“ řady progresivních firem a laboratoří, které v oblasti ADC přípravků působí.
Antibody-drug conjugates are a promising therapeutic class of cytostatic agents. They consist of a monoclonal antibody, usually of the IgG type, to which one or more molecules of a cytotoxic drug are conjugated via a suitable linker (covalent chain). The conjugated form of the drug has significantly lower toxicity than the free form, which cannot be used alone in therapy. The Czech State Institute for Drug Control database has so far registered 9 antibody-drug conjugates for use in oncology. Given the vast number of possible combinations of antibody-drug pairs, aboom in sales is expected by 2025. In the blood circulation, the antibody-drug conjugate specifically targets the antigen expressed on the surface of the tumor cell. The linker between the antibody and the drug must be stable in the blood circulation environment and only be completely degraded in the target tumor cell after internalization of the conjugate or remain bound to the drug after degradation of the protein part of the conjugate (the so-called cleavable or non-cleavable linkers). Subsequently, the drug causes apoptosis of the tumor cell by various mechanisms. The drugs used in the 2nd generation antibody-drug conjugates were up to 1,000 times more toxic than the chemotherapeutic drug doxorubicin and were mainly auristatin and maytansine derivatives. Less toxic drugs based on camptothecins, amanitins, etc. are also being tested in the current 3rd generation of conjugates under development. The linker drug is attached to the antibody by various bioconjugation methods. Here, a range of synthetic chemistry techniques are applied, and bioconjugation can be either non-specific or specific. In particular, the peripheral amino acids of the antibody – cysteine, lysine, histidine, tyrosine, glutamine and reduced disulfide bridges between the two heavy chains or between the heavy and light chains – are conjugated. For a specific conjugation, e.g. glycoengineering techniques based on N-glycosylation of antibody on asparagine (N297) have been developed. Conjugation techniques, as well as the synthesis of "naked" human antibodies are the subject of classified "know-how" of a number of commercial progressive companies and laboratories.
Biocompatibility is one of the key issues for implants, especially in the case of stainless steel with medium to low biocompatibility, which may lead to a lack of osseointegration and consequently to implant failure or rejection. To precisely control preferential cell growth sites and, consequently, the biocompatibility of prosthetic devices, two types of surfaces were analyzed, containing periodic nanogrooves laser induced periodic surface structure (LIPSS) and square-shaped micropillars. For the fast and efficient production of these surfaces, the unique combination of high energy ultrashort pulsed laser system with multi-beam and beamshaping technology was applied, resulting in increased productivity by 526% for micropillars and 14 570% for LIPSS compared to single beam methods.In vitroanalysis revealed that micro and nanostructured surfaces provide a better environment for cell attachment and proliferation compared to untreated ones, showing an increase of up to 496% in the number of cells compared to the reference. Moreover, the combination of LIPSS and micropillars resulted in a precise cell orientation along the periodic microgroove pattern. The combination of these results demonstrates the possibility of mass production of functionalized implants with control over cell organization and growth. Thus, reducing the risk of implant failure due to low biocompatibility.
Cíle: Hodnocení dlouhodobého vlivu léčby rheoferézou na suchou formu věkem podmíněné makuární degenerace. Materiál a metody: Do hodnocení jsme zařadili 65 pacientů a do kontrolní skupiny 55 pacientů s minimální dobou sledování 60 měsíců. Základní léčba se skládala z 8 procedur rheoferézy, přídatná léčba (booster therapy) ze 2 rheoferéz za 1,5-2 roky po základní léčbě. Hodnotili jsme změny nejlépe korigované zrakové ostrosti, anatomických poměrů a elektrické aktivity sítnice, hematologické, biochemické a imunologické parametry. Výsledky: Léčba rheoferézou významně přispěla: 1) Ke stabilizaci nejlépe korigované zrakové ostrosti léčených pacientů, která se nejprve nevýznamně zvyšovala do 2 let sledování a následně jen mírně klesala. Naproti tomu v kontrolní skupině se zraková ostrost snižovala, do 4 let nevýznamně, poté již statisticky významně. 2) Ke zlepšení morfologického nálezu u 62,4 % léčených pacientů v porovnání se 7,5 % kontrol, naproti tomu k progresi onemocnění do 3. stadia (vlhká forma onemocnění nebo geografická atrofie) s významným poklesem zrakových funkcí došlo jen u 7,1 % léčených pacientů oproti 37,0 % kontrol. 3) K regresi, dokonce i k přiložení drúzového odchlípení retinálního pigmentového epitelu (DPED). Ke zmenšení plochy DPED u 80,4 % léčených pacientů, naproti tomu ke zvětšení plochy DPED u 47,1 % kontrol a rozvoji nového DPED jen u 2 očí léčených pacientů oproti 16 očím kontrol. 4) K udržení integrity vrstvy elipsoidů ve fovee u 68,2 % léčených pacientů, naproti tomu defektní elipsoidní vrstvu ve fovee jsme zaznamenali u 66,6 % kontrol. 5) Ke stabilizaci aktivity gangliových buněk, čípkového systému a aktivity centrální oblasti sítnice s excentricitou mezi 1,8° a 30° u léčených pacientů v porovnání s její alterací v kontrolní skupině projevující se zejména od 3,5 let sledování. 6) K statisticky významnému zlepšení rheologických ukazatelů, a tím ke zvýšení průtoku v mikrocirkulaci a pozitivnímu ovlivnění metabolizmu v sítnici. K pozitivnímu vlivu na klasickou, alternativní i lektinovou cestu aktivace komplementu, snížení hladiny PCSK9 (proprotein konvertáza subticilin kexin 9), a tím i hladiny LDL-cholesterolu a 7) Přídatná léčba 2 procedurami RHF (tzv. „booster therapy“) se zdá být bezpečnou a vhodnou metodou prodloužení fáze stabilizace, či dokonce zlepšení zrakové ostrosti, anatomického a funkčního nálezu. Závěr: Prokázali jsme pozitivní změny anatomických, funkčních i humorálních ukazatelů při léčbě VPMD rheoferézou. Jejich korelace skýtá reálnou možnost ohrožené nemocné vytipovat a řídit individualizovanou intenzitu rheoterapie. Metodika je účinná a bezpečná s nízkým procentem nezávažných vedlejších účinků.
Purpose: Evaluation of the long-term effect of rheopheresis treatment of dry form of age-related macular degeneration (AMD). Materials and Methods: The treatment group consisted of 65 patients and 55 patients in the control group, with a minimum follow-up period of 60 months. The basic treatment consisted of 8 rheopheresis procedures, and the additional treatment (booster therapy) of 2 rheopheresis procedures 1.5–2 years after the basic treatment. We evaluated changes in best corrected visual acuity, anatomical effect, electrical activity of the retina, haematological, biochemical and immunological parameters. Results: Rheopheresis treatment contributed significantly: 1) to stabilisation of best corrected visual acuity of the treated patients, which initially showed an insignificant increased during the 2-years follow-up period, and then slightly decreased. By contrast, visual acuity decreased in the control group, to an insignificant degree up to 4 years, then statistically significantly. 2) to an improvement of the morphological findings in 62.4% of treated patients compared to 7.5% in the control group, while disease progression to stage 3 (neovascular form of the disease or geographic atrophy) with a significant decrease of visual acuity occurred in only 7.1% of treated patients, versus 37.0% in the control group. 3) to regression, even to the attachment of drusenoid pigment epithelial detachment (DPED). To a reduction of the area of DPED in 80.4% of treated patients, in contrast with an steaincrease in the area of DPED in 47.1% of patients in the control group, and the development of new DPED in only 2 eyes of treated patients compared with 16 eyes of patients in the control group. 4) to a preservation of the integrity of the ellipsoid layer in the fovea in 68.2% of the treated patients, while by contrast we found a damaged ellipsoid layer in the fovea in 66.6% of the control patients. 5) to a stabilisation of the activity of ganglion cells, the pineal system and the activity of the central area of the retina, with eccentricity between 1.8° and 30° in the treated patients, compared to alteration in the control group manifested mainly after 3.5 years of the follow-up period. 6) to a statistically significant improvement in rheological parameters, thereby increasing flow in microcirculation and positively influencing the metabolism in the retina. Also to a positive effect on the classical, alternative and lectin pathway of complement activation, a reduction in the level of proprotein convertase subtilisin kexin 9 (PCSK9), and thus also the level of LDLcholesterol, and 7) Additional treatment with 2 RHF procedures (so-called "booster therapy") seems to be a safe and suitable method of prolonging the stabilisation phase, or even improving visual acuity, anatomical and functional findings. Conclusion: We demonstrated positive changes in anatomical, functional and humoral parameters upon rheopheresis treatment of AMD. Their correlation provides a real possibility to identify patients at risk and to manage an individualised regime of rheopheresis therapy. This method of treatment is effective and safe, with a low percentage of non-serious adverse effects.
- MeSH
- Geographic Atrophy pathology therapy MeSH
- Humans MeSH
- Macular Degeneration pathology therapy MeSH
- Plasmapheresis * methods adverse effects MeSH
- Proprotein Convertase 9 therapeutic use MeSH
- Retina pathology MeSH
- Blood Component Removal methods adverse effects MeSH
- Research MeSH
- Check Tag
- Humans MeSH
INTRODUCTION: The formation of diabetic ulcers (DU) is a common complication for diabetic patients resulting in serious chronic wounds. There is therefore, an urgent need for complex treatment of this problem. This study examines a bioactive wound dressing of a biodegradable electrospun nanofibrous blend of poly(L-lactide-co-ε-caprolactone) and poly(ε-caprolactone) (PLCL/PCL) covered by a thin fibrin layer for sustained delivery of bioactive molecules. METHODS: Electrospun PLCL/PCL nanofibers were coated with fibrin-based coating prepared by a controlled technique and enriched with human platelet lysate (hPL), fibroblast growth factor 2 (FGF), and vascular endothelial growth factor (VEGF). The coating was characterized by scanning electron microscopy and fluorescent microscopy. Protein content and its release rate and the effect on human saphenous vein endothelial cells (HSVEC) were evaluated. RESULTS: The highest protein amount is achieved by the coating of PLCL/PCL with a fibrin mesh containing 20% v/v hPL (NF20). The fibrin coating serves as an excellent scaffold to accumulate bioactive molecules from hPL such as PDGF-BB, fibronectin (Fn), and α-2 antiplasmin. The NF20 coating shows both fast and a sustained release of the attached bioactive molecules (Fn, VEGF, FGF). The dressing significantly increases the viability of human saphenous vein endothelial cells (HSVECs) cultivated on a collagen-based wound model. The exogenous addition of FGF and VEGF during the coating procedure further increases the HSVECs viability. In addition, the presence of α-2 antiplasmin significantly stabilizes the fibrin mesh and prevents its cleavage by plasmin. DISCUSSION: The NF20 coating supplemented with FGF and VEGF provides a promising wound dressing for the complex treatment of DU. The incorporation of various bioactive molecules from hPL and growth factors has great potential to support the healing processes by providing appropriate stimuli in the chronic wound.
- MeSH
- alpha-2-Antiplasmin MeSH
- Endothelial Cells MeSH
- Wound Healing MeSH
- Humans MeSH
- Nanofibers * MeSH
- Bandages MeSH
- Polyesters pharmacology MeSH
- Vascular Endothelial Growth Factor A * MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Elongated protein-based micro- and nanostructures are of great interest for a wide range of biomedical applications, where they can serve as a backbone for surface functionalization and as vehicles for drug delivery. Current production methods for protein constructs lack precise control of either shape and dimensions or render structures fixed to substrates. This work demonstrates production of recombinant spider silk nanowires suspended in solution, starting with liquid bridge induced assembly (LBIA) on a substrate, followed by release using ultrasonication, and concentration by centrifugation. The significance of this method lies in that it provides i) reproducability (standard deviation of length <13% and of diameter <38%), ii) scalability of fabrication, iii) compatibility with autoclavation with retained shape and function, iv) retention of bioactivity, and v) easy functionalization both pre- and post-formation. This work demonstrates how altering the function and nanotopography of a surface by nanowire coating supports the attachment and growth of human mesenchymal stem cells (hMSCs). Cell compatibility is further studied through integration of nanowires during aggregate formation of hMSCs and the breast cancer cell line MCF7. The herein-presented industrial-compatible process enables silk nanowires for use as functionalizing agents in a variety of cell culture applications and medical research.
- MeSH
- Cell Culture Techniques MeSH
- Silk chemistry MeSH
- Humans MeSH
- Nanowires * MeSH
- Nanostructures * MeSH
- Spiders * MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
