Cellular Uptake and Release of Intact Lactoferrin and Its Derivatives in an Intestinal Enterocyte Model of Caco-2 Cells

Matsuzaki, Takumi; Nakamura, Masao; Nogita, Takehide; Sato, Atsushi

doi:10.1248/bpb.b19-00011

Cited by 21 publications

(13 citation statements)

References 18 publications

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“…For MSN@PLA–PEG-CPP, sodium heparin showed the highest inhibition in cellular uptake, and sodium azide, 4 °C, filipin, protamine sulfate, sodium heparin, and sodium chlorate also showed inhibition to different degrees. Heparin limits ligand binding with the heparan sulfate proteoglycan (HSPG) receptor, the sulfated glycosaminoglycan polysaccharides on outer cell membranes. Due to carboxyl and sulfate groups, the HSPG receptor often presents a polyanionic property and can bind with cationic CPPs or other macromolecular cargos via electrostatic interactions , and affects cellular uptake of CPPs with the polyarginine, polylysine, or arginine/lysine hybrid structure. , In addition, protamine sulfate leads to the blockade of anionic sites like glycoproteins and their receptors, , and sodium chlorate also inhibits glycosaminoglycan sulfation.…”

Section: Resultsmentioning

confidence: 99%

Hydrophilic and Electroneutral Nanoparticles to Overcome Mucus Trapping and Enhance Oral Delivery of Insulin

et al. 2020

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The oral delivery of macromolecules using nanoparticles is limited by secreted mucus, resulting in low contact or internalization via intestinal cells and, thus, both mucus trapping and further low cellular uptake need to be overcome. Here, hydrophilic and electroneutral nanoparticles were developed to overcome mucus trapping and enhance the oral delivery of macromolecules. Mesoporous silica nanoparticles (MSNs) were synthesized and modified with a hydrophilic block polymer (poly(lactic acid)−methoxy poly(ethylene glycol), PLA−PEG), and then an overall electroneutrality and promoted cellular uptake were achieved by sequential modification with cell-penetrating peptides (CPPs). Reduced hydrophobic and electrostatic interactions of MSN@PLA−PEG-CPP with mucus decreased mucus trapping by 36.0%, increased the cellular uptake of MSN@PLA−PEG-CPP by 2.3-folds in mucous conditions via active heparan sulfate proteoglycan receptor (HSPG)-mediated and caveolae-mediated endocytosis and electrostatic interactions. Furthermore, insulin, a model macromolecular drug, was successfully loaded into the nanoparticles (INS@MSN@PLA−PEG-CPP). Compared with insulin solution, in vitro cellular uptake in mucous conditions and in vivo pharmacodynamic effects were significantly increased by 9.1-and 14.2-folds, respectively. As well, all nanoparticles with or without insulin loading presented negligible in vitro and in vivo toxicity. Herein, hydrophilic and electroneutral nanoparticles with sequential PEG and CPP modification could promote cellular uptake against mucus trapping and finally show good prospects for oral insulin delivery.

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Section: Resultsmentioning

confidence: 99%

Hydrophilic and Electroneutral Nanoparticles to Overcome Mucus Trapping and Enhance Oral Delivery of Insulin

et al. 2020

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“…In particular, there is evidence that it can bind to at least some of the receptors used by coronaviruses and thereby block their entry [ 75 ]. LF makes use of Heparan Sulfate Proteoglycans (HSPGs) on the surface of epithelial cells of the stomach to facilitate entry into the bloodstream via endocytosis [ 77 , 78 ]. Coronaviruses bind to host cells by first attaching to HSPGs utilizing them as preliminary docking sites to assist with viral entry and LF is able to interfere with some of the receptors used by coronaviruses.…”

Section: Discussionmentioning

confidence: 99%

Proteomics of fibrin amyloid microclots in long COVID/post-acute sequelae of COVID-19 (PASC) shows many entrapped pro-inflammatory molecules that may also contribute to a failed fibrinolytic system

Kruger

Vlok

Turner

et al. 2022

Cardiovasc Diabetol

105

131

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Background Post-acute sequelae of COVID-19 (PASC), also now known as long COVID, has become a major global health and economic burden. Previously, we provided evidence that there is a significant insoluble fibrin amyloid microclot load in the circulation of individuals with long COVID, and that these microclots entrap a substantial number of inflammatory molecules, including those that might prevent clot breakdown. Scientifically, the most challenging aspect of this debilitating condition is that traditional pathology tests such as a serum CRP (C-reactive protein) may not show any significant abnormal inflammatory markers, albeit these tests measure only the soluble inflammatory molecules. Elevated, or abnormal soluble biomarkers such as IL-6, D-Dimer or fibrinogen indicate an increased risk for thrombosis or a host immune response in COVID-19. The absence of biomarkers in standard pathology tests, result in a significant amount of confusion for patients and clinicians, as patients are extremely sick or even bed-ridden but with no regular identifiable reason for their disease. Biomarkers that are currently available cannot detect the molecules present in the microclots we identified and are therefore unable to confirm their presence or the mechanisms that drive their formation. Methods Here we analysed the protein content of double-digested microclots of 99 long COVID patients and 29 healthy controls. The patients suffering from long COVID reported their symptoms through a questionnaire completed by themselves or their attending physician. Results Our long COVID cohort’s symptoms were found to be in line with global findings, where the most prevalent symptoms were constant fatigue (74%,) cognitive impairment (71%) and depression and anxiety (30%). Our most noteworthy findings were a reduced level of plasma Kallikrein compared to our controls, an increased level of platelet factor 4 (PF4) von Willebrand factor (VWF), and a marginally increased level of α-2 antiplasmin (α-2-AP). We also found a significant presence of antibodies entrapped inside these microclots. Conclusion Our results confirm the presence of pro-inflammatory molecules that may also contribute to a failed fibrinolysis phenomenon, which could possibly explain why individuals with long COVID suffer from chronic fatigue, dyspnoea, or cognitive impairment. In addition, significant platelet hyperactivation was noted. Hyperactivation will result in the granular content of platelets being shed into the circulation, including PF4. Overall, our results provide further evidence of both a failed fibrinolytic system in long COVID/PASC and the entrapment of many proteins whose presence might otherwise go unrecorded. These findings might have significant implications for individuals with pre-existing comorbidities, including cardiovascular disease and type 2 diabetes.

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“…Small molecules, including pharmaceutical drugs, require solute carriers of the SLC family (116) to effect their uptake (117)(118)(119)(120)(121)(122)(123)(124). Lactoferrin, as a protein, is far too large to exploit such a route, and instead passes from the stomach via epithelial cells and into the blood using endocytosis (125,126), especially via Peyer's patches (127), and when it is encapsulated ("enterically formulated") in liposomes (128)(129)(130). This uptake then occurs mostly via the lymphatic rather than the portal circulation (131,132).…”

Section: Lactoferrin Transportmentioning

confidence: 99%

The Biology of Lactoferrin, an Iron-Binding Protein That Can Help Defend Against Viruses and Bacteria

Heyden²,

2020

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Lactoferrin is a nutrient classically found in mammalian milk. It binds iron and is transferred via a variety of receptors into and between cells, serum, bile, and cerebrospinal fluid. It has important immunological properties, and is both antibacterial and antiviral. In particular, there is evidence that it can bind to at least some of the receptors used by coronaviruses and thereby block their entry. Of importance are Heparan Sulfate Proteoglycans (HSPGs) and the host receptor angiotensin-converting enzyme 2 (ACE2), as based on other activities lactoferrin might prevent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from attaching to the host cells. Lactoferrin (and more specifically enteric-coated LF because of increased bioavailability) may consequently be of preventive and therapeutic value during the present COVID-19 pandemic.

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Cellular Uptake and Release of Intact Lactoferrin and Its Derivatives in an Intestinal Enterocyte Model of Caco-2 Cells

Cited by 21 publications

References 18 publications

Hydrophilic and Electroneutral Nanoparticles to Overcome Mucus Trapping and Enhance Oral Delivery of Insulin

Hydrophilic and Electroneutral Nanoparticles to Overcome Mucus Trapping and Enhance Oral Delivery of Insulin

Proteomics of fibrin amyloid microclots in long COVID/post-acute sequelae of COVID-19 (PASC) shows many entrapped pro-inflammatory molecules that may also contribute to a failed fibrinolytic system

The Biology of Lactoferrin, an Iron-Binding Protein That Can Help Defend Against Viruses and Bacteria

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