lactoferrin and Necrosis

This review mainly summarizes results of recent studies on the anticancer activity of the multifunctional protein lactoferrin (Lf) and its derived peptide lactoferricin (Lfcin). The basic information on Lf and Lfcin, such as their sources, structures, and biological properties which favor their antitumor activity is introduced. The major anticancer mechanisms of Lf and Lfcin including cell cycle arrest, apoptosis, anti-angiogenesis, antimetastasis, immune modulation and necrosis are discussed. Other information from in vivo studies employing a mouse model is also provided. In addition, the roles of talatoferrin and delta lactoferrin, as well as improvement in drug delivery will be covered.

Topics: Angiogenesis Inhibitors; Animals; Antineoplastic Agents; Apoptosis; Cell Cycle; Humans; Lactoferrin; Necrosis; Neoplasms; Neovascularization, Pathologic

Osteonecrosis of the jaws is an emerging pathological condition characterized by un-exposure or exposure of the necrotic bone, independently from the etiology. This term is usually referred to medication-related osteonecrosis of the jaws due to severe adverse reaction to certain medicines, as bisphosphonates, used for the treatment of cancer and osteoporosis. The management of patients with Bisphosphonate-Related Osteonecrosis of the Jaws (BRONJ) remains challenging because surgical and medical interventions may not eradicate this pathology. The goal of treatment of patients at risk of developing BRONJ or of those who have active disease is the preservation of quality of life by controlling pain, managing infection, and preventing the development of new areas of necrosis. The treatment of osteonecrosis consists in the surgical removal of necrotic bone followed by antibiotic therapy and application of sterile greasy gauze until the wound closure. The classical medical treatment has been compared with the innovative one consisting in the application of sterile greasy gauze soaked with bovine lactoferrin (bLf) after surgery. Here, for the first time, bLf efficacy on wound repair in subjects suffering from BRONJ with the progressive destruction of bone in the mandible or maxilla has been demonstrated. The positive results consist in a significant shorter time of wound closure (1 or 2 weeks) compared to that observed with classical surgical treatment (2-3 months). These promising results are an interesting tool for the innovative treatment of this pathology and for increasing the quality of life of these patients.

Topics: Administration, Oral; Aged; Animals; Bisphosphonate-Associated Osteonecrosis of the Jaw; Bone Density Conservation Agents; Cattle; Diphosphonates; Female; Humans; Lactoferrin; Male; Middle Aged; Necrosis; Quality of Life

Liver cancer and leukemia are the fourth and first causes, respectively, of cancer death in children and adults worldwide. Moreover, cancer treatments, although beneficial, remain expensive, invasive, toxic, and affect the patient's quality of life. Therefore, new anticancer agents are needed to improve existing agents. Because bovine lactoferrin (bLF) and its derived peptides have antitumor properties, we investigated the anticancer effect of bLF and LF peptides (LFcin17-30, LFampin265-284 and LFchimera) on liver cancer HepG2 cells and leukemia Jurkat cells. HepG2 and Jurkat cells were incubated with bLF and LF peptides. Cell proliferation was quantified by an MTT assay, and cell morphology and damage were visualized by light microscopy or by phalloidin-TRITC/DAPI staining. The discrimination between apoptosis/necrosis was performed by staining with Annexin V-Alexa Fluor 488 and propidium iodide, and the expression of genes related to apoptosis was analyzed in Jurkat cells. Finally, the synergistic interaction of bLF and LF peptides with cisplatin or etoposide was assessed by an MTT assay and the combination index. The present study demonstrated that bLF and LF peptides inhibited the viability of HepG2 and Jurkat cells, inducing damage to the cell monolayer of HepG2 cells and morphological changes in both cell lines. bLF, LFcin17-30, and LFampin265-284 triggered apoptosis in both cell lines, whereas LFchimera induced necrosis. These results suggested that bLF and LF peptides activate apoptosis by increasing the expression of genes of the intrinsic pathway. Additionally, bLF and LF peptides synergistically interacted with cisplatin and etoposide. In conclusion, bLF and LF peptides display anticancer activity against liver cancer and leukemia cells, representing an alternative or improvement in cancer treatment.

Topics: Child; Cisplatin; Etoposide; Hep G2 Cells; Humans; Jurkat Cells; Lactoferrin; Liver Neoplasms; Necrosis; Peptides; Quality of Life

LF11-322 (PFWRIRIRR-NH2) (PFR peptide), a nine amino acid-residue peptide fragment derived from human lactoferricin, possesses potent cytotoxicity against bacteria. We report here the discovery and characterization of its antitumor activity in leukemia cells. PFR peptide inhibited the proliferation of MEL and HL-60 leukemia cells by inducing cell death in the absence of the classical features of apoptosis, including chromatin condensation, Annexin V staining, Caspase activation and increase of abundance of pro-apoptotic proteins. Instead, necrotic cell death as evidenced by increasing intracellular PI staining and LDH release, inducing membrane disruption and up-regulating intracellular calcium level, was observed following PFR peptide treatment. In addition to necrotic cell death, PFR peptide also induced G0/G1 cell cycle arrest. Moreover, PFR peptide exhibited favorable antitumor activity and tolerability in vivo. These findings thus provide a new clue of antimicrobial peptides as a potential novel therapy for leukemia.

Topics: Animals; Antimicrobial Cationic Peptides; Antineoplastic Agents; Apoptosis; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Disease Models, Animal; Female; Hemolysis; HL-60 Cells; Humans; Lactoferrin; Leukemia; Mice; Necrosis; Peptides

Several short linear peptides derived from cyclic bovine lactoferricin were synthesized and tested for their cytotoxic effect against the oral cavity squamous-cell carcinoma (OSCC) cell lines CAL27 and SCC15. As a control, an immortalized and nontumorigenic cell line, Het-1A, was used. Linear peptides based on the RRWQWR core sequence showed a moderate cytotoxic effect and specificity towards tumorigenic cells. A tetrameric peptide, LfcinB(20-25)4, containing the RRWQWR motif, exhibited greater cytotoxic activity (>90%) in both OSCC cell lines compared to the linear lactoferricin peptide or the lactoferrin protein. Additionally, this tetrameric peptide showed the highest specificity towards tumorigenic cells among the tested peptides. Interestingly, this effect was very fast, with cell shrinkage, severe damage to cell membrane permeability, and lysis within one hour of treatment. Our results are consistent with a necrotic effect rather than an apoptotic one and suggest that this tetrameric peptide could be considered as a new candidate for the therapeutic treatment of OSCC.

Topics: Animals; Carcinoma, Squamous Cell; Cattle; Cell Line, Tumor; Cell Membrane Permeability; Cytotoxins; Humans; Lactoferrin; Mouth Neoplasms; Necrosis; Peptides

Polyethylene glycol (PEG) is attached to proteins in order to increase their half-life in circulation and reduce their immunogenicity in vivo. The present study was conducted to examine whether two different sizes of PEGylated bovine lactoferrin (40k-PEG-bLf and 20k-PEG-bLf) would enhance the protective effect of native bLf on liver injury induced by carbon tetrachloride (CCl(4)) in rats. Silymarin, a known hepatoprotective drug was used as a positive control. Compared to native bLf, the treatment of PEGylated bLf more markedly prevented the elevation of serum levels of hepatic enzyme markers and inhibited fatty degeneration and the hepatic necrosis induced by CCl(4). 40k-PEG-bLf showed a more significant suppressive effect on CCl(4)-induced hepatic injury than 20k-PEG-bLf. The treatment with PEGylated bLf elevated serum SOD activity reduced by CCl(4) more significantly than native bLf. 40k-PEG-bLf enhanced serum SOD activity more significantly than 20k-PEG-bLf. Immunohistochemical study showed that the PEGylation of bLf enhanced its intracellular transportation to hepatocytes. The increases in intracellular transportation of the PEGylated bLf in order were: 40k-PEG-bLf>20k-PEG-bLf>native bLf. These findings suggested that the mechanism of the enhancement of hepatoprotective effect by PEGylated bLf was associated with an increase in the intracellular transportation of PEGylated bLf in hepatocytes.

Topics: Animals; Apoptosis; Carbon Tetrachloride Poisoning; Chemical and Drug Induced Liver Injury; Fatty Liver; Hepatocytes; Immunohistochemistry; Lactoferrin; Lipid Peroxidation; Liver; Male; Necrosis; Polyethylene Glycols; Protective Agents; Rats; Rats, Wistar; Silymarin; Superoxide Dismutase

Plasma lactoferrin dynamics was investigated in 120 of the 2250 patients with local and generalized soft tissue infections. Systemic symptoms were observed in 15% of the patients with soft tissue infections, syndrome of systemic inflammatory response in 13%, and sepsis in 42%. In 89% of the patients with systemic inflammatory reactions the blood lactoferrin level was 1.1-1.3 times the normal one within 72 hours after the onset of therapy; it dropped to the normal value in 12-15 days. Normalization of blood lactoferrin in patients with mild and moderate systemic inflammatory reactions roughly coincided with the disappearance of symptoms of generalized infection. It occurred 5-6 days after the septic process was resolved in patients with severe inflammatory reactions. Normal blood lactoferrin levels were characteristic of a mild inflammatory reaction and local forms of infection. A rise in blood lactoferrin above 1400 ng/ml combined with the syndrome of systemic inflammatory reaction over 72 hr in duration was regarded as a diagnostic criterion of sepsis. It is suggested that monitoring blood lactoferrin during treatment of systemic inflammatory reactions and sepsis be used for the choice of therapeutic strategy and the assessment of efficiency of its efficiency.

Topics: Biomarkers; Diagnosis, Differential; Follow-Up Studies; Humans; Immunoenzyme Techniques; Lactoferrin; Necrosis; Prospective Studies; Sepsis; Soft Tissue Infections

Model studies have shown that peptides derived from the N-terminal region of bovine lactoferrin (Lf-B) exhibit antitumor activity against certain cell lines. This activity is due primarily to the peptides' apoptogenic effect. Several reports indicate that cationic residues clustered in two regions of the peptide sequence can be shuffled into one region and thereby increase cytotoxic activity, although the mechanism of this enhanced cytotoxic effect has not been clarified. In this paper, we considered several parameters that determine the mode of cell death after exposure to a native Lf-B derived peptide (Pep1, residues 17-34), and a modified peptide (mPep1) wherein the cationic residues of Pep1 are clustered in a single region of its helical structure. We found that the cytotoxic activity of mPep1 was about 9.6 fold-higher than that of Pep1 against HL-60 cells, as determined by the 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethonyphenol)-2-(4-sulfophenyl)-2 H-tetrazolium (MTS) assay. In investigating the expression of phosphatidylserine, we observed that the native peptide (Pep1) caused both apoptotic cell death and necrotic cell death, depending on the concentration of the peptide. In contrast, the action of mPep1 was exclusively characteristic of necrotic cell death. This observation was further confirmed by agarose gel electrophoresis, in which clear ladder-like DNA bands were observed from cells exposed to Pep1, whereas DNA from cells treated with mPep1 produced a smeared pattern. We extended the study by investigating the release of mitochondrial cytochrome c into the cytosol, and the activation of caspase-3; both peptides caused the release of cytochrome c into the cytosol, and the activation of caspase-3.These results suggest that Pep1 may kill cancer cells by activating an apoptosis-inducing pathway, whereas mPep1 causes necrotic cell death by destroying cellular membrane structure notwithstanding sharing some cellular events with apoptotic cell death.

Topics: Amino Acid Sequence; Apoptosis; Blotting, Western; Caspase 3; Cell Survival; Cytochromes c; Flow Cytometry; HL-60 Cells; Humans; Lactoferrin; Leukemia; Molecular Sequence Data; Necrosis; Peptides

Listeria monocytogenes is a food-borne pathogen that causes serious listeriosis in humans. Antimicrobial effects of human lactoferrin (hLF) against L. monocytogenes have been clearly demonstrated in in vitro studies. However, in vivo studies have not been reported yet. This study investigated whether the oral administration of hLF could inhibit oral infection of listeria in BALB/c mice. The MICs for several strains of L. monocytogenes were determined, and the most sensitive strain was used for the animal work. hLF was administered to BALB/c mice for 7 days, commencing 4 days before oral infection. The effect of hLF was determined by bacterial enumeration and histopathological analysis of the liver and spleen, which are well-known as the major targets of oral listeria infection in mice. In bacterial enumeration, hLF decreased the number of L. monocytogenes cells in the liver. Histopathologically, the size and frequency of necrotic foci in the liver samples decreased with hLF administration. However, these changes were not observed in the spleen samples. The mRNA levels of inflammatory cytokines, such as interleukin (IL)-1beta, tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma, decreased in the liver of mice receiving hLF. This study has shown that hLF decreases the hepatic colonization of L. monocytogenes, hepatic necrosis and expression of inflammatory cytokines. It revealed that perorally given hLF could mediate antimicrobial and anti-inflammatory activities remote from the gut (i.e. in the liver) of mice challenged with L. monocytogenes.

Topics: Animals; Anti-Bacterial Agents; Colony Count, Microbial; Cytokines; Disease Models, Animal; Female; Lactoferrin; Listeria monocytogenes; Listeriosis; Liver; Mice; Mice, Inbred BALB C; Microbial Sensitivity Tests; Necrosis; RNA, Messenger; Spleen

The efficacy of prophylactic treatment with human lactoferrin 1-11 (hLF1-11), a broad-spectrum antimicrobial peptide, was studied in a rabbit model of femur infection.. Calcium phosphate cement with 50 mg/g hLF1-11 or gentamicin was injected into the femoral canal, after inoculation with Staphylococcus aureus. Three weeks later, slices of the proximal femora were sawn for quantitative bacterial culture and histology.. Treatment with hLF1-11 (P<0.038) or gentamicin (P<0.008) caused a reduction of cfu compared with the untreated control rabbits. The number of sterile cultures was higher in hLF1-11- (3/7) and gentamicin- (5/6) treated animals than in controls (1/7). Radiological and histological analysis showed early bone ingrowth into the cement cracks, and only moderate pathological changes in rabbits with positive cultures.. Local prophylaxis with hLF1-11 effectively reduced development of osteomyelitis in a rabbit model, but gentamicin resulted in a larger number of sterile femora.

Topics: Abscess; Animals; Anti-Bacterial Agents; Anti-Infective Agents; Bone Cements; Calcium Phosphates; Colony Count, Microbial; Drug Carriers; Female; Femur; Gentamicins; Lactoferrin; Methicillin Resistance; Necrosis; Osteogenesis; Osteomyelitis; Peptide Fragments; Rabbits

Isoproterenol hydrochloride (ISO), a beta adrenergic agonist, is known to cause ischemic necrosis in rats. Cardiotoxicity of three different doses of ISO were studied using physiological, biochemical and histopathological parameters. The effects of single and double dose of ISO were analysed, which illustrated that single ISO dose was more cardiotoxic than double ISO dose due to ischemic preconditioning. The tetrapeptide derivatives L-lysine-L-arginine-L-aspartic acid-L-serine (tetrapeptide A) and di-tert.butyloxycarbonyl-L-lysine-L-arginine-L-aspartic acid-tert.butyl O-tert.butyl-L-serinate (tetrapeptide B) along with acetylsalicylic acid as positive control were analysed at different time points for their cardioprotective effect. The results demonstrated that optimal protective effects were observed by pretreatment with 5 mg/kg of tetrapeptide B and this was found to be slightly better than that of acetylsalicylic acid. A lesser degree of cardioprotection was noticed when low doses of tetrapeptide B were administered. This study clearly showed that single dose of ISO (50 mg/kg, s.c.) induced myocardial necrosis could be used as a model to assess cardiovascular drugs and in this model, it was demonstrated that the tetrapeptide B could exhibit optimal cardioprotective effect.

Topics: Adrenergic beta-Agonists; Animals; Aspirin; Body Weight; Creatine Kinase; Disease Models, Animal; Female; Hemodynamics; Hemorrhage; Isoproterenol; L-Lactate Dehydrogenase; Lactoferrin; Myocardial Ischemia; Myocardium; Necrosis; Oligopeptides; Organ Size; Peptide Fragments; Platelet Aggregation Inhibitors; Rats; Rats, Wistar

Lactoferrin, as measured in the pancreatic juice, has been thought to be of diagnostic value in chronic pancreatitis, but due to the hazards in cannulation of the pancreatic duct in the acute phase of pancreatitis the behavior of lactoferrin has remained obscure. In this study, lactoferrin levels were studied in pancreas tissue specimens obtained in ablative surgery for acute necrotizing pancreatitis (ANP) and in serum samples. A higher pancreatic lactoferrin content was found in ANP than in normal pancreas. Lactoferrin seemed not to leak from a necrotic pancreas in any considerable amounts into the circulation, as no differences were found in serum lactoferrin concentrations between ANP and controls. It remains an open question whether the lactoferrin increase is only an unspecific reaction in inflammation or is something specific for pancreatitis.

Topics: Acute Disease; Alkaline Phosphatase; Amylases; Humans; Lactoferrin; Lactoglobulins; Necrosis; Pancreas; Pancreatitis

35 specimens of human parotid gland and 37 of submandibular gland were transplanted into athymic nude mice. At distinct time intervals, from 1 day to 8 months the transplants were collected and examined. The transplanted glands were studied by light microscopy, immunohistology and autoradiography. The following changes were detectable: acute injury to the xenograft and inflammatory reaction (day 1-7), regeneration of the transplant and the beginning of adaptation to the "mouse milieu" (day 8-30), completion of adaptation (day 30 and later). The presence of the following substances was analysed: amylase, lactoferrin, secretory component, tissue polypeptide antigen (TPA). Amylase was only detected in the early transplants. Lactoferrin was seen only in the small duct system. TPA was present during all transplantation periods and was quantitatively correlated with the 3H thymidine labeling index. From our observations we can say that the salivary glands show two different reacting compartments: a large and a small duct system. The histogenesis of the xenografts, and the relationships of the changes observed to human salivary gland diseases were discussed.

Topics: Amylases; Animals; Autoradiography; Cell Division; Connective Tissue; Epithelium; Female; Histocytochemistry; Humans; Inflammation; Lactoferrin; Mice; Mice, Nude; Mitotic Index; Necrosis; Parotid Gland; Salivary Glands; Secretory Component; Submandibular Gland; Time Factors; Transplantation, Heterologous