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Research Article | DOI: https://doi.org/10.31579/2690-4861/589
1 Discipline of Pharmaceutical Sciences, College of Health Sciences, University of KwaZulu-Natal, Private Bag X54001, Durban, South Africa
2 Discipline of Medical Biochemistry, College of Health Sciences, University of KwaZulu-Natal, Private Bag X54001, Durban, South Africa
3 School of Laboratory Medicine and Medical Sciences, College of Health Sciences, University of KwaZulu-Natal, Private Bag X54001, Durban, South Africa
*Corresponding Author: Mbuso Faya, Discipline of Pharmaceutical Sciences, College of Health Sciences, University of KwaZulu-Natal, Private Bag X54001, Durban, South Africa
Citation: Lusanda Mtetwa, Nkeiruka Igbokwe, Eman I. Abdallah, Makabongwe Mazibuko, Aviwe Ntsethe, et al, (2025), Design and Development of Novel Anticancer Peptide Encapsulated Liposomes for Targeting of Solid Tumours In Vitro, International Journal of Clinical Case Reports and Reviews, 25(5); DOI:10.31579/2690-4861/589
Copyright: © 2025, Mbuso Faya. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Received: 25 April 2025 | Accepted: 07 May 2025 | Published: 15 May 2025
Keywords: anticancer peptides; liposomes; formulation; solid tumours
Conventional cancer treatments possess limitations for solid tumours, such as lack of selectivity, thus nanomedicine is explored as an efficient tool in anticancer drug development. Therefore, this study aimed to design a novel anticancer peptide (ACP) and encapsulate it into a lipid-based nanoparticle for efficient delivery to tumours. ACP was designed using in silico methods and thereafter encapsulated into a liposomal formulation (P1CF1) using a thin-film hydration method. The shape, size, polydispersity index (PDI), and zeta potential (ZP) of the formulation were evaluated using cryo-transmission electron microscopy, and zetasizer, while the percentage drug encapsulation efficiency (%EE) and drug release were investigated using the ultra-filtration and the dialysis methods. The biocompatibility, cytotoxicity, and apoptosis activity of P1CF1 were evaluated using hemolysis, 3-[(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] and annexin V-FITC/PI assays. P1CF1 was spherical, had a size of 193.46 ± 0.10 nm, a PDI of 0.342 ± 0.12, a ZP of -7.67 ± 0.04 mV, and an %EE of 91.23 ± 0,01%. Furthermore, P1CF1 was biocompatible at low concentrations, showed a controlled in vitro drug release, induced high cytotoxic (IC50 value of 2.967 µg/ml) and apoptosis effects on the cancer cells MCF-7 and was well tolerated by non-cancer HEK293 cells (IC50 value of 135.3 µg/ml). Overall, P1CF1 showed efficient encapsulation capacity, enhanced biocompatibility, and significant anticancer activity in tumour cells with minimum effect on healthy cells. These positive characteristics indicate potential in vivo applicability. Thus, future research can include in vivo evaluation of this novel formulation.
Cancer is a significant public health challenge as it is the second-leading cause of death in the world [1]. From the many available cancer treatment modalities, chemotherapy is regarded with greater significance in cancer treatment options and will most likely remain so for many decades to come [2]. However, though cancer therapy with conventional chemotherapeutic drugs has some benefits, the direct administration of chem-drugs is associated with severe limitations, such as rapid elimination, low bioavailability, lack of specificity, which can result in systemic toxicity and adverse consequences; and multidrug resistance (MDR), which can lead to recurring, unresponsive tumors and inadequate drug dose, which can impede certain apoptotic pathways and hinder cell death processes [3-6]. Thus, methods to address these challenges are still required.
The use of lipid-based delivery systems such as liposomes as drug carriers in nanomedicine has gained popularity due to their sophistication and attractive traits [7]. These include flexibility, ease of synthesis, reliability, versatility, nano-size (50-500 nm), biocompatibility, enhanced bioavailability, biodegradability, the capacity to shield payloads from intracellular enzyme degradation; targeted drug delivery to tumor tissues; enhanced effectiveness and therapeutic index; the capacity to encapsulate drugs that are both hydrophilic and hydrophobic; high levels of drug loading and entrapment efficiency; regulated drug release profiles; reduced toxicity of the encapsulated drugs; enhanced pharmacokinetic effects including reduced excretion, prolonged circulation lifespans; and ability to be tailor-designed for targeted drug delivery [8-12]. Because of the leaky nature of tumor tissue capillaries, these vesicles' spherical form and micro size allow them to extravasate and passively accumulate in cancer regions, a phenomenon known as the increased permeability and retention (EPR) effect [13]. To date, numerous liposome formulations have been approved for cancer therapy, including OnivydeTM, Marqibo®, Doxil®, Visudyne®, and Depocyt® [14]. Future research and innovation in liposomal drug delivery systems hold considerable potential for the advancement of pharmaceuticals and nanomedicine.
The concept of peptide-targeted liposomes is a significant advancement in the usage of liposomal formulations in cancer treatment. Peptides have been identified as attractive and viable targeting ligands for directing liposomes to target tumours and sequentially enhancing the selectivity and specificity of drug-loaded liposomes, minimizing off-target delivery [15, 16]. This is supported by many in vivo and in vitro research that have been documented over the years [16-24]. However, there is a huge gap in the literature and drug development focussing on liposomal encapsulation of anticancer peptides (APCs) that target solid tumours.
Recently, ACPs have vastly become a developing approach in nanomedicine, particularly, in drug development. These are a series of short amino acids, with anticancer properties which are alternatives to chemo-drugs used in cancer treatments. These ACPs cause cell death by different mechanisms, such as mediated immunity, membrane disruption apoptosis, DNA synthesis inhibition, hormonal/membrane receptors, and anti-angiogenic [25, 26]. Though these drugs exhibit great anticancer properties, limited studies have been done on them as therapeutic payloads, and this could be due to their lack of bioavailability because of intracellularly degraded following administration. Therefore, extensive research is required to establish whether liposomes can encapsulate these ACPs and deliver them safely and efficiently to tumour sites. Thus, this study aimed to design and advance a novel anti-cancer peptide using computational-aided drug design tools, encapsulate it into a liposome and evaluate its physicochemical features as well as its biological activity in vitro.
2.1 Materials
Novel anticancer peptides (sequence: FKKLLAKLAK) were designed in-house using the lead anticancer peptide purchased from ChinaPeptides (QYAOBIO) Ltd, China, by a solid phase peptide synthesis protocol. Cholesterol and phosphatidylcholine that were purchased from Sigma-Aldrich (USA) were used for liposomal preparations. Trifluoracetic acid (TFA) acetonitrile, phosphate-buffered saline (PBS), dichloromethane, and a dialysis bag with a molecular weight cut-off (MWCO) size of 10,000 Dalton (Da) was purchased from Sigma-Aldrich (USA). All other solvents used were of High-Performance Liquid Chromatography (HPLC) analytical grade and were used without additional purification. Distilled water was used throughout this study and was purified in the laboratory with a Milli-Q purification system (Millipore Corp., USA).
2.2 Methods
2.2.1 Anticancer Peptide Design
The amino acid sequences of the purified peptides were determined using the automated online software CancerPPD (Database of Anticancer Peptides and Proteins) (http://crdd.osdd.net/raghava/cancerppd/) [27]. The peptide sequence FAKLLAKLAK with ID 1854 and a 10 amino acid chain, was used as the anticancer reference sequence that targeted solid tumours - breast cancer. The reference amino acid sequence was incorporated into CellPPD: Designing of Cell Penetrating Peptides (http://crdd.osdd.net/raghava/cellppd) online software to select mutants of the reference compound. A mutant peptide was generated with a 10 amino acid length, FKKLLAKLAK. The lead anticancer peptide was then purchased from ChinaPeptides (QYAOBIO) Ltd., China The sequence FKKLLAKLAK (Phe-Lys-Lys-Leu-Leu-Ala-Lys-Leu-Ala-Lys) was identified with a purity of 98.28% and was soluble at 1 mg/ml in 17
Over the years, therapeutic peptides have seemingly received significant attention from scientists as potential drug candidates [36]. Peptides as therapeutic agents have proved to be favourable for many diseases, including cancer, and their applications have been highly advantageous due to their size, high biocompatibility, simplicity in production, and/or modification together with their capacity to penetrate tumours [37]. Much research has been conducted on novel therapeutic ACPs using in silico tools to target various diseases. Consequently, there have been many ACPs that have entered clinical trials but still only a few have been approved.
3.1 Identification and Design of Novel Anti-Cancer Peptides (ACPs)
The ACP was designed using the module CancerPPD database (Database of Anticancer Peptides and Proteins) which has already been validated with anticancer activity [38]. For each peptide on the CancerPPD database, adequate details on the specific assays utilized and the experimentally measured activity of the peptides against different cancer cell lines are provided [39]. During selection, the peptide sequences needed to be short, thus lengths ranged from 5 - 10 amino acids due to the consideration of synthesis and cost of the peptide. The initially selected sequence was FLAK50 T1 (FAKLLAKLAK), and the strategy for novelty was to adjust the amino acids in the given sequence. The sequence FLAK50 T1 originates from FLAK peptides that are highly rich in Phe, Leu, Ala, and Lys and were compiled in the Owen patent dataset [40]. The peptide targets breast cancer and has chirality L, the sequence was linear and the reported activity was LD50 = 615 µg/ml. The retrieved sequence from CancerPPD was then taken to CellPPD to develop mutant peptides of the sequence. CellPPD database, which is a support vector machine (SMV), assists by developing and designing cell-penetrating peptides (CPPs), which was essential when looking for mutants of the retrieved sequence [27, 41]. CellPPD allows for the design of single mutant analogues of given peptide sequences and identifies whether they are penetrating cells or not. Additionally, CellPPD also provides the physicochemical properties of the generated mutant peptide. From the retrieved sequence, Ala amino acid was substituted with Lys to generate the mutant peptide. The newly generated peptide had the sequence FKKLLAKLAK (Phe-Lys-Lys-Leu-Leu-Ala-Lys-Leu-Ala-Lys) (Fig 1). The peptide had a molecular weight of 1159.67 and a +4 net charge, which advantageously leads to the destruction of cancer cells by engaging with their anionic cell membrane components [42]. The SVM score was 0.30 and the peptide had a clear CPP prediction. For ACPs, hydrophobicity plays an important role in the peptide's ability to cross membrane barriers and hence exert their anticancer effects [43]. The ACP had a hydrophobicity of -0.17, which was ideal as the peptide showed to be mostly hydrophilic. The hydrophilicity was 0.31 and the hydropathicity was 0.22.
Figure 1: 3D structure (a) and 2D structure (b) of the designed ACP - FKKLLAKLAK (Phe-Lys-Lys-Leu-Leu-Ala-Lys-Leu-Ala-Lys). Images drawn using ChemDraw Software
3.2 Characterisation of ACP Encapsulated Liposomes
3.2.1 Cryo-TEM, DLS-Zeta Sizer
The physicochemical properties of the drug delivery systems have a substantial impact on their tumour permeability, biodistribution, and blood circulation half-life [44]. The ACP-loaded liposome formulations and blank liposomes (without ACP) were successfully formulated using the thin-film hydration method. A small volume of cholesterol was included in the formulation to increase the stability of the lipid bilayer in biological fluids such as blood plasma [45].
Cryo-TEM images revealed that all formulated liposomes formed spherical structures with similar homogenous sizes (Fig 2). The DLS-Zeta Sizer revealed that the sizes of the liposome formulations ranged from 164 nm – 194 nm, with the blank liposome having a mean diameter size of 164.1 ± 0.11nm, while the ACP liposome has a slightly larger mean diameter size of 193.4 ± 0.10 nm (Table 1). The increase in size could have been due to the presence of the ACP within the liposome. Studies have shown that NPs larger than 200 nm in diameter tend to activate the complement system, which causes them to leave the bloodstream fast and accumulate in the liver and spleen [44]. Therefore, the formulated liposome sizes fell within the standard size range which is between 100 nm and 200 nm needed for drug delivery through non-specific or receptor-specific endocytosis cellular uptake [46-49]. The use of probe sonication could have had a major influence in obtaining the ideal particle size, as this technique can rearrange and reassemble the lipid content of the liposome to create favourable particle sizes [50]. The PDI value which is correlated to the distribution stability of the formulation, is an important indicator of the overall liposome size distribution [51]. According to literature, PDI values, close to 1.0 are considered not ideal as they may indicate the inverse distribution of the particles and/or the presence of large particles [52]. Ideally, PDI values should range between 0.30 and low, which indicates that over 60% of the nano-formulations are within the same particle size range and are evenly distributed [51-53]. From the results obtained, the PDI of the blank was below the optimum 0.30, however, the ACP encapsulated liposomes ranged close to the optimum (0.294 ± 0,12 nm to 0.394 ± 0,12 nm) thus showing high-to-medium homogeneity of the liposome mean sizes.
To investigate the physical stability of liposomes, zeta potential was measured. Zeta potential distinguishes the particle surface charge, provides data on the repulsive forces that exist between the particles, and aids in colloidal dispersion stability estimations [54]. Good colloidal stability of nanoparticles is related to ZP values greater than +/- 25 mV [55, 56]. All the liposome formulations in the study presented negative ZP values (negative charge), which could be due to the phosphatidylcholine’s (used in the thin film) headgroup orientation located at the vesicles surface, and the position of the phosphate group above the choline group plane [57, 58]. The blank liposomes exhibited a ZP of -7.73 ± 0.47 mV, while the ACP liposomes showed a ZP of -7.67 ± 0.04 mV. These findings suggested that the ACP liposome formulation displayed low colloidal stability, which has no profound impact in in vitro studies. However, before the application of the formulation in vivo, the inclusion of a cationic phospholipid or cationic polymer, helper lipid, and stealth polymer (e.g., polyethylene glycol, PEG) is mandatory for improved colloidal stability and longer circulation half-life of this system.
Figure 2: TEM micrographs of the P1CF1 formulation at different resolutions showing efficient encapsulation of the peptide in the liposome. Scale bars = 0.2 µm; 100 nm.
Table 1: Particle size, polydispersity Index (PDI), and zeta potential (ZP) of blank liposomes and ACP encapsulated liposomes. Data displayed as mean ± SD (n = 3).
3.3 Determination of Encapsulation Efficiency Percentage
The analysis of liposome encapsulation efficiency (EE) is an extremely crucial parameter since liposomes are utilized in pharmaceuticals as drug carriers that facilitate drug absorption, targeting effects, and protection of drugs [59]. According to the literature, the EE% is calculated from the difference between the entrapped drug (the total amount of drug added to the liposome formulation) and the unentrapped drug (the amount of drug found in the supernatant of the resulting liposome formulation) divided by the total drug added (EE% = total drug added – free non-entrapped drug / total drug added) [60]. In this study, the encapsulation efficiency of the liposome was determined using RP-HPLC at wavelength 220 nm. The standard curve of the formulation was made by plotting HPLC peak areas against the concentration. The steps indirect method used for the entrapped and unentrapped values were reported by Suleiman and coworkers [61], however, the dilutions used in this study were with acetonitrile. The concentrations of the entrapped and unentrapped drugs were calculated using the calibration curve with the equation y=5045,2x. To measure the amount of active drug loaded in a liposome, the drug would have had to be fully encapsulated by the lipid of the liposome formulation. To determine the EE, the mass ratio between the amount of the drug integrated into the liposome and this ratio was employed in the liposome preparation. The overall entrapment yield was calculated to be 91.23 ± 0.01%, which is relatively high; this was expected as many liposomes have been reported to possess high EE because of their high volume to surface area ratio [9]. The relatively high EE% indicates that the ratio of the used lipids to the ACP was an optimal choice for the formulation. The obtained results from the study revealed high EE and this may be due to many factors, one of them being the effect of probe sonication. According to the literature, liposomal formulations that have been probe-sonicated for longer tend to be more homogenized and hence more susceptible to interacting with surrounding molecules, which greatly influences their enhanced encapsulation efficiency [62]. Additionally, the hydrophilic nature of the peptide used herein could have further facilitated the observed high EE%. This is according to previous studies which have shown that the encapsulation of hydrophilic peptides (as with the one used in this study) in liposomes is expected to be the most efficient due to the electrostatic interaction between the peptide and the liposome surface [61, 63].
3.4 In vitro Hemolysis Testing
The in vivo application of drugs/drug-loaded nanoformulations entails their transportation via the bloodstream which could lead to adverse effects, such as immunological responses, complex formation with macromolecules, and cell damage [64]. Exposure to chemicals, such as drugs or drug-loaded nanoformulations, can result in an early breakdown of erythrocytes of red blood cells releasing the hemoglobin (Hb), which can disrupt normal oxygen transport and induce hemolytic anemia/hemolysis [65, 66]. The drugs/drug-loaded nanoformulations may either adsorb on the membrane of the erythrocyte, causing the membrane to distort and become damaged [67]; or they have the ability to cause osmotic lysis by causing holes in the erythrocyte membrane [68]. Thus, it is important to determine if drug-loaded nanoformulations with therapeutic effects induce hemolysis in erythrocytes in vitro [69]. To establish this, erythrocytes of sheep red blood cells were exposed to different doses of the ACP encapsulated liposome formulation (P1CF1) and evaluated for possible hemolysis [70].
The percentage (%) of erythrocyte hemolysis induced by the P1CF1 formulation is shown in Fig 3. Both the graph and the image insert depicted that the formulation was non-toxic to erythrocytes at doses 0.05 and 0.1 mg/ml, suggesting its biocompatibility at these low therapeutic doses. Hemolysis was neglectable at a dose of 0.2 mg/ml and increased with increasing concentration of the P1CF1 formulation from doses greater than 0.2 mg/ml up to 0.5 mg/ml. This can be credited to the agglomeration feature of ACP liposomes at higher doses. Moreover, compared to untreated cells (C2), an 88% increase in hemolysis was observed at a dose of 0.5 mg/ml post-treatment, indicating that the formulation had an adverse effect on erythrocytes at higher doses. Overall, these findings indicated that the formulation was non-toxic at low therapeutic concentrations equal to or less than 0.2 mg/ml.
Figure 3: Hemolysis toxicity of the P1CF1 formulation towards red blood cells at varying concentrations (0.05 to 0.5 mg/ml). Data is shown as means ± SD (n = 3). The image insert shows the representation of the haemolytic behaviour of RBCs after treatment with P1CF1 formulation. C1 denotes control 1 = positive control (+) = RBCs + distilled water; C2 denotes control 2 = negative control (-) = RBCs + PBS. ∗∗∗∗p < 0>.0001 vs. C2; ns indicates non-statistical significance from C2.
3.5 In vitro Drug Release Analysis
To predict the quantity of a drug's accumulation into the bloodstream and tumour sites over time, in vitro drug release studies are usually conducted. The in vitro drug release profile of the ACP encapsulated liposomes (P1CF1 formulation) was assessed using a dialysis bag technique with PBS solutions at pH 6.0 and 7.4 prepared at 37°C for 72 hrs. These pH buffer solutions mimic the basic physiological state and the acidic endosomal cancer cell microenvironment (pH 4-6) [71]. Bare peptide was used as a reference or negative control. There was only 5.1% and 3.7% of
bare peptide released at pH of 7.4 and pH 6.0 respectively at 72 hrs (Figure 8 in the Supplementary Information). This was expected since the peptide was not encapsulated in any nanoparticle, so, its large size could have restricted its release or diffusion across the semipermeable dialysis bag membrane.
Figure 4 displays the release profile of P1CF1 formulations and the accumulative release of loaded ACP, which was sustained/controlled, acid-dependent, and exhibited a biphasic release pattern across the time studied. During the first 10 hours, there was a quick release of the APC, followed by a slow controlled release of the APC for the remaining duration of 72 hrs. A higher ACP release rate of 88% was seen at an acidic pH of 6.0 whereas a slower drug release rate of 79% was seen at a healthy pH of 7.4. The initial burst ACP release may have been caused by the release of the unentrapped ACP on the liposome periphery, while the subsequent slow and controlled release may have been caused by the release of the ACP encapsulated inside the liposome [71, 72]. The protonation of the amine groups of the encapsulated ACP (see Fig 1) at acidic pHs results in a conformational change of the liposome, causing swelling, bursting, and releasing of the encapsulated ACP into the buffer solution [73]. Sustained drug release is of high importance for cancer therapeutics as it enables the drugs to be released for longer durations, ensuring a continuous stable dosage of the drug to the tumour sites [74]. The cholesterol present within the liposome also helps to regulate the properties of the lipid bilayer of the liposomes as well as the release of water-soluble compounds (i.e. the ACP peptide) from liposomes by influencing the fluidity and permeability of the lipid bilayer [1, 75]. The pH-dependent drug release property raises the possibility that this P1CF1 formulation could be employed to deliver anticancer drugs specifically to tumours (with an acidic microenvironment) [74, 76]. Additionally, the results indicate that most of the ACP encapsulated within the liposome will remain intact in the plasma (pH 7.4) after injection, greatly decreasing the possibility of any systemic adverse effects on the healthy tissues. Furthermore, the results showed that once inside the tumour cells, the cytosolic release of the ACP-encapsulated liposomes/ P1CF1 from the endosome is expected to be quicker due to the lower pH than physiological pH conditions. Consequently, it is hoped that the encapsulation of ACP into liposomes will improve the effectiveness of targeted cancer treatment.
Figure 4: In vitro drug release profile of ACP encapsulated liposome in pH 6.0 and 7.4 over 72 hours. Data is shown as mean ± SD (n = 3).
3.6 In vitro Cytotoxicity Analysis
The initial step in assessing a drug delivery system's biocompatibility usually includes the use of cell-culture-based research such as in vitro cell viability studies, which typically starts with the assessment of its cytotoxicity profile. The MTT test that was used in this study quantifies metabolically active cells by measuring yellow formazan products produced from the conversion of purple MTT salt by mitochondrial reductase enzyme found in live cells [34]. Untreated cells served as negative controls, azacitidine, a known anticancer drug [77], was used as a positive control.
MCF-7 cancer cells and HEK293 non-cancer cells were treated with ACP-encapsulated liposomes (P1CF1), bare peptide, and azacitidine at varying doses ranging from 10 to 200 µg/ml (Figure 5-6). Following treatment, a drastic decrease in
A novel anticancer peptide-encapsulated liposome formulation (P1CF1) was successfully designed and formulated using in silico and thin-film layer rehydration methods, respectively. The formulation had optimal physiochemical characteristics for efficient drug delivery in vitro, as well as improved encapsulation efficiency. Encapsulating the novel anticancer peptide in a liposome delivery system improved its in vitro bioavailability and biocompatibility. At acidic pH conditions, the formulation also demonstrated increased controlled and sustained in vitro anticancer peptide release, implying that treatment at the target tumour sites might be prolonged. The P1CF1 formulation had higher cytotoxicity than the known anticancer drug, azacitidine in cancer cells, with a percentage cell inhibition of 98.2%. However, it was ineffective in non-cancer cells (
project conceptualization and designing, Lusanda Mtetwa, Nkeiruka Igbokwe, and Mbuso Faya.; resources, Mbuso Faya and Anil Chuturgoon.; data collection.; Lusanda Mtetwa.; data curation and software, Lusanda Mtetwa, Nkeiruka Igbokwe, Aviwe Ntsethe, Makabongwe Mazibuko and Terisha Ghazi.; writing - initial draft preparation, Lusanda Mtetwa.; writing - final draft preparation, Lusanda Mtetwa, Nkeiruka Igbokwe and Londiwe Simphiwe Mbatha.; final draft reviewing and editing, Londiwe Simphiwe Mbatha and Mbuso Faya.; funding acquisition, Mbuso Faya and Anil Chuturgoon.; project supervision and administration, Mbuso Faya. All authors have read and agreed to the published version of the manuscript.
The University of KwaZulu Natal, College of Health Science is acknowledged for financial support as well as the National Research Foundation (grant number: 138291). Any opinions, findings, conclusions, or recommendations expressed in this article are those of the authors.
Supporting data are available from the corresponding author upon request.
Conflicts of interest: The authors declare no conflict of interest.
Declarations: Ethical approval, is not applicable.
Fig 8. In vitro drug release profile of bare ACP in pH 7.4 and pH 6.0 over 72 hours. Data is shown as mean ± SD (n = 3); Fig 9(a-d). IC50 estimated values of P1CF1 (a), bare ACP (b), and azacitidine (c) in MCF-7 cell lines; IC50 estimated value of P1CF1 (d) in HEK293; Table 3. Flow cytometry analysis; gate total percentages of MCF-7 only, P1CF1, Bare ACP, and Azacitidine.
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I am very pleased to serve as EBM of the journal, I hope many years of my experience in stem cells can help the journal from one way or another. As we know, stem cells hold great potential for regenerative medicine, which are mostly used to promote the repair response of diseased, dysfunctional or injured tissue using stem cells or their derivatives. I think Stem Cell Research and Therapeutics International is a great platform to publish and share the understanding towards the biology and translational or clinical application of stem cells.
I would like to give my testimony in the support I have got by the peer review process and to support the editorial office where they were of asset to support young author like me to be encouraged to publish their work in your respected journal and globalize and share knowledge across the globe. I really give my great gratitude to your journal and the peer review including the editorial office.
I am delighted to publish our manuscript entitled "A Perspective on Cocaine Induced Stroke - Its Mechanisms and Management" in the Journal of Neuroscience and Neurological Surgery. The peer review process, support from the editorial office, and quality of the journal are excellent. The manuscripts published are of high quality and of excellent scientific value. I recommend this journal very much to colleagues.
Dr.Tania Muñoz, My experience as researcher and author of a review article in The Journal Clinical Cardiology and Interventions has been very enriching and stimulating. The editorial team is excellent, performs its work with absolute responsibility and delivery. They are proactive, dynamic and receptive to all proposals. Supporting at all times the vast universe of authors who choose them as an option for publication. The team of review specialists, members of the editorial board, are brilliant professionals, with remarkable performance in medical research and scientific methodology. Together they form a frontline team that consolidates the JCCI as a magnificent option for the publication and review of high-level medical articles and broad collective interest. I am honored to be able to share my review article and open to receive all your comments.
“The peer review process of JPMHC is quick and effective. Authors are benefited by good and professional reviewers with huge experience in the field of psychology and mental health. The support from the editorial office is very professional. People to contact to are friendly and happy to help and assist any query authors might have. Quality of the Journal is scientific and publishes ground-breaking research on mental health that is useful for other professionals in the field”.
Dear editorial department: On behalf of our team, I hereby certify the reliability and superiority of the International Journal of Clinical Case Reports and Reviews in the peer review process, editorial support, and journal quality. Firstly, the peer review process of the International Journal of Clinical Case Reports and Reviews is rigorous, fair, transparent, fast, and of high quality. The editorial department invites experts from relevant fields as anonymous reviewers to review all submitted manuscripts. These experts have rich academic backgrounds and experience, and can accurately evaluate the academic quality, originality, and suitability of manuscripts. The editorial department is committed to ensuring the rigor of the peer review process, while also making every effort to ensure a fast review cycle to meet the needs of authors and the academic community. Secondly, the editorial team of the International Journal of Clinical Case Reports and Reviews is composed of a group of senior scholars and professionals with rich experience and professional knowledge in related fields. The editorial department is committed to assisting authors in improving their manuscripts, ensuring their academic accuracy, clarity, and completeness. Editors actively collaborate with authors, providing useful suggestions and feedback to promote the improvement and development of the manuscript. We believe that the support of the editorial department is one of the key factors in ensuring the quality of the journal. Finally, the International Journal of Clinical Case Reports and Reviews is renowned for its high- quality articles and strict academic standards. The editorial department is committed to publishing innovative and academically valuable research results to promote the development and progress of related fields. The International Journal of Clinical Case Reports and Reviews is reasonably priced and ensures excellent service and quality ratio, allowing authors to obtain high-level academic publishing opportunities in an affordable manner. I hereby solemnly declare that the International Journal of Clinical Case Reports and Reviews has a high level of credibility and superiority in terms of peer review process, editorial support, reasonable fees, and journal quality. Sincerely, Rui Tao.
Clinical Cardiology and Cardiovascular Interventions I testity the covering of the peer review process, support from the editorial office, and quality of the journal.
Clinical Cardiology and Cardiovascular Interventions, we deeply appreciate the interest shown in our work and its publication. It has been a true pleasure to collaborate with you. The peer review process, as well as the support provided by the editorial office, have been exceptional, and the quality of the journal is very high, which was a determining factor in our decision to publish with you.
The peer reviewers process is quick and effective, the supports from editorial office is excellent, the quality of journal is high. I would like to collabroate with Internatioanl journal of Clinical Case Reports and Reviews journal clinically in the future time.
Clinical Cardiology and Cardiovascular Interventions, I would like to express my sincerest gratitude for the trust placed in our team for the publication in your journal. It has been a true pleasure to collaborate with you on this project. I am pleased to inform you that both the peer review process and the attention from the editorial coordination have been excellent. Your team has worked with dedication and professionalism to ensure that your publication meets the highest standards of quality. We are confident that this collaboration will result in mutual success, and we are eager to see the fruits of this shared effort.
Dear Dr. Jessica Magne, Editorial Coordinator 0f Clinical Cardiology and Cardiovascular Interventions, I hope this message finds you well. I want to express my utmost gratitude for your excellent work and for the dedication and speed in the publication process of my article titled "Navigating Innovation: Qualitative Insights on Using Technology for Health Education in Acute Coronary Syndrome Patients." I am very satisfied with the peer review process, the support from the editorial office, and the quality of the journal. I hope we can maintain our scientific relationship in the long term.
Dear Monica Gissare, - Editorial Coordinator of Nutrition and Food Processing. ¨My testimony with you is truly professional, with a positive response regarding the follow-up of the article and its review, you took into account my qualities and the importance of the topic¨.
Dear Dr. Jessica Magne, Editorial Coordinator 0f Clinical Cardiology and Cardiovascular Interventions, The review process for the article “The Handling of Anti-aggregants and Anticoagulants in the Oncologic Heart Patient Submitted to Surgery” was extremely rigorous and detailed. From the initial submission to the final acceptance, the editorial team at the “Journal of Clinical Cardiology and Cardiovascular Interventions” demonstrated a high level of professionalism and dedication. The reviewers provided constructive and detailed feedback, which was essential for improving the quality of our work. Communication was always clear and efficient, ensuring that all our questions were promptly addressed. The quality of the “Journal of Clinical Cardiology and Cardiovascular Interventions” is undeniable. It is a peer-reviewed, open-access publication dedicated exclusively to disseminating high-quality research in the field of clinical cardiology and cardiovascular interventions. The journal's impact factor is currently under evaluation, and it is indexed in reputable databases, which further reinforces its credibility and relevance in the scientific field. I highly recommend this journal to researchers looking for a reputable platform to publish their studies.
Dear Editorial Coordinator of the Journal of Nutrition and Food Processing! "I would like to thank the Journal of Nutrition and Food Processing for including and publishing my article. The peer review process was very quick, movement and precise. The Editorial Board has done an extremely conscientious job with much help, valuable comments and advices. I find the journal very valuable from a professional point of view, thank you very much for allowing me to be part of it and I would like to participate in the future!”
Dealing with The Journal of Neurology and Neurological Surgery was very smooth and comprehensive. The office staff took time to address my needs and the response from editors and the office was prompt and fair. I certainly hope to publish with this journal again.Their professionalism is apparent and more than satisfactory. Susan Weiner
My Testimonial Covering as fellowing: Lin-Show Chin. The peer reviewers process is quick and effective, the supports from editorial office is excellent, the quality of journal is high. I would like to collabroate with Internatioanl journal of Clinical Case Reports and Reviews.
My experience publishing in Psychology and Mental Health Care was exceptional. The peer review process was rigorous and constructive, with reviewers providing valuable insights that helped enhance the quality of our work. The editorial team was highly supportive and responsive, making the submission process smooth and efficient. The journal's commitment to high standards and academic rigor makes it a respected platform for quality research. I am grateful for the opportunity to publish in such a reputable journal.
My experience publishing in International Journal of Clinical Case Reports and Reviews was exceptional. I Come forth to Provide a Testimonial Covering the Peer Review Process and the editorial office for the Professional and Impartial Evaluation of the Manuscript.
I would like to offer my testimony in the support. I have received through the peer review process and support the editorial office where they are to support young authors like me, encourage them to publish their work in your esteemed journals, and globalize and share knowledge globally. I really appreciate your journal, peer review, and editorial office.
Dear Agrippa Hilda- Editorial Coordinator of Journal of Neuroscience and Neurological Surgery, "The peer review process was very quick and of high quality, which can also be seen in the articles in the journal. The collaboration with the editorial office was very good."
I would like to express my sincere gratitude for the support and efficiency provided by the editorial office throughout the publication process of my article, “Delayed Vulvar Metastases from Rectal Carcinoma: A Case Report.” I greatly appreciate the assistance and guidance I received from your team, which made the entire process smooth and efficient. The peer review process was thorough and constructive, contributing to the overall quality of the final article. I am very grateful for the high level of professionalism and commitment shown by the editorial staff, and I look forward to maintaining a long-term collaboration with the International Journal of Clinical Case Reports and Reviews.
To Dear Erin Aust, I would like to express my heartfelt appreciation for the opportunity to have my work published in this esteemed journal. The entire publication process was smooth and well-organized, and I am extremely satisfied with the final result. The Editorial Team demonstrated the utmost professionalism, providing prompt and insightful feedback throughout the review process. Their clear communication and constructive suggestions were invaluable in enhancing my manuscript, and their meticulous attention to detail and dedication to quality are truly commendable. Additionally, the support from the Editorial Office was exceptional. From the initial submission to the final publication, I was guided through every step of the process with great care and professionalism. The team's responsiveness and assistance made the entire experience both easy and stress-free. I am also deeply impressed by the quality and reputation of the journal. It is an honor to have my research featured in such a respected publication, and I am confident that it will make a meaningful contribution to the field.
"I am grateful for the opportunity of contributing to [International Journal of Clinical Case Reports and Reviews] and for the rigorous review process that enhances the quality of research published in your esteemed journal. I sincerely appreciate the time and effort of your team who have dedicatedly helped me in improvising changes and modifying my manuscript. The insightful comments and constructive feedback provided have been invaluable in refining and strengthening my work".
I thank the ‘Journal of Clinical Research and Reports’ for accepting this article for publication. This is a rigorously peer reviewed journal which is on all major global scientific data bases. I note the review process was prompt, thorough and professionally critical. It gave us an insight into a number of important scientific/statistical issues. The review prompted us to review the relevant literature again and look at the limitations of the study. The peer reviewers were open, clear in the instructions and the editorial team was very prompt in their communication. This journal certainly publishes quality research articles. I would recommend the journal for any future publications.
Dear Jessica Magne, with gratitude for the joint work. Fast process of receiving and processing the submitted scientific materials in “Clinical Cardiology and Cardiovascular Interventions”. High level of competence of the editors with clear and correct recommendations and ideas for enriching the article.
We found the peer review process quick and positive in its input. The support from the editorial officer has been very agile, always with the intention of improving the article and taking into account our subsequent corrections.
My article, titled 'No Way Out of the Smartphone Epidemic Without Considering the Insights of Brain Research,' has been republished in the International Journal of Clinical Case Reports and Reviews. The review process was seamless and professional, with the editors being both friendly and supportive. I am deeply grateful for their efforts.
To Dear Erin Aust – Editorial Coordinator of Journal of General Medicine and Clinical Practice! I declare that I am absolutely satisfied with your work carried out with great competence in following the manuscript during the various stages from its receipt, during the revision process to the final acceptance for publication. Thank Prof. Elvira Farina
Dear Jessica, and the super professional team of the ‘Clinical Cardiology and Cardiovascular Interventions’ I am sincerely grateful to the coordinated work of the journal team for the no problem with the submission of my manuscript: “Cardiometabolic Disorders in A Pregnant Woman with Severe Preeclampsia on the Background of Morbid Obesity (Case Report).” The review process by 5 experts was fast, and the comments were professional, which made it more specific and academic, and the process of publication and presentation of the article was excellent. I recommend that my colleagues publish articles in this journal, and I am interested in further scientific cooperation. Sincerely and best wishes, Dr. Oleg Golyanovskiy.
Dear Ashley Rosa, Editorial Coordinator of the journal - Psychology and Mental Health Care. " The process of obtaining publication of my article in the Psychology and Mental Health Journal was positive in all areas. The peer review process resulted in a number of valuable comments, the editorial process was collaborative and timely, and the quality of this journal has been quickly noticed, resulting in alternative journals contacting me to publish with them." Warm regards, Susan Anne Smith, PhD. Australian Breastfeeding Association.
