Pub Date : 2024-11-07DOI: 10.1016/j.ijpharm.2024.124928
Fiona Fernandes, Indrani Talukdar, Meenal Kowshik
Efficient and safe gene delivery is vital for genetic manipulation of stem cells for regenerative medicine. Gold nanoparticles have been used for various biomedical applications in the past, and are currently being researched as transfection agents. In this study, we report a simple one-pot synthesis of positively charged gold nanoparticles functionalized with cysteamine. The nanoparticles exhibit no cytotoxicity and can bind to both plasmid DNA (pDNA) as well as small interference RNA (siRNA). We observed that a five fold lower concentration of pDNA was sufficient for achieving comparable overexpression as that of a commercial transfection agent. We also observed that about 70 % transient silencing of the target gene was achieved with only 25 nM siRNA delivered by our nano-vehicle. To better understand the fate of the nanoparticle, we attempted to identify its uptake mechanism. The results indicate that while all the mechanisms contribute to the uptake, the clathrin-dependent pathway plays a major role. This is the first study on understanding the mechanism of uptake of CA-AuNPs conjugated to pDNA by embryonic stem cells. This is also the first study, where a successful transfection using gold based nanoparticles has been achieved in ESCs at a concentration as low as 0.5 µg/ml for pDNA and 25ƞM siRNA.
{"title":"Cysteamine functionalized gold nanoparticles exhibit high efficiency delivery of genetic materials in embryonic stem cells majorly via clathrin mediated endocytosis","authors":"Fiona Fernandes, Indrani Talukdar, Meenal Kowshik","doi":"10.1016/j.ijpharm.2024.124928","DOIUrl":"10.1016/j.ijpharm.2024.124928","url":null,"abstract":"<div><div>Efficient and safe gene delivery is vital for genetic manipulation of stem cells for regenerative medicine. Gold nanoparticles have been used for various biomedical applications in the past, and are currently being researched as transfection agents. In this study, we report a simple one-pot synthesis of positively charged gold nanoparticles functionalized with cysteamine. The nanoparticles exhibit no cytotoxicity and can bind to both plasmid DNA (pDNA) as well as small interference RNA (siRNA). We observed that a five fold lower concentration of pDNA was sufficient for achieving comparable overexpression as that of a commercial transfection agent. We also observed that about 70 % transient silencing of the target gene was achieved with only 25 nM siRNA delivered by our nano-vehicle. To better understand the fate of the nanoparticle, we attempted to identify its uptake mechanism. The results indicate that while all the mechanisms contribute to the uptake, the clathrin-dependent pathway plays a major role. This is the first study on understanding the mechanism of uptake of CA-AuNPs conjugated to pDNA by embryonic stem cells. This is also the first study, where a successful transfection using gold based nanoparticles has been achieved in ESCs at a concentration as low as 0.5 µg/ml for pDNA and 25ƞM siRNA.</div></div>","PeriodicalId":14187,"journal":{"name":"International Journal of Pharmaceutics","volume":"667 ","pages":"Article 124928"},"PeriodicalIF":5.3,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142619833","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-07DOI: 10.1016/j.ijpharm.2024.124880
Sohaib Mahri , Céline Cassiers , Sandra Gracin , Donatienne Tyteca , Francine Uwambayinema , François Huaux , Mariam Ibrahim , Sian Piper , Antonio Llinas , Markus Fridén , Rita Vanbever
Spray drying is a widely employed method for generating dry powder formulations for inhalation. Yet, it presents substantial challenges when applied to therapeutic proteins due to stability issues. The formation of protein aggregates during the atomization and the heating steps can diminish protein activity and raise immunogenicity concerns. Here, we assessed the impact of varying levels of protein aggregates generated during spray-drying on the fate and the immunogenicity of the human monoclonal antibody NIP228 following intratracheal administration in mice. Aggregate-free rhodamine labelled NIP228 was spray-dried with or without 1% polysorbate 80 surfactant, resulting in the generation of powder formulations with associated low and high protein aggregate levels, respectively. Confocal imaging highlighted the presence of aggregates in the lungs for both powders but not for the solution following a single dose administration. Flow cytometry analysis designated alveolar macrophages as the main immune cells taking up rhod-NIP228 in the lungs with very little involvement of dendritic cells and interstitial macrophages. Notably, repeated intratracheal administration of the three formulations in mice did not impact the magnitude of the anti-drug antibody response in sera or broncho-alveolar lavages. Furthermore, the pulmonary route appeared to evoke a more robust immune response when compared to subcutaneous administration. Overall, the level of NIP228 aggregation in this study did not appear to be the primary driver of NIP228 immunogenicity following delivery to the lungs in mice.These findings shed new light on the interplay between protein aggregation and immunogenicity in the context of the pulmonary delivery of therapeutic proteins.
{"title":"Impact of protein aggregation on the immunogenicity of a human monoclonal antibody following pulmonary administration in mice","authors":"Sohaib Mahri , Céline Cassiers , Sandra Gracin , Donatienne Tyteca , Francine Uwambayinema , François Huaux , Mariam Ibrahim , Sian Piper , Antonio Llinas , Markus Fridén , Rita Vanbever","doi":"10.1016/j.ijpharm.2024.124880","DOIUrl":"10.1016/j.ijpharm.2024.124880","url":null,"abstract":"<div><div>Spray drying is a widely employed method for generating dry powder formulations for inhalation. Yet, it presents substantial challenges when applied to therapeutic proteins due to stability issues. The formation of protein aggregates during the atomization and the heating steps can diminish protein activity and raise immunogenicity concerns. Here, we assessed the impact of varying levels of protein aggregates generated during spray-drying on the fate and the immunogenicity of the human monoclonal antibody NIP228 following intratracheal administration in mice. Aggregate-free rhodamine labelled NIP228 was spray-dried with or without 1% polysorbate 80 surfactant, resulting in the generation of powder formulations with associated low and high protein aggregate levels, respectively. Confocal imaging highlighted the presence of aggregates in the lungs for both powders but not for the solution following a single dose administration. Flow cytometry analysis designated alveolar macrophages as the main immune cells taking up rhod-NIP228 in the lungs with very little involvement of dendritic cells and interstitial macrophages. Notably, repeated intratracheal administration of the three formulations in mice did not impact the magnitude of the anti-drug antibody response in sera or broncho-alveolar lavages. Furthermore, the pulmonary route appeared to evoke a more robust immune response when compared to subcutaneous administration. Overall, the level of NIP228 aggregation in this study did not appear to be the primary driver of NIP228 immunogenicity following delivery to the lungs in mice.These findings shed new light on the interplay between protein aggregation and immunogenicity in the context of the pulmonary delivery of therapeutic proteins.</div></div>","PeriodicalId":14187,"journal":{"name":"International Journal of Pharmaceutics","volume":"667 ","pages":"Article 124880"},"PeriodicalIF":5.3,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142620550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-07DOI: 10.1016/j.ijpharm.2024.124917
Xin Wang , Shurong Zhang , Xiaoyue Wang , Liping Zhou , Yang Tang , Yan Xiao , Yu Zhang , Wei Li
Advanced nanosized drug delivery systems can significantly improve efficacy and safety of first-line chemotherapeutics by enhancing tumor targeting. Herein, one-pot covalent crosslinking approach was developed to generate biodegradable tumor-targeted composite Nanogels from carboxymethyl chitosan, hyaluronic acid, cystamine and 6-ethylene-diamine-6-deoxy-β-cyclodextrin loaded with doxorubicin (DOX) for controlled intracellular DOX release. The optimized synthetic procedures generated Nanogels of about 190 nm in size and 28.3 % drug loading capability. DOX-loaded Nanogels was effectively internalized into tumor cells mainly by CD44 receptor-mediated endocytosis and rapidly released DOX in response to the high level of GSH in cytoplasm and acidic intracellular environments. DOX-loaded Nanogels significantly inhibited the tumor growth especially without appreciable side toxicities in 4 T1 tumor-bearing mice model owing to CD44 receptor-mediated active targeting and the passive targeting of Nanogels by enhanced permeation and retention effect. Overall, our newly developed composite Nanogels might be employed as a potentially effective therapeutic strategy for tumor therapy.
{"title":"β-cyclodextrin-modified carboxymethyl chitosan/hyaluronic acid-based crosslinked composite nanogels as a dual responsive carrier for targeting anti-tumor therapy","authors":"Xin Wang , Shurong Zhang , Xiaoyue Wang , Liping Zhou , Yang Tang , Yan Xiao , Yu Zhang , Wei Li","doi":"10.1016/j.ijpharm.2024.124917","DOIUrl":"10.1016/j.ijpharm.2024.124917","url":null,"abstract":"<div><div>Advanced nanosized drug delivery systems can significantly improve efficacy and safety of first-line chemotherapeutics by enhancing tumor targeting. Herein, one-pot covalent crosslinking approach was developed to generate biodegradable tumor-targeted composite Nanogels from carboxymethyl chitosan, hyaluronic acid, cystamine and 6-ethylene-diamine-6-deoxy-β-cyclodextrin loaded with doxorubicin (DOX) for controlled intracellular DOX release. The optimized synthetic procedures generated Nanogels of about 190 nm in size and 28.3 % drug loading capability. DOX-loaded Nanogels was effectively internalized into tumor cells mainly by CD44 receptor-mediated endocytosis and rapidly released DOX in response to the high level of GSH in cytoplasm and acidic intracellular environments. DOX-loaded Nanogels significantly inhibited the tumor growth especially without appreciable side toxicities in 4 T1 tumor-bearing mice model owing to CD44 receptor-mediated active targeting and the passive targeting of Nanogels by enhanced permeation and retention effect. Overall, our newly developed composite Nanogels might be employed as a potentially effective therapeutic strategy for tumor therapy.</div></div>","PeriodicalId":14187,"journal":{"name":"International Journal of Pharmaceutics","volume":"667 ","pages":"Article 124917"},"PeriodicalIF":5.3,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142619614","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-07DOI: 10.1016/j.ijpharm.2024.124899
Salama A. Al Yabhouni , Mohammad Sayem Mozumder , Nurudeen Hassan , Abdel-Hamid I. Mourad , Tareq M.A Issa MD
Human eyes have the most complex and advanced physiological defense barriers. Due to these barriers, efficient delivery of ocular drugs is a major challenge in the treatment of eye diseases and disorders. Posterior eye diseases such as retinopathy are the leading causes of impaired vision and blindness globally. The topical and systemic administration of drugs such as eye drops, ointments, intravitreal injections, intraocular implants, contact lenses, and emulsions are the perennial approaches employed to treat ocular diseases. However, these modalities are inefficient due to the low bioavailability of the active drug and the potential for drug-related cytotoxicity to the ocular tissue. In this review, the conventional approaches in ocular drug delivery systems (DDSs) are explored and the limitations associated with each technique are elucidated. A comparison between the different DDSs is presented, showing the most effective treatment techniques available to date. In addition, this review presents recent advances in the field of nanocarriers and microcarriers used in ocular drug delivery systems such as nanoparticles, nano-suspensions, nanofibers, nanogels, nano-liposomes, nano micelles, dendrimers, contact lens, microneedle, and implants. Further, this review identifies the utility of nano-carriers in enabling the development of new-generation ocular DDSs with low toxicity, high efficiency, and high stability of targeted drug delivery systems to overcome the limitations observed with conventional ocular DDSs. In addition, this manuscript sheds light on the incidence and unique landscape of ocular diseases in the United Arab Emirates (UAE), and the potential for employing novel ocular DDSs for targeted treatment of conditions such as diabetic retinopathy in the UAE. It also discusses the putative role genetic variants of the VEGF gene may play in predisposing the local population in the UAE to developing posterior eye segment diseases such as retinopathy.
{"title":"Nanocarrier-Based, ocular drug delivery: Challenges, prospects, and the therapeutic landscape in the United Arab Emirates","authors":"Salama A. Al Yabhouni , Mohammad Sayem Mozumder , Nurudeen Hassan , Abdel-Hamid I. Mourad , Tareq M.A Issa MD","doi":"10.1016/j.ijpharm.2024.124899","DOIUrl":"10.1016/j.ijpharm.2024.124899","url":null,"abstract":"<div><div>Human eyes have the most complex and advanced physiological defense barriers. Due to these barriers, efficient delivery of ocular drugs is a major challenge in the treatment of eye diseases and disorders. Posterior eye diseases such as retinopathy are the leading causes of impaired vision and blindness globally. The topical and systemic administration of drugs such as eye drops, ointments, intravitreal injections, intraocular implants, contact lenses, and emulsions are the perennial approaches employed to treat ocular diseases. However, these modalities are inefficient due to the low bioavailability of the active drug and the potential for drug-related cytotoxicity to the ocular tissue. In this review, the conventional approaches in ocular drug delivery systems (DDSs) are explored and the limitations associated with each technique are elucidated. A comparison between the different DDSs is presented, showing the most effective treatment techniques available to date. In addition, this review presents recent advances in the field of nanocarriers and microcarriers used in ocular drug delivery systems such as nanoparticles, nano-suspensions, nanofibers, nanogels, nano-liposomes, nano micelles, dendrimers, contact lens, microneedle, and implants. Further, this review identifies the utility of nano-carriers in enabling the development of new-generation ocular DDSs with low toxicity, high efficiency, and high stability of targeted drug delivery systems to overcome the limitations observed with conventional ocular DDSs. In addition, this manuscript sheds light on the incidence and unique landscape of ocular diseases in the United Arab Emirates (UAE), and the potential for employing novel ocular DDSs for targeted treatment of conditions such as diabetic retinopathy in the UAE. It also discusses the putative role genetic variants of the VEGF gene may play in predisposing the local population in the UAE to developing posterior eye segment diseases such as retinopathy.</div></div>","PeriodicalId":14187,"journal":{"name":"International Journal of Pharmaceutics","volume":"667 ","pages":"Article 124899"},"PeriodicalIF":5.3,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142620554","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Chemotherapy in the management of cancer is constrained by limitations like off-target effects, poor bioavailability, and dose-dependent toxicity. Nutraceuticals have been explored as an innovative strategy to overcome chemotherapy drawbacks. However, the clinical utility of nutraceuticals is restricted due to their complex structures, less water solubility, reduced stability, decreased bioavailability and more obstacles in the gastrointestinal tract. Nanonutraceuticals are nanosized nutraceutical particles having enhanced solubility, improved bioavailability, stability, and targeted delivery to specific cells. Nutraceuticals can be co-delivered with other chemotherapeutic drugs in nanocarriers to elicit synergistic effects. The targeting of nutraceuticals against cancer cells can be enabled by coupling ligands with the nanocarriers, which direct to the overexpressed receptors found at the surface of the cancer cells. Transitioning a nanonutraceutical from pre-clinical research to clinical trials is a pivotal step. This focus on advancing their application holds great potential for impacting clinical research and improving the treatment landscape for cancer patients. This review focuses on the role of nutraceuticals for cancer treatment, various nanocarriers for the efficient delivery of nutraceuticals along with co-administration of nutraceuticals with chemotherapeutic drugs using nanocarriers. Also, emphasize the targeting of ligands coupled nanocarriers to the cancer cells along with patents and clinical trials for nanonutraceuticals.
{"title":"Transforming cancer treatment: The potential of nanonutraceuticals","authors":"Girish Kumar , Tarun Virmani , Vaishnavi Chhabra , Reshu Virmani , Kamla Pathak , Md Sayeed Akhtar , Mulazim Hussain Asim , Shumaila Arshad , Farzana Siddique , Pedro Fonte","doi":"10.1016/j.ijpharm.2024.124919","DOIUrl":"10.1016/j.ijpharm.2024.124919","url":null,"abstract":"<div><div>Chemotherapy in the management of cancer is constrained by limitations like off-target effects, poor bioavailability, and dose-dependent toxicity. Nutraceuticals have been explored as an innovative strategy to overcome chemotherapy drawbacks.<!--> <!-->However, the clinical utility of nutraceuticals is restricted due to their complex structures, less water solubility, reduced stability, decreased bioavailability and more obstacles in the gastrointestinal tract. Nanonutraceuticals are nanosized nutraceutical particles having enhanced solubility, improved bioavailability, stability, and targeted delivery to specific cells. Nutraceuticals can be co-delivered with other chemotherapeutic drugs in nanocarriers to elicit synergistic effects. The targeting of nutraceuticals against cancer cells can be enabled by coupling ligands with the nanocarriers, which direct to the overexpressed receptors found at the surface of the cancer cells. Transitioning a nanonutraceutical from pre-clinical research to clinical trials is a pivotal step. This focus on advancing their application holds great potential for impacting clinical research and improving the treatment landscape for cancer patients. This review focuses on the role of nutraceuticals for cancer treatment, various nanocarriers for the efficient delivery of nutraceuticals along with co-administration of nutraceuticals with chemotherapeutic drugs using nanocarriers. Also, emphasize the targeting of ligands coupled nanocarriers to the cancer cells along with patents and clinical trials for nanonutraceuticals.</div></div>","PeriodicalId":14187,"journal":{"name":"International Journal of Pharmaceutics","volume":"667 ","pages":"Article 124919"},"PeriodicalIF":5.3,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142620156","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-07DOI: 10.1016/j.ijpharm.2024.124918
Xinmin Wang , Lizhen Huang , Qing Du , Jing Li , Qin Zheng , Yingchong Chen , Pengfei Yue
To improve the retention time and skin-whitening efficacy of Atractylodes macrocephale essential oil (AMO), a novel Pickering emulsion based nanogel loaded with AMO (AMO-PEG) was successfully developed. This formulation employed nano-pearl powder (NPP) as the particle stabilizer for the Pickering emulsion and Bletilla striata polysaccharide (BSP) as the gel matrix. The pH, rheological properties, hardness, and elasticity of AMO-PEG were affected by the ratio of AMO-Pickering emulsion (AMO-PE) to BSP gel matrix. The results showed that AMO-PEG exhibited solid-like behavior and was capable of forming nanogels when the ratio of AMO-PE to BSP was 1:1. AMO-PE and AMO-PEG are two different dosage forms in the preparation of AMO. The effects of varying dosage forms on AMO were evaluated by in vitro transdermal release, skin irritation test, and skin-whitening effect. AMO-PEG conforms to the zero-order kinetic equation (R2 = 0.9189). The skin retention rate of AMO-PEG was 1.37 times higher than that of AMO-PE, indicating that AMO-PEG could continuously and slowly exert the whitening effect of the drugs. Compared with AMO-PE, AMO-PEG significantly increased the inhibition of tyrosinase activity and melanogenesis in B16F10 cells. AMO-PEG can promote the inhibition of B16F10 cells and improve the whitening effect of AMO and BSP. In conclusion, the Pickering emulsion based nanogel appears to be a promising strategy for enhancing the skin-whitening efficacy of both AMO and BSP.
{"title":"Pickering emulsions embedded in Bletilla striata polysaccharide based nanogel for enhancing skin-whitening effect of essential oils","authors":"Xinmin Wang , Lizhen Huang , Qing Du , Jing Li , Qin Zheng , Yingchong Chen , Pengfei Yue","doi":"10.1016/j.ijpharm.2024.124918","DOIUrl":"10.1016/j.ijpharm.2024.124918","url":null,"abstract":"<div><div>To improve the retention time and skin-whitening efficacy of <em>Atractylodes macrocephale</em> essential oil (AMO), a novel Pickering emulsion based nanogel loaded with AMO (AMO-PEG) was successfully developed. This formulation employed nano-pearl powder (NPP) as the particle stabilizer for the Pickering emulsion and <em>Bletilla striata</em> polysaccharide (BSP) as the gel matrix. The pH, rheological properties, hardness, and elasticity of AMO-PEG were affected by the ratio of AMO-Pickering emulsion (AMO-PE) to BSP gel matrix. The results showed that AMO-PEG exhibited solid-like behavior and was capable of forming nanogels when the ratio of AMO-PE to BSP was 1:1. AMO-PE and AMO-PEG are two different dosage forms in the preparation of AMO. The effects of varying dosage forms on AMO were evaluated by <em>in vitro</em> transdermal release, skin irritation test, and skin-whitening effect. AMO-PEG conforms to the zero-order kinetic equation (R<sup>2</sup> = 0.9189). The skin retention rate of AMO-PEG was 1.37 times higher than that of AMO-PE, indicating that AMO-PEG could continuously and slowly exert the whitening effect of the drugs. Compared with AMO-PE, AMO-PEG significantly increased the inhibition of tyrosinase activity and melanogenesis in B16F10 cells. AMO-PEG can promote the inhibition of B16F10 cells and improve the whitening effect of AMO and BSP. In conclusion, the Pickering emulsion based nanogel appears to be a promising strategy for enhancing the skin-whitening efficacy of both AMO and BSP.</div></div>","PeriodicalId":14187,"journal":{"name":"International Journal of Pharmaceutics","volume":"667 ","pages":"Article 124918"},"PeriodicalIF":5.3,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142620558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-07DOI: 10.1016/j.ijpharm.2024.124914
Claudia Moscheni , Patrizia Sartori , Kaiyue Hu , Silvia Zecchini , Luigi Brambilla , Alessandro Arcari , Alessandra Napoli , Emanuele Mocciaro , Marco Uboldi , Lucia Zema , Cristiana Perrotta , Chiara Castiglioni
Thanks to an environmentally friendly physical treatment of high purity graphite, a good control of the structure of graphene nanoparticles (GNPs) has been obtained with the production of stable and reproducible GNPs water dispersions. The preparation protocol entailed ball-milling of synthetic graphite followed by sonication in water and centrifugation/separation procedures. This way, two different GNPs samples with slightly different structural characteristics were harvested: TOP60, showing an average lateral size of the graphene layers <L> = 70 nm and average number of stacked layers <N> = 4, and BOTTOM60, with <L> = 120 nm and <N> = 6. A detailed structural characterization of GNPs was performed as mandatory pre-requisite to build reliable structure/properties correlations, in terms of both biomedical efficacy and toxicity, aiming at a rationale design of tailored materials for applications in biological environments.
To this end, in this study GNPs were thoroughly characterized, focusing on cytotoxicity, cellular uptake, and inflammatory response, by testing their effect in different cell lines. BOTTOM60 GNPs in culture medium and in the presence of cells showed a tendency to form big aggregates, phenomenon that was probably responsible for their cytotoxicity at high concentrations. On the other hand, TOP60 GNPs showed a diverse behavior depending on the cell type under investigation. Indeed, the nanoparticles were internalized by cells specialized in endo/phagocytosis, such as astrocytoma cells, but not by carcinoma cells of epithelial origin. Moreover, TOP60 GNPs caused a reduction of proliferation only at high concentration and did not trigger an inflammatory response in THP-1-derived macrophages.
The evidence here collected paves the way for further investigations towards the development of GNPs-based drug delivery systems.
{"title":"Tailored graphene nanoparticles for biomedical application: preliminary in vitro characterization of the functionality in model cell lines","authors":"Claudia Moscheni , Patrizia Sartori , Kaiyue Hu , Silvia Zecchini , Luigi Brambilla , Alessandro Arcari , Alessandra Napoli , Emanuele Mocciaro , Marco Uboldi , Lucia Zema , Cristiana Perrotta , Chiara Castiglioni","doi":"10.1016/j.ijpharm.2024.124914","DOIUrl":"10.1016/j.ijpharm.2024.124914","url":null,"abstract":"<div><div>Thanks to an environmentally friendly physical treatment of high purity graphite, a good control of the structure of graphene nanoparticles (GNPs) has been obtained with the production of stable and reproducible GNPs water dispersions. The preparation protocol entailed ball-milling of synthetic graphite followed by sonication in water and centrifugation/separation procedures. This way, two different GNPs samples with slightly different structural characteristics were harvested: TOP60, showing an average lateral size of the graphene layers <L> = 70 nm and average number of stacked layers <N> = 4, and BOTTOM60, with <L> = 120 nm and <N> = 6. A detailed structural characterization of GNPs was performed as mandatory pre-requisite to build reliable structure/properties correlations, in terms of both biomedical efficacy and toxicity, aiming at a rationale design of tailored materials for applications in biological environments.</div><div>To this end, in this study GNPs were thoroughly characterized, focusing on cytotoxicity, cellular uptake, and inflammatory response, by testing their effect in different cell lines. BOTTOM60 GNPs in culture medium and in the presence of cells showed a tendency to form big aggregates, phenomenon that was probably responsible for their cytotoxicity at high concentrations. On the other hand, TOP60 GNPs showed a diverse behavior depending on the cell type under investigation. Indeed, the nanoparticles were internalized by cells specialized in endo/phagocytosis, such as astrocytoma cells, but not by carcinoma cells of epithelial origin. Moreover, TOP60 GNPs caused a reduction of proliferation only at high concentration and did not trigger an inflammatory response in THP-1-derived macrophages.</div><div>The evidence here collected paves the way for further investigations towards the development of GNPs-based drug delivery systems.</div></div>","PeriodicalId":14187,"journal":{"name":"International Journal of Pharmaceutics","volume":"667 ","pages":"Article 124914"},"PeriodicalIF":5.3,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142619413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A formerly developed mathematical model describing drug release from hydrophilic matrices (HMs) took into account resistance to drug release given by its dissolution and by the presence of a growing gel layer. Such a model was applied to previously reported release data obtained from HMs made of hydroxypropyl methylcellulose (HPMC), where acetaminophen was used as model drug and a cellulolytic product was added as “active” excipient to attain zero-order release kinetics. The Levich theory applied to acetaminophen intrinsic dissolution rate (IDR) data highlighted the suitability of such a drug for modeling purposes, given its good surface wettability. First assessment of the model ability to describe drug release from the abovementioned systems was carried out on partially coated matrices, representing a simplified physical frame, but results were then confirmed on uncoated systems. Experimental and model release data showed good agreement; therefore, the release-describing equation was combined with that of the global mass balance to obtain two new equations related to erosion and diffusion fronts time evolution. Changes over time in the dissolution and gel contributions to total resistance, calculated using model output parameters, highlighted that the enzyme, through its hydrolytic activity on HPMC, was responsible for a time-dependent reduction of the resistance component related to gel layer.
{"title":"Modeling of drug release, erosion and diffusion fronts movement in high viscosity HPMC matrices containing a cellulolytic enzyme","authors":"Ilaria Filippin , Saliha Moutaharrik , Michela Abrami , Lucia Grassi , Andrea Gazzaniga , Alessandra Maroni , Mario Grassi , Anastasia Foppoli","doi":"10.1016/j.ijpharm.2024.124902","DOIUrl":"10.1016/j.ijpharm.2024.124902","url":null,"abstract":"<div><div>A formerly developed mathematical model describing drug release from hydrophilic matrices (HMs) took into account resistance to drug release given by its dissolution and by the presence of a growing gel layer. Such a model was applied to previously reported release data obtained from HMs made of hydroxypropyl methylcellulose (HPMC), where acetaminophen was used as model drug and a cellulolytic product was added as “active” excipient to attain zero-order release kinetics. The Levich theory applied to acetaminophen intrinsic dissolution rate (IDR) data highlighted the suitability of such a drug for modeling purposes, given its good surface wettability. First assessment of the model ability to describe drug release from the abovementioned systems was carried out on partially coated matrices, representing a simplified physical frame, but results were then confirmed on uncoated systems. Experimental and model release data showed good agreement; therefore, the release-describing equation was combined with that of the global mass balance to obtain two new equations related to erosion and diffusion fronts time evolution. Changes over time in the dissolution and gel contributions to total resistance, calculated using model output parameters, highlighted that the enzyme, through its hydrolytic activity on HPMC, was responsible for a time-dependent reduction of the resistance component related to gel layer.</div></div>","PeriodicalId":14187,"journal":{"name":"International Journal of Pharmaceutics","volume":"667 ","pages":"Article 124902"},"PeriodicalIF":5.3,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142620150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-07DOI: 10.1016/j.ijpharm.2024.124921
Maria José Sánchez , Pablo Leivar , Salvador Borrós , Cristina Fornaguera , Martí Lecina
Extracellular Vesicles (EVs) are nanosized particles with significant role in disease pathogenesis and as therapeutic potential. However, the lack of reliable and efficient methods for the characterization, quantification and tracking of EVs, combined with the limitations of detection techniques in differentiating specific EVs subtypes with beneficial properties, makes these process complex and time-consuming. To address this challenge, EVs were engineered using a tricistronic plasmid that encodes fluorescent proteins fused to tetraspanins (eGFP-CD63 and mCherry-CD9), with both fluorophores localized within the luminal space. Double fluorescently labelled small EVs (sEVs) were then produced in a stably transfected HEK293SF-3F6 cell line. The fluorescently labelled sEVs were characterized using a variety of techniques. Protein expression analysis showed that the fused proteins were efficiently produced and incorporated in sEVs, as evidenced by clear fluorescence signal detected. Comparisons of the size distribution and concentration of modified sEVs with controls indicated that sEVs engineering did not affect their biogenesis and morphology. Fluorescently labelled sEVs were then quantified by flow cytometry, allowing to distinguish sEVs from other EVs subtypes or sample particles. The values were then compared to fluorometry measurements, obtaining a linear correlation what enabled a novel sEVs quantification method. The functionality of engineered sEVs was assessed by monitoring their uptake and trafficking in recipient cells, obtaining an efficient internalisation by target cells. Overall, these results demonstrate that the implementation of dual fluorescent methodology is feasible for sEVs characterization, quantification, for in vitro study of EVs interaction with cells, and intercellular communication, as well as a valuable tool in the in vitro development of targeted therapeutic EVs delivery systems.
{"title":"Enhanced quantification and cell tracking of dual fluorescent labeled extracellular vesicles","authors":"Maria José Sánchez , Pablo Leivar , Salvador Borrós , Cristina Fornaguera , Martí Lecina","doi":"10.1016/j.ijpharm.2024.124921","DOIUrl":"10.1016/j.ijpharm.2024.124921","url":null,"abstract":"<div><div>Extracellular Vesicles (EVs) are nanosized particles with significant role in disease pathogenesis and as therapeutic potential. However, the lack of reliable and efficient methods for the characterization, quantification and tracking of EVs, combined with the limitations of detection techniques in differentiating specific EVs subtypes with beneficial properties, makes these process complex and time-consuming. To address this challenge, EVs were engineered using a tricistronic plasmid that encodes fluorescent proteins fused to tetraspanins (eGFP-CD63 and mCherry-CD9), with both fluorophores localized within the luminal space. Double fluorescently labelled small EVs (sEVs) were then produced in a stably transfected HEK293SF-3F6 cell line. The fluorescently labelled sEVs were characterized using a variety of techniques. Protein expression analysis showed that the fused proteins were efficiently produced and incorporated in sEVs, as evidenced by clear fluorescence signal detected. Comparisons of the size distribution and concentration of modified sEVs with controls indicated that sEVs engineering did not affect their biogenesis and morphology. Fluorescently labelled sEVs were then quantified by flow cytometry, allowing to distinguish sEVs from other EVs subtypes or sample particles. The values were then compared to fluorometry measurements, obtaining a linear correlation what enabled a novel sEVs quantification method. The functionality of engineered sEVs was assessed by monitoring their uptake and trafficking in recipient cells, obtaining an efficient internalisation by target cells. Overall, these results demonstrate that the implementation of dual fluorescent methodology is feasible for sEVs characterization, quantification, <strong>for <em>in vitro</em></strong> study of EVs interaction with cells, and intercellular communication, as well as a valuable tool in the <strong><em>in vitro</em></strong> development of targeted therapeutic EVs delivery systems.</div></div>","PeriodicalId":14187,"journal":{"name":"International Journal of Pharmaceutics","volume":"667 ","pages":"Article 124921"},"PeriodicalIF":5.3,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142619918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Gold nanoparticles (AuNPs) have become a focus of interest in biomedicine due to their unique properties. By attaching peptides to these nanoparticles (NPs), they can be utilized for a wide range of applications. Peptides, which are short chains of amino acids, can be customized for specific molecular interactions, making them ideal for delivering AuNPs to particular cells or tissues. One of the peptide-AuNP-based bio-nano technological approaches involves targeted drug delivery. Including peptides as targeting agents, these NPs can be designed to bind to specific cell receptors or biomarkers. This allows for the direct delivery of therapeutic agents to diseased cells while minimizing unwanted side effects, improving the effectiveness of treatments. Additionally, peptide-functionalized AuNPs (PAuNPs) are crucial for imaging and diagnostics. By functionalizing the NPs with peptides that bind to specific molecular targets, such as cancer biomarkers, these NPs can be used to visualize diseased tissues. This enables the early detection of diseases and helps in determining the severity of conditions for better diagnosis and treatment outcomes. Moreover, PAuNPs have displayed promising potential in photothermal therapy. Once PAuNPs uptake and penetrate target cancer cells effectively, these NPs generate heat when exposed to specific wavelengths of light, efficiently eliminating tumors while preserving healthy surrounding tissues. Therefore, in this paper, we systematically review the potential of PAuNPs in various biomedical applications, including therapy and diagnosis, providing a future perspective.
{"title":"Biomedical applications of peptide-gold nanoarchitectonics","authors":"Samane Maghsoudian , Esmat Sajjadi , Niloufar Hadavi , Mobina Soltani , Zahra Karami , Alwan Abed Hamadi Al Qushawi , Mohammad Akrami , Farid Kalantari","doi":"10.1016/j.ijpharm.2024.124920","DOIUrl":"10.1016/j.ijpharm.2024.124920","url":null,"abstract":"<div><div>Gold nanoparticles (AuNPs) have become a focus of interest in biomedicine due to their unique properties. By attaching peptides to these nanoparticles (NPs), they can be utilized for a wide range of applications. Peptides, which are short chains of amino acids, can be customized for specific molecular interactions, making them ideal for delivering AuNPs to particular cells or tissues. One of the peptide-AuNP-based bio-nano technological approaches involves targeted drug delivery. Including peptides as targeting agents, these NPs can be designed to bind to specific cell receptors or biomarkers. This allows for the direct delivery of therapeutic agents to diseased cells while minimizing unwanted side effects, improving the effectiveness of treatments. Additionally, peptide-functionalized AuNPs (PAuNPs) are crucial for imaging and diagnostics. By functionalizing the NPs with peptides that bind to specific molecular targets, such as cancer biomarkers, these NPs can be used to visualize diseased tissues. This enables the early detection of diseases and helps in determining the severity of conditions for better diagnosis and treatment outcomes. Moreover, PAuNPs have displayed promising potential in photothermal therapy. Once PAuNPs uptake and penetrate target cancer cells effectively, these NPs generate heat when exposed to specific wavelengths of light, efficiently eliminating tumors while preserving healthy surrounding tissues. Therefore, in this paper, we systematically review the potential of PAuNPs in various biomedical applications, including therapy and diagnosis, providing a future perspective.</div></div>","PeriodicalId":14187,"journal":{"name":"International Journal of Pharmaceutics","volume":"667 ","pages":"Article 124920"},"PeriodicalIF":5.3,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142619740","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号