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1.
Nephron ; 147(11): 650-654, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37423209

RESUMO

The kidneys are the body's main excretion organ with several additional functions, and the nephron represents their central structural unit. It is comprised of endothelial, mesangial, glomerular, and tubular epithelial cells, as well as podocytes. Treatment of acute kidney injury or chronic kidney disease (CKD) is complex due to broad etiopathogenic mechanisms and limited regeneration potential as kidney cells finish their differentiation after 34 weeks of gestation. Despite the ever-increasing prevalence of CKD, very limited treatment modalities are available. The medical community should therefore strive to improve existing treatments and develop new ones. Furthermore, polypharmacy is present in most CKD patients, while current pharmacologic study designs lack effectiveness in predicting potential drug-drug interactions and the resulting clinically relevant complications. An opportunity for addressing these issues lies in developing in vitro cell models based on patient-derived renal cells. Currently, several protocols have been described for isolating desired kidney cells, of which the most isolated are the proximal tubular epithelial cells. These play a significant role in water homeostasis, acid-base control, reabsorption of compounds, and secretion of xenobiotics and endogenous metabolites. When developing a protocol for the isolation and culture of such cells, one must focus on several steps. These include harvesting cells from biopsy specimens or after nephrectomies, using different digestion enzymes and culture mediums to facilitate the selective growth of only the desired cells. The literature reports several existing models, from simple 2D in vitro cultures to more complex ones created with bioengineering methods, such as kidney-on-a-chip models. While their creation and use depend on the target research, one should consider factors such as equipment, cost, and, even more importantly, source tissue quality and availability.


Assuntos
Podócitos , Insuficiência Renal Crônica , Humanos , Rim/patologia , Células Epiteliais/patologia , Insuficiência Renal Crônica/patologia , Glomérulos Renais/patologia , Podócitos/patologia
2.
Int J Biol Macromol ; 236: 123951, 2023 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-36898451

RESUMO

Masks proved to be necessary protective measure during the COVID-19 pandemic, but they provided a physical barrier rather than inactivating viruses, increasing the risk of cross-infection. In this study, high-molecular weight chitosan and cationised cellulose nanofibrils were screen-printed individually or as a mixture onto the inner surface of the first polypropylene (PP) layer. First, biopolymers were evaluated by various physicochemical methods for their suitability for screen-printing and antiviral activity. Second, the effect of the coatings was evaluated by analysing the morphology, surface chemistry, charge of the modified PP layer, air permeability, water-vapour retention, add-on, contact angle, antiviral activity against the model virus phi6 and cytotoxicity. Finally, the functional PP layers were integrated into face masks, and resulting masks were tested for wettability, air permeability, and viral filtration efficiency (VFE). Air permeability was reduced for modified PP layers (43 % reduction for kat-CNF) and face masks (52 % reduction of kat-CNF layer). The antiviral potential of the modified PP layers against phi6 showed inhibition of 0.08 to 0.97 log (pH 7.5) and cytotoxicity assay showed cell viability above 70 %. VFE of the masks remained the same (~99.9 %), even after applying the biopolymers, confirming that these masks provided high level of protection against viruses.


Assuntos
COVID-19 , Quitosana , Humanos , COVID-19/prevenção & controle , Antivirais/farmacologia , Pandemias/prevenção & controle , Celulose/farmacologia , Máscaras
3.
J Funct Biomater ; 13(3)2022 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-36135591

RESUMO

BACKGROUND: Biomaterials and biotechnology are becoming increasingly important fields in modern medicine. For cranial bone defects of various aetiologies, artificial materials, such as poly-methyl-methacrylate, are often used. We report our clinical experience with poly-methyl-methacrylate for a novel in vivo bone defect closure and artificial bone flap development in various neurosurgical operations. METHODS: The experimental study included 12 patients at a single centre in 2018. They presented with cranial bone defects after various neurosurgical procedures, including tumour, traumatic brain injury and vascular pathologies. The patients underwent an in vivo bone reconstruction from poly-methyl-methacrylate, which was performed immediately after the tumour removal in the tumour group, whereas the trauma and vascular patients required a second surgery for cranial bone reconstruction due to the bone decompression. The artificial bone flap was modelled in vivo just before the skin closure. Clinical and surgical data were reviewed. RESULTS: All patients had significant bony destruction or unusable bone flap. The tumour group included five patients with meningiomas destruction and the trauma group comprised four patients, all with severe traumatic brain injury. In the vascular group, there were three patients. The average modelling time for the artificial flap modelling was approximately 10 min. The convenient location of the bone defect enabled a relatively straightforward and fast reconstruction procedure. No deformations of flaps or other complications were encountered, except in one patient, who suffered a postoperative infection. CONCLUSIONS: Poly-methyl-methacrylate can be used as a suitable material to deliver good cranioplasty cosmesis. It offers an optimal dural covering and brain protection and allows fast intraoperative reconstruction with excellent cosmetic effect during the one-stage procedure. The observations of our study support the use of poly-methyl-methacrylate for the ad hoc reconstruction of cranial bone defects.

4.
Pharmaceutics ; 14(4)2022 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-35456523

RESUMO

Despite medical advances, skin-associated disorders continue to pose a unique challenge to physicians worldwide. Skin cancer is one of the most common forms of cancer, with more than one million new cases reported each year. Currently, surgical excision is its primary treatment; however, this can be impractical or even contradictory in certain situations. An interesting potential alternative could lie in topical treatment solutions. The goal of our study was to develop novel multilayer nanofilms consisting of a combination of polyhydroxyethyl methacrylate (PHEMA), polyhydroxypropyl methacrylate (PHPMA), sodium deoxycholate (NaDOC) with incorporated superparamagnetic iron-platinum nanoparticles (FePt NPs), and the potent anticancer drug (5-fluorouracil), for theranostic skin cancer treatment. All multilayer systems were prepared by spin-coating and characterised by atomic force microscopy, infrared spectroscopy, and contact angle measurement. The magnetic properties of the incorporated FePt NPs were evaluated using magnetisation measurement, while their size was determined using transmission electron microscopy (TEM). Drug release performance was tested in vitro, and formulation safety was evaluated on human-skin-derived fibroblasts. Finally, the efficacy for skin cancer treatment was tested on our own basal-cell carcinoma cell line.

5.
Biomedicines ; 10(2)2022 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-35203587

RESUMO

Mesenchymal stem cells (MSCs) represent the basis of novel clinical concepts in cellular therapy and tissue regeneration. Therefore, the isolation of MSCs from various tissues has become an important endeavour for stem cell biobanking and the development of regenerative therapies. Paravertebral adipose tissue is readily exposed during spinal procedures in children and could be a viable source of stem cells for therapeutic applications. Here, we describe the first case of MSCs isolated from paravertebral adipose tissue (PV-ADMSCs), obtained during a routine spinal surgery on a child. Using quantitative real-time PCR and flow cytometry, we show that PV-ADMSCs have different levels of stem marker expression compared to the MSCs from other sources while having the highest proliferation rate. Furthermore, we evaluate the multipotency of PV-ADMSCs by the three-lineage (adipogenic, osteogenic and chondrogenic) differentiation and compare it to the multipotency of MSCs from other sources. It was found that the PV-ADMSCs have a strong osteogenic potential in particular. Taken together, our data indicate that PV-ADMSCs meet the criteria for successful cell therapy, defined by the International Society for Cellular Therapy (ISCT), and thus, could provide a source of MSCs that is relatively easy to isolate and expand in culture. Due to their strong osteogenic potential, these cells provide a promising basis, especially for orthopaedic applications.

6.
Plants (Basel) ; 10(3)2021 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-33802757

RESUMO

Cannabis sativa is one of the oldest medicinal plants used by humans, containing hundreds of bioactive compounds. The biological effects and interplay of these compounds are far from fully understood, although the plant's therapeutic effects are beyond doubt. Extraction methods for these compounds are becoming an integral part of modern Cannabis-based medicine. Still, little is known about how different methods affect the final composition of Cannabis extracts and thus, their therapeutic effects. In this study, different extraction methods were tested, namely maceration, Soxhlet, ultrasound-assisted extraction (UAE), and supercritical CO2 extraction methods. The obtained extracts were evaluated for their cannabinoid content, antioxidant properties, and in vitro bioactivity on human colon cancer and healthy colon cells. Our data suggest that Cannabis extracts, when properly prepared, can significantly decrease cancer cell viability while protecting healthy cells from cytotoxic effects. However, post-processing of extracts poses a significant limitation in predicting therapeutic response based on the composition of the crude extract, as it affects not only the actual amounts of the respective cannabinoids but also their relative ratio to the primary extracts. These effects must be carefully considered in the future preparations of new therapeutic extracts.

7.
J Cell Mol Med ; 25(8): 3680-3698, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33650759

RESUMO

Cell lines are widely used for various research purposes including cancer and drug research. Recently, there have been studies that pointed to discrepancies in the literature and usage of cell lines. That is why we have prepared a comprehensive overview of the most common gynaecological cancer cell lines, their literature, a list of currently available cell lines, and new findings compared with the original studies. A literature review was conducted via MEDLINE, PubMed and ScienceDirect for reviews in the last 5 years to identify research and other studies related to gynaecological cancer cell lines. We present an overview of the current literature with reference to the original studies and pointed to certain inconsistencies in the literature. The adherence to culturing rulesets and the international guidelines helps in minimizing replication failure between institutions. Evidence from the latest research suggests that despite certain drawbacks, variations of cancer cell lines can also be useful in regard to a more diverse genomic landscape.


Assuntos
Técnicas de Cultura de Células/métodos , Neoplasias dos Genitais Femininos/patologia , Linhagem Celular Tumoral , Feminino , Humanos
8.
Cells ; 11(1)2021 12 30.
Artigo em Inglês | MEDLINE | ID: mdl-35011679

RESUMO

Triple-negative breast cancer (TNBC) is a breast cancer (BC) subtype that accounts for approximately 15-20% of all BC cases. Cancer cell lines (CLs) provide an efficient way to model the disease. We have recently isolated a patient-derived triple-negative BC CL MFUM-BrTNBC-1 and performed a detailed morphological and molecular characterisation and a comprehensive comparison with three commercial BC CLs (MCF-7, MDA-MB-231, MDA-MB-453). Light and fluorescence microscopy were used for morphological studies; immunocytochemical staining for hormone receptor, p53 and Ki67 status; RNA sequencing, qRT-PCR and STR analysis for molecular characterisation; and biomedical image analysis for comparative phenotypical analysis. The patient tissue-derived MFUM-BrTNBC-1 maintained the primary triple-negative receptor status. STR analysis showed a stable and unique STR profile up to the 6th passage. MFUM-BrTNBC-1 expressed EMT transition markers and displayed changes in several cancer-related pathways (MAPK, Wnt and PI3K signalling; nucleotide excision repair; and SWI/SNF chromatin remodelling). Morphologically, MFUM-BrTNBC-1 differed from the commercial TNBC CL MDA-MB-231. The advantages of MFUM-BrTNBC-1 are its isolation from a primary tumour, rather than a metastatic site; good growth characteristics; phenotype identical to primary tissue; complete records of origin; a unique identifier; complete, unique STR profile; quantifiable morphological properties; and genetic stability up to (at least) the 6th passage.


Assuntos
Neoplasias de Mama Triplo Negativas/genética , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Humanos , Transdução de Sinais
9.
Mol Biol Rep ; 47(9): 6929-6940, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32876844

RESUMO

Macrophages are among the most important cells of the immune system. Among other functions, they take part in almost all defense actions against foreign bodies and bacteria, being particularly important in infections, wound healing, and foreign body reactions. Considering their importance for the health of the human body, as well as their important role in several diseases, the in vitro studies based on these cells, are a crucial research field. Taking all mentioned into account, this study describes a simple isolation method of human macrophages (MFUM-HMP-001 and MFUM-HMP-002 cell lines) from peripheral blood. For this purpose, the morphology, the viability, and the phagocytotic activity of the isolated cells were tested. The Immunostaining of MFUM-HMP-001 and MFUM-HMP-002 cells confirmed the macrophage cell markers CD68, CD80, and CD163/M130. The phagocytotic activity was marked in both MFUM-HMP-001 and MFUM-HMP-002 cells, as was the phagocytosis of the pHrodo green Escherichia coli bioparticles conjugates, which was enhanced with the addition of lipopolysaccharide. The cells were stable and exhibited good growth. According to our results, both cell lines are useful for the development of novel macrophage cell-based in vitro models.


Assuntos
Técnicas de Cultura de Células/métodos , Macrófagos/citologia , Macrófagos/metabolismo , Fagocitose , Antígenos CD/sangue , Antígenos de Diferenciação Mielomonocítica/sangue , Antígeno B7-1/sangue , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Escherichia coli/metabolismo , Humanos , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Fagocitose/efeitos dos fármacos , Receptores de Superfície Celular/sangue
10.
Acta Med Acad ; 49 Suppl 1: 4-13, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33543625

RESUMO

OBJECTIVE: The aim of this study was to establish an optimised protocol for glioblastoma (GBM) cell isolation from brain resection samples, with a high yield and low risk for contamination. METHODS: Human GBM cells can be obtained following cranial tumour operations. In sterile conditions, the fragments of viable tissue removed during surgery were collected. The tissue was cut and mechanically coarsely decomposed. The sediment was harvested after centrifugation, the cells were seeded in suspension, and supplemented with a special medium (Advanced DMEM) containing high level nutrients and antibiotics. RESULTS: In an appropriate environment, the isolated cells retained viability and proliferated quickly. Attachments were observed after ten hours, and proliferation after two days. The time to full confluence was about one week. The cells were stable. Under standard culture conditions, cell proliferation and cluster formation were observed. Cell viability was 95%. CONCLUSION: The protocol described for isolation is easy, quick and affordable, leading to stable GBM cells. The isolation technique provides sufficient quantities of isolated cells that may be used as an important new tool for in vitro research. The availability of this system will permit the study of cell properties, biochemical aspects, and provides the potential of therapeutic candidates for pathological disorders in a well-controlled environment.


Assuntos
Glioblastoma , Proliferação de Células , Separação Celular , Humanos
12.
Mol Biol Rep ; 47(2): 1399-1411, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31848918

RESUMO

Endometrial cancer is one of the most common gynaecological malignancies worldwide. One type of research in this field is the growing of cell lines (CLs) and cultures, which can be used to explore the biological mechanisms of cancer. The purpose of this review is to offer an overview of the current literature and highlight the importance of correct CL studies. We carried out a literature analysis of more than 60 articles from the Pubmed, Medline databases that were almost exclusively published in indexed journals in the last 10 years as well as the primary originating scientific studies of specific CLs. We then summarized the newest findings and recommendations. Cell lines are becoming widely used as in vitro tumour models. Recent work has shown inconsistencies in nomenclature and culturing of CLs. Their genomic evolution leads to a high degree of variation across CL strains therefore it is of the utmost importance to recognize the variability within laboratory cancer models. Laboratories must adapt, incorporate additional characterisation techniques and view this situation as an opportunity to improve the reproducibility of pre-clinical cancer research. The authors offer a comprehensive literature review about endometrial cancer CLs, a review of the current literature and advice on culturing CLs.


Assuntos
Neoplasias do Endométrio/patologia , Técnicas de Cultura de Células , Linhagem Celular Tumoral , Neoplasias do Endométrio/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos
13.
Bosn J Basic Med Sci ; 19(3): 297-303, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-30877836

RESUMO

The meninges are involved in various pathologies and are often directly or indirectly severed during surgical procedures, especially the dura mater. This can pose a real challenge for the surgeon, as a proper reconstruction of the meninges is important to prevent complications such as cerebrospinal fluid leak (CSF). A variety of techniques for dural reconstruction have been described, employing natural and artificial materials. A novel technique for dural reconstruction involves soft tissue grafts in the form of fibrous or fibromuscular flaps, which are placed on the dural defects to seal the gaps. These soft tissue grafts represent an appropriate scaffold for cell ingrowth and fibrosis, thus preventing CSF. In this pilot study, we described the application of soft tissue grafts for dural reconstruction in 10 patients who underwent convexity meningioma surgery.


Assuntos
Neoplasias Encefálicas/cirurgia , Dura-Máter/cirurgia , Neoplasias Meníngeas/cirurgia , Meninges/cirurgia , Meningioma/cirurgia , Procedimentos Neurocirúrgicos/métodos , Procedimentos de Cirurgia Plástica/métodos , Terapia de Tecidos Moles/métodos , Adulto , Idoso , Vazamento de Líquido Cefalorraquidiano/prevenção & controle , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Complicações Pós-Operatórias/prevenção & controle , Complicações Pós-Operatórias/terapia , Estudos Prospectivos , Retalhos Cirúrgicos , Alicerces Teciduais
14.
Technol Cancer Res Treat ; 18: 1533033818821093, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30803358

RESUMO

Macrophages play key role in host defense and tissue repair, and thus understanding regulation of their function is important. For instance, our previous results have shown that in chicken macrophage system (CoMA cell line), application of a pulse of electromagnetic fields of frequencies 0.618, 1.054, 5.229, and 100.414 kHz induces production of interferon γ-like molecules. In this study, we have shown that the electromagnetic field of 100.414 kHz is the most effective in inducing synthesis of chicken interferon γ and chicken interferon γ-like molecules in CoMA cells, especially when combined with Lens culinaris agglutinin and 10% phosphate-buffered saline washouts of different Holocene minerals. A 2-minute pulse of electromagnetic field was produced by Defender's pulse generator. Both chicken interferon γ and chicken interferon γ-like molecules from the cell supernatant were evaluated by an antiviral assay and were also analyzed with reverse-phase high-performance liquid chromatography on Phenomenex, Aeris peptide columns. Our results show that application of a single inducing factor ( Lens culinaris agglutinin, 100.414 kHz electromagnetic field, 10% phosphate buffer saline washout) or combined usage of 2 of them moderately stimulated production of chicken interferon γ-like molecules (from 1.550 to 48.028 IU/mL), whereas the combination of 10% phosphate-buffered saline washout of Koprivnica rock + Lens culinaris agglutinin + 100.414 kHz/9 V resulted in an output of 162.122 IU/mL. Hence, we may conclude that a combined use of electromagnetic field, Holocene minerals, and Lens culinaris agglutinin greatly stimulates synthesis of chicken interferon γ-like molecules in CoMA cells.


Assuntos
Fontes de Energia Elétrica , Campos Eletromagnéticos , Interferon gama/metabolismo , Macrófagos/metabolismo , Minerais/farmacologia , Lectinas de Plantas/farmacologia , Animais , Células Cultivadas , Galinhas , Macrófagos/efeitos dos fármacos , Macrófagos/efeitos da radiação
15.
Eur J Pharm Biopharm ; 128: 230-246, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29730258

RESUMO

In this comprehensive study several analytical techniques were used in order to evaluate multi-layered biomedical surface coatings composed of a drug (diclofenac) and a polymer (chitosan). Such a thorough examination is of paramount importance in order to assure safety and prove efficiency of potential biomedical materials already at the in vitro level, hence leading to their potentially faster introduction to clinical trials. For the first time a novel technique based on thermal diffusivity and conductivity measurements (photothermal beam deflection spectroscopy - BDS) was employed in order to analyse in a non-destructive way the thickness of respective layers, together with their thermal diffusivity and conductivity. In addition to attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR), BDS confirmed successive surface layers of the prepared coatings. Scanning electron microscopy and atomic force microscopy were used to examine structural information on the macro- and micro/nano-scale, respectively. Surface hydrophobicity was measured with the contact angle analysis, which clearly showed differences in hydrophobicity between coated and non-coated samples. Considering the targeted application of the prepared coatings (as implant in orthopaedic treatments), the in vitro drug release was analysed spectrophotometrically to examine the coatings potential for a controlled drug release. Furthermore, the material was also tested by electrochemical impedance spectroscopy and cyclic polarisation techniques, which were able to detect even minor differences between the performance of the coated and non-coated materials. As the final test, the biocompatibility of the coatings with human osteoblasts was determined.


Assuntos
Materiais Revestidos Biocompatíveis/química , Teste de Materiais/métodos , Aço Inoxidável/química , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Quitosana/química , Diclofenaco/química , Diclofenaco/farmacologia , Espectroscopia Dielétrica , Humanos , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura , Osteoblastos , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de Superfície
16.
Bioengineering (Basel) ; 5(1)2018 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-29315214

RESUMO

Cytocompatible polysaccharide-based functional scaffolds are potential extracellular matrix candidates for soft and hard tissue engineering. This paper describes a facile approach to design cytocompatible, non-toxic, and multifunctional chitosan-cellulose based hydrogel beads utilising polysaccharide dissolution in sodium hydroxide-urea-water solvent system and coagulation under three different acidic conditions, namely 2 M acetic acid, 2 M hydrochloric acid, and 2 M sulfuric acid. The effect of coagulating medium on the final chemical composition of the hydrogel beads is investigated by spectroscopic techniques (ATR-FTIR, Raman, NMR), and elemental analysis. The beads coagulated in 2 M acetic acid displayed an unchanged chitosan composition with free amino groups, while the beads coagulated in 2 M hydrochloric and sulfuric acid showed protonation of amino groups and ionic interaction with the counterions. The ultrastructural morphological study of lyophilized beads showed that increased chitosan content enhanced the porosity of the hydrogel beads. Furthermore, cytocompatibility evaluation of the hydrogel beads with human breast adenocarcinoma cells (soft tissue) showed that the beads coagulated in 2 M acetic acid are the most suitable for this type of cells in comparison to other coagulating systems. The acetic acid fabricated hydrogel beads also support osteoblast growth and adhesion over 192 h. Thus, in future, these hydrogel beads can be tested in the in vitro studies related to breast cancer and for bone regeneration.

17.
Med Arch ; 72(6): 444-448, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30814778

RESUMO

INTRODUCTION: Wounds and their complications present a frequent cause of morbidity and mortality in everyday clinical practice. In order to reduce the wound burden, much effort has been directed into the physiology of healing and new therapeutic approaches. AIM: This paper provides an overview from the literature about the role of endothelial and epithelial cells in tissue filler employment for wound healing. MATERIAL AND METHODS: The scientific literature was reviewed through PubMed, Medline and Science Direct. The articles were chosen in correlation with the study objective and their scientific relevance. RESULTS: Successful wound healing depends on many diverse processes, cell types and molecular mediators. The definitive aim of wound healing is a properly healed wound. Tissue fillers are becoming an important alternative in wound management, although augmentation of soft tissue can present a demanding problem due to the difficulties in tissue survival. In order to prevent its failure, an optimal vascular network needs to form from wound edges into the filler. CONCLUSIONS: Because of the importance of chemotaxis and angiogenesis in various physiological and pathological processes, both events present an extensive area of intense research. Additionally, epithelial cells are needed to cover the wound defect and sealing the wound environment from outer world.


Assuntos
Proliferação de Células/fisiologia , Células Endoteliais/fisiologia , Células Epiteliais/fisiologia , Neovascularização Patológica , Cicatrização/fisiologia , Ferimentos e Lesões/patologia , Materiais Biocompatíveis/farmacologia , Movimento Celular , Preenchedores Dérmicos/farmacologia , Humanos , Ferimentos e Lesões/terapia
18.
Biomed Pharmacother ; 94: 1176-1185, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28830068

RESUMO

Over the past decade, it has become clear that specific probiotic lactobacilli are valuable in the prevention and treatment of infectious and inflammatory diseases of gastrointestinal tract but their successful application would benefit greatly from a better understanding of the mechanisms of individual strains. Hence, each probiotic strain should be characterized for their immune activity before being proposed for clinical applications. The aim of the study was to characterize the immunomodulatory activity of the strain Lactobacillus (L.) plantarum LS/07 in vitro using functional gut model and to study its anti-inflammatory potential in dextran sulphate sodium (DSS)-induced colitis in rats. We showed that L. plantarum LS/07 induced production of IL-10 in macrophages derived from blood monocytes as well as monocyte/macrophages cell line stimulated indirectly via enterocytes in vitro. In rat model of colitis, L. plantarum LS/07 attenuated the DSS-induced signs of inflammatory process in colon such as weight loss, diarrhoea, infiltration of inflammatory cells associated with decreased colon weight/length ratio, inhibited gut mucosa destruction and depletion of goblet cells. Moreover, the strain increased the concentration of anti-inflammatory cytokine IL-10 in mucosal tissue. In conclusion, the protective effects of L. plantarum LS/07 in the DSS-induced colitis model seem to be related to the stimulation of IL-10 and the restoration of goblet cells and indicate it as a good candidate to prevent and treat diseases associated with inflammation.


Assuntos
Colite/tratamento farmacológico , Citocinas/metabolismo , Lactobacillus plantarum/fisiologia , Probióticos/farmacologia , Adulto , Animais , Anti-Inflamatórios/farmacologia , Linhagem Celular , Colite/induzido quimicamente , Colite/metabolismo , Colo/efeitos dos fármacos , Colo/metabolismo , Sulfato de Dextrana/farmacologia , Modelos Animais de Doenças , Células Caliciformes/efeitos dos fármacos , Células Caliciformes/metabolismo , Humanos , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Interleucina-10/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Lactobacillus plantarum/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Pessoa de Meia-Idade , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Ratos , Ratos Sprague-Dawley
19.
PeerJ ; 5: e3079, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28344902

RESUMO

BACKGROUND: Cartilage tissue engineering is a fast-evolving field of biomedical engineering, in which the chondrocytes represent the most commonly used cell type. Since research in tissue engineering always consumes a lot of cells, simple and cheap isolation methods could form a powerful basis to boost such studies and enable their faster progress to the clinics. Isolated chondrocytes can be used for autologous chondrocyte implantation in cartilage repair, and are the base for valuable models to investigate cartilage phenotype preservation, as well as enable studies of molecular features, nature and scales of cellular responses to alterations in the cartilage tissue. METHODS: Isolation and consequent cultivation of primary human adult articular chondrocytes from the surgical waste obtained during total knee arthroplasty (TKA) was performed. To evaluate the chondrogenic potential of the isolated cells, gene expression of collagen type 2 (COL2), collagen 1 (COL1) and aggrecan (ACAN) was evaluated. Immunocytochemical staining of all mentioned proteins was performed to evaluate chondrocyte specific production. RESULTS: Cartilage specific gene expression of COL2 and ACAN has been shown that the proposed protocol leads to isolation of cells with a high chondrogenic potential, possibly even specific phenotype preservation up to the second passage. COL1 expression has confirmed the tendency of the isolated cells dedifferentiation into a fibroblast-like phenotype already in the second passage, which confirms previous findings that higher passages should be used with care in cartilage tissue engineering. To evaluate the effectiveness of our approach, immunocytochemical staining of the evaluated chondrocyte specific products was performed as well. DISCUSSION: In this study, we developed a protocol for isolation and consequent cultivation of primary human adult articular chondrocytes with the desired phenotype from the surgical waste obtained during TKA. TKA is a common and very frequently performed orthopaedic surgery during which both femoral condyles are removed. The latter present the ideal source for a simple and relatively cheap isolation of chondrocytes as was confirmed in our study.

20.
Sci Rep ; 6: 26653, 2016 05 24.
Artigo em Inglês | MEDLINE | ID: mdl-27215333

RESUMO

Corrosion resistance, biocompatibility, improved osteointegration, as well the prevention of inflammation and pain are the most desired characteristics of hip replacement implants. In this study we introduce a novel multi-layered coating on AISI 316LVM stainless steel that shows promise with regard to all mentioned characteristics. The coating is prepared from alternating layers of the biocompatible polysaccharide chitosan and the non-steroid anti-inflammatory drug (NSAID), diclofenac. Electrochemical methods were employed to characterize the corrosion behavior of coated and uncoated samples in physiological solution. It is shown that these coatings improve corrosion resistance. It was also found that these coatings release the incorporated drug in controlled, multi-mechanism manner. Adding additional layers on top of the as-prepared samples, has potential for further tailoring of the release profile and increasing the drug dose. Biocompatibility was proven on human-derived osteoblasts in several experiments. Only viable cells were found on the sample surface after incubation of the samples with the same cell line. This novel coating could prove important for prolongation of the application potential of steel-based hip replacements, which are these days often replaced by more expensive ceramic or other metal alloys.


Assuntos
Artroplastia de Quadril , Quitosana/química , Materiais Revestidos Biocompatíveis/química , Diclofenaco/química , Prótese de Quadril , Teste de Materiais , Osteoblastos/metabolismo , Aço Inoxidável/química , Humanos , Masculino
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