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1.
Acta cir. bras ; 39: e390524, 2024. tab, graf, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1533358

ABSTRACT

Purpose: We aimed to reveal the effects of rosmarinic acid (RA), which has come to the forefront with its antitumor and antioxidant properties in many studies recently in the ovarian adenocarcinoma cell line, on the epidermal growth factor receptor (EFGR) signaling pathway in the presence of doxorubicin (DOX). Methods: Ovarian adenocarcinoma cell line (OVCAR3) and human skin keratinocyte cell line human skin keratinocyte cell line (HaCaT) were used as control. (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was applied to determine the effect of RA and DOX on the proliferation of OVCAR3 and HaCaT cells. Bcl2 expression and epidermal growth factor receptor (EGFR) and western blot analysis were performed to determine the expression levels of the markers. Results: It was determined that RA (IC50 = 437.6 µM) and DOX (IC50 = 0.08 µM) have the ability to inhibit the proliferation of OVCAR3 cells and induce apoptosis in a 72-hour time and dose-dependent manner. Western blot showed that the expression level of Bcl-2 and EGFR in OVCAR3 cells was down-regulated by RA and DOX. Conclusions: Apoptosis in OVCAR3 cells can potentially be induced by RA via the EGFR pathway, and RA may be a potent agent for cancer therapy.


Subject(s)
Ovarian Neoplasms , Adenocarcinoma , Doxorubicin/administration & dosage , ErbB Receptors
2.
International Eye Science ; (12): 686-690, 2024.
Article in Chinese | WPRIM | ID: wpr-1016578

ABSTRACT

AIM: To explore the synthesis of thermo-sensitive poly N-isopropylacry-lamide(PNIPAAm)and the petri dish grafted with PNIPAAm hydrogels by the electron accelerator, as well as the growth conditions and the biological characteristics of rabbit corneal stromal cells on thermo-sensitive PNIPAAm hydrogels, and the cell sheets obtained from the PNIPAAm hydrogels.METHODS: NIPAAm monomer was dissolved in 2-propanol at concentrations of 55% with 0.5% N,N'-Methylenebisacry-lamide(MBA). Solution(70 μL)was added and spread uniformly over 35 mm petri dish. These dishes were immediately subjected to irradiation. After follow-up treatment, rabbit corneal stromal cells were cultured on thermo-sensitive petri dish in vitro.RESULTS: According to the monomer formula and radiation synthesis scheme in this experiment, PNIPAAm can be synthesized on the surface of the petri dish. Rabbit corneal stromal cells grew well in the thermo- sensitive surface and can be separated into sheets.CONCLUSION: The single and multilayer carrier-free cell sheets can be obtained from the use of thermo-sensitive petri dish.

3.
Chinese Journal of Biologicals ; (12): 335-342, 2024.
Article in Chinese | WPRIM | ID: wpr-1013398

ABSTRACT

@#Objective To develop and verify a multiplex fluorescent quantitative PCR method for detection of common bacterial and fungal contaminants in cell culture medium.Methods According to NUC gene of Staphylococcus aureus,COLA gene of Clostridium spore,ITS-2 segment sequence of Candida albicans,a set of primers and probes were designed for each respectively,and using UBI3 gene of capsicum introduced as external standard gene,a triple reaction system of Staphylococcus aureus,Clostridium spore and external standard gene and a double reaction system of Candida albicans and external standard gene were established.The primer specificity,linear range,limit of detection,specificity,anti-interference performance and precision of the method were verified.Finally,100 samples of 293T cell culture medium were detected by using the developed method,which was compared with the common PCR method.Results Three pairs of primers all amplified about 100 bp specific gene bands corresponding to the three strains at different annealing temperatures(56,57,58 and59 ℃),and the size was consistent with the expected.In the range of 5.80 × 10~6 — 5.80 × 10~2 copies/μL,the standard plasmids of the three strains showed a good linear relationship with the Ct values.The standard curve equations were:Y=-3.373 X+37.48,Y=-3.557X+36.59 and Y=-3.536 X+39.78,each R~2> 0.99,respectively,and the amplification efficiency was in the range of 90%—110%.All the limits of detection of the three strains were 10~1 CFU/mL.The primers and probes of the three strains showed no specific amplification on the genomic DNA of six kinds of cells that were prone to cross-reaction.The genomic DNA of 293T cells,Yeast,Escherichia coli and Mycoplasma sp.had no effect on the detection.The CVs of repeatability and intermediate precision verification were both less than 15%.Among 100 cell culture medium samples,14 positive and 86 negative samples were detected,and the results of common PCR method for three positive and two negative samples randomly selected were consistent with the developed method.Conclusion The multiplex fluorescent quantitative PCR method developed in this study for the detection of bacteria and fungi in cell culture medium has good specificity,anti-interference performance and precision,and is simple to operate with low cost and high sensitivity,which can quickly detect the contaminants during cell culture.

4.
Chinese Journal of Biologicals ; (12): 26-31, 2024.
Article in Chinese | WPRIM | ID: wpr-1006192

ABSTRACT

@#Objective To investigate the surface properties of different fibracel carriers and their culture effects on different cells.Methods Three fibracel carriers(A,B,C)were selected to analyze the chemical element composition of their materials,and the contact angles of the carriers before and after pretreatment with 0. 1 mol/L NaOH solution were tested. By measuring the adhesion effect and glucose consumption of Vero,MDCK and MRC-5 cells on the carrier,and observing the cell growth state by fluorescent staining,the cell adhesion efficiency and culture effect of the three carriers were compared and analyzed.Results The three carriers were mainly composed of C,H,O,and contained a small amount of N and S elements. Before pretreatment,the contact angle of carrier B was 0°,which was significantly lower than that of A[(109 ± 3. 13)°]and C[(121 ± 6. 82)°](each F = 709. 1,each P < 0. 000 1),and the hydrophilicity was stronger. Carriers A and C had poor hydrophilicity. After pretreatment,the contact angles of the surfaces of the three carriers A,B,and C were all 0°,with no significant difference(F = 0. 069 4,P > 0. 05),all of which were hydrophilic. The adherence rates of the three types of carriers within 3 h of cell culture were all above 80%. The cells were dense and evenly grown on the carrier fibers,the glucose consumption curves tended to“S”type,and the continuous cell culture effect was good. The total glucose consumption of carrier A and carrier C was basically the same,and carrier B was lower than carrier A and carrier C.Conclusion The chemical element composition and the relationship between the hydrophilic and hydrophobic properties of the three fibracel carriers were analyzed,and the adhesion rate and culture effect of Vero,MDCK and MRC-5 cells were evaluated,which provide reference for the subsequent research and production application of fibracel carriers.

5.
Rev. Ciênc. Saúde ; 13(3): 23-30, 20230921.
Article in English | LILACS | ID: biblio-1510835

ABSTRACT

Objective: The present study aims to evaluate the viability of adult human neural cells in culture obtained from traumatized brain tissues collected in emergency surgery procedures. Methods: Exploratory, descriptive, quantitative and cross-sectional study evaluating samples obtained from patients who underwent traumatic brain injury with extrusion of brain tissue submitted to cell culture in a standardized medium, being preserved during 168h. After observation under phase contrast microscopy and immunohistochemical processing for neuronal (MAP-2) and glial (GFAP) markers, morphometric parameters of neural cells (cell body area, dendritic field length and fractal dimension) were evaluated using ImageJ software, with data obtained after 24, 72 and 168h being compared using non-parametric Kruskal Wallis test, followed by Dunn's post hoc test. Results: The explant of the nervous tissue revealed a consolidated pattern of cell migration into the culture medium. Cell proliferation, upon reaching confluence, presented an aspect of cellular distribution juxtaposed along the culture medium at all time points analyzed. Both neurons and glial cells remained viable after 168h in culture, with their morphologies not varying significantly throughout the time points evaluated. Immunohistochemistry for MAP-2 showed a relatively well-preserved cytoskeletal organization. GFAP immunoreactivity revealed activated astrocytes especially at the later time point. Conclusions: Our results point out the viability of cell culture from traumatized human nervous tissue, opening up perspectives for the use of substances of natural origin that may contribute neuroprotectively to neuronal maintenance in culture, allowing future translational approach.


Subject(s)
Humans , Male , Adult , Brain Injuries , Cell Culture Techniques , Neurons , Wounds and Injuries , Traumatology , Immunohistochemistry
6.
Rev. peru. med. exp. salud publica ; 40(3): 297-306, jul. 2023. ilus, graf
Article in Spanish | LILACS, INS-PERU | ID: biblio-1522787

ABSTRACT

Objetivo . Desarrollar y validar un método de suspensión celular utilizando células Vero 76 para el cultivo del virus Zika (ZIKV) basado en la infección de células recién sembradas no adheridas. Material y métodos . Se utilizaron tres multiplicidades de infección diferentes del ZIKV para desarrollar y comparar este novedoso método con el método estándar de monocapa de células confluentes. Además, validamos preliminarmente el método de suspensión utilizando muestras clínicas caracterizadas como positivas o negativas para el ZIKV. El método estándar de monocapa se utilizó como método de referencia, y el aislamiento viral se confirmó mediante un RT-PCR específico del ZIKV. Se estimó la sensibilidad e intervalos de confianza del 95% para el método de suspensión. Asimismo, se realizó una comparación técnica del método de suspensión contra el método de monocapa. Resultados . Nuestros hallazgos sugieren que tanto la carga viral como la replicación del ZIKV fueron comparables entre los métodos de infección en monocapa y en suspensión. Aunque ambos métodos fueron adecuados para cultivar y aislar el ZIKV, el método de suspensión se caracterizó por ser más fácil, barato y rápido, así como una técnica de aislamiento sensible. En comparación con el método de monocapa, el método de suspensión fue cuatro veces más sensible en la detección del ZIKV en casos inconclusos por RT-PCR. Conclusiones . El método de suspensión tiene el potencial de ser un método eficaz para cultivar y aislar el ZIKV y su uso es potencialmente útil tanto en la investigación como en entornos clínicos.


Objective. To develop and validate a cell suspension method using Vero 76 cells for culturing Zika virus (ZIKV) based on infection of detached freshly seeded cells. Material and methods. Three different multiplicities of infection of ZIKV were used to develop and compare this novel method to the standard confluent cell monolayer method. In addition, we preliminary validated the cell suspension method using well-characterized ZIKV positive and negative clinical samples. The standard confluent cell monolayer method was used as the reference method, and viral isolation was confirmed by a ZIKV-specific RT-PCR. The sensitivity and its 95% confidence intervals for the cell suspension method were estimated. Also, a technical comparison of the cell suspension method against the cell monolayer method was performed. Results. Our findings suggested that both the viral load and replication of ZIKV were comparable between both monolayer- and suspension-infection methods. Although both methods were suitable for culturing and isolating ZIKV, the cell suspension method was easier, cheaper, and quicker as well as a sensitive isolation technique. The cell suspension method was significantly more sensitive in detecting Zika in inconclusive cases by RT-PCR, with a fourfold increase compared to the confluent cell monolayer method. Conclusion. The cell suspension method has the potential to be an effective method for cultivating and isolating ZIKV and its application is potentially useful in both research and clinical settings.


Subject(s)
Zika Virus Infection , Cell Culture Techniques , Public Health Surveillance
7.
Mem. Inst. Oswaldo Cruz ; 118: e230090, 2023. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1506730

ABSTRACT

BACKGROUND According to the last 2023 Monkeypox (Mpox) Outbreak Global Map from the Centres for Disease Control and Prevention (CDC), more than 100 countries with no Mpox infection report cases. Brazil stands out in this group and is the second country with the highest number of cases in the last outbreak. OBJECTIVE To contribute to knowledge of the virus infection effects in a cellular model, which is important for diagnosis infections not yet included in a provider´s differential diagnosis and for developing viral inhibition strategies. METHODS We describe a virus isolation protocol for a human clinical sample from a patient from Brazil, the viral growth in a cell model through plaque forming units (PFU) assay, reverse transcriptase polymerase chain reaction (RT-PCR) and transmission electron microscopy (TEM). FINDINGS We follow the viral isolation in Vero cell culture from a Mpox positive clinically diagnosed sample and show the infection effects on cellular structures using a TEM. MAIN CONCLUSIONS Understanding the impact of viral growth on cellular structures and its replication kinetics may offer better strategies for the development of new drugs with antiviral properties.

8.
Braz. j. med. biol. res ; 56: e12713, 2023. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1447680

ABSTRACT

Mesenchymal stromal/stem cells stem (MSC) have been widely studied due to their great potential for application in tissue engineering and regenerative and translational medicine. In MSC-based therapy for human diseases, cell proliferation is required to obtain a large and adequate number of cells to ensure therapeutic efficacy. During in vitro culture, cells are under an artificial environment and manipulative stress that can affect genetic stability. Several regulatory agencies have established guidelines to ensure greater safety in cell-based regenerative and translational medicine, but there is no specific definition about the maximum number of passages that ensure the lowest possible risk in MSC-based regenerative medicine. In this context, the aim of this study was to analyze DNA damage and chromosome alterations in adipose-derived mesenchymal stromal cells (ADMSC) until the eleventh passage and to provide additional subsidies to regulatory agencies related to number of passages in these cells. Thus, two methods in genetic toxicology were adopted: comet assay and micronucleus test. The comet assay results showed an increase in DNA damage from the fifth passage onwards. The micronucleus test showed a statistically significant increase of micronucleus from the seventh passage onwards, indicating a possible mutagenic effect associated with the increase in the number of passages. Based on these results, it is important to emphasize the need to assess genetic toxicology and inclusion of new guidelines by regulatory agencies to guarantee the safety of MSC-based therapies for human diseases.

9.
J. appl. oral sci ; 31: e20230032, 2023. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1448548

ABSTRACT

Abstract Simulating a bacterial-induced pulpitis environment in vitro may contribute to exploring mechanisms and bioactive molecules to counteract these adverse effects. Objective To investigate the chronic exposure of human dental pulp cells (HDPCs) to lipopolysaccharides (LPS) aiming to establish a cell culture protocol to simulate the impaired odontogenic potential under pulpitis conditions. Methodology HDPCs were isolated from four healthy molars of different donors and seeded in culture plates in a growth medium. After 24 h, the medium was changed to an odontogenic differentiation medium (DM) supplemented or not with E. coli LPS (0 - control, 0.1, 1, or 10 µg/mL) (n=8). The medium was renewed every two days for up to seven days, then replaced with LPS-free DM for up to 21 days. The activation of NF-κB and F-actin expression were assessed (immunofluorescence) after one and seven days. On day 7, cells were evaluated for both the gene expression (RT-qPCR) of odontogenic markers (COL1A1, ALPL, DSPP, and DMP1) and cytokines (TNF, IL1B, IL8, and IL6) and the production of reactive nitrogen (Griess) and oxygen species (Carboxy-H2DCFDA). Cell viability (alamarBlue) was evaluated weekly, and mineralization was assessed (Alizarin Red) at 14 and 21 days. Data were analyzed with ANOVA and post-hoc tests (α=5%). Results After one and seven days of exposure to LPS, NF-κB was activated in a dose-dependent fashion. LPS at 1 and 10 µg/mL concentrations down-regulated the gene expression of odontogenic markers and up-regulated cytokines. LPS at 10 µg/mL increased both the production of reactive nitrogen and oxygen species. LPS decreased cell viability seven days after the end of exposure. LPS at 1 and 10 µg/mL decreased hDPCs mineralization in a dose-dependent fashion. Conclusion The exposure to 10 µg/mL LPS for seven days creates an inflammatory environment that is able to impair by more than half the odontogenic potential of HDPCs in vitro, simulating a pulpitis-like condition.

10.
Einstein (Säo Paulo) ; 21: eAO0160, 2023. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1440063

ABSTRACT

ABSTRACT Objective This study verified the replication efficiency of the Rocio virus in a primary culture of mouse neural cells. Methods Mixed primary cultures (neurons/glia) obtained from the brains of newborn isogenic BALB/c mice were inoculated with Rocio virus on the 7 th day of culture, and the development of cytopathogenic effects was monitored. The infection was confirmed via immunocytochemistry (anti-ROCV), while viral replication was quantified in infected primary cultures. The titration method used depended on the infection period. Results Rocio virus efficiently infected primary cultured neural cells, with the highest viral titer causing cytopathic changes was observed at 2 days post infection. The virus-infected primary culture survived for up to 7 days post infection, and viral load quantitation showed viral replication kinetics compatible with the cell death kinetics of cultures. Conclusion The findings of this study suggest that mouse neural cell primary cultures support Rocio virus replication and could be used as an alternative system for studying Flavivirus infection in the central nervous system.

11.
Journal of Biomedical Engineering ; (6): 602-608, 2023.
Article in Chinese | WPRIM | ID: wpr-981582

ABSTRACT

Three-dimensional (3D) cell culture model is a system that co-culture carriers with 3D structural materials and different types of cells in vitro to simulate the microenvironment in vivo. This novel cell culture model has been proved to be close to the natural system in vivo. In the process of cell attachment, migration, mitosis and apoptosis, it could produce biological reactions different from that of monolayer cell culture. Therefore, it can be used as an ideal model to evaluate the dynamic pharmacological effects of active substances and the metastasis process of cancer cells. This paper compared and analyzed the different characteristics of cell growth and development under two-dimensional (2D) and 3D model culture and introduced the establishment method of 3D cell model. The application progress of 3D cell culture technology in tumor model and intestinal absorption model was summarized. Finally, the application prospect of 3D cell model in the evaluation and screening of active substance was revealed. This review is expected to provide reference for the development and application of new 3D cell culture models.


Subject(s)
Cell Culture Techniques, Three Dimensional , Cell Culture Techniques , Apoptosis , Cell Proliferation , Technology
12.
Chinese Journal of Biologicals ; (12): 631-635, 2023.
Article in Chinese | WPRIM | ID: wpr-996382

ABSTRACT

@#Cell culture medium is one of the essential raw materials in the field of life and health.In recent years,the performance and quality of domestic cell culture media have been improving,and the market share of domestic vendors has steadily increased from 19.2% in 2017 to 33.7% in 2021.However,there are also some problems and shortcomings in industrial development,mainly including:technology and process accumulation need to be strengthened;product quality need to be improved;lack of industry standards and norms.Based on literature research,special topic discussion and expert interview,this paper reviews the development history of cell culture medium,deeply analyzes and systematically combs the opportunities and challenges faced by the industry development from the basic situation,current situation and trend of the development of cell culture medium industry in China,and puts forward relevant countermeasures and suggestions.

13.
Chinese Journal of Biotechnology ; (12): 74-85, 2023.
Article in Chinese | WPRIM | ID: wpr-970360

ABSTRACT

Droplet microfluidics technology offers refined control over the flows of multiple fluids in micro/nano-scale, enabling fabrication of micro/nano-droplets with precisely adjustable structures and compositions in a high-throughput manner. With the combination of proper hydrogel materials and preparation methods, single or multiple cells can be efficiently encapsulated into hydrogels to produce cell-loaded hydrogel microspheres. The cell-loaded hydrogel microspheres can provide a three-dimensional, relatively independent and controllable microenvironment for cell proliferation and differentiation, which is of great value for three-dimensional cell culture, tissue engineering and regenerative medicine, stem cell research, single cell study and many other biological science fields. In this review, the preparation methods of cell-loaded hydrogel microspheres based on droplet microfluidics and its applications in biomedical field are summarized and future prospects are proposed.


Subject(s)
Hydrogels/chemistry , Microfluidics/methods , Microspheres , Regenerative Medicine , Tissue Engineering/methods
14.
Chinese Journal of Biotechnology ; (12): 3724-3737, 2023.
Article in Chinese | WPRIM | ID: wpr-1007988

ABSTRACT

Gelatin microspheres were discussed as a scaffold material for bone tissue engineering, with the advantages of its porosity, biodegradability, biocompatibility, and biosafety highlighted. This review discusses how bone regeneration is aided by the three fundamental components of bone tissue engineering-seed cells, bioactive substances, and scaffold materials-and how gelatin microspheres can be employed for in vitro seed cell cultivation to ensure efficient expansion. This review also points out that gelatin microspheres are advantageous as drug delivery systems because of their multifunctional nature, which slows drug release and improves overall effectiveness. Although gelatin microspheres are useful for bone tissue creation, the scaffolds that take into account their porous structure and mechanical characteristics might be difficult to be created. This review then discusses typical techniques for creating gelatin microspheres, their recent application in bone tissue engineering, as well as possible future research directions.


Subject(s)
Tissue Engineering/methods , Tissue Scaffolds/chemistry , Gelatin/chemistry , Microspheres , Bone and Bones , Porosity
15.
Chinese Journal of Biologicals ; (12): 1515-1521+1529, 2023.
Article in Chinese | WPRIM | ID: wpr-1005878

ABSTRACT

@#The productivity of cells per unit area determines the scale-up potential of the cell culture process,and the largescale application of the microcarrier system provides space for the high-yield culture of anchorage-dependent animal cells. The microcarrier animal cell culture technology is suitable for efficient production process development and optimized amplification. In recent years,microcarrier-based culture technology has been widely used in various types of animal cell culture to produce many important biological products such as vaccines,enzymes,hormones,antibodies,interferons and other probiotics. In this paper,the research progress of domestic and foreign microcarrier cell culture technology,the comparison of new microcarriers and traditional microcarriers and their applications were reviewed,so as to provide reference for the in-depth research and application of large-scale cell culture technology based on new microcarriers.

16.
Chinese Journal of Blood Transfusion ; (12): 642-646, 2023.
Article in Chinese | WPRIM | ID: wpr-1004803

ABSTRACT

Human platelet lysates(HPL), as a new type of biomaterial, can promote tissue repair, cell proliferation and inflammation control. This paper introduced the development of HPL in the field of regenerative medicine and cell therapy and summarizes their application. The potential of HPL to promote cell proliferation was used as an entry point to show its advantages as a supplement of cell culture medium. Since there is currently no standard procedure for HPL preparation, this paper sorts out the standardization elements such as raw materials source, donor variability and preparation technology, in order to provide reference for the establishment of standards of relevant industry in the future.

17.
Braz. j. otorhinolaryngol. (Impr.) ; 88(supl.5): 69-74, Nov.-Dec. 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1420910

ABSTRACT

Abstract Objectives: Three-dimensional (3D) cell cultures have many applications such as stem cell biology research, new drug discovery, cancer, and Chronic Rhinosinusitis with Nasal Polyps (CRSwNP). This disease is characterized by a significant impact on quality of life and productivity. The diversity of factors that act in the progression of CRSwNP point to the creation of a cell culture model that allows the integration of different cell types with extracellular matrix. This work aimed to create a cell culture model in 3 dimensions (spheroids) for the study of Nasal Polyposis. Methods: Nasal polyp tissue from patients diagnosed with CRSwNP was mechanically dissociated using tweezers and a scalpel and the solution containing cells and small aggregates of nasal polyps was transferred to a Petri dish containing 5 mL of culture medium at the concentration of 106 cells/mL. Results: The spheroids were cultivated for 20 days, fixed and analyzed using confocal microscopy. In a 3D culture environment, the spheroids were formed both by clustering cells and from small tissue fragments. In the cultures analyzed, the ciliary beat was present from the dissociation of the cells up to 20 days in culture. Conclusion: Our findings also point to these characteristics showing the environment generated in our study, the cells remained differentiated for a longer time and with ciliary beating. Thus, this work shows that nasal polyp-derived cells can be maintained in a 3D environment, enabling better strategies for understanding CRSwNP in situations similar to those found in vivo. Level of evidence: Laboratory studies.

18.
Odontoestomatol ; 24(40)dic. 2022.
Article in Spanish | LILACS-Express | LILACS | ID: biblio-1431006

ABSTRACT

Objetivo: Determinar el efecto de la intensidad de dos unidades de fotopolimerización sobre la biocompatibilidad, resistencia flexural y módulo elástico de una resina compuesta. Metodología: Se crearon dos grupos de resina compuesta Filtek Z250XT cada uno fotopolimerizado con intensidades diferentes (800 mW/cm2 por 20s). La viabilidad celular fue analizada mediante ensayo de MTT a las 24 y 48 horas siguiendo la normativa ISO 10993-5. La resistencia flexural y módulo elástico fueron analizadas siguiendo la normativa ISO 4049. Resultados: En el grupo fotopolimerizado con una intensidad <400 mW/cm2, la citotoxicidad fue estadísticamente mayor tanto a las 24 como a las 48 horas y la resistencia flexural y módulo elástico fueron estadísticamente menores Conclusión: Una intensidad de polimerización <400 mW/cm2, aumenta los niveles de citotoxicidad y disminuye las propiedades mecánicas de las resinas compuestas. Se destaca la importancia del control periódico de las unidades de fotopolimerización.


Objetivo: Determinar o efeito da intensidade de duas unidades de fotopolimerização na biocompatibilidade, resistência à flexão e módulo de elasticidade de uma resina composta. Metodologia: Foram fabricados dois grupos de resina composta Filtek Z250XT, cada um deles foi fotopolimerizado com intensidades diferentes ( 800 mW/cm2 por 20s). A viabilidade celular foi analisada por ensaio de MTT em 24 e 48 horas seguindo a norma ISO 10993-5. A resistência à flexão e o módulo de elasticidade foram analisados ​​seguindo a norma ISO 4049. Resultados: No grupo fotopolimerizado com intensidade <400mW/cm2, a citotoxicidade foi estatisticamente maior nas 24 e 48 horas e a resistência à flexão e o módulo de elasticidade foram estatisticamente menores. Conclusão: Uma intensidade de polimerização <400 mW/cm2 aumenta os níveis de citotoxicidade e diminui as propriedades mecânicas das resinas compostas. Destaca-se a importância do controle periódico das unidades de fotopolimerização.


Objective: To determine the effect of the intensity of two light curing units on the biocompatibility, flexural strength and elastic modulus of a composite resin. Methodology: Two groups of Filtek Z250XT (3M ESPE) composite resin were created, each one photopolymerized using different intensities ( 800 mW/cm2 for 20s). Cell viability was analyzed by MTT assay at 24 and 48 hours following the ISO 10993-5 standard. The flexural strength and elastic modulus were analyzed following the ISO 4049 standard. Results: In the group photopolymerized with an intensity <400 mW/cm2, cytotoxicity was statistically higher both at 24 and 48 hours and flexural strength and elastic modulus were statistically lower. Conclusion: A polymerization intensity <400 mW/cm2 increases the levels of cytotoxicity and decreases the mechanical properties of composite resins. The importance of the periodic control of the light curing units is emphasized.

19.
Hematol., Transfus. Cell Ther. (Impr.) ; 44(4): 560-566, Oct.-dec. 2022. tab, graf
Article in English | LILACS | ID: biblio-1421532

ABSTRACT

ABSTRACT Introduction: Culturing bone marrow mesenchymal stem cells (BM-MSCs) is a key point in different fields of research, including tissue engineering and regenerative medicine and studies of the bone marrow microenvironment. However, isolating and expanding murine BM-MSCs in vitro has challenged researchers due to the low purity and yield of obtained cells. In this study, we aimed to evaluate five different protocols to culture murine BM-MSCs in vitro. Methods: All protocols were based on the adhesion capacity of BM-MSCs to the tissue culture plastic surface and varied in the types of plate, culture media, serum, additional supplementation and initial cell density. Flow cytometry analysis was used to investigate lineage purity after expansion. Results: The expression of CD45 and CD11b was detected in the cultures generated according to all protocols, indicating low purity with the presence of hematopoietic cells and macrophages. The cellular growth rate and morphology varied between the cultures performed according to each protocol. Cells cultured according to protocol 5 (8 × 107cells/plate, Roswell Park Memorial Institute (RPMI) culture medium during first passage and then Iscove's Modified Delbecco's Medium (IMDM) culture medium, both supplemented with 9% fetal bovine serum, 9% horse serum, 12μM L-glutamine) presented the best performance, with a satisfactory growth rate and spindle-shape morphology. Conclusion: Our results point out that the purity and satisfactory growth rate of murine BM-MSC cultures are not easily achieved and additional approaches must be tested for a proper cell expansion.


Subject(s)
Animals , Male , Rats , Mesenchymal Stem Cells , Bone Marrow , In Vitro Techniques , Cell Culture Techniques , Mice
20.
Indian J Ophthalmol ; 2022 Oct; 70(10): 3693-3697
Article | IMSEAR | ID: sea-224642

ABSTRACT

The shortage of donor corneal tissue worldwide has led to extensive research for alternate corneal equivalents utilizing tissue engineering methods. We conducted experiments using Poly D, L lactic acid polymer along with a copolymer (Eudragit) in varying concentrations to create a biodegradable scaffold suitable for in vitro growth of corneal epithelial stem cells. It was found that stable, spherical, and porous microparticles can be prepared by combining PDLLA and Eudragit RL100 polymers in the ratio of 90:10 and 70:30. The microparticles can then be fused to form scaffold membranes with porous architecture and good water retention capacity at room temperature using methanol, which can withstand handling during transplantation procedures. The scaffolds made using a 70:30 ratio were found to be suitable for the promotion of growth of laboratory corneal epithelial stem cell lines (SIRC cell lines). This innovation can pave way for further developments in corneal stem cell research and growth, thus providing for viable laboratory-derived corneal substitutes.

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