scholarly journals Evaluación in vitro de la adhesión de células troncales mesenquimales a matrices dentales impresas en tercera dimensión / In Vitro Evaluation of the Adhesion of Mesenchymal Stem Cells to Three-Dimensional Printed Matrices

2018 ◽  
Vol 36 (77) ◽  
Author(s):  
Álvaro Andrés Rodríguez Sáenz ◽  
María Alejandra Lozano Macías ◽  
Astrid Eugenia Benedetti Canabal ◽  
Juan Carlos Munévar Niño ◽  
Jorge Alberto Sarmiento O'Meara ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Three Dimensional ◽  
Cellular Adhesion ◽  
Poly Lactic Acid ◽  
Time Period ◽  
The Matrix ◽  
Human Dental Pulp ◽  
Dental Stem Cell

RESUMEN. Antecedentes: En la ingeniería de tejidos es fundamental estudiar el sinergismo entre las células troncales mesenquimales y el biomaterial para tener un mayor control sobre los biomiméticos. De esto depende el éxito de tratamientos de lesiones de tamaño crítico. Objetivo: Evaluar la adhesión celular in vitro de células troncales de la pulpa dental humana (hDPSCs) en matrices impresas con ácido poliláctico (APL). Métodos: Se utilizaron muestras de hDPSCs criopreservadas y expandidas, cultivadas sobre 24 matrices dentales impresas 3D en APL durante 1, 7 y 15 días. Se evaluó la fenotipificacion de la hDPSCs por citometría de flujo y la adhesión celular a la matriz por medio de microscopio electrónico de barrido (SEM). Los datos fueron reportados en porcentajes, tanto para marcador analizado, como para la cantidad de células adheridas. Resultados: hDPSCs expresaron positivamente anticuerpos CD73 y CD90 de casi 100 % y CD105 de 56,7 %. Así mismo, expresaron negativamente, anticuerpos CD34 y CD45 mayor a 98 %. Se observó en SEM que a los 15 días el 99,88 % de las hDPSCs presentaron forma fusiforme o estrellada lo que significa que estas células se adhirieron a la matriz de APL. Conclusión: El APL no es citotóxico para las hDPSCs por su composición y características biocompatibles, lo que proporcionó que las células se adhirieron y proliferaron sobre la matriz dental impresa en 3D demostrando ser un método in vitro efectivo para emplear en futuros estudios de regeneración de tejidos en odontología.ABSTRACT. Background: The study of synergy between mesenchymal stem cells and biomaterial in tissue engineering is fundamental in order to have greater control over the biomimetics for the success of critical clinical treatments depends on this. Objective: to evaluate in vitro cellular adhesion of human dental pulp stem cells (hDPSCs) in three-dimensional printed matrices synthesized with poly-lactic acid (PLA). Method: The study used passage-expanded cryopreserved dental stem cell samples cultivated on 24 three-dimensional dental matrices of PLA during 1, 7 and 15 days. Phenotypification of the DMSC was carried out with flow cytometry and the cellular adhesion to the matrix with morphological analysis using an SEM electron microscope. Data were reported in percentages for each marker in the phenotypification as well as for the amount of adhered cells per time period. Results: The hDPSCs expressed positively, CD73 and CD 90 antibodies of almost 100 % while the CD 105 only had 56, 7 %. The cells presented negative expression of the CD34 and CD45 antibodies in more than 98 %. The SEM showed that 99, 88 % of the DMSC had a fusiform or star shape after 15 days meaning that they had adhered to the matrix. Conclusion: PLA biomaterial is not cytotoxic for the DMSC due to its composition and biocompatible characteristics, which helped the cells, adhere themselves and proliferate on the matrix. This proved to be an effective in vitro method, which may be used in future studies of critical- size tissue lesion studies in dentistry. 

scholarly journals Comparison of Immunosuppressive and Angiogenic Properties of Human Amnion-Derived Mesenchymal Stem Cells between 2D and 3D Culture Systems

2019 ◽  
Vol 2019 ◽  
pp. 1-16 ◽  
Author(s):  
Vitale Miceli ◽  
Mariangela Pampalone ◽  
Serena Vella ◽  
Anna Paola Carreca ◽  
Giandomenico Amico ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Three Dimensional ◽  
3D Culture ◽  
Culture Method ◽  
Paracrine Factors ◽  
Human Amnion ◽  
Tissue Repair And Regeneration ◽  
3D Culture System

The secretion of potential therapeutic factors by mesenchymal stem cells (MSCs) has aroused much interest given the benefits that it can bring in the field of regenerative medicine. Indeed, the in vitro multipotency of these cells and the secretive capacity of both angiogenic and immunomodulatory factors suggest a role in tissue repair and regeneration. However, during culture, MSCs rapidly lose the expression of key transcription factors associated with multipotency and self-renewal, as well as the ability to produce functional paracrine factors. In our study, we show that a three-dimensional (3D) culture method is effective to induce MSC spheroid formation, to maintain the multipotency and to improve the paracrine activity of a specific population of human amnion-derived MSCs (hAMSCs). The regenerative potential of both 3D culture-derived conditioned medium (3D CM) and their exosomes (EXO) was assessed against 2D culture products. In particular, tubulogenesis assays revealed increased capillary maturation in the presence of 3D CM compared with both 2D CM and 2D EXO. Furthermore, 3D CM had a greater effect on inhibition of PBMC proliferation than both 2D CM and 2D EXO. To support this data, hAMSC spheroids kept in our 3D culture system remained viable and multipotent and secreted considerable amounts of both angiogenic and immunosuppressive factors, which were detected at lower levels in 2D cultures. This work reveals the placenta as an important source of MSCs that can be used for eventual clinical applications as cell-free therapies.

scholarly journals Three-Dimensional Aggregates Enhance the Therapeutic Effects of Adipose Mesenchymal Stem Cells for Ischemia-Reperfusion Induced Kidney Injury in Rats

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Xiaozhi Zhao ◽  
Xuefeng Qiu ◽  
Yanting Zhang ◽  
Shiwei Zhang ◽  
Xiaoping Gu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Ischemia Reperfusion ◽  
Three Dimensional ◽  
Kidney Injury ◽  
Therapeutic Effects ◽  
Adipose Mesenchymal Stem Cells ◽  
In Vitro Data

It has been shown that administration of adipose derived mesenchymal stem cells (AdMSCs) enhanced structural and functional recovery of renal ischemia-reperfusion (IR) injury. Low engraftment of stem cells, however, limits the therapeutic effects of AdMSCs. The present study was designed to enhance the therapeutic effects of AdMSCs by delivering AdMSCs in a three-dimensional (3D) aggregates form. Microwell was used to produce 3D AdMSCs aggregates. In vitro data indicated that AdMSCs in 3D aggregates were less susceptible to oxidative and hypoxia stress induced by 200 μM peroxide and hypoxia/reoxygenation, respectively, compared with those cultured in two-dimensional (2D) monolayer. Furthermore, AdMSCs in 3D aggregates secreted more proangiogenic factors than those cultured in 2D monolayer. 2D AdMSCs or 3D AdMSCs aggregates were injected into renal cortex immediately after induction of renal IR injury. In vivo data revealed that 3D aggregates enhanced the effects of AdMSCs in recovering function and structure after renal IR injury. Improved grafted AdMSCs were observed in kidney injected with 3D aggregates compared with AdMSCs cultured in 2D monolayer. Our results demonstrated that 3D AdMSCs aggregated produced by microwell enhanced the retention and therapeutic effects of AdMSCs for renal IR injury.

scholarly journals Differential chondro- and osteo-stimulation in three-dimensional porous scaffolds with different topological surfaces provides a design strategy for biphasic osteochondral engineering

2019 ◽  
Vol 10 ◽  
pp. 204173141982643 ◽  
Author(s):  
Chinmaya Mahapatra ◽  
Jung-Ju Kim ◽  
Jung-Hwan Lee ◽  
Guang-Zhen Jin ◽  
Jonathan C Knowles ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Three Dimensional ◽  
Poly Lactic Acid ◽  
Porous Scaffolds ◽  
Salt Leaching ◽  
Matrix Adhesion ◽  
Nanofibrous Scaffolds ◽  
Differential Properties

Bone/cartilage interfacial tissue engineering needs to satisfy the differential properties and architectures of the osteochondral region. Therefore, biphasic or multiphasic scaffolds that aim to mimic the gradient hierarchy are widely used. Here, we find that two differently structured (topographically) three-dimensional scaffolds, namely, “dense” and “nanofibrous” surfaces, show differential stimulation in osteo- and chondro-responses of cells. While the nanofibrous scaffolds accelerate the osteogenesis of mesenchymal stem cells, the dense scaffolds are better in preserving the phenotypes of chondrocytes. Two types of porous scaffolds, generated by a salt-leaching method combined with a phase-separation process using the poly(lactic acid) composition, had a similar level of porosity (~90%) and pore size (~150 μm). The major difference in the surface nanostructure led to substantial changes in the surface area and water hydrophilicity (nanofibrous ≫ dense); as a result, the nanofibrous scaffolds increased the cell-to-matrix adhesion of mesenchymal stem cells significantly while decreasing the cell-to-cell contracts. Importantly, the chondrocytes, when cultured on nanofibrous scaffolds, were prone to lose their phenotype, including reduced chondrogenic expressions (SOX-9, collagen type II, and Aggrecan) and glycosaminoglycan content, which was ascribed to the enhanced cell–matrix adhesion with reduced cell–cell contacts. On the contrary, the osteogenesis of mesenchymal stem cells was significantly accelerated by the improved cell-to-matrix adhesion, as evidenced in the enhanced osteogenic expressions (RUNX2, bone sialoprotein, and osteopontin) and cellular mineralization. Based on these findings, we consider that the dense scaffold is preferentially used for the chondral-part, whereas the nanofibrous structure is suitable for osteo-part, to provide an optimal biphasic matrix environment for osteochondral tissue engineering.

scholarly journals Three-dimensional printed bone scaffolds: The role of nano/micro-hydroxyapatite particles on the adhesion and differentiation of human mesenchymal stem cells

2017 ◽  
Vol 231 (6) ◽  
pp. 555-564 ◽  
Author(s):  
Marco Domingos ◽  
Antonio Gloria ◽  
Jorge Coelho ◽  
Paulo Bartolo ◽  
Joaquim Ciurana
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Calcium Phosphates ◽  
Three Dimensional ◽  
Human Mesenchymal Stem Cells ◽  
Mechanical Tests ◽  
Engineering Research

Bone tissue engineering is strongly dependent on the use of three-dimensional scaffolds that can act as templates to accommodate cells and support tissue ingrowth. Despite its wide application in tissue engineering research, polycaprolactone presents a very limited ability to induce adhesion, proliferation and osteogenic cell differentiation. To overcome some of these limitations, different calcium phosphates, such as hydroxyapatite and tricalcium phosphate, have been employed with relative success. This work investigates the influence of nano-hydroxyapatite and micro-hydroxyapatite (nHA and mHA, respectively) particles on the in vitro biomechanical performance of polycaprolactone/hydroxyapatite scaffolds. Morphological analysis performed with scanning electron microscopy allowed us to confirm the production of polycaprolactone/hydroxyapatite constructs with square interconnected pores of approximately 350 µm and to assess the distribution of hydroxyapatite particles within the polymer matrix. Compression mechanical tests showed an increase in polycaprolactone compressive modulus ( E) from 105.5 ± 11.2 to 138.8 ± 12.9 MPa (PCL_nHA) and 217.2 ± 21.8 MPa (PCL_mHA). In comparison to PCL_mHA scaffolds, the addition of nano-hydroxyapatite enhanced the adhesion and viability of human mesenchymal stem cells as confirmed by Alamar Blue assay. In addition, after 14 days of incubation, PCL_nHA scaffolds showed higher levels of alkaline phosphatase activity compared to polycaprolactone or PCL_mHA structures.

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