Modulation of the inflammatory response using a guided bone regeneration membrane with dual drug delivery capacity
Bone is a complex and dynamic tissue that fulfills several critical functions such as protecting vital organs including the brain, heart and lungs; providing sites of attachment for muscles to allow movement and maintaining ion homeostasis. Moreover, bone has a remarkable regenerative capacity which...
| Author: | |
|---|---|
| Format: | doctoral thesis |
| Status: | Published version |
| Publication Date: | 2021 |
| Country: | España |
| Institution: | CBUC, CESCA |
| Repository: | TDR. Tesis Doctorales en Red |
| OAI Identifier: | oai:www.tdx.cat:10803/671867 |
| Online Access: | http://hdl.handle.net/10803/671867 |
| Access Level: | Open access |
| Keyword: | Macrophage Inflammation Guided Bone Regeneration Ion Drug Release Bioingeniería 61 |
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Modulation of the inflammatory response using a guided bone regeneration membrane with dual drug delivery capacity |
| title |
Modulation of the inflammatory response using a guided bone regeneration membrane with dual drug delivery capacity |
| spellingShingle |
Modulation of the inflammatory response using a guided bone regeneration membrane with dual drug delivery capacity Díez Tercero, Leire Macrophage Inflammation Guided Bone Regeneration Ion Drug Release Bioingeniería 61 |
| title_short |
Modulation of the inflammatory response using a guided bone regeneration membrane with dual drug delivery capacity |
| title_full |
Modulation of the inflammatory response using a guided bone regeneration membrane with dual drug delivery capacity |
| title_fullStr |
Modulation of the inflammatory response using a guided bone regeneration membrane with dual drug delivery capacity |
| title_full_unstemmed |
Modulation of the inflammatory response using a guided bone regeneration membrane with dual drug delivery capacity |
| title_sort |
Modulation of the inflammatory response using a guided bone regeneration membrane with dual drug delivery capacity |
| dc.creator.none.fl_str_mv |
Díez Tercero, Leire |
| author |
Díez Tercero, Leire |
| author_facet |
Díez Tercero, Leire |
| author_role |
author |
| dc.contributor.none.fl_str_mv |
Pérez Antoñanzas, Román Delgado Garoña, Luis María Universitat Internacional de Catalunya. Departament de Ciències Bàsiques |
| dc.subject.none.fl_str_mv |
Macrophage Inflammation Guided Bone Regeneration Ion Drug Release Bioingeniería 61 |
| topic |
Macrophage Inflammation Guided Bone Regeneration Ion Drug Release Bioingeniería 61 |
| description |
Bone is a complex and dynamic tissue that fulfills several critical functions such as protecting vital organs including the brain, heart and lungs; providing sites of attachment for muscles to allow movement and maintaining ion homeostasis. Moreover, bone has a remarkable regenerative capacity which allows the complete healing of the tissue upon damage. However, this capacity can be exceeded when the size of the defect is too large due to clinical procedures such as tumor resection or the presence of traumatic fracture or osteolysis, which constitute a significant clinical challenge nowadays. Autologous grafts, as well as allografts and xenografts present several limitations to their clinical application such as limited bone supply, disease transmission and ethical issues. Therefore, tissue engineering combining biomaterials and stimulatory molecules to guide bone regeneration presents as an alternative to these methods. Recent advances in bone biology have shown that osteogenesis occurs due to the interaction of multiple systems and not only by the actions of the bone tissue. In this sense, the immune system has gained great importance since the inflammatory response promoted by either tissue damage or the immune recognition of the implanted biomaterial can direct the outcome of the bone healing response. More precisely, macrophages have been described to have a central role in bone regeneration. Their differentiation to a pro-inflammatory phenotype (M1) phenotype can lead to the development of chronic inflammation, which impairs bone healing, whereas their differentiation to an anti-inflammatory phenotype (M2) can lead to enhanced biomaterial integration and improved bone regeneration. Therefore, the design of biomaterials has focused on modulating macrophage differentiation to M2 phenotype to improve bone regeneration. One of the approaches to modulate macrophage response has been the release of antiinflammatory modulators such as cytokines, viral vectors or siRNAs. However, their short half-life and concerns in their efficiency in cellular uptake, as well as long term safety limit VI their clinical application. Ions have risen as a promising alternative since they are stable cues which are present at low concentrations in the body and have already shown benefits on angiogenesis and bone regeneration when delivered from scaffolds. However, there are limited evidences showing their immunomodulatory potential. This thesis is focused on modulating macrophage response by developing a dual drug delivery system with the ability to release ions, as well as small drugs, to promote the M2 macrophage phenotype. First, an initial screening of three bioactive ions was performed to determine the cytotoxic and therapeutic concentrations in macrophages. Those concentrations that induced macrophage differentiation towards the M2 phenotype were tested in presence of different concentrations of a pro-inflammatory stimulus, which allowed to determine the anti-inflammatory potential of these ions. Then, the effect of the ions combined with an anti-inflammatory drug was tested in macrophages to observe a possible synergistic effect between both molecules, although no major differences were observed compared to the effect of the drug alone. Following these assays, the dual drug delivery system was developed, which consisted of a collagen film with ion loaded microparticles and drug loaded microspheres. The results showed that these films not only were able to release controlled concentrations of the ion, but they were also able to perform a sustained release of the drug. Finally, macrophages and mesenchymal stem cells (MSCs) were exposed to the films, showing that they were able to induce M2 macrophage differentiation and osteogenesis. Moreover, treating MSCs with conditioned media from film-induced M2 macrophages further improved the osteogenic differentiation. |
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2021 |
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2021 2021 2023 |
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info:eu-repo/semantics/doctoralThesis info:eu-repo/semantics/publishedVersion |
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doctoralThesis |
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publishedVersion |
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http://hdl.handle.net/10803/671867 |
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http://hdl.handle.net/10803/671867 |
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Inglés |
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Inglés |
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info:eu-repo/semantics/openAccess |
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openAccess |
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255 p. application/pdf application/pdf |
| dc.publisher.none.fl_str_mv |
Universitat Internacional de Catalunya |
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Universitat Internacional de Catalunya |
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TDX (Tesis Doctorals en Xarxa) reponame:TDR. Tesis Doctorales en Red instname:CBUC, CESCA |
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CBUC, CESCA |
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TDR. Tesis Doctorales en Red |
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TDR. Tesis Doctorales en Red |
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1869425738603560960 |
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Modulation of the inflammatory response using a guided bone regeneration membrane with dual drug delivery capacityDíez Tercero, LeireMacrophageInflammationGuided Bone RegenerationIonDrug ReleaseBioingeniería61Bone is a complex and dynamic tissue that fulfills several critical functions such as protecting vital organs including the brain, heart and lungs; providing sites of attachment for muscles to allow movement and maintaining ion homeostasis. Moreover, bone has a remarkable regenerative capacity which allows the complete healing of the tissue upon damage. However, this capacity can be exceeded when the size of the defect is too large due to clinical procedures such as tumor resection or the presence of traumatic fracture or osteolysis, which constitute a significant clinical challenge nowadays. Autologous grafts, as well as allografts and xenografts present several limitations to their clinical application such as limited bone supply, disease transmission and ethical issues. Therefore, tissue engineering combining biomaterials and stimulatory molecules to guide bone regeneration presents as an alternative to these methods. Recent advances in bone biology have shown that osteogenesis occurs due to the interaction of multiple systems and not only by the actions of the bone tissue. In this sense, the immune system has gained great importance since the inflammatory response promoted by either tissue damage or the immune recognition of the implanted biomaterial can direct the outcome of the bone healing response. More precisely, macrophages have been described to have a central role in bone regeneration. Their differentiation to a pro-inflammatory phenotype (M1) phenotype can lead to the development of chronic inflammation, which impairs bone healing, whereas their differentiation to an anti-inflammatory phenotype (M2) can lead to enhanced biomaterial integration and improved bone regeneration. Therefore, the design of biomaterials has focused on modulating macrophage differentiation to M2 phenotype to improve bone regeneration. One of the approaches to modulate macrophage response has been the release of antiinflammatory modulators such as cytokines, viral vectors or siRNAs. However, their short half-life and concerns in their efficiency in cellular uptake, as well as long term safety limit VI their clinical application. Ions have risen as a promising alternative since they are stable cues which are present at low concentrations in the body and have already shown benefits on angiogenesis and bone regeneration when delivered from scaffolds. However, there are limited evidences showing their immunomodulatory potential. This thesis is focused on modulating macrophage response by developing a dual drug delivery system with the ability to release ions, as well as small drugs, to promote the M2 macrophage phenotype. First, an initial screening of three bioactive ions was performed to determine the cytotoxic and therapeutic concentrations in macrophages. Those concentrations that induced macrophage differentiation towards the M2 phenotype were tested in presence of different concentrations of a pro-inflammatory stimulus, which allowed to determine the anti-inflammatory potential of these ions. Then, the effect of the ions combined with an anti-inflammatory drug was tested in macrophages to observe a possible synergistic effect between both molecules, although no major differences were observed compared to the effect of the drug alone. Following these assays, the dual drug delivery system was developed, which consisted of a collagen film with ion loaded microparticles and drug loaded microspheres. The results showed that these films not only were able to release controlled concentrations of the ion, but they were also able to perform a sustained release of the drug. Finally, macrophages and mesenchymal stem cells (MSCs) were exposed to the films, showing that they were able to induce M2 macrophage differentiation and osteogenesis. Moreover, treating MSCs with conditioned media from film-induced M2 macrophages further improved the osteogenic differentiation.El hueso es un tejido complejo y dinámico que cumple varias funciones críticas como la protección de órganos vitales como el cerebro, el corazón y los pulmones; permite el movimiento del cuerpo humano proporcionando sitios de unión para los músculos; y el mantenimiento de la homeostasis iónica. Además, el hueso tiene una notable capacidad regenerativa que permite la curación completa del tejido en caso de que haya habido alguna lesión. Sin embargo, la capacidad regenerativa del hueso puede verse sobrepasada cuando el tamaño del defecto es demasiado grande, lo que ocurre en procedimientos clínicos como la resección tumoral, ciertas fracturas o en situaciones en las que hay osteolisis. Los injertos autólogos, así como los aloinjertos y los xenoinjertos presentan varias limitaciones para su aplicación clínica, entre las que se encuentran la disponibilidad de suficiente cantidad de hueso, la transmisión de enfermedades y los problemas éticos. Por tanto, la ingeniería de tejidos se presenta como una alternativa a estos métodos, combinando biomateriales y moléculas que guíen la regeneración ósea. Avances recientes en biología ósea han demostrado que la osteogénesis se produce debido a la interacción de múltiples sistemas y no solo por las acciones del tejido óseo. En este sentido, el sistema inmune ha ganado una gran importancia, ya que la respuesta inflamatoria causada por el daño tisular o el reconocimiento del biomaterial implantado por parte del sistema inmune pueden influenciar el resultado de la regeneración ósea. Concretamente, se ha descrito que los macrófagos tienen un papel central en la regeneración ósea. Su diferenciación a un fenotipo pro-inflamatorio (M1) puede conducir al desarrollo de inflamación crónica, que afecta negativamente a la regeneración ósea, mientras que su diferenciación a un fenotipo anti-inflamatorio (M2) puede derivar en una mayor integración del biomaterial y en una mejora de la regeneración ósea. Por tanto, el diseño de biomateriales se ha centrado en favorecer la diferenciación de macrófagos hacia el fenotipo M2 con el fin de promover la regeneración ósea. Una de las estrategias para lograrlo ha sido la liberación de moléculas anti-inflamatorias como citocinas, vectores virales o siRNA. Sin embargo, su corta vida media, así como las VIII dudas en cuanto a la eficiencia en la absorción celular y su seguridad a largo plazo de estos sistemas, limitan su aplicación clínica. Los iones han surgido como una alternativa prometedora, ya que son señales estables que están presentes en concentraciones bajas en el cuerpo y ya han mostrado beneficios en la angiogénesis y la regeneración ósea cuando se incorporan dentro de biomateriales. Sin embargo, existen evidencias limitadas que muestran su potencial inmunomodulador. Esta tesis se centra en el desarrollo de un sistema de administración de fármacos dual con la capacidad de liberar iones, así como fármacos pequeños, para promover el fenotipo M2 en macrófagos, modulando así la respuesta inmune. En primer lugar, se realizó una selección inicial de tres iones bioactivos y se determinaron las concentraciones citotóxicas y terapéuticas en macrófagos. Aquellas concentraciones que indujeron la diferenciación de macrófagos hacia el fenotipo M2 se probaron en presencia de diferentes intensidades de un estímulo pro-inflamatorio, lo que permitió determinar el potencial anti-inflamatorio de estos iones. Luego, se trató de determinar si podría haber un efecto sinérgico en la diferenciación de los macrófagos cuando estos iones se combinaran con un fármaco antiinflamatorio, aunque no se observaron diferencias importantes en comparación con el efecto del fármaco solo. Después de estos ensayos, se desarrolló el sistema de administración dual de fármacos, que consistía en una membrana de colágeno con micropartículas cargadas de iones y microesferas cargadas de fármaco. Los resultados mostraron que estas membranas no solo pudieron liberar concentraciones controladas del ión, sino que también pudieron realizar una liberación sostenida del fármaco. Finalmente, se cultivaron macrófagos y células madre mesenquimales (MSC) en contacto con las membranas, y se pudo demostrar que eran capaces de inducir la diferenciación de los macrófagos hacia fenotipo M2 y de promover la osteogénesis. Además, el tratamiento de las MSC con medios condicionados procedentes de macrófagos M2 mejoró aún más la diferenciación osteogénica.Universitat Internacional de CatalunyaPérez Antoñanzas, RománDelgado Garoña, Luis MaríaUniversitat Internacional de Catalunya. Departament de Ciències Bàsiques202120232021info:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/publishedVersion255 p.application/pdfapplication/pdfhttp://hdl.handle.net/10803/671867TDX (Tesis Doctorals en Xarxa)reponame:TDR. Tesis Doctorales en Redinstname:CBUC, CESCAInglésADVERTIMENT. Tots els drets reservats. L'accés als continguts d'aquesta tesi doctoral i la seva utilització ha de respectar els drets de la persona autora. Pot ser utilitzada per a consulta o estudi personal, així com en activitats o materials d'investigació i docència en els termes establerts a l'art. 32 del Text Refós de la Llei de Propietat Intel·lectual (RDL 1/1996). Per altres utilitzacions es requereix l'autorització prèvia i expressa de la persona autora. En qualsevol cas, en la utilització dels seus continguts caldrà indicar de forma clara el nom i cognoms de la persona autora i el títol de la tesi doctoral. No s'autoritza la seva reproducció o altres formes d'explotació efectuades amb finalitats de lucre ni la seva comunicació pública des d'un lloc aliè al servei TDX. Tampoc s'autoritza la presentació del seu contingut en una finestra o marc aliè a TDX (framing). Aquesta reserva de drets afecta tant als continguts de la tesi com als seus resums i índexs.info:eu-repo/semantics/openAccessoai:www.tdx.cat:10803/6718672026-06-14T12:46:07Z |
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15,301603 |