Climate change, in particular the increase in sea surface temperature, is causing severe and progressive degradation of coral reefs. One of the main threats to these habitats is coral bleaching, a phenomenon related to oxidative stress and the excessive production of reactive oxygen species (ROS). In recent decades, thermal stress events have intensified both in frequency and duration, leading to increasingly severe and widespread bleaching episodes. This process profoundly compromises the symbiosis between corals and their symbiotic microalgae – zooxanthellae – triggering a series of intracellular molecular cascades that can ultimately lead to coral death. In light of this, there is a growing urgency to develop active intervention strategies to mitigate the effects of bleaching. The main objective of this PhD research was the design and development of an efficient, environmentally friendly, and fully biodegradable drug delivery system capable of ensuring a controlled, localized, and prolonged administration of natural antioxidant compounds. In the first part of the work, curcumin was selected as a model antioxidant compound, and two different materials were developed and tested: a multilayer film system and protein-based microparticles intended for coral application. Morphological, chemical, and mechanical analyses performed on the multilayer film confirmed its structural integrity, water absorption capacity, and hydrogel-like behavior – key characteristics for underwater applications. At the same time, the morphological, physicochemical properties, release kinetics, and biocompatibility of microparticles were carefully evaluated. Ingestion studies conducted on Heliofungia fralinae confirmed that the microparticles penetrate the gastric cavity and locally release curcumin within the coral tissues. After the complete characterization of both systems, the second part of the PhD focused on in vivo tests carried out on corals of the species Stylophora pistillata through a bleaching simulation. Coral fragments were exposed to 31 °C for one week to induce oxidative stress, and the two materials were applied to evaluate their effectiveness in increasing the thermal tolerance of corals. Without the need for direct contact, the film system effectively mitigated oxidative stress, similarly to the microparticles. In both cases, the treated corals showed higher chlorophyll content, greater Symbiodiniaceae density, and reduced lipid peroxidation compared to the controls. These results confirm the potential of both materials as innovative, biocompatible, and sustainable therapies to improve coral resilience. Finally, in the third part of the PhD – carried out at the Scripps Institution of Oceanography – the two systems were combined into a single integrated drug delivery platform. Using a scalable 3D bioprinting technique, antioxidant microparticles were directly encapsulated within a hydrogel-like matrix, achieving a higher level of release control. Along with the microparticles, symbiotic algae of the species Breviolum psygmophilum were also incorporated into the scaffold to mimic the internal structure of the coral–alga symbiosis. The system was successfully characterized and tested in vivo, demonstrating its effectiveness in preventing oxidative stress. This final therapeutic system represents not only the culmination of the PhD project but also a concrete prospect for future post-bleaching treatments, offering a promising technological approach to support global strategies for coral reef conservation and restoration.
Il cambiamento climatico, in particolare l’aumento della temperatura della superficie marina, sta causando un grave e progressivo degrado delle barriere coralline. Una delle principali minacce per questi habitat è rappresentata dallo sbiancamento dei coralli, fenomeno legato allo stress ossidativo e all’eccessiva produzione di specie reattive dell’ossigeno (ROS). Negli ultimi decenni, gli episodi di stress termico si sono intensificati sia in frequenza sia in durata, provocando eventi di sbiancamento sempre più gravi ed estesi. Questo processo compromette profondamente la simbiosi tra i coralli e le microalghe simbionti (zooxantelle) attivando una serie di cascate molecolari intracellulari che possono condurre alla morte del corallo. Alla luce di ciò emerge con forza l’urgenza di sviluppare strategie di intervento attivo per mitigare gli effetti dello sbiancamento. L’obiettivo principale di questa ricerca di dottorato è stato la progettazione e lo sviluppo di un sistema di rilascio di farmaci efficiente, ecocompatibile e completamente biodegradabile, capace di garantire una somministrazione controllata, localizzata e prolungata di composti antiossidanti naturali. Nella prima parte del lavoro, la curcumina è stata selezionata come composto antiossidante modello e sono stati sviluppati e testati due diversi materiali: un sistema multistrato in forma di film e microparticelle a base proteica destinate all’applicazione sui coralli. Le analisi morfologiche, chimiche e meccaniche condotte sul film multistrato hanno confermato l’integrità strutturale, la capacità di assorbimento d’acqua e il comportamento simil-idrogel del materiale. Parallelamente, le proprietà morfologiche, chimico-fisiche, la cinetica di rilascio e la biocompatibilità delle particelle sono state accuratamente valutate. Dopo la completa caratterizzazione di entrambi i sistemi, la seconda parte del dottorato si è concentrata su test in vivo condotti su coralli della specie Stylophora pistillata tramite una simulazione di sbiancamento. I frammenti di corallo sono stati esposti a 31 °C per una settimana per indurre stress ossidativo, e i due materiali sono stati applicati per valutarne l’efficacia nell’aumentare la tolleranza termica dei coralli. Senza necessità di contatto diretto, il sistema a film ha dimostrato di mitigare efficacemente lo stress ossidativo, analogamente alle microparticelle di seta. In entrambi i casi i coralli trattati hanno mostrato un maggiore contenuto di clorofilla, una più elevata densità di Symbiodiniaceae e una ridotta perossidazione lipidica rispetto ai controlli. Questi risultati confermano il potenziale di entrambi i materiali come terapie innovative, biocompatibili e sostenibili per migliorare la resilienza dei coralli. Infine, nella terza parte del dottorato (presso Scripps Institution of Oceanography) i due sistemi sono stati combinati in un’unica piattaforma di rilascio di farmaci. Mediante una tecnica di biostampa 3D, le microparticelle antiossidanti sono state direttamente incapsulate all’interno di una matrice simil-idrogel, ottenendo un livello superiore di controllo del rilascio. Insieme alle microparticelle, sono state inglobate nello scaffold anche alghe simbionti della specie Breviolum psygmophilum, per mimare la struttura interna della simbiosi corallo–alga. Il sistema è stato caratterizzato e testato con successo in vivo, dimostrando la sua efficacia nel prevenire lo stress ossidativo. Questo sistema terapeutico finale rappresenta non solo il culmine del progetto di dottorato, ma anche una prospettiva concreta per futuri trattamenti post-sbiancamento, offrendo un approccio tecnologico promettente a supporto delle strategie globali di conservazione e recupero delle barriere coralline.
Rinaldi, C (2026). Materiali sostenibili e innovativi per la protezione e il recupero dei coralli dagli eventi di sbiancamento. (Tesi di dottorato, , 2026).
Materiali sostenibili e innovativi per la protezione e il recupero dei coralli dagli eventi di sbiancamento
RINALDI, CAMILLA
2026
Abstract
Climate change, in particular the increase in sea surface temperature, is causing severe and progressive degradation of coral reefs. One of the main threats to these habitats is coral bleaching, a phenomenon related to oxidative stress and the excessive production of reactive oxygen species (ROS). In recent decades, thermal stress events have intensified both in frequency and duration, leading to increasingly severe and widespread bleaching episodes. This process profoundly compromises the symbiosis between corals and their symbiotic microalgae – zooxanthellae – triggering a series of intracellular molecular cascades that can ultimately lead to coral death. In light of this, there is a growing urgency to develop active intervention strategies to mitigate the effects of bleaching. The main objective of this PhD research was the design and development of an efficient, environmentally friendly, and fully biodegradable drug delivery system capable of ensuring a controlled, localized, and prolonged administration of natural antioxidant compounds. In the first part of the work, curcumin was selected as a model antioxidant compound, and two different materials were developed and tested: a multilayer film system and protein-based microparticles intended for coral application. Morphological, chemical, and mechanical analyses performed on the multilayer film confirmed its structural integrity, water absorption capacity, and hydrogel-like behavior – key characteristics for underwater applications. At the same time, the morphological, physicochemical properties, release kinetics, and biocompatibility of microparticles were carefully evaluated. Ingestion studies conducted on Heliofungia fralinae confirmed that the microparticles penetrate the gastric cavity and locally release curcumin within the coral tissues. After the complete characterization of both systems, the second part of the PhD focused on in vivo tests carried out on corals of the species Stylophora pistillata through a bleaching simulation. Coral fragments were exposed to 31 °C for one week to induce oxidative stress, and the two materials were applied to evaluate their effectiveness in increasing the thermal tolerance of corals. Without the need for direct contact, the film system effectively mitigated oxidative stress, similarly to the microparticles. In both cases, the treated corals showed higher chlorophyll content, greater Symbiodiniaceae density, and reduced lipid peroxidation compared to the controls. These results confirm the potential of both materials as innovative, biocompatible, and sustainable therapies to improve coral resilience. Finally, in the third part of the PhD – carried out at the Scripps Institution of Oceanography – the two systems were combined into a single integrated drug delivery platform. Using a scalable 3D bioprinting technique, antioxidant microparticles were directly encapsulated within a hydrogel-like matrix, achieving a higher level of release control. Along with the microparticles, symbiotic algae of the species Breviolum psygmophilum were also incorporated into the scaffold to mimic the internal structure of the coral–alga symbiosis. The system was successfully characterized and tested in vivo, demonstrating its effectiveness in preventing oxidative stress. This final therapeutic system represents not only the culmination of the PhD project but also a concrete prospect for future post-bleaching treatments, offering a promising technological approach to support global strategies for coral reef conservation and restoration.| File | Dimensione | Formato | |
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phd_unimib_815758.pdf
accesso aperto
Descrizione: Advanced Sustainable Materials to Protect and Heal Reefs from Coral Bleaching Events
Tipologia di allegato:
Doctoral thesis
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11.9 MB
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