Lorenza Garau Paganella

All Publications

• Cyclic jetting enables microbubble-mediated drug delivery NATURE PHYSICS Cattaneo, M., Guerriero, G., Shakya, G., Krattiger, L. A., Paganella, L. G., Narciso, M. L., Supponen, O. 2025

  View details for DOI 10.1038/s41567-025-02785-0

  View details for Web of Science ID 001427346100001

• Injectable Senolytic Hydrogel Depot for the Clearance of Senescent Cells BIOMACROMOLECULES Paganella, L., Bovone, G., Cuni, F., Labouesse, C., Cui, Y., Giampietro, C., Tibbitt, M. W. 2025; 26 (2): 814-824

  Abstract

  Small molecules are frontline therapeutics for many diseases; however, they are often limited by their poor solubility. Therefore, hydrophobic small molecules are often encapsulated or prepared as pure drug nanoparticles. Navitoclax, used to eliminate senescent cells, is one such small molecule that faces challenges in translation due to its hydrophobicity and toxic side effects. Further, as senescent cells exhibit context-dependent pathologic or beneficial properties, it is preferable to eliminate senescent cells locally. To formulate navitoclax and enable local treatment, we designed an injectable hydrogel loaded with navitoclax nanoparticles as a senolytic delivery vehicle. Navitoclax nanoparticles (Ø ∼ 110 nm) were prepared via solvent-antisolvent nanoprecipitation and formulated in an injectable polymer-nanoparticle (PNP) hydrogel to create a local senolytic depot. Navitoclax-loaded PNP hydrogels selectively cleared senescent cells in vitro in senescent endothelial monolayers. This work demonstrates the value of formulating lipophilic small molecules and the potential of localized drug delivery strategies to improve senolytic therapies.

  View details for DOI 10.1021/acs.biomac.4c00851

  View details for Web of Science ID 001393299900001

  View details for PubMedID 39783796

  View details for PubMedCentralID PMC11815846

• Variations in fluid chemical potential induce fibroblast mechano-response in 3D hydrogels. Biomaterials advances Garau Paganella, L., Badolato, A., Labouesse, C., Fischer, G., Sänger, C. S., Kourouklis, A., Giampietro, C., Werner, S., Mazza, E., Tibbitt, M. W. 2024; 163: 213933

  Abstract

  Mechanical deformation of skin creates variations in fluid chemical potential, leading to local changes in hydrostatic and osmotic pressure, whose effects on mechanobiology remain poorly understood. To study these effects, we investigate the specific influences of hydrostatic and osmotic pressure on primary human dermal fibroblasts in three-dimensional hydrogel culture models. Cyclic hydrostatic pressure and hyperosmotic stress enhanced the percentage of cells expressing the proliferation marker Ki67 in both collagen and PEG-based hydrogels. Osmotic pressure also activated the p38 MAPK stress response pathway and increased the expression of the osmoresponsive genes PRSS35 and NFAT5. When cells were cultured in two-dimension (2D), no change in proliferation was observed with either hydrostatic or osmotic pressure. Furthermore, basal, and osmotic pressure-induced expression of osmoresponsive genes differed in 2D culture versus 3D hydrogels, highlighting the role of dimensionality in skin cell mechanotransduction and stressing the importance of 3D tissue-like models that better replicate in vivo conditions. Overall, these results indicate that fluid chemical potential changes affect dermal fibroblast mechanobiology, which has implications for skin function and for tissue regeneration strategies.

  View details for DOI 10.1016/j.bioadv.2024.213933

  View details for PubMedID 38972277

• Protein Isolation from 3D Hydrogel Scaffolds. Current protocols Da Silva André, G., Paganella, L. G., Badolato, A., Sander, S., Giampietro, C., Tibbitt, M. W., Dengjel, J., Labouesse, C. 2024; 4 (1): e966

  Abstract

  Protein isolation is an essential tool in cell biology to characterize protein abundance under various experimental conditions. Several protocols exist, tailored to cell culture or tissue sections, and have been adapted to particular downstream analyses (e.g., western blotting or mass spectrometry). An increasing trend in bioengineering and cell biology is to use three-dimensional (3D) hydrogel-based scaffolds for cell culture. In principle, the same protocols can be used to extract protein from hydrogel-based cell and tissue constructs. However, in practice the yield and quality of the recovered protein pellet is often substantially lower when using standard protocols and requires tuning of multiple steps, including the selected lysis buffer and the scaffold homogenization strategy, as well as the methods for protein purification and reconstitution. We present here specific protocols tailored to common 3D hydrogels to help researchers using hydrogel-based 3D cell culture improve the quantity and quality of their extracted protein. We focus on three materials: protease-degradable PEG-based hydrogels, collagen hydrogels, and alginate hydrogels. We discuss how the protein extraction procedure can be adapted to the scaffold of interest (degradable or non-degradable gels), proteins of interests (soluble, matrix-bound, or phosphoproteins), and downstream biochemical assays (western blotting or mass spectrometry). With the growing interest in 3D cell culture, the protocols presented should be useful to many researchers in cell biology, protein science, biomaterials, and bioengineering communities. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Isolating proteins from PEG-based hydrogels Basic Protocol 2: Isolating proteins from collagen hydrogels Basic Protocol 3: Isolating proteins from alginate hydrogels Alternate Protocol: Isolating protein from alginate gels using EDTA to dissolve the gel Support Protocol: Isolating protein and RNA simultaneously from the same samples.

  View details for DOI 10.1002/cpz1.966

  View details for PubMedID 38206582

• Serine protease 35 regulates the fibroblast matrisome in response to hyperosmotic stress. Science advances Sänger, C. S., Cernakova, M., Wietecha, M. S., Garau Paganella, L., Labouesse, C., Dudaryeva, O. Y., Roubaty, C., Stumpe, M., Mazza, E., Tibbitt, M. W., Dengjel, J., Werner, S. 2023; 9 (35): eadh9219

  Abstract

  Hyperosmotic stress occurs in several diseases, but its long-term effects are largely unknown. We used sorbitol-treated human fibroblasts in 3D culture to study the consequences of hyperosmotic stress in the skin. Sorbitol regulated many genes, which help cells cope with the stress condition. The most robustly regulated gene encodes serine protease 35 (PRSS35). Its regulation by hyperosmotic stress was dependent on the kinases p38 and JNK and the transcription factors NFAT5 and ATF2. We identified different collagens and collagen-associated proteins as putative PRSS35 binding partners. This is functionally important because PRSS35 affected the extracellular matrix proteome, which limited cell proliferation. The in vivo relevance of these findings is reflected by the coexpression of PRSS35 and its binding partners in human skin wounds, where hyperosmotic stress occurs as a consequence of excessive water loss. These results identify PRSS35 as a key regulator of the matrisome under hyperosmotic stress conditions.

  View details for DOI 10.1126/sciadv.adh9219

  View details for PubMedID 37647410

  View details for PubMedCentralID PMC10468140

• Control of hydrostatic pressure and osmotic stress in 3D cell culture for mechanobiological studies. Biomaterials advances Kourouklis, A. P., Wahlsten, A., Stracuzzi, A., Martyts, A., Paganella, L. G., Labouesse, C., Al-Nuaimi, D., Giampietro, C., Ehret, A. E., Tibbitt, M. W., Mazza, E. 2023; 145: 213241

  Abstract

  Hydrostatic pressure (HP) and osmotic stress (OS) play an important role in various biological processes, such as cell proliferation and differentiation. In contrast to canonical mechanical signals transmitted through the anchoring points of the cells with the extracellular matrix, the physical and molecular mechanisms that transduce HP and OS into cellular functions remain elusive. Three-dimensional cell cultures show great promise to replicate physiologically relevant signals in well-defined host bioreactors with the goal of shedding light on hidden aspects of the mechanobiology of HP and OS. This review starts by introducing prevalent mechanisms for the generation of HP and OS signals in biological tissues that are subject to pathophysiological mechanical loading. We then revisit various mechanisms in the mechanotransduction of HP and OS, and describe the current state of the art in bioreactors and biomaterials for the control of the corresponding physical signals.

  View details for DOI 10.1016/j.bioadv.2022.213241

  View details for PubMedID 36529095