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Gawlitta, D.

Publications and source records attributed to Gawlitta, D..

6 recordsLinked to original sources

Self-feeding living materials enabled by cell responsive glycogen nanoparticles as metabolic batteries

Scaling engineered living materials to clinically relevant dimensions is limited by diffusion-dependent depletion of oxygen and nutrients, which rapidly induces metabolic failure. We introduce glycogen as nutritional nanoparticle that provides cell-mediated, autonomous nutrient release to support long-term survival under extreme metabolic stress. We demonstrate that human mesenchymal stromal cells (hMSCs) survive for weeks in anoxia and serum deprivation when provided extracellular glycogen. Contrary to long-held assumptions, hMSCs secrete glycogen-degrading enzymes, enabling cell-density controlled extracellular glycogenolysis and sustained release of glucose and metabolic intermediates, positioning glycogen as the first-of-its-kind metabolic battery. This cell-responsive process maintains metabolic activity, limits glycolytic acidosis, and enhances pro-angiogenic signaling. To translate this mechanism into a versatile materials platform, we engineered core-shell dextran-tyramine microcapsules that stably encapsulate glycogen while permitting diffusion of enzymes and degradation products. Integrated into centimeter-scale GelMA constructs, these microcapsules maintained hMSC viability and function for at least one month under anoxia. In vivo, glycogen-loaded implants promote deep cellular infiltration, enhanced matrix remodeling, increased M2 macrophage polarization, and orchestrated accelerated vascularization. This work establishes the novel concept of glycogen-based nutritional nanoparticles as metabolic batteries to endow engineered tissues with autonomous self-feeding capacity, enabling scalable and functional living materials for regenerative medicine and related technologies.

bioengineering↗

Osteoimmunomodulatory Stem Cell Nanoghosts as a Novel Nanotherapeutic for Bone Regeneration

Critical-sized bone defects and implant-associated complications are often exacerbated by chronic inflammation, which compromises tissue repair and implant integration. Mesenchymal stromal cell (MSC)-derived extracellular vesicles have emerged as promising immunomodulatory nanotherapeutics; however, their clinical translation remains constrained by low yield, heterogeneity, and poor scalability. Here we present a bioengineered MSC-derived nanoghosts platform designed to overcome these translational barriers while enabling tunable osteoimmunomodulatory function. By coupling high-yield nanoghost fabrication with biomimetic MSC conditioning, we demonstrate that oxygen tension (5 or 21% O2) and 3D culture substrates (5 or 15 wt-% GelMA) can reprogram MSC immunophenotype. Nanoghosts generated under hypoxic and 3D conditions displayed enriched anti-inflammatory cargo, preserved MSC viability under inflammatory stress, and partially rescued osteogenic mineralization in the presence of pro-inflammatory cytokines. Together, these findings showcase MSC nanoghosts as scalable and bioactive immunoregulatory nanotherapeutic capable of modulating immune-bone crosstalk, providing a translational strategy to mitigate inflammation-driven impairment of bone regeneration and implant integration. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=90 SRC="FIGDIR/small/724218v1_ufig1.gif" ALT="Figure 1"> View larger version (24K): org.highwire.dtl.DTLVardef@f965deorg.highwire.dtl.DTLVardef@18f007borg.highwire.dtl.DTLVardef@168a194org.highwire.dtl.DTLVardef@4818fe_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

Refining the Bio-manufacturing of Microalgae-derived Extracellular Vesicles as a Potential Nanotherapeutic for Osteoarthritis

Osteoarthritis (OA) is a degenerative joint disease marked by oxidative stress, chronic inflammation, and cartilage degradation. Current treatments are limited and fail to address the underlying disease mechanisms. Extracellular vesicles (EVs) have emerged as promising nanotherapeutics; however, mammalian-derived EVs face cost and scalability challenges. Microalgae represent a sustainable alternative, yet their potential as EV biofactories for regenerative medicine remains unknown. This study aims to refine the bio-manufacturing of microalgae EVs as a next-generation nanotherapeutic for OA. Microalgae species (Chlorella sorokiniana, Synechococcus sp., Leptolyngbya sp., Chlamydomonas reinhardtii CC1690) were screened under varying photoperiods (0h, 16h, 24h light/day) to assess the influence on viability, growth and EV production. EVs were characterized by transmission electron microscopy, nanoparticle tracking analysis, protein quantification and immunoblotting. Their antioxidant capacity, cellular recruitment, and therapeutic efficacy were evaluated in a cytokine-induced OA-like in vitro model. Our findings demonstrated that microalgae growth and EV yield were highly light-dependent, with all species maintaining high viability (>80%) across different photoperiods. Notably, Leptolyngbya sp. (Leptolyngbya) exhibited the fastest growth and highest EV yield under extended illumination, producing EVs with strong antioxidant activity. Leptolyngbya-derived EVs (Lepto-EVs) enhanced the proliferation and migration of human bone marrow-derived mesenchymal stromal cells and provided protection against matrix degradation within a cytokine-induced OA-like model. These findings position microalgae, particularly Leptolyngbya, as a highly scalable and sustainable producer of therapeutic EVs. Lepto-EVs offer a potent, cell-free nanotherapeutic exhibiting anti-catabolic properties alleviating cytokine-induced matrix degradation in an OA-like milieu, establishing microalgae EVs as a promising, cost-effective frontier in regenerative nanomedicine. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=98 SRC="FIGDIR/small/685512v1_ufig1.gif" ALT="Figure 1"> View larger version (29K): org.highwire.dtl.DTLVardef@ac6400org.highwire.dtl.DTLVardef@1d948a7org.highwire.dtl.DTLVardef@30fc13org.highwire.dtl.DTLVardef@1f576c8_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

Bioengineering Developmentally Inspired Matrix Vesicles as Designer Nanotherapeutics for Bone Regeneration

Extracellular vesicles (EVs) are emerging as promising acellular nanotherapeutics for musculoskeletal repair. Matrix vesicles, a matrix-bound subset of EVs, are essential mediators of endochondral ossification in bone development and fracture repair. This study aims to design bioengineered matrix vesicles from hypertrophic cartilage microtissues to drive endochondral ossification for bone repair. Human mesenchymal stromal cell (hBMSC) microtissues were differentiated with/without BMP2 in chondrogenic or hypertrophic medium. Isolated matrix vesicles were characterized for physiochemical properties and biological functionality. BMP2 and hypertrophic conditioning significantly increased vesicle yield (1.5-fold), alkaline phosphatase activity (3.24-fold), calcium binding capacity (8.82-fold), and growth factor content (BMP2, VEGF). These vesicles promoted proliferation, migration, and mineralization of hBMSCs and enhanced angiogenesis in human endothelial colony forming cells (hECFCs), with BMP2 and hypertrophically conditioned vesicles showing the most pronounced effects. Proteomics analysis confirmed the enrichment of proteins involved in extracellular matrix remodelling, mineral deposition and vascularization within these hypertrophically engineered vesicles. These findings demonstrate that hypertrophic induction of cartilaginous microtissues substantially improves the yield and therapeutic potential of matrix vesicles. Taken together, this research unveils a powerful strategy to bioengineer developmentally inspired vesicles that not only recapitulate key cues of endochondral ossification but offers a tailorable, multifunctional nanotherapeutic platform for improved bone regeneration strategies. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/684111v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@17327a9org.highwire.dtl.DTLVardef@1310583org.highwire.dtl.DTLVardef@160f095org.highwire.dtl.DTLVardef@152eff0_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

Evaluating material-driven regeneration in a tissue engineered human in vitro bone defect model

Advanced in vitro human bone defect models can contribute to the evaluation of materials for in situ bone regeneration, addressing both translational and ethical concerns regarding animal models. In this study, we attempted to develop such a model to study material-driven regeneration, using a tissue engineering approach. By co-culturing human umbilical vein endothelial cells (HUVECs) with human bone marrow-derived mesenchymal stromal cells (hBMSCs) on silk fibroin scaffolds with in vitro critically sized defects, the growth of vascular-like networks and three-dimensional bone-like tissue was facilitated. After a model build-up phase of 28 days, materials were artificially implanted and HUVEC and hBMSC migration, cell-material interactions, and osteoinduction were evaluated 14 days after implantation. The materials physiologically relevant for bone regeneration included a platelet gel as blood clot mimic, cartilage spheres as soft callus mimics, and a fibrin gel as control. Although the in vitro model was limited in the evaluation of immune responses, hallmarks of physiological bone regeneration were observed in vitro. These included the endothelial cell chemotaxis induced by the blood clot mimic and the mineralization of the soft callus mimic. Therefore, the present in vitro model could contribute to an improved pre-clinical evaluation of biomaterials while reducing the need for animal experiments. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=91 SRC="FIGDIR/small/502914v1_ufig1.gif" ALT="Figure 1"> View larger version (24K): org.highwire.dtl.DTLVardef@ff8a7borg.highwire.dtl.DTLVardef@dcbddcorg.highwire.dtl.DTLVardef@34c865org.highwire.dtl.DTLVardef@15084f2_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

Impact of endotoxins in gelatine hydrogels on chondrogenic differentiation and inflammatory cytokine secretion in vitro

Gelatine methacryloyl (GelMA) hydrogels are widely used in studies aiming at cartilage regeneration. However, the endotoxin content of commercially available GelMAs and gelatines used in these studies is often overlooked, even though endotoxins may influence several cellular functions. Moreover, regulations for clinical use of biomaterials dictate a stringent endotoxin limit. We determined the endotoxin level of five different GelMAs and evaluated the effect on the chondrogenic differentiation of equine mesenchymal stromal cells (MSCs). Cartilage-like matrix production was evaluated by biochemical assays and immunohistochemistry. Furthermore, equine peripheral blood mononuclear cells (PBMCs) were cultured on the hydrogels for 24 hours, followed by the assessment of tumour necrosis factor (TNF)- and C-C motif chemokine ligand (CCL)2 as inflammatory markers. The GelMAs were found to have widely varying endotoxin content (two with >1000 EU/ml and three with <10 EU/ml), however, this was not a critical factor determining in vitro cartilage-like matrix production of embedded MSCs. PBMCs did produce significantly higher TNF- and CCL2 in response to the GelMA with the highest endotoxin level compared to the other GelMAs. Although limited effects on chondrogenic differentiation were found in this study, caution with the use of commercial hydrogels is warranted in the translation from in vitro to in vivo studies because of regulatory constraints and potential inflammatory effects of the content of these hydrogels.

bioengineering↗