Fabrication of 3D cell-laden hydrogel microstructures through photo-mold patterning

Occhetta, Paola; Sadr, Nasser; Piraino, Francesco; Redaelli, Alberto; Moretti, Matteo; Rasponi, Marco

doi:10.1088/1758-5082/5/3/035002

Cited by 56 publications

(42 citation statements)

References 45 publications

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“…Viability decreased as the intensity of oxidative stress experienced increases in photocrosslinking conditions. Other researchers have also observed cell specific sensitivity to photoinitiator toxicity and different toxicity of various photoinitiators 1,5,37,38,50 , and have attributed encapsulated cell death to free radical damage 5 . Additionally, other researchers have shown free radical scavengers such as hydroquinone 1 and lactic acid 32 , and protective antioxidant encapsulating nanoparticles 43 can mitigate radical cytotoxicity.…”

Section: Discussionmentioning

confidence: 95%

“…(2% versus 82%) 9 at the weight ratios used to get comparable mechanical properties in 3D. 1 to 30 Irgacure 2959 to VA086 gave comparable viability between the two photoinitiator types in a 2D study using human umbilical vein endothelial cells and bone marrow stromal cells 37 . Cytotoxicity in 3D encapsulation conditions may be less than in 2D culture because in the encapsulation conditions the cells, polymer precursors, and additives are all competing targets for the photoinitiator radicals 5,9,37,50 .…”

Section: Discussionmentioning

confidence: 97%

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Optimizing Photo-Encapsulation Viability of Heart Valve Cell Types in 3D Printable Composite Hydrogels

et al. 2016

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Photocrosslinking hydrogel technologies are attractive for the biofabrication of cardiovascular soft tissues, but 3D printing success is dependent on multiple variables. In this study we systematically test variables associated with photocrosslinking hydrogels (photoinitiator type, photoinitiator concentration, and light intensity) for their effects on encapsulated cells in an extrusion 3D printable mixture of methacrylated gelatin/poly-ethylene glycol diacrylate/alginate (MEGEL/PEGDA3350/alginate). The fabrication conditions that produced desired hydrogel mechanical properties were compared against those that optimize aortic valve or mesenchymal stem cell viability. In the 3D hydrogel culture environment and fabrication setting studied, Irgacure can increase hydrogel stiffness with a lower proportional decrease in encapsulated cell viability compared to VA086. Human adipose derived mesenchymal stem cells (HADMSC) survived increasing photoinitiator concentrations in photo-encapsulation conditions better than aortic valve interstitial cells (HAVIC) and aortic valve sinus smooth muscle cells (HASSMC). Within the range of photo-encapsulation fabrication conditions tested with MEGEL/PEGDA/alginate (0.25–1.0% w/v VA086, 0.025–0.1% w/v Irgacure 2959, and 365 nm light intensity 2–136 mW/cm2), the highest viabilities achieved were 95%, 93%, and 93% live for HASSMC, HAVIC, and HADMSC respectively. These results identify parameter combinations that optimize cell viability during 3D printing for multiple cell types. These results also indicate that general oxidative stress is higher in photocrosslinking conditions that induce lower cell viability. However, suppressing this increase in intracellular oxidative stress did not improve cell viability, which suggests that other stress mechanisms also contribute.

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Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 97%

Optimizing Photo-Encapsulation Viability of Heart Valve Cell Types in 3D Printable Composite Hydrogels

et al. 2016

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“…Hydrogels such as poly(ethylene glycol) diacrylate (PEGDA) are widely used in 3D cell culture to encapsulate cells and mimic the tissue-like microenvironment around the cell during in-vitro culture (Tibbitt and Anseth 2009, Durst, Cuchiara et al 2011). So far, the patterning of such 3D cultures are limited to UV curable hydrogels due to the ease and accessibility of photolithography (Occhetta, Sadr et al 2013). The ability of the vacuum method to inject solutions in complex, long, and dead-end reversibly sealed microchannels can be used to obtain 3D patterned cell cultures by injecting a hydrogel precursor solution mixed with a cell suspension into the PDMS microchannels.…”

Section: Discussionmentioning

confidence: 99%

Vacuum-assisted fluid flow in microchannels to pattern substrates and cells

et al. 2014

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Substrate and cell patterning are widely used techniques in cell biology to study cell-to-cell and cell-to-substrate interactions. Conventional patterning techniques work well only with simple shapes, small areas and selected bio-materials. This paper describes a method to distribute cell suspensions as well as substrate solutions into complex, long, closed (dead-end) polydimethylsiloxane (PDMS) microchannels using negative pressure. Our method builds upon a previous vacuum-assisted method used for micromolding (Jeon, Choi et al. 1999) and successfully patterned collagen-I, fibronectin and Sal-1 substrates on glass and polystyrene surfaces, filling microchannels with lengths up to 120 mm and covering areas up to 13 × 10 mm2. Vacuum-patterned substrates were subsequently used to culture mammalian PC12 and fibroblast cells and amphibian neurons. Cells were also patterned directly by injecting cell suspensions into microchannels using vacuum. Fibroblast and neuronal cells patterned using vacuum showed normal growth and minimal cell death indicating no adverse effects of vacuum on cells. Our method fills reversibly sealed PDMS microchannels. This enables the user to remove the PDMS microchannel cast and access the patterned biomaterial or cells for further experimental purposes. Overall, this is a straightforward technique that has broad applicability for cell biology.

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“…This report showed that by spatially patterning cells in 3D hydrogels, their viability and functionality can be improved. There have been a number of other studies that have encapsulated cells in patterned 3D hydrogel scaffolds and studied cell migration (31), cell proliferation/spreading (135), and cell elongation/fibril formation (136). …”

Section: Current Strategies For 3d Biofabricationmentioning

confidence: 99%

3D Biofabrication Strategies for Tissue Engineering and Regenerative Medicine

Bajaj¹,

Schweller

Khademhosseini

et al. 2014

Annu. Rev. Biomed. Eng.

538

418

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Over the past several decades, there has been an ever-increasing demand for organ transplants. However, there is a severe shortage of donor organs, and as a result of the increasing demand, the gap between supply and demand continues to widen. A potential solution to this problem is to grow or fabricate organs using biomaterial scaffolds and a person’s own cells. Although the realization of this solution has been limited, the development of new biofabrication approaches has made it more realistic. This review provides an overview of natural and synthetic biomaterials that have been used for organ/tissue development. It then discusses past and current biofabrication techniques, with a brief explanation of the state of the art. Finally, the review highlights the need for combining vascularization strategies with current biofabrication techniques. Given the multitude of applications of biofabrication technologies, from organ/tissue development to drug discovery/screening to development of complex in vitro models of human diseases, these manufacturing technologies can have a significant impact on the future of medicine and health care.

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Fabrication of 3D cell-laden hydrogel microstructures through photo-mold patterning

Cited by 56 publications

References 45 publications

Optimizing Photo-Encapsulation Viability of Heart Valve Cell Types in 3D Printable Composite Hydrogels

Optimizing Photo-Encapsulation Viability of Heart Valve Cell Types in 3D Printable Composite Hydrogels

Vacuum-assisted fluid flow in microchannels to pattern substrates and cells

3D Biofabrication Strategies for Tissue Engineering and Regenerative Medicine

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