Fibre-infused gel scaffolds guide cardiomyocyte alignment in 3D-printed ventricles
Open accessAbstract
Hydrogels are attractive materials for tissue engineering, but efforts to date have shown limited ability to produce the microstructural features necessary to promote cellular self-organization into hierarchical three-dimensional (3D) organ models. Here we develop a hydrogel ink containing prefabricated gelatin fibres to print 3D organ-level scaffolds that recapitulate the intra- and intercellular organization of the heart. The addition of prefabricated gelatin fibres to hydrogels enables the tailoring of the ink rheology, allowing for a controlled sol-gel transition to achieve precise printing of free-standing 3D structures without additional supporting materials. Shear-induced alignment of fibres during ink extrusion provides microscale geometric cues that promote the self-organization of cultured human cardiomyocytes into anisotropic muscular tissues in vitro. The resulting 3D-printed ventricle in vitro model exhibited biomimetic anisotropic electrophysiological and contractile properties.
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Claims the paper makes
32 of 55 cited sources checked · 9 supported · 0 flagged · accuracy not yet measured
- Premisecitation
Prior hydrogel tissue engineering approaches have had limited capacity to create the microstructural features required for cells to self-organize into hierarchical three-dimensional organ models.
“Hydrogels are attractive materials for tissue engineering, but efforts to date have shown limited ability to produce the microstructural features necessary to promote cellular self-organization into hierarchical 3D organ models.”
Quote in the paperSubstantial claim4 of 7 sources checked- cites [6] Park, S. J. et al. Insights into the pathogenesis of catecholaminergic polymorphic ventricular tachycardia from engineered human heart tissue. Circulation 140, 390–404 (2019). source
The Methods describe micro-molded gelatin substrates, and the Results report that cardiomyocytes assembled on them with parallel alignment. Neither passage assesses whether prior hydrogel approaches could produce the microstructural features needed for hierarchical 3D organ models. Searches for those features and for limitations of hydrogel approaches found no passage addressing that claim; page numbers were unavailable. (LLM, conf=0.76)
Experimental — accuracy not yet measured - cites [7] Wang, G. et al. Modeling the mitochondrial cardiomyopathy of barth syndrome with induced pluripotent stem cell and heart-on-chip technologies. Nat. Med. 20, 616–623 (2014). source
The paper describes cardiomyocytes self-organizing on micropatterned elastomers (Abstract; page not provided) and details PDMS microcontact printing (Microcontact Printing; page not provided). Searches for hydrogels, microstructural limitations, and hierarchical 3D organ models found no passage assessing whether prior hydrogel approaches could create those features. (LLM, conf=0.77)
Experimental — accuracy not yet measured - cites [8] McCain, M. L., Agarwal, A., Nesmith, H. W., Nesmith, A. P. & Parker, K. K. Micromolded gelatin hydrogels for extended culture of engineered cardiac tissues. Biomaterials 35, 5462–5471 (2014). source
The Results (sections 3.1–3.2; page not provided) show that micromolded hydrogel features aligned cardiac cells, supporting the use of hydrogel microstructures to organize tissue. The Discussion (section 4; page not provided) describes moving toward 3D systems as future work, but the paper does not establish that prior hydrogel approaches generally lacked the microstructural features needed for hierarchical 3D organ models. (LLM, conf=0.78)
Experimental — accuracy not yet measured - cites [9] Feinberg, A. W. et al. Muscular thin films for building actuators and powering devices. Science 317, 1366–1370 (2007). source
- cites [10] Li, R. A. et al. Bioengineering an electro-mechanically functional miniature ventricular heart chamber from human pluripotent stem cells. Biomaterials 163, 116–127 (2018). source
The Introduction describes limitations of earlier cardiac constructs in measuring pump performance, not an inability of hydrogels to produce microstructural features. The Results describe self-assembly of the authors’ hydrogel-based chamber, while the Discussion notes that their simplified chamber lacks the native heart’s hierarchical muscle-fiber organization. Neither passage establishes the claimed general limitation of prior hydrogel approaches; searches for microstructural features and hierarchical self-organization found no such statement. (LLM, conf=0.72)
Experimental — accuracy not yet measured - cites [11] Karp, J. M. et al. A photolithographic method to create cellular micropatterns. Biomaterials 27, 4755–4764 (2006). source
- cites [12] Wu, X. et al. Preparation of aligned porous gelatin scaffolds by unidirectional freeze-drying method. Acta Biomater. 6, 1167–1177 (2010). source
- Premisereasoning
Bridging length scales from the micro- and nanometer scale to the centimeter scale to induce intra- and intercellular organization within engineered heart models remains a major challenge in 3D printing functional organs and tissues.
“As a result, bridging the length scales from the micro- and nanometer scale to the centimeter scale to induce both intra- and intercellular organization within engineered heart models remains a major challenge in 3D printing functional organs and tissues32, 34, 35.”
Quote in the paperSubstantial claim2 of 3 sources checked- cites [32] Kupfer, M. E. et al. In situ expansion, differentiation, and electromechanical coupling of human cardiac muscle in a 3D bioprinted, chambered organoid. Circ. Res. 127, 207–224 (2020). source
The Discussion describes the challenge of expanding microscale cardiac models to macroscale structures while preserving cell–cell interactions and geometric complexity. The Introduction likewise discusses difficulties forming functional cell–cell junctions in 3D-printed chambered models. The paper does not specifically frame this as bridging nanometer-to-centimeter scales to induce both intracellular and intercellular organization; page numbers are unavailable in our copy. (LLM, conf=0.83)
Experimental — accuracy not yet measured - cites [34] Campostrini, G., Windt, L. M., van Meer, B. J., Bellin, M. & Mummery, C. L. Cardiac tissues from stem cells: new routes to maturation and cardiac regeneration. Circ. Res. 128, 775–801 (2021). source
In “3D Cardiac Models > Scaffold-Free Cardiac Tissues” (page unavailable), the review describes challenges in bioprinting cardiac tissues and says printing can create complex architectures with specific cell organization. It does not identify bridging nanometer-to-centimeter length scales to achieve intra- and intercellular organization as the challenge. (LLM, conf=0.83)
Experimental — accuracy not yet measured - cites [35] Ahrens, J. H. et al. Programming cellular alignment in engineered cardiac tissue via bioprinting anisotropic organ building blocks. Adv. Mater. 34, e2200217 (2022). source
- Premisecitation
The hierarchical organization of cardiac tissues is needed to regulate the spatiotemporal dynamics of excitation-contraction coupling that produce the cardiac cycle.
“This is particularly important in the heart, where the hierarchical organization of cardiac tissues3 is needed to regulate the spatiotemporal dynamics of excitation-contraction coupling, resulting in the cardiac cycle4, 5.”
Quote in the paperSubstantial claim1 of 3 sources checked- cites [3] LeGrice, I., Pope, A. & Smaill, B. in Interstitial Fibrosis in Heart Failure (ed. Villarreal, F. J.) 3–21 (Springer New York, 2005).
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
- cites [4] Ronaldson-Bouchard, K. et al. Advanced maturation of human cardiac tissue grown from pluripotent stem cells. Nature 556, 239–243 (2018). source
- cites [5] Kleber, A. G. & Rudy, Y. Basic mechanisms of cardiac impulse propagation and associated arrhythmias. Physiol. Rev. 84, 431–488 (2004). source
- Premisecitation
In physiologically healthy cardiac tissues, cytoskeletal structures including sarcomeres and actin filaments are highly aligned with orientational order parameter values close to unity.
“In physiologically healthy cardiac tissues, cytoskeletal structures including sarcomeres42, 43 and actin filaments44, 45, are highly aligned with OOP values close to unity.”
Quote in the paperSubstantial claim3 of 4 sources checked- cites [42] Bray, M. A., Sheehy, S. P. & Parker, K. K. Sarcomere alignment is regulated by myocyte shape. Cell Motil. Cytoskeleton 65, 641–651 (2008). source
The Results (page unavailable) show that sarcomeres become more aligned as the aspect ratio of cultured, patterned cardiac myocytes increases. The paper does not establish that sarcomeres in physiologically healthy cardiac tissue have orientational order parameter (OOP) values close to unity; its Methods (page unavailable) use a different anisotropy measure. (LLM, conf=0.88)
Experimental — accuracy not yet measured - cites [43] Kuo, P. L. et al. Myocyte shape regulates lateral registry of sarcomeres and contractility. Am. J. Pathol. 181, 2030–2037 (2012). source
The Discussion describes tightly registered myofibrils in well-differentiated cardiomyocytes, supporting the qualitative point about sarcomere organization. The Results examine sarcomere alignment in engineered single myocytes, but the paper does not report OOP values close to unity for healthy cardiac tissue. Citations 42 and 43 attach to sarcomeres, not to the separate actin-filament claim. (LLM, conf=0.84)
Experimental — accuracy not yet measured - cites [44] Geisse, N. A., Sheehy, S. P. & Parker, K. K. Control of myocyte remodeling in vitro with engineered substrates. Vitr. Cell. Dev. Biol. Anim. 45, 343–350 (2009). source
- cites [45] Grosberg, A. et al. Self-organization of muscle cell structure and function. PLoS Comput. Biol. 7, e1001088 (2011). source
The Results section (page unavailable) reports that actin fibers in cardiac myocytes cultured on square and triangular ECM islands aligned along the cells’ longest axes, with punctuations indicating sarcomeres. The Methods section defines an OOP of one as perfect alignment, but the paper does not report OOP values close to one for physiologically healthy cardiac tissue; its alignment findings concern cultured, geometrically patterned cells. (LLM, conf=0.86)
Experimental — accuracy not yet measured
- Premisecitation
Cardiac impulses propagate in aligned cardiac tissues approximately two times faster in the longitudinal direction than in the transverse direction due to rapid conduction along the longitudinal axis and intercellular conduction at gap junctions.
“Cardiac impulses propagate in aligned cardiac tissues approximately two times faster in the longitudinal (Vlong) than the transverse direction (Vtrans,Vlong/Vtrans=∼2) due to rapid conduction along the longitudinal axis and intercellular conduction at gap junctions49.”
Quote in the paperSubstantial claim1 of 1 sources checked- cites [49] Kleber, A. G. & Jin, Q. Coupling between cardiac cells—an important determinant of electrical impulse propagation and arrhythmogenesis. Biophys. Rev. 2, 031301 (2021). source
Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.
Experimental — accuracy not yet measured
- Premisecitation
Methods such as microcontact printing, photolithography, directional freeze drying, fiber spinning, and microphysiological chip fabrication are limited in producing tissues on complex geometries that fully replicate cardiac function.
“These methods are however limited in their ability to produce tissues on complex geometries that fully replicate cardiac function.”
Quote in the paperSubstantial claim4 of 7 sources checked- cites [6] Park, S. J. et al. Insights into the pathogenesis of catecholaminergic polymorphic ventricular tachycardia from engineered human heart tissue. Circulation 140, 390–404 (2019). source
The Introduction (page unavailable) describes engineered cardiac microphysiological systems that produce native-like laminar tissue, while the Discussion (page unavailable) identifies limitations of this paper’s own tissue model. Neither passage addresses whether the listed fabrication methods are limited in producing tissue on complex geometries that fully replicates cardiac function. Searches for the named methods and for the complex-geometries limitation found no supporting passage. (LLM, conf=0.74)
Experimental — accuracy not yet measured - cites [7] Wang, G. et al. Modeling the mitochondrial cardiomyopathy of barth syndrome with induced pluripotent stem cell and heart-on-chip technologies. Nat. Med. 20, 616–623 (2014). source
The Discussion (page unavailable) says that modeling contractile function at the myocardial-tissue level has been challenging. It does not say that the listed fabrication methods are limited by complex geometries or assess whether they fully replicate cardiac function. (LLM, conf=0.78)
Experimental — accuracy not yet measured - cites [8] McCain, M. L., Agarwal, A., Nesmith, H. W., Nesmith, A. P. & Parker, K. K. Micromolded gelatin hydrogels for extended culture of engineered cardiac tissues. Biomaterials 35, 5462–5471 (2014). source
The Introduction (page not provided) describes limitations of earlier heart-on-a-chip substrates: PDMS is much stiffer than native myocardium, while alginate fabrication lacked control over cantilever shape. It does not establish that the listed methods as a group cannot produce tissues on complex geometries that fully replicate cardiac function. (LLM, conf=0.78)
Experimental — accuracy not yet measured - cites [9] Feinberg, A. W. et al. Muscular thin films for building actuators and powering devices. Science 317, 1366–1370 (2007). source
- cites [10] Li, R. A. et al. Bioengineering an electro-mechanically functional miniature ventricular heart chamber from human pluripotent stem cells. Biomaterials 163, 116–127 (2018). source
The Introduction (page not provided) says earlier cardiac monolayers, patches, thin films, micro-tissues and strips could not provide direct pressure- and volume-based measures of heart-pump performance. It does not assess the five named fabrication methods or establish that they are limited in producing tissues on complex geometries. The paper discusses recapitulating key cardiac features, not fully replicating cardiac function. (LLM, conf=0.85)
Experimental — accuracy not yet measured - cites [11] Karp, J. M. et al. A photolithographic method to create cellular micropatterns. Biomaterials 27, 4755–4764 (2006). source
- cites [12] Wu, X. et al. Preparation of aligned porous gelatin scaffolds by unidirectional freeze-drying method. Acta Biomater. 6, 1167–1177 (2010). source
- Findingreasoning
Shear-induced alignment of fibers during ink extrusion provides microscale geometric cues that promote self-organization of cultured human cardiomyocytes into anisotropic muscular tissues in vitro.
“Shear-induced alignment of fibers during ink extrusion provides microscale geometric cues that promote self-organization of cultured human cardiomyocytes into anisotropic muscular tissues in vitro.”
Quote in the paperSubstantial claim - Findingreasoning
The resulting 3D printed ventricle in vitro model exhibited biomimetic anisotropic electrophysiological and contractile properties.
“The resulting 3D printed ventricle in vitro model exhibited biomimetic anisotropic electrophysiological and contractile properties.”
Quote in the paperSubstantial claim - Findingown data
By ionic crosslinking of alginate with 1 wt% calcium chloride and enzymatic crosslinking of gelatin with about 8 wt% microbial transglutaminase, the printed scaffolds and their microscale architecture remained intact in water.
“By ionic crosslinking of alginate (1 wt% calcium chloride) and enzymatic crosslinking of gelatin (~8 wt% microbial transglutaminase), the printed scaffolds and their microscale architecture remained intact in water (Supplementary Fig. 4b).”
Quote in the paperSubstantial claim - Findingown data
Each FIG orientation resulted in distinct contractile patterns such as rolling, twisting, and folding.
“Each FIG orientation resulted in distinct contractile patterns such as rolling, twisting, and folding, respectively (Fig. 3c–e).”
Quote in the paperSubstantial claim - Findingown data
Following point electrical stimulation of NRVM cultured ventricle models, Ca2+ transient propagations were faster in the transverse direction, the direction of printing, compared to the longitudinal direction.
“Following point electrical stimulation of NRVMs cultured ventricle models, we observed that Ca2+ transient propagations were faster in the transverse (direction of printing) compared to the longitudinal direction (Supplementary Fig. 12a–b, and Supplementary Video. 9).”
Quote in the paperSubstantial claim - Findingown data
Preferential electromechanical coupling of cardiomyocytes along the printing direction enabled cyclic deformations in response to electrical stimulation.
“Preferential electromechanical coupling of cardiomyocytes along the printing direction enabled cyclic deformations in response to electrical stimulation (Fig. 3e–f).”
Quote in the paperSubstantial claim - Findingown data
Cardiac tissues on FIG scaffolds propagated Ca2+ waves over a 6 cm serpentine pattern, demonstrating robust electrical coupling along the direction of printing.
“Cardiac tissues on FIG scaffolds also propagated Ca2+ waves over a 6 cm serpentine pattern, demonstrating robust electrical coupling along the direction of printing (Supplementary Fig. 9 and Supplementary Video. 5).”
Quote in the paperSubstantial claim - Findingown data
Cardiac tissues on FIG scaffolds using NRVMs and hiPSC-CMs exhibited longitudinal-to-transverse propagation speed ratios of 1.67 ± 0.085 and 1.62 ± 0.18, respectively.
“Cardiac tissues on FIG scaffolds using NRVMs and human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CM, Supplementary Fig. 8, Supplementary Video. 2) exhibited Vlong/Vtrans of 1.67 ± 0.085 and 1.62 ± 0.18, respectively (mean ± SEM, Fig. 3b, Supplementary Fig. 6f–g and 8, and Supplementary Video. 3–4).”
Quote in the paperSubstantial claim - Findingown data
The high elastic modulus of gel-like FIG inks provided sufficient gel stability to print self-supportive walls with thicknesses between 200 μm and 350 μm without scaffolds or sacrificial baths.
“The high elastic modulus of gel-like FIG inks provided sufficient gel stability to print self-supportive walls with thicknesses ranging between 200 μm and 350 μm without the need for scaffolds or sacrificial baths (Supplementary Fig. 2a–g, and Rheology of FIG inks in Supplementary information).”
Quote in the paperSubstantial claim - Findingreasoning
Shear-induced fiber alignment on the micron scale was achieved across the entire length scale of the printed structure.
“Following this process, shear induced fiber alignment on the micron scale was achieved across the entire length scale of the printed structure (Fig. 1i–j).”
Quote in the paperSubstantial claim - Findingown data
Incorporating 8 wt% pre-fabricated gelatin fibers into Gel-Alg hydrogels rendered the hydrogels into a predominantly elastic gel.
“Unlike Gel-Alg hydrogels where 3D printing is limited due to their fluidity, incorporating 8 wt% pre-fabricated gelatin fibers into Gel-Alg hydrogels rendered the hydrogels into a predominantly elastic gel (8 wt% fiber, 2.4 wt% gelatin, 2.4% alginate in PBS, Fig. 1d–f, Supplementary Fig. 2a–d).”
Quote in the paperSubstantial claim - Findingown data
NRVMs and hiPSC-CMs cultured on a 3D printed ventricle scaffold with circumferential fiber alignment formed laminar anisotropic cardiac tissues in a 3D ventricular shape.
“We cultured NRVMs and hiPSC-CMs on a 3D printed ventricle scaffold (8 wt% fiber, 2.4 wt% gelatin, and 2.4 wt% alginate) with circumferential fiber alignment, forming laminar anisotropic cardiac tissues in a 3D ventricular shape (Fig. 4a–b, Supplementary Fig. 11, and Supplementary Video. 7–8).”
Quote in the paperSubstantial claim - Findingown data
NRVM tissues on control scaffolds without fibers exhibited a propagation speed ratio close to unity at Day 5, indicating their isotropy.
“In contrast, NRVM tissues on control scaffolds without fibers exhibited a propagation speed ratio close to unity (0.96 ± 0.067 at Day 5, mean ± SEM) indicating their isotropy.”
Quote in the paperSubstantial claim - Findingown data
The resulting in vitro ventricle models showed cyclic contractile motions of the heart.
“Resulting in vitro ventricle models showed cyclic contractile motions of the heart, however, to enhance cardiac performance, multilayered tissue ventricle models are required in the future.”
Quote in the paperSubstantial claim - Findingnone given
Infused gelatin fibers acted as a rheological modifier in the hydrogel inks, allowing printing of complex 3D objects without sacrificial baths.
“Here, infused gelatin fibers acted as a rheological modifier in our hydrogel inks, allowing for the printing of complex 3D objects without the use of sacrificial baths.”
Quote in the paperSubstantial claim - Findingown data
Fluorescent microparticle flow corresponded with fluid movement out of and into the in vitro ventricle models.
“Fluorescent microparticles flow corresponded with the fluid movement out of and into the in vitro ventricle models (Fig. 4e, Supplementary Fig. 12d, and Supplementary Video. 12).”
Quote in the paperSubstantial claim - Findingown data
Synchronized and coordinated contractions of hiPSC-CM cultured ventricle models were demonstrated by deformation mapping analysis measuring displacement of the in vitro ventricle models.
“This led to synchronized and coordinated contractions of hiPSC-CM cultured ventricle models, which were demonstrated by deformation mapping analysis measuring displacement of the in vitro ventricle models (Fig. 4c–d and Supplementary Video. 11).”
Quote in the paperSubstantial claim - Findingown data
The paper demonstrated the ability to 3D print tissue-engineered ventricle models with hydrogel-based FIG ink that simultaneously recapitulates the heart's microstructural ECM architecture and macrostructural organ-level geometry.
“We have demonstrated the ability to 3D print tissue-engineered ventricle models with hydrogel-based FIG ink, which simultaneously recapitulates the heart’s microstructural ECM architecture and macrostructural organ-level geometry.”
Quote in the paperSubstantial claim - Interpretationown data
This enables control of local dynamic contractile motions similar to those seen in laminar muscles around the ventricle.
“This enables control of local dynamic contractile motions similar to those seen in laminar muscles around the ventricle50.”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [50] Buckberg, G., Hoffman, J. I., Mahajan, A., Saleh, S. & Coghlan, C. Cardiac mechanics revisited: the relationship of cardiac architecture to ventricular function. Circulation 118, 2571–2587 (2008). source
- Interpretationcitation
The pre-stress generated by intra- and inter-cellular cytoskeletal structures determined nuclear elongation and orientation.
“Consequently, the pre-stress generated by intra- and inter-cellular cytoskeletal structures, determined nuclear elongation and orientation46, 47.”
Quote in the paperSubstantial claim0 of 2 sources checked- cites [46] Bray, M. A. et al. Nuclear morphology and deformation in engineered cardiac myocytes and tissues. Biomaterials 31, 5143–5150 (2010). source
- cites [47] Lee, H. et al. Cytoskeletal prestress regulates nuclear shape and stiffness in cardiac myocytes. Exp. Biol. Med. 240, 1543–1554 (2015). source
- Interpretationreasoning
The scaffolds were durable over about 800,000 mechanical load cycles without visible degradation.
“This suggested that the scaffolds were durable over ~800,000 mechanical load cycles without visible degradation.”
Quote in the paperSubstantial claim
- Classified as probably not a substantial claim. 19
- May combine two claims. 27
Classified as probably not a substantial claim. May combine two claims.
PremisecitationExtracellular matrix proteins act as topological and biochemical cues for cells and are crucial for tissue development.
“To achieve these organized structures, extracellular matrix (ECM) proteins act as topological and biochemical cues for the cells, which are crucial for tissue development1, 2.”
Quote in the paperSubstantial claim1 of 2 sources checked- cites [1] Frantz, C., Stewart, K. M. & Weaver, V. M. The extracellular matrix at a glance. J. Cell Sci. 123, 4195–4200 (2010). source
- cites [2] Blazeski, A. et al. Functional properties of engineered heart slices incorporating human induced pluripotent stem cell-derived cardiomyocytes. Stem Cell Rep. 12, 982–995 (2019). source
The Introduction (page unavailable) describes decellularized myocardial matrix as a source of biochemical and topographical cues that direct cardiomyocyte differentiation and maturation. The Discussion (page unavailable) also says its topographical cues guide cellular organization. These passages support the cue function, but do not establish that ECM proteins are crucial for tissue development generally. (LLM, conf=0.83)
Experimental — accuracy not yet measured
Probably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationEfforts to develop hydrogel-based inks or printing techniques to enhance 3D stability and shape fidelity include multimaterial inks, photo-crosslinkable hydrogels, viscosifiers, and sacrificial supporting materials.
“To this end, considerable efforts were made in developing hydrogel-based inks or printing techniques to enhance 3D stability and shape fidelity, such as printing multimaterial inks with combined nozzle system24, 25, photo-crosslinkable hydrogels17, 26, 27, adding viscosifiers28–30, or printing in sacrificial supporting materials31–33.”
Quote in the paperNot a substantial claim5 of 8 sources checked- cites [24] Jia, W. et al. Direct 3D bioprinting of perfusable vascular constructs using a blend bioink. Biomaterials 106, 58–68 (2016). source
- cites [25] Ashammakhi, N. et al. Bioinks and bioprinting technologies to make heterogeneous and biomimetic tissue constructs. Mater. Today Bio 1, 100008 (2019). source
The Multicomponent bioinks section (page unavailable) describes removable alginate support that maintains GelMA's shape, and the Digital light projector section links photocrosslinking to high shape fidelity. The paper also describes multimaterial nozzle printing and viscosity-increasing additives, but does not establish that the nozzle approach improves shape fidelity; it reports that one viscosity-increasing additive made deposited structures difficult to maintain. Its sacrificial-template example is not clearly printing *in* a sacrificial support material. (LLM, conf=0.83)
Experimental — accuracy not yet measured - cites [17] Yue, K. et al. Synthesis, properties, and biomedical applications of gelatin methacryloyl (GelMA) hydrogels. Biomaterials 73, 254–271 (2015). source
- cites [26] Ouyang, L. et al. Expanding and optimizing 3D bioprinting capabilities using complementary network bioinks. Sci. Adv. 6, abc5529 (2020). source
- cites [27] Choi, J. R., Yong, K. W., Choi, J. Y. & Cowie, A. C. Recent advances in photo-crosslinkable hydrogels for biomedical applications. BioTechniques 66, 40–53 (2019). source
- cites [28] Markstedt, K. et al. 3D bioprinting human chondrocytes with nanocellulose-alginate bioink for cartilage tissue engineering applications. Biomacromolecules 16, 1489–1496 (2015). source
- cites [29] Nadernezhad, A. et al. Nanocomposite bioinks based on agarose and 2D nanosilicates with tunable flow properties and bioactivity for 3D bioprinting. ACS Appl. Bio Mater. 2, 796–806 (2019). source
- cites [30] Schwab, A. et al. Printability and shape fidelity of bioinks in 3D bioprinting. Chem. Rev. 120, 11028–11055 (2020). source
May combine two claims.
Findingown dataGelatin fibers produced by rotary jet spinning and truncated by ultrasonication had a length of 85.9 ± 2.53 μm and a diameter of 4.20 ± 0.23 μm.
“Gelatin fibers produced by rotary jet spinning (Supplementary Fig. 1b) 14, 15 were truncated by ultrasonication to a length of 85.9 ± 2.53 μm with a diameter of 4.20 ± 0.23 μm (mean ± SEM), reducing variance in length and enabling passage through the printing nozzle with a diameter of 200 μm (Supplementary Fig. 1c–d).”
Quote in the paperSubstantial claim2 of 2 sources checked- cites [14] Capulli, A. K. et al. Jetvalve: rapid manufacturing of biohybrid scaffolds for biomimetic heart valve replacement. Biomaterials 133, 229–241 (2017). source
Materials and Methods §2.2 describes rotary jet spinning of P4HB/gelatin blends, supporting the fiber-production method. It does not report ultrasonic truncation or the stated fiber length and diameter; Results §3.1 instead reports diameters for the blended JetValve scaffold fibers. No page numbers were supplied for these passages. (LLM, conf=0.85)
Experimental — accuracy not yet measured - cites [15] Chang, H. et al. Recreating the heart’s helical structure-function relationship with focused rotary jet spinning. Science 377, 180–185 (2022). source
May combine two claims.
PremisecitationFibronectin-coated gelatin fibers contain RGD peptide binding domains that promote matrix-cell adhesion through integrins, unlike cellulose nanofibers and carbon nanotubes previously used in inks.
“In contrast to cellulose nanofiber and carbon nanotubes, previously used in inks 36–38, fibronectin-coated gelatin fibers contain arginine–glycine–aspartic (RGD) acid peptide binding domains that promote matrix-cell adhesion through integrins39.”
Quote in the paperSubstantial claim2 of 4 sources checked- cites [36] Gladman, A. S., Matsumoto, E. A., Nuzzo, R. G., Mahadevan, L. & Lewis, J. A. Biomimetic 4D printing. Nat. Mater. 15, 413–418 (2016). source
- cites [37] Siqueira, G. et al. Cellulose nanocrystal inks for 3D printing of textured cellular architectures. Adv. Funct. Mater. 27, 1604619 (2017). source
- cites [38] Jinkins, K. R., Chan, J., Jacobberger, R. M., Berson, A. & Arnold, M. S. Substrate‐wide confined shear alignment of carbon nanotubes for thin film transistors. Adv. Electron. Mater. 5, 1800593 (2018). source
- cites [39] Caliari, S. R. & Burdick, J. A. A practical guide to hydrogels for cell culture. Nat. Methods 13, 405–414 (2016). source
The Alginate section (page unavailable) identifies RGD as an adhesive ligand that enables cell attachment, and “Selecting a hydrogel for cell culture” says some materials interact with cells through integrin–ligand interactions. The paper does not specifically say that fibronectin-coated gelatin fibers contain RGD domains or demonstrate adhesion through those fibers; the cellulose-nanofiber and carbon-nanotube comparison is cited separately. (LLM, conf=0.85)
Experimental — accuracy not yet measured
Classified as probably not a substantial claim.
PremisecitationBuilding organ models that mimic 3D geometry requires additive manufacturing strategies.
“Specifically, to build organ models that mimic the 3D geometry requires additive manufacturing strategies15, 20.”
Quote in the paperSubstantial claim2 of 2 sources checked- cites [15] Chang, H. et al. Recreating the heart’s helical structure-function relationship with focused rotary jet spinning. Science 377, 180–185 (2022). source
The Abstract describes focused rotary jet spinning as an additive manufacturing approach for fabricating scaffolds in three-dimensional geometries. The section on dual-chambered ventricles and full-scale heart models reports using it to build a full-size heart model. This supports additive manufacturing as a strategy for such models, but the paper does not establish that it is required. (LLM, conf=0.83)
Experimental — accuracy not yet measured - cites [20] Capulli, A. K., MacQueen, L. A., Sheehy, S. P. & Parker, K. K. Fibrous scaffolds for building hearts and heart parts. Adv. Drug Deliv. Rev. 96, 83–102 (2016). source
Section 3.2 says that building three-dimensional cardiac scaffold geometries from synthetic fibers requires innovative, cumulative fiber-collection techniques. It does not say that organ models generally require additive manufacturing; Section 3 also describes decellularized tissues and organs as providing 3D substrates. No page number is available in our copy. (LLM, conf=0.86)
Experimental — accuracy not yet measured
Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Premisecitation3D printing has been increasingly used for engineering tissue models because it enables facile design and editing of 3D structure before printing and fabrication of complex 3D structures with flexible biomaterial ink formulation.
“Particularly, 3D printing has been increasingly used for engineering tissue models for enabling facile design and editing of 3D structure prior to printing, and fabrication of complex 3D structures with flexible formulation of biomaterial inks21–23.”
Quote in the paperNot a substantial claim1 of 3 sources checked- cites [21] Yi, H. G. et al. A bioprinted human-glioblastoma-on-a-chip for the identification of patient-specific responses to chemoradiotherapy. Nat. Biomed. Eng. 3, 509–519 (2019). source
- cites [22] Daly, A. C., Davidson, M. D. & Burdick, J. A. 3D bioprinting of high cell-density heterogeneous tissue models through spheroid fusion within self-healing hydrogels. Nat. Commun. 12, 753 (2021). source
The Introduction (page unavailable) says bioprinting enables programmed assembly into complex 3D geometries, and the Methods describe designing molds with CAD before printing. The paper does not establish that tissue structures are easily edited before printing or that biomaterial inks can be formulated flexibly; the Results and discussion instead describe bioink requirements as a significant challenge. (LLM, conf=0.83)
Experimental — accuracy not yet measured - cites [23] Brassard, J. A., Nikolaev, M., Hubscher, T., Hofer, M. & Lutolf, M. P. Recapitulating macro-scale tissue self-organization through organoid bioprinting. Nat. Mater. 20, 22–29 (2021). source
May combine two claims.
Findingown dataThe hiPSC-CM based ventricular volumetric change between peak contraction and peak relaxation was 5.94 ± 1.66 % (n = 4), which is 2~5 and 8~20 times higher than values previously reported as ejection fraction for NRVM and hiPSC-CM in vitro ventricle models.
“We calculated our hiPSC-CM based ventricular volumetric change between peak contraction and peak relaxation to be 5.94 ± 1.66 % (n = 4, mean ± SEM, Fig. 4g), which is 2~5 and 8~20 times higher than values previously reported (ejection fraction) for NRVM and hiPSC-CM in vitro ventricle models, respectively13, 15, 32.”
Quote in the paperSubstantial claim3 of 3 sources checked- cites [13] MacQueen, L. A. et al. A tissue-engineered scale model of the heart ventricle. Nat. Biomed. Eng. 2, 930–941 (2018). source
Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.
Experimental — accuracy not yet measured - cites [15] Chang, H. et al. Recreating the heart’s helical structure-function relationship with focused rotary jet spinning. Science 377, 180–185 (2022). source
Split into 2 atoms — 1 confirmed, 1 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.
Experimental — accuracy not yet measured - cites [32] Kupfer, M. E. et al. In situ expansion, differentiation, and electromechanical coupling of human cardiac muscle in a 3D bioprinted, chambered organoid. Circ. Res. 127, 207–224 (2020). source
- Rests on an assumption: The volumetric change measured by particle imaging velocimetry in the hiPSC-CM ventricle model is comparable to ejection fraction values reported for other in vitro ventricle models.
Probably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductcitationhiPSC-CM differentiation was conducted for 15 days following published protocols.
“The differentiation process for hiPSC-CMs was conducted for 15 days following published protocols52.”
Quote in the paperNot a substantial claim1 of 1 sources checked- cites [52] Lian, X. et al. Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/β-catenin signaling under fully defined conditions. Nat. Protoc. 8, 162–175 (2013). source
May combine two claims.
Findingnone givenAdding pre-fabricated gelatin fibers to hydrogels enables tailoring of ink rheology and a controlled sol-gel transition, allowing precise printing of free-standing 3D structures without additional supporting materials.
“The addition of pre-fabricated gelatin fibers to hydrogels enables tailoring ink rheology, allowing for a controlled sol-gel transition to achieve precise printing of free-standing 3D structures without additional supporting materials.”
Quote in the paperSubstantial claimMay combine two claims.
FindingcitationThe shear thinning property of FIG inks is essential for printing and for inducing fiber alignment along the direction of printing.
“To achieve this, we exploited the shear thinning property of FIG inks (Supplementary Fig. 2h–i), which is not only essential for printing, but also for inducing fiber alignment along the direction of printing40.”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [40] Hausmann, M. K. et al. Dynamics of cellulose nanocrystal alignment during 3D printing. ACS Nano 12, 6926–6937 (2018). source
May combine two claims.
FindingreasoningIncorporating fibers into the hydrogel modifies ink rheology, allowing creation of accurate and complex 3D scaffolds without supporting structures or materials.
“Furthermore, incorporating fibers into the hydrogel modifies the ink rheology, allowing for creating accurate and complex 3D scaffolds without any supporting structures or materials.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataNuclear orientation of NRVMs on FIG scaffolds aligned along the printing direction of 0°, whereas those on control Gel-Alg scaffolds showed no specific orientation.
“Nuclear orientation of NRVMs on FIG scaffolds also aligned along the printing direction (0°) in contrast to those on control Gel-Alg scaffolds that showed no specific orientation (Fig. 2d and f, and Supplementary Fig. 6d).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataAlternating alignment directions in one scaffold showed that cardiomyocyte organization follows the local fiber direction, resulting in corresponding local tissue contractile patterns.
“Alternating alignment directions in one scaffold also showed that cardiomyocyte organization follows the local fiber direction, resulting in corresponding local tissue contractile patterns (Supplementary Fig. 10b).”
Quote in the paperSubstantial claimMay combine two claims.
FindingreasoningThe resulting printed scaffolds had a resolution closer to the fiber diameter of about 5 μm, which acted as geometric cues for cell alignment despite the printing resolution being limited by the nozzle diameter of 200-300 μm.
“The resulting printed scaffolds had a resolution closer to the fiber diameter (~5 μm) which acted as geometric cues for cell alignment despite the printing resolution being limited by the nozzle diameter (200–300 μm).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataThe contractile stress of hiPSC-CM tissues on FIG MTFs was 455.13 ± 130.33 Pa at day 14, significantly lower than the more mature NRVM tissues.
“We found the contractile stress of hiPSC-CM tissues to be 455.13 ± 130.33 Pa (mean ± SEM) at day 14 (Supplementary Fig. 10d–f), which was significantly lower than the more mature NRVM tissues.”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataDuring printing, shear stresses in the nozzle were sufficient to turn the elastic FIG ink into a liquid, and its elastic stability was restored upon extrusion.
“During printing, shear stresses in the nozzle was sufficient to turn the elastic FIG ink into a liquid, and its elastic stability was restored upon extrusion (Supplementary Fig. 2e).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataSeeding neonatal rat ventricular cardiomyocytes on printed 2D FIG scaffolds resulted in highly aligned cardiac tissues that retained cell viabilities comparable to pleomorphic culture conditions.
“This resulted in the formation of highly aligned cardiac tissues (Fig. 2a–b, Supplementary Fig. 6b), which retained comparable cell viabilities relative to pleomorphic culture conditions (Supplementary Fig. 7).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataAlignment of incorporated fibers was well-maintained after extrusion due to the gel stability of FIG inks, and was further enhanced by lowering the printing bed temperature to 6°C compared to room temperature.
“Alignment of the incorporated fibers was well-maintained post extrusion due to the gel stability of FIG inks (Fig. 1k and Supplementary Fig. 5), which was further enhanced by lowering the printing bed temperature to 6°C compared to printing at room temperature (Supplementary Fig. 2f).”
Quote in the paperSubstantial claimMay combine two claims.
FindingreasoningThese nonlinear rheological properties allow continuous extrusion of FIG inks with minimal post-extrusion swelling and spreading and enable printing of layered structures for increasingly complex geometries without sacrificial layers or baths.
“These nonlinear rheological properties allow for continuous extrusion of FIG inks with minimal post-extrusion swelling and spreading (Fig. 1d and e and Supplementary Fig. 3) and enables printing of layered structures for increasingly complex geometries without sacrificial layers/bath as supporting structures (Fig. 1f).”
Quote in the paperSubstantial claimMay combine two claims.
FindingreasoningFIG inks allowed printing of self-supportive hollow 3D structures such as a cardiac ventricle with a broad range of design parameters, reproducibility, and accuracy.
“This allowed us to print self-supportive hollow 3D structures such as a cardiac ventricle with a broad range of design parameters, reproducibility, and accuracy (Fig. 1g, Supplementary Fig. 4a–b, and Supplementary Video. 1).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataNuclei were elongated on FIG scaffolds with a mean eccentricity ratio of 2.37 ± 0.17, compared to nuclei on control Gel-Alg scaffolds that remained relatively round with an eccentricity ratio of 1.40 ± 0.02.
“Nuclei were elongated on FIG scaffolds with a mean eccentricity ratio (length of major axis/length of minor axis) of 2.37± 0.17 compared to nuclei on control Gel-Alg scaffolds that remained relatively round (eccentricity ratio = 1.40±0.02, mean ± SEM, Fig. 2d and e).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataUltrasonication reduced variance in fiber length and enabled the fibers to pass through a 200 μm printing nozzle.
“Gelatin fibers produced by rotary jet spinning (Supplementary Fig. 1b) 14, 15 were truncated by ultrasonication to a length of 85.9 ± 2.53 μm with a diameter of 4.20 ± 0.23 μm (mean ± SEM), reducing variance in length and enabling passage through the printing nozzle with a diameter of 200 μm (Supplementary Fig. 1c–d).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataThe average contractile force of Day 7 NRVM tissues on FIG MTFs was 1661.48 ± 209.46 Pa, comparable to NRVMs on polydimethylsiloxane MTFs at 1801.66 ± 428.94 Pa, and they responded to increasing pacing frequencies up to 3 Hz.
“The average contractile force of Day 7 NRVM tissues on FIG MTFs was 1661.48 ± 209.46 Pa, comparable to NRVMs on polydimethylsiloxane MTFs (1801.66 ± 428.94 Pa, mean ± SEM, Supplementary Fig. 10e–f), and they responded to increasing pacing frequencies up to 3 Hz (Supplementary Fig. 10g).”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
FindingreasoningMore complex structures, such as heart valves, dual chambered hearts, and left ventricles with varying angular orientations, could also be printed using FIG inks.
“More complex structures, such as heart valves, dual chambered hearts, and left ventricles with varying angular orientations, could also be printed using FIG inks (Fig. 1h and Supplementary Fig. 4c–d).”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataThe in vitro ventricle models were maintained for 14 days in culture with a spontaneous beat rate of about 0.71 beats per second.
“The in vitro ventricle models were maintained for 14 days in culture with a spontaneous beat rate of ~0.71 beats per second.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Findingown datahiPSC-CMs were isolated after visual confirmation of cardiomyocyte contractility.
“After visual confirmation of cardiomyocytes contractility (Supplementary Video. S2), hiPSC-CMs were isolated”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataOptical mapping detected Ca2+ activity in NRVM tissues on the printed scaffolds at Day 5 and in iPSC-CM tissues at Day 7.
“we observed Ca2+ activities of cardiac tissues on the 3D printed scaffolds using optical mapping system at Day 5 and Day 7, respectively.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Findingown dataTissue on PDMS MTFs averaged 5 μm in thickness.
“the average PDMS thickness and tissue thickness for PDMS MTFs were 20 μm and 5 μm, respectively.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Findingown dataFIG MTF scaffolds averaged 70 μm in thickness.
“The average FIG scaffold thickness and tissue thickness for FIG MTFs were 70 μm, and 10 μm, respectively”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Findingown dataFIG's elastic modulus, measured by DMA compression along the fiber direction at 1 Hz, was 51 kPa.
“The elastic modulus of FIG was 51 kPa measured by compressing in fiber direction at 1 Hz oscillation using a dynamic mechanical analyzer (DMA, DMA1-STAR, Mettler Toledo, USA).”
Quote in the paperNot a substantial claimMay combine two claims.
FindingreasoningThe fibers provided biochemical and microstructural cues that promoted cell adhesion and self-organization into a functional syncytium.
“In addition, these fibers also provided biochemical and microstructural cues, which promoted cell adhesion and self-organization into a functional syncytium.”
Quote in the paperSubstantial claim- Rests on an assumption: The fibronectin coating on gelatin fibers remains functional for integrin-mediated cell adhesion after EDC/NHS crosslinking and the printing process.
Probably not a substantial claim: may not pass S3 (load-bearing).
Findingown dataPDMS MTFs had an average thickness of 20 μm.
“the average PDMS thickness and tissue thickness for PDMS MTFs were 20 μm and 5 μm, respectively.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Findingown dataOscillation amplitude sweeps failed to quantify yield stress for the 0% fiber hydrogel inks because their values were too low.
“The yield stress for the 0% fiber hydrogel inks could not be quantified using the oscillation amplitude sweep tests as the value ranges were too low.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Findingown dataTissue on FIG MTFs averaged 10 μm in thickness.
“The average FIG scaffold thickness and tissue thickness for FIG MTFs were 70 μm, and 10 μm, respectively”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Findingown dataFor the 0% fiber inks, yield stress was instead derived from stress-shear rate plots that displayed a plateau as shear rate fell.
“Therefore, the yield stress for the 0% fiber hydrogel inks was calculated using the stress-shear rate graphs (Supplementary Fig. 2c) which showed a plateau region as the shear rate decreased.”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataNRVM tissue began spontaneous beating by Day 3, and iPSC-CM tissue by Day 5.
“After observing spontaneous beating of NRVM tissue at Day 3 and iPSC-CM tissue at Day 5”
Quote in the paperNot a substantial claimMay combine two claims.
Interpretationown dataImpulse propagation in the 3D printed ventricle scaffolds occurred along tissue alignment, which is determined by the printing direction.
“This confirms that impulse propagation in the 3D printed ventricle scaffolds also occurred along tissue alignment, which is determined by the printing direction.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Study design and conductown dataAt least three independent experiments were conducted for representative data and images, showing similar results.
“For representative data and images given in the figures, at least three or more independent experiments were conducted, showing similar results.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S2 (checkable).
InterpretationreasoningThis work suggests that 3D FIG ink printing will play an important role in promoting standardized and accessible bioprinting processes to recapitulate native organ geometries with microstructural precision.
“This work suggests that 3D FIG ink printing will play an important role in promoting standardized and accessible bioprinting processes to recapitulate native organ geometries with microstructural precision.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationThe animal protocol was based on previously published methods.
“and is based on previously published methods9”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [9] Feinberg, A. W. et al. Muscular thin films for building actuators and powering devices. Science 317, 1366–1370 (2007). source
Probably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone givenHarvard's Animal Care and Use Committee approved animal protocol 24-01-2.
“The animal protocol numbered 24–01-2 is approved by the Harvard Animal Care and Use Committee”
Quote in the paperNot a substantial claimClassified as probably not a substantial claim.
Study design and conductown dataThe study statistically compared viability, alignment, nuclear aspect ratio and angle distribution, contractility, and Ca2+ conduction anisotropy (longitudinal vs transverse) between Gel-Alg and FIG tissues.
“Statistical analysis was conducted to compare cell viability, tissue alignment, nucleus aspect ratio, nucleus angle distribution, tissue contractility, Ca2+ conduction anisotropy ratio in the longitudinal direction versus the transverse direction between tissues on the printed Gel-Alg hydrogel and the FIG scaffolds.”
Quote in the paperSubstantial claim
The paper relies on this without support.
The volumetric change measured by particle imaging velocimetry in the hiPSC-CM ventricle model is comparable to ejection fraction values reported for other in vitro ventricle models.
“We calculated our hiPSC-CM based ventricular volumetric change between peak contraction and peak relaxation to be 5.94 ± 1.66 % (n = 4, mean ± SEM, Fig. 4g), which is 2~5 and 8~20 times higher than values previously reported (ejection fraction) for NRVM and hiPSC-CM in vitro ventricle models, respectively13, 15, 32.”
The paper relies on this without support.
The fibronectin coating on gelatin fibers remains functional for integrin-mediated cell adhesion after EDC/NHS crosslinking and the printing process.
“Afterwards, we chemically crosslinked the fibers with N-(3-Dimethylaminopropyl)-Nʹ-ethylcarbodiimidehydrochlorideN-hydroxysuccinimide (EDC/NHS) and fibronectin (Supplementary Fig. 1e).”
The authors state this as an assumption.
The starting volume of the 3D printed ventricle can be estimated by assuming the shape of a half-oblate ellipsoid.
“assuming the shape of a half-oblate ellipsoid.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Fiber-reinforced composites show enhanced mechanical strength and fiber-induced cell alignment in various cell studies.
“Fiber-reinforced composites show enhanced mechanical strength and fiber-induced cell alignment in various cell studies[23, 26–30].”
From Development of silk microfiber-reinforced bioink for muscle tissue engineering and in situ printing by a handheld 3D printer (2025) · cites this paper as [27]
The Results sections “Printed 3D hydrogel scaffolds provide microscale anisotropy” and “Fiber infused gel scaffolds potentiate cellular alignment” report improved gel elasticity and greater cardiomyocyte alignment on fiber-infused scaffolds than on controls (page numbers unavailable). They do not establish enhanced mechanical strength specifically, or findings across various cell studies. (LLM, conf=0.85)
Experimental — accuracy not yet measuredThese methods oftentimes involve scaffolds which might interfere with tissue properties.
“In addition, these methods oftentimes involve scaffolds which might interfere with tissue properties (Patel and Birla, 2018; Chang et al., 2022; Choi et al., 2023; MacQueen et al., 2018; Michas et al., 2022; Xie et al., 2023; Williams et al., 2025).”
From Mini-hearts for disease modeling and drug testing – process optimization versus biological functionality (2026) · cites this paper as Choi et al., 2023
The Results sections on cellular alignment and functional anisotropy show that this method uses scaffolds and that scaffold design affects cardiac tissue organization, electrical propagation, and contraction. The paper presents those effects as beneficial guidance, not as evidence that scaffolds adversely interfere with tissue properties; that implication is unsupported. (LLM, conf=0.84)
Experimental — accuracy not yet measuredThe highest performing model currently reports an EF of about 6%, well below physiological levels (50%-70%).
“The highest performing model currently reports an EF of about 6%, well below physiological levels (50%–70%) (Choi et al., 2023).”
From Mini-hearts for disease modeling and drug testing – process optimization versus biological functionality (2026) · cites this paper as Choi et al., 2023
The Results section (‘Engineered 3D ventricle models exhibit cyclic contractility’; page unavailable) reports 5.94 ± 1.66% ventricular volumetric change for the authors’ model and says this exceeds values previously reported for comparable in vitro ventricle models. The copy does not establish that it is the highest-performing model overall or give the claimed physiological EF range of 50%–70%. (LLM, conf=0.86)
Experimental — accuracy not yet measuredPrevious studies demonstrated similar outcomes, including fiber-oriented cell alignment.
“This finding is consistent with previous studies that have demonstrated similar outcomes, including fiber-oriented cell alignment [23, 27].”
From Development of silk microfiber-reinforced bioink for muscle tissue engineering and in situ printing by a handheld 3D printer (2025) · cites this paper as [27]
Kupfer et al., Chang et al. and Choi et al. reported volumetric performance data, with Kupfer et al. additionally demonstrating Pressure Difference (ΔP) measurements.
“Nonetheless, Kupfer et al., Chang et al. and Choi et al. reported volumetric performance data, with Kupfer et al. additionally demonstrating Pressure Difference (ΔP) measurements (Kupfer et al., 2020; Chang et al., 2022; Choi et al., 2023).”
From Mini-hearts for disease modeling and drug testing – process optimization versus biological functionality (2026) · cites this paper as Choi et al., 2023
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
NRVM thin-film measurement is dated to both day 7 and day 5
MinorThe results identify the FIG muscular thin-film measurement as day 7, while the methods say the NRVM thin-film experiment was performed after five days of culture.
“The average contractile force of Day 7 NRVM tissues on FIG MTFs was 1661.48 ± 209.46 Pa”
“MTF experiment was performed after 5 days of NRVMs culture in a 3.5 mm petri dish.”
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Self-citationFibrous scaffolds for building hearts and heart parts
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Self-citationMyocyte Shape Regulates Lateral Registry of Sarcomeres and Contractility
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Self-citationSelf-Organization of Muscle Cell Structure and Function
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Self-citationJetValve: Rapid manufacturing of biohybrid scaffolds for biomimetic heart valve replacement
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Self-citationNuclear morphology and deformation in engineered cardiac myocytes and tissues
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Self-citationStructural Phenotyping of Stem Cell-Derived Cardiomyocytes
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Self-citationCardiac Tissues From Stem Cells: New Routes to Maturation and Cardiac Regeneration
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Self-citationCytoskeletal prestress regulates nuclear shape and stiffness in cardiac myocytes
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Self-citationSubstrate‐Wide Confined Shear Alignment of Carbon Nanotubes for Thin Film Transistors
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Conduction in cardiac tissue. Historical reflections
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Self-citationOpenPIV/openpiv-python: fixed windows conda-forge failure with encoding
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A Versatile Microporous Design toward Toughened yet Softened Self‐Healing Materials
Sun, F., Zhang, J., Liu, T., Yao, H., Wang, L., Meng, H., Gao, Y., Cao, Y., Yao, B., Xu, J., et al. (2024) · Advanced Materials · doi:10.1002/adma.202410650 · Cited by 157 · see citing works
Mechanical Regulation of Polymer Gels
Xu, C., Chen, Y., Zhao, S., Li, D., Tang, X., Zhang, H., Huang, J., Guo, Z., & Liu, W. (2024) · Chemical Reviews · doi:10.1021/acs.chemrev.3c00498 · Cited by 152 · see citing works
4D bioprinting of programmed dynamic tissues
Lai, J., Liu, Y., Lü, G., Yung, P. S., Wang, X., Tuan, R. S., & Li, Z. A. (2024) · Bioactive Materials · doi:10.1016/j.bioactmat.2024.03.033 · Cited by 117 · see citing works
“Enabling the production of shape-morphing cardiac patches that match well with the curved topology of the heart; production of dynamic ventricles recapitulating the dynamic motion of native counterparts [79,203,204] functionality of 4D-bioprinted constructs.”
BackgroundOh, B., Baek, S., Nam, K. S., et al. (2024) · Nature Communications · doi:10.1038/s41467-024-50264-6 · Cited by 84 · see citing works
“Such properties of 3D printing have led to its wide usage in biomedical applications for the multi-material patterning of tissue-like materials 6,7 , such as conducting polymers and hydrogels, into complex anatomical structures 8 and bioelectronic devices 9 .”
Self-citationDirect 3D‐Bioprinting of hiPSC‐Derived Cardiomyocytes to Generate Functional Cardiac Tissues
Esser, T. U., Anspach, A., Muenzebrock, K. A., Kah, D., Schrüfer, S., Schenk, J., Heinze, K. G., Schubert, D. W., Fabry, B., & Engel, F. B. (2023) · Advanced Materials · doi:10.1002/adma.202305911 · Cited by 75 · see citing works
Multimaterial cryogenic printing of three-dimensional soft hydrogel machines
Li, J., Cao, J., Bian, R., et al. (2025) · Nature Communications · doi:10.1038/s41467-024-55323-6 · Cited by 65 · see citing works
“Towards this goal, many interesting works have been reported, generally focusing on ink-by-ink solute regulation to cater to conventional solute-triggered formation mechanisms, such as polymerization 9 – 11 , sol-gel transition 12,13 , self-assembly 14 , and phase separation 15,16 .”
Zhou, W., Rahman, M. S. U., Sun, C., Li, S., Zhang, N., Chen, H., Han, C. C., Xu, S., & Liu, Y. (2024) · Advanced Materials · doi:10.1002/adma.202307805 · Cited by 65 · see citing works
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Sun, Y., Cui, J. T., Feng, S., Cui, J., Guo, Y., Chen, L., Gao, W., Lu, Z., Liu, F., & Zhang, B. (2024) · Advanced Materials · doi:10.1002/adma.202400103 · Cited by 57 · see citing works
Anderson–Fabry disease management: role of the cardiologist
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Instant assembly of collagen for tissue engineering and bioprinting
Gong, X., Wen, Z., Liang, Z., Xiao, H., Lee, S., Rossello‐Martinez, A., Xing, Q., Wright, T., Nguyen, R. Y., & Mak, M. (2025) · Nature Materials · doi:10.1038/s41563-025-02241-7 · Cited by 73 · see citing works
“Earlier bioprinting-based cardiac ventricles to achieve contractile pumping motions primarily relied on a cardiac cell layer scaf-folded on a pre-printed hydrogel shell 10,41 or used a cell-laden gelatin microgel-based bioink that hinders whole-tissue activities 42 .”
4D printing for biomedical applications
Mandal, A., & Chatterjee, K. (2024) · Journal of Materials Chemistry B · doi:10.1039/d4tb00006d · Cited by 55 · see citing works
Kumi, M., Ejeromedoghene, O., Sudane, W. D., & Zhang, Z. (2024) · European Polymer Journal · doi:10.1016/j.eurpolymj.2024.112906 · Cited by 49 · see citing works
Tough Hydrogels with Different Toughening Mechanisms and Applications
Xu, Z., Chen, Y., Cao, Y., & Xue, B. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms25052675 · Cited by 48 · see citing works
“It is worth noting that this supercapacitor has a stretchability of 900% with slightly reduced performance, even if it has defects.”
BackgroundMachine Learning in Gel-Based Additive Manufacturing: From Material Design to Process Optimization
Zhang, Z., Wang, Y., & Wang, W. (2025) · Gels · doi:10.3390/gels11080582 · Cited by 52 · see citing works
“These attributes enable applications ranging from scaffolds for tissue growth [10] and regenerative medicine [11] to controlled drug [12] release platforms and flexible wearables [13].”
Ren, C., Chen, W., Liao, Y. J., Huang, Y., Yu, C., Chen, T., Zeng, Q., Yang, Y., Huang, R., Liu, T., et al. (2024) · Advanced Materials · doi:10.1002/adma.202416432 · Cited by 49 · see citing works
Biomedical Application of Enzymatically Crosslinked Injectable Hydrogels
Nam, M., Lee, J.-W., & Cha, G. D. (2024) · Gels · doi:10.3390/gels10100640 · Cited by 50 · see citing works
“…hydrogels have been spotlighted as matrices for use in various biomedical applications [2,3], e.g., medical device interfaces [4–6], 3D cell culture platforms [7], bio-ink materials for use in 3D printing [8–10], conductive contact electrodes [11–14], and filler delivery reservoirs [15,16].”
BackgroundOrganoid bioinks: construction and application
Wang, F., Song, P., Wang, J., Wang, S., Liu, Y., Bai, L., & Su, J. (2024) · Biofabrication · doi:10.1088/1758-5090/ad467c · Cited by 42 · see citing works
“Encouragingly, certain findings suggest that ventricular models produced with this organoid bioink showcase biomimetic anisotropic electrophysiological and contractile attributes [102] (figures 10(F)–(I)).”
BackgroundAdvances of conductive hydrogel designed for flexible electronics: A review
Zhu, G., Javanmardia, N., Qian, L., Jin, F., Li, T., Zhang, S., He, Y., Wang, Y., Xu, X., Wang, T., et al. (2024) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2024.136115 · Cited by 47 · see citing works
Li, Y., Yang, S., You, Y., et al. (2024) · Chemical Engineering Journal · doi:10.1016/j.cej.2024.154357 · Cited by 40 · see citing works
Advanced Nanomaterials in Medical 3D Printing
Liu, H., He, L., Kuzmanović, M., Huang, Y., Zhang, L., Zhang, Y., Zhu, Q., Ren, Y., Dong, Y., Cardon, L., et al. (2023) · Small Methods · doi:10.1002/smtd.202301121 · Cited by 41 · see citing works
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4D Printed Protein‐AuNR Nanocomposites with Photothermal Shape Recovery
Yu, S., Sadaba, N., Sanchez‐Rexach, E., Hilburg, S. L., Pozzo, L. D., Altın, G., Liz‐Marzán, L. M., de Aberasturi, D. J., Sardón, H., & Nelson, A. (2023) · Advanced Functional Materials · doi:10.1002/adfm.202311209 · Cited by 37 · see citing works
SonoPrint: Acoustically Assisted Volumetric 3D Printing for Composites
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Ionic Hydrogel Sensors toward Next‐Gen Personalized Healthcare
Sun, Y., Tian, G., Deng, W., & Yang, W. (2025) · Advanced Materials · doi:10.1002/adma.202509122 · Cited by 50 · see citing works
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Tunable Macroscopic Alignment of Self-Assembling Peptide Nanofibers
Farsheed, A. C., Delgado, C. Z., Yu, L. T., Saeidifard, S., Swain, J. W. R., Makhoul, J. T., Thomas, A. J., Cole, C. C., Huitron, E. I. G., Grande-Allen, K. J., et al. (2024) · ACS Nano · doi:10.1021/acsnano.4c02030 · Cited by 34 · see citing works
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“…technology is the inclusion of 3D printing into tissue engineering and regenerative medicine applications, allowing the rapid and reproducible fabrication of complex biomimetic tissues or organs in vitro, such as 3D-bioprinted ventricles, corneal stroma, skin, bone, and cartilage tissue [130–133].”
BackgroundAnti-swelling and ROS-scavenging nanofiber hydrogel to prevent postoperative abdominal adhesion
Luo, W., Pan, X., Ren, C., Wang, T., Li, G., Jin, L., & Zhang, D. (2025) · Chemical Engineering Journal · doi:10.1016/j.cej.2025.164202 · Cited by 35 · see citing works
Hydrogel-based inks for extrusion 3D printing: A rheological viewpoint
Thareja, P., Swarupa, S., Ahmad, S., & Jinugu, M. E. (2025) · Current Opinion in Colloid & Interface Science · doi:10.1016/j.cocis.2025.101918 · Cited by 33 · see citing works
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Recent Development of Fibrous Hydrogels: Properties, Applications and Perspectives
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Kim, M. S., Choi, Y., & Lee, K. Y. (2025) · Polymers · doi:10.3390/polym17172337 · Cited by 35 · see citing works
“Conventional 3D printing, although acellular, has played a crucial role in creating patient-specific anatomical models, vascular grafts, and structural scaffolds that aid in pre-surgical planning, device testing, and the development of biomedical implants [5–7] (Figure 1b).”
Self-citationWu, Q., Xu, Y., Han, S., et al. (2024) · ACS Nano · doi:10.1021/acsnano.4c08896 · Cited by 23 · see citing works
Local governance and occupational choice among young people: First evidence from Vietnam
Tran, T. Q., Tran, A. L., Pham, T. M., & Vu, H. V. (2018) · Children and Youth Services Review · doi:10.1016/j.childyouth.2018.01.019 · Cited by 24 · see citing works
Towards more realistic cultivated meat by rethinking bioengineering approaches
Piantino, M., Muller, Q., Nakadozono, C., YAMADA, A., & Matsusaki, M. (2024) · Trends in biotechnology · doi:10.1016/j.tibtech.2024.08.008 · Cited by 24 · see citing works
Dynamic bioinks for tissue/organ bioprinting: Principle, challenge, and perspective
Ma, D., Lu, W. W., Liu, W., Liu, W., & Ruan, C. (2025) · Progress in Materials Science · doi:10.1016/j.pmatsci.2025.101527 · Cited by 24 · see citing works
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Morphology-guided cellular behavior modulation with 3D-printed engineered ECM
Liu, N. B., Shi, Y., Li, J., et al. (2025) · Cell Biomaterials · doi:10.1016/j.celbio.2025.100090 · Cited by 19 · see citing works
Harnessing native blueprints for designing bioinks to bioprint functional cardiac tissue
Akter, M. Z., Tufail, F., Ahmad, A., Oh, Y. W., Kim, J. M., Kim, S., Hasan, M. M., Li, L., Lee, D., Kim, Y. S., et al. (2025) · iScience · doi:10.1016/j.isci.2025.111882 · Cited by 20 · see citing works
“70 These scaffolds, printed in different patterns (parallel, angled, and perpendicular) createdistinct contractile patterns.”
Self-citationBioprinted Scaffolds for Biomimetic Applications: A State-of-the-Art Technology
Gebeshuber, I. C., Khawas, S., Sharma, R., & Sharma, N. (2025) · Biomimetics · doi:10.3390/biomimetics10090595 · Cited by 21 · see citing works
“Moreover, bilayer hydrogels embedded with fibroblasts and magnetic particles have enabled cyclic folding movements, simulating the contractile behavior of native tissues and offering platforms to study biomechanical cell responses [125].”
Shi, B., Zhu, T., Luo, Y., et al. (2024) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2024.136106 · Cited by 18 · see citing works
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Electrospun microfibers to enhance nutrient supply in bioinks and 3D-bioprinted tissue precursors
Neuhäusler, A., Rogg, K., Schröder, S., Spiehl, D., Zora, H., Arefaine, E., Schettler, J., Hartmann, H., & Blaeser, A. (2024) · Biofabrication · doi:10.1088/1758-5090/ad9d7a · Cited by 16 · see citing works
Zhang, H., Wang, P., Zhang, H., et al. (2024) · ACS Applied Materials & Interfaces · doi:10.1021/acsami.4c18098 · Cited by 16 · see citing works
Volume‐Metallization 3D‐Printed Polymer Composites
De-hai, Y., Chi, G., Mao, X., et al. (2024) · Advanced Materials · doi:10.1002/adma.202403088 · Cited by 16 · see citing works
Designed wrinkles for optical encryption and flexible integrated circuit carrier board
Zhong, S., Zhu, Z., Huo, Q., Long, Y., Gong, L., Ma, Z., Yu, D., Zhang, Y., Liang, W., Liu, W., et al. (2024) · Nature Communications · doi:10.1038/s41467-024-50069-7 · Cited by 18 · see citing works
“34 – 38 and Supplementary Movies2, 3).”
Yu, K., Yang, L., Zhang, S., et al. (2024) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2024.133417 · Cited by 16 · see citing works
Bakırcı, E., Adib, A. A., Ashraf, S. F., & Feinberg, A. W. (2025) · Biofabrication · doi:10.1088/1758-5090/adb7c3 · Cited by 17 · see citing works
“For example, the myofibers in the ventricular wall of the heart form a helical architecture that interweaves around both chambers [33–36].”
Multidirectional Filamented Light Biofabrication Creates Aligned and Contractile Cardiac Tissues
Jones, L. S., Filippi, M., Michelis, M. Y., Balciunaite, A., Yaşa, Ö., Aviel, G., Narciso, M., Freedrich, S., Generali, M., Tzahor, E., et al. (2024) · Advanced Science · doi:10.1002/advs.202404509 · Cited by 15 · see citing works
Yao, Z., Kuang, Y., Wu, H., Lundqvist, E. M., Fu, X., Celt, N., Pei, J., Yee, A. F., & Ardoña, H. A. M. (2024) · Advanced Materials · doi:10.1002/adma.202312231 · Cited by 15 · see citing works
Self-citationAdvancements in robotic arm-based 3D bioprinting for biomedical applications
Li, K., Huang, W., Guo, H., Liu, Y., Chen, S., Liu, H., & Gu, Q. (2023) · Life Medicine · doi:10.1093/lifemedi/lnad046 · Cited by 16 · see citing works
Advances and application potential in the research of silicate mineral-based 3D printing materials
Zhao, Q., Gao, C., Zhang, Y., & Zhang, Y. (2025) · Progress in Materials Science · doi:10.1016/j.pmatsci.2025.101450 · Cited by 17 · see citing works
3D printing in biofabrication: From surface textures to biological engineering
Ma, Z., Wang, J., Qin, L., & Chortos, A. (2024) · Chemical Engineering Journal · doi:10.1016/j.cej.2024.156477 · Cited by 15 · see citing works
A critical review on advances and challenges of bioprinted cardiac patches
Zhang, X., Zhao, G., Ma, T., Simmons, C. A., & Santerre, J. P. (2024) · Acta Biomaterialia · doi:10.1016/j.actbio.2024.09.056 · Cited by 17 · see citing works
Programmable Anisotropic Hydrogels for Biomedicine: From Precise Design to Advanced Applications
Zhang, F., Gao, Z., Sun, L., Jin, Z., Liu, Y., Tong, M., Zhao, Y., Jiang, X., & Gu, X. (2025) · ACS Applied Materials & Interfaces · doi:10.1021/acsami.5c12804 · Cited by 17 · see citing works
Zeng, Q., Yang, Y., Wang, H., Tan, Y., Wang, Z., Chai, M., Lu, Z., He, S., Yang, H., Zhang, J. Z., et al. (2025) · Biomaterials · doi:10.1016/j.biomaterials.2025.123573 · Cited by 19 · see citing works
Liu, S., Wang, S., Sang, M., et al. (2024) · Chemical Engineering Journal · doi:10.1016/j.cej.2024.152109 · Cited by 13 · see citing works
Digital light 3D printed fast- and controlled degradation of covalent hydrogel networks
Chen, L., Cui, J., Cui, J., et al. (2023) · Chemical Engineering Journal · doi:10.1016/j.cej.2023.148027 · Cited by 13 · see citing works
Injectable Immune-Regulating Mesenchymal Stem Cells Hydrogel for Localized Scleroderma Treatment
Liu, R., Kong, B., Lu, J., Zhao, Y., & Sun, L. (2025) · ACS Nano · doi:10.1021/acsnano.5c10112 · Cited by 13 · see citing works
Thomas, D., & Wu, J. C. (2025) · iScience · doi:10.1016/j.isci.2025.113003 · Cited by 14 · see citing works
“Despite the success in obtaining high degree of anisotropy and stiffness using electrospinning and 3D-printed free-standing structures, 173 one disadvantage of scaffold pre-fabrication using synthetic matrices crosslinked prior to cell seeding is that it may impede progressive stretch yielding…”
Nanocomposite Hydrogels and Micro/Nanostructures for Printing Organoids
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“(Choi et al., 2023).”
Influential
- Authors
- Choi, S., Lee, K. Y., Kim, S. L., MacQueen, L. A., Chang, H., Zimmerman, J. F., Jin, Q., Peters, M. M., Ardoña, H. A. M., Liu, X., Heiler, A.-C., Gabardi, R., Richardson, C., Pu, W. T., Bausch, A. R., & Parker, K. K.
- Affiliations
- Harvard University (US)
- Boston Children's Hospital (US)
- Technical University of Munich (DE)
- Max Planck School Matter to Life (DE)
- Corresponding author
- Parker, Kevin Kit
- ORCID
- Venue
- Nature Materials
- Published
- 2023-07-27
- Volume
- 22
- Issue
- 8
- Pages
- 1039-1046
- ISSN
- 1476-1122 (ISSN-L)
- 1476-4660
- PMC
- PMC10686196
- OpenAlex
- W4385330852
- Cited by
- 212
- References
- 55
- Citations by year
- 2026: 55 · 2025: 75 · 2024: 69 · 2023: 12 · 2018: 1
- FWCI
- 16.62
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Nature Materials (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- U.S. Department of Health & Human Services | NIH | National Center for Advancing Translational Sciences (UH3HL141798, UG3TR003279)
- NSF | Directorate for Mathematical & Physical Sciences | Division of Materials Research (DMR-1420570, DMR-2011754)
- American Chemical Society
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Hydrogels: synthesis, properties, applications
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- 3D bioprinting, hydrogel scaffolds, cardiomyocyte alignment, human cardiomyocytes, alginate-gelatin hydrogel, sol-gel transition, ink rheology, electrophysiological properties, cardiac tissue engineering
- MeSH terms
- Gelatin, Humans, Hydrogels, Tissue Engineering, Myocytes, Cardiac, Tissue Scaffolds, Printing, Three-Dimensional
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