#ml s6 criticism

[fernsnouveau]

The design """update""" makes sense when you consider that the showrunners don't want the audiences to like Chat Noir anymore.

Sharp Cheddar

Vs.

This

The round boy feels too safe...

yeah

[miraculous-lesbians]

Me, who knows nothing about ML but knows about RGU: "Yes, this makes sense..."

But! in all seriousness love to see new blogs! Lookin' forward to your critical look at ML!

Thank you so much! I'm excited to dive into ML more as well~ I'm debating on sitting down and watching ML (because most/all of my knowledge come from Tumblr), buuut that's a lot for me to watch lol.

Btw, Chloe and Nanami just generally fit the "mean, rich blonde popular girl the narrative tends to make fun of." But where Nanami is generally sympathized by the writers, who understand she's a teen girl who's still developing her frontal lobe, Chloe does not get this at all. Her treatment is so over the top and silly, it's hard to genuinely believe what the writers are doing imo.

[fernsnouveau]

Anyway maybe people should be allowed to be upset that this show, which spent the first 4+ seasons setting itself up as a narrative where a controlling child abuser is the main villain, and lack of agency as a victim of parental abuse was a major problem to overcome for one of the title characters, did a jarring thematic 180° near the end of the first major story arc (that was originally supposed to be the whole show), and turned into blatant victim-blaming and abuse victim grooming propaganda material.

And the showrunner is also engaging in this victim-blaming and abuse apologism rhetoric and insulting people on Twitter if they question it. Which is... kind of really damning, at this point.

People who are upset about this aren't just "hating on the show" for the heck of it. This is, in fact, reasonable basis to be upset. People are allowed to voice their disappointment after the show wasted their sincere emotional attachment (yes, you should be allowed to have sincere emotional attachments even if it's something silly like a mediocre magical girl cartoon) and spat in the face of its own story and themes and characters.

Also a worrisomely significant segment of the fanbase has gone very mask off in starting to regurgigate and agree with this abuse apologia rhetoric, encouraged by the canon validating it.

[miraculousalt]

So when S6 starts Adrien wont know yet that Ladybug didn get the Butterfly miraculous cuz she'll probably just tell suhan, Alya n maybe Luka. So Chat Noir will likly ask her for a reveal in ep 1 cause thats wha they always said, they reveal when its save.

Is there evn any point in hoping tha she ll go bout that any other way than jus lyin her ass off again, stall for time with excuse afta excuse until Lila will attack as Butterfly & Chat has to find out the worst way again tha she lied to him n keepin his hopes up? Her jus keepin him 'happy' by telling him they ll finlly getta know each other soon just for her to never have had that intention in the first place?

Wow, that sounds like Ephemeral all over again. Great, cant wait.

[actually-factually-anonymous]

Indeed, people overly praising Marinette for giving Adrien his free will back are missing that not enslaving another person is hardly an exceptional feat of morality.

What, is Adrien not entitled to basic free will? If he is, then Marinette passing his rings back to him is the bare minimum for any character who isn't a villain.

Except...

There's actually reason why she shouldn't. As you point out: she hasn't told him about those rings, about what they mean to him. About how easily someone could use them against him to turn him into their puppet for life, or how those relatively fragile bits of metal could end his life instantly if they were broken somehow.

But choosing to give them to him without telling him what they are: Marinette has chosen the worst option between keeping them herself and giving Adrien he needs: giving him the rings without the knowledge of what makes them vital or why he needs to protect them. The only worse possible option here I can think of would be for her to go full Yandere and use them herself.

Like so many of Marinette's choices since S5, it's utterly indefensible. Giving Adrien he needs to know as well as the rings is understandable and ideal. Not giving Adrien any knowledge is questionable at best, but as long as Marinette is holding onto the rings for whatever reason (protection maybe)- it's not careless to the point of potential manslaughter.

But giving him them rings without the knowledge about what they are and why he needs to protect them is just... insanity. The only viable reason and defense I could find for it across the internet was that Marinette simply doesn't know what the rings are, and that she either failed to cotton on to the full details of Felix's bullshit play or that she doesn't know what Adrien's Amok is...

Though with Nathalie and Felix easily contactable for her now, even that excuse really fails to hold water unless we assume "find out where my 'beloved' boyfriend's vital two in one lifesupport and free will is" was never once a priority for her during those months between her getting the rings and... any after that really.

Marinette is praised for alot of unwarranted things, but to this day the thing that takes the cake for me is people praising Marinette into the heavens for giving Adrien his rings back after Gabriel's death

People act like that's Marinette acting on god-like benevolence and the purest heart ever, when, like, what is she even being praised for?

For not withholding Adrien's literal control remote from him? Not wearing it herself and right away enslaving him?

Praising Marinette into the heavens for giving the rings back implies that people think that it would have been fine if Marinette had cried over this "hard decision" for the next season. And that's just pathetic at this point.

Not to mention that she objectively did nothing in the final battle to save Adrien. It was Gabriel who used the wish for Adrien's good, Marinette instead said he has to face whatever consequences as orphan now that's coming his way. So Gabriel helped Adrien, Marinette was willing to have him pay every price cuz it aint her.

Then Marinette being so uninterested in the plot lead to Bug Noire almost killing Adrien because she didn't know that she would have destroyed Adrien's amok rings alongside the Miraculous rings on Gabriel's hand. So once again, Gabriel had to save Adrien from Marinette by keeping her from using Cataclysm.

Marinette also isn't the one to take the amok rings away from Gabriel before he dies. Gabriel himself had to give Adrien free and prevent that his amoks will vanish with him.

What the hell is Marinette being praised for in the s5 finale for being Adrien's White Knight? She didn't save him in anyway. She almost killed him, she was willing to sign him up to entirely for his family's sins as orphan because "it's the right thing to do", and she didn't take Adrien's ring from Gabriel

Gabriel did all the saving and protecting, Marinette is just as always taking all the credit.

So what's Marinette being glorified for? For not instantly enslaving her abused and now orphaned boyfriend? Cause that's all the saving of Adrien Marinette did in the end, I guess: saving him from her herself enslaving him for no fucking reason

And is that REALLY something Marinette needs to be praised and made out into a saint for? Cause that's beyond pathetic, even downside offensive at this point

---

At this point, I think the show could show Marinette successfully wiping her ass and the stans would praise her for how talented she is. Everything Marinette does has to be the most amazing show of skill or the most kindhearted thing anyone has ever done in the stans’ eyes because the pedestal they’ve built for Marinette has grown so insanely tall. At this point, there’s nothing she could do that would break that illusion of perfection, I think. Even if she had kept those rings, the stans would have defended her noble decision to keep Adrien safe from people who’d take advantage of him, like Marinette herself hasn’t been taking advantage of him for free therapy for as long as they’ve been dating, no remote controls needed.

And that’s the thing with Marinette not keeping Adrien’s remote control; she doesn’t need it for Adrien to do whatever she wants. Adrien is so dependent on her that his life’s goal is just to be with her. Marinette knows Adrien is completely devoted to her, it's why she was so willing to send Zoé to confess her love to Adrien, because she knew Adrien would refuse Zoé and stay with her instead. Marinette gave up nothing when she gave those rings to Adrien, her control over him is still absolute.

Meanwhile, the fandom is celebrating and romanticizing this shit. I have seen more than one person consider Marinette giving those rings to Adrien to be like a proposal/promise ring situation, where Marinette doing the bare minimum of giving the sapient mind control robot servant his remote control is treated as an ultimate gesture of love. Because truly Marinette must love Adrien if, instead of ordering him to please every single one of her whims and needs, she grants him the magnificent honor of choosing to do so instead, while already knowing he will. How loving, how caring, how selfless.

Please ignore the reality that this entire setup relies on Marinette keeping guard over Adrien and his amoks indefinitely, because Adrien lacks the knowledge to do so himself. Please ignore the reality that, if Adrien, Marinette or both ever fell out of love and ended the relationship, Adrien would be left incredibly vulnerable because he doesn’t know about the rings being amoks. Marinette is counting on Adrien staying with her for the rest of his life, because she’s taken conservatorship over him and his free will. She has made it her job to be her boyfriend’s keeper.

[ct-multifandom]

Miraculous s6 Trailers Breakdown

Before I get into it, if it calms anyone’s nerves, I think this poster is a case of questionable graphic design. The weird official render style makes the characters look different from normal, plus Chat, Carapace, and Rena are all squished to match Ladybug’s proportions on the poster. This is an interesting choice from the designer: making the smallest character in the group take up the entire height and corresponding width of the poster and then awkwardly smushing the others in next to her. That’s why Carapace looks more short and stout than usual, and why Chat and Rena are kinda elongated. It doesn’t help that they shrunk Rena to accommodate her fox ears.

I’m liking the little updates to the hero suits. I honestly wasn’t expecting to see change for anyone but Ladybug and Chat, so now I’m excited to see everyone else. I love the changes on Carapace. They took his old, pretty simple suit and added a lot of new textures like the chest plate and wrist guards, and they gave him fingerless gloves and neck speakers. This gives me hope that they’ll update other designs that get criticized for just being a bodysuit with a decal on it like Ryuko and Viperion’s into something more interesting.

Rena is very similar, but her ears are bigger and pointier, the flute has been redesigned, her palms are white now, she has big poppin’ eyeliner, and her tail seems to be dark. Some people noted that they seem to be making Alya’s skin look darker after accusations of whitewashing her, so I hope they’ll finally give Pegasus the same treatment.

I’ve seen people say LB and CN must’ve taken influence from their reverse selves for their current designs, and CN seems to have some PV nods like his new shirt collar. Ladybug’s design feels more mature and is probably a response to criticism of her looking too simple as a character with a fashion interest among other superheroes with more detailed designs.

I didn’t include the poster of Marinette’s outfit, but overall I like it. I agree with people saying the bow on her shirt feels extra, but I like her emonette jacket. Her tights under jorts remind me of Mylene’s outfit, making me wonder if there’s some character influence there, or if they’ll be changing Mylene’s clothes so they don’t look too similar.

First look at Alya! Yay! I love her new haircut. It reminds me of Haru from persona 5. She seems to be wearing the same shirt, which is lame. Maybe the rest of her outfit has changed, or maybe they’re not fixing what isn’t broke. It’s disappointing, though, that Adrien’s outfit looks the same. I can only dream that they’re leading up to a makeover scene where he realizes he’s allowed to pick out his own clothes now, but alas.

Here Adrien is jogging with a new character (?) who has a prosthetic leg. There’s no good look at their face and even this screenshot sucks. If this is the same scene as Marinette with binoculars, then we might be getting a rare moment of a new character being introduced through Adrien rather than Marinette, perhaps someone he knows from rich people stuff or whatever nebulous and dubiously canon sports he participates in.

This frame appears after a camera pan right from a modernized remodel of the school. Before, there were average city blocks around the school, so whatever this is might be part of an expanded campus built for Bustier’s utopian academy.

I think I’ve written in a post before that I hope ML does something to commemorate the Paris 2024 Olympics, so I’m thrilled to see an Olympian akuma in the trailer! Perhaps this character is a celebrity cameo, but idk anything about Team France and its stars. It’s possible this scenario will be a special episode as well, but maybe more like the plastic special than the world specials. I hope they have to beat her by competing in games like Penalteam but with less cheating. It’d be awesome if the other heroes helped represent events that fit their characters. An episode like that would be a fun and light way to introduce the new animation and hero designs without diving into deep overarching drama and lore.

Perhaps the character jogging with Adrien is connected to this episode since they have a prosthetic running leg and seem athletic.

Here’s our first look at Lila’s akumatization. Her mask thing is very pretty and soft looking compared to Gabriel’s sharp, geometric, simple one. It reminds me of a masquerade mask with all the swirls. Maybe there’s some visual symbolism here of Lila being a more complex butterfly user than Hawkmoth. We also see her shooting some sort of beam at CN from the mask, maybe separate from the akuma’s power. I hope they’ll revisit the somewhat-retconned bloodbending power we saw Hawkmoth use on Evillustrator and Pixelator and explore the full extent of the butterfly’s power over their akumas.

Tomoe? Astruc said he wants Kagami to fight her and take her down further down the road, and I believe she may be a bigger villain than they’re making her out to be now. Lila is great and all, but an adult threat who owns a tech megacorp and is one of the only surviving members of Gabriel’s creepy billionaire sentibaby cult seems like a scarier enemy than the pathological liar from class. We don’t know Lila’s deal yet, but I imagine her villain motives are probably something deeply personal. Meanwhile, Tomoe might be trying to reach some greater goal that Gabriel failed to achieve. Both she and Lila practically had all the fruits of his evil labor fall into their laps without lifting a finger, so they might have an interesting dynamic as rival villains or fake allies.

Okay this is from the London special. The synopsis says someone found Ladybug’s identity to take her down, and she and Bunnyx have to solve the mystery and reverse the events. This masked person looks like they may be the enemy, and their design is themed after the rabbit miraculous. People have been kinda assuming that it’s just Lila, but everyone in the audience knows she’s the next big villain right now, so it feels like a cop-out to frame this special as a mystery. It could still be her, and it’s just a mystery to the heroes, but what if the enemy is someone else? If they’re using the rabbit miraculous, Lila wouldn’t have access to it during season 5 unless she stole it somehow and gave it back unnoticed.

My theory is that the villain is Tomoe who got the miraculous from Gabriel. The trailer shows catbug looking in on Adrien and Kagami in their white rooms, so maybe during that time, Gabriel sent rabbit!Tomoe after Ladybug to ensure his victory in the upcoming finale, and maybe she has her own motives as well.

I recall Zag releasing concepts of some hero or spy characters from London a long time ago, and I thought they’d be for the London world special like Eagle and Lady Dragon, but maybe they’re for a different show or got scrapped in favor of focusing on the main miraculous plot (which honestly, good. Please focus on the existing characters)

Finally, poor Nooroo in Lila’s creepy catacomb bunker. I hope her jumpscare reaction at the end of s5 wasn’t just the sight of him. It would feel odd after the latest of Marinette’s hero friends were totally chill seeing kwamis for the first time. He’s in some bubble here. Maybe it’s stopping him from sharing things he’s not allowed to say, or maybe Lila is forcing him to stay put? I’m also curious about the angle and what that metal bar is a part of. Is Lila sitting under a table?

Anyway! TLDR I like the new designs overall and I’m excited to see everyone else. I’m also excited at the prospect of an Olympics episode and I think Tomoe may be the next big villain next to or even surpassing Lila.

[yeet-noir]

hey, i just wanted to ask what you think of s5 so far? are you liking, does it meet ur expectations or are you underwhelmed?

It's hard to have conclusive thoughts about certain aspecrs without the whole season being out. I will say it's been frustrating to have networks air episodes so far out of order and no consistent schedule, but that's always been ML 🫠 The thing really hurting from that is the activity from the fandom on Tumblr imo 🥲

SPOILERS FOR S5 EPISODES 1-11!!!!!

With that said I have been enjoying the first 11 episodes (I believe it's been 11 at least in order right hahaa)!. I was one of the people that was not very critical to how episodic it was previous, but I am delighted to see the big plot points unfold! My expectations for season 5 (and please if somebody who has read the leaks or watch the episodes out of order keep comments to yourself about my speculations PLEASE) is that we will have a reveal and Monarch will be defeated. Soooooooo hopefully those will play out well 😬

Honestly the only thing the show has done this that I was skeptical about was have Adrienette confession. I always hoped we would have the identity reveal before that and s6 would have post reveal pre-relationship shenanigans. BUT what is making up for it is how slow burn it is I do not want them to rush and date. I want them to slowly build their comfort level in their relationship so I am happy that is happening. I predict (again please NO LEAK COMMENTS sorry but anytime I speculate somebody gotta mention "well according to the bible 🤓") that Marinette will only be able to say I love you to Adrien when she realizes he is Chat Noir which would be very cute.

And I lied there is a thing I wasn't very happy about and it was Marinette and Adrien having their Miraculous revoked by their Kwamis and them giving it to Alya and Zoé. It felt too weird... and forced? Ladynoir never really addressed it after... Nobody seemed bothered that their fav superheros just left.... Marinette never questions why Chat Noir had to go... and Zoé was such a random person to receive it and I am just not invested in her storyline so it took up too much time when the focus should of been the Love Square. I am happy part 2 wrapped that up and we got to see Marinette in action with no suit THAT WAS AWESOME. And Adrien taking the earrings ICONIC. So while I didn't enjoy part 1 the ending of part 2 really made me feel good.

Sorry I am rambling I appreciate being asked 🥹 I usually keep most my thoughts in a private GC especially with bible leakers sniping commentary posts it is hard to share. I love this show so much it's my guilty pleasure and I still have a positive outlook on it! I just hope in my soul that the identity reveal is god-tier and that their relationship start is to my taste. I also look forward to the Agreste plotline and my little monster Adrien senti-plot cause whether people like it or not it is very clear he is one and love that for him <3

Again thanks for the ask!! Sorry is this was all over the place too

[flightfoot]

Tough ask, maybe one that isn't fair but hey you are the ML guru, I apologize if it is.

What makes someone an ML 'Salter'?

I had someone call me a salter before, but then I the 'They would write Marinette as Velma' ask and essh, no.

My concern has always been how the show seemed to deviate from its own one-time core values or treat characters as disposable props for singular points. Is that salty?

I know all shows have slight OOC moments for plot, but degree varies. I think it is also safe two say prople approach these moments differently. Dome people acknowledge them, others weave narrative justification. One camp will accuse the other of being too narrow and looking for any excise at all to bash the characters/writers. The other will say the counterpart will do anything to justify the text, even read it while squinting with one eye covered and standing on their head.

But when I see something like the homework situation- something made ludicrous by existing since kindergarten unchecked, and we have Chloé(yes I know discourse lightning rod) about to be saddled with 9 years of makeup work, that seems for ML to be the perfect blend of comedic commuppanance. Chloé buried under a pile of books and papers is a great image, and perfect kharma for dodging so long (even if enabled by literally everyone)

So when Marinette -the protagonist- jumps up and starts railing about how it isn't enough. It feels... Gross? It doesn't feel like Marinette. Is she having guilt issues about facing zero consequences for losing the miraculous? We get nothing, but it's nails on a chalkboard and very much feels like someone else speaking from Marinette's mouth.

Am I going to go out and write Marinette-as-Velms fic? No. I *like* Marinette. Marinette is not mean spirited. That's the very problem I have with the interaction here.

If I were to write anything it would focus forward. Chloé buried under her extra work, visibly struggling because while the school assigned all the work and tests, they gave no plan or structure for the learning (now that is a good school critique)

Marinette feels for her, because Chloé is in full on Diktator-roof-meltdown. Sure there is satisfaction at first, but Marinette doesn't revel in Misery. She inquires, which leads to a chain of events. Chloé fobbing her off->admitting she needs help->Marinette tries to build her a study plan-> Chloé trying to get Marinette to do some of her work->Marinette realizing and rejecting this->Chloé coming back defeated again, submissive, just asking Marinette to check her work->Marinette realizing quickly that Chloé has been following her plan(though she gets a lot wrong) and studying->Marinette offering to help Chloé study for the subject-aptitude tests only, Chloé has to do all the coursework on her own.->Chloé finally coming to terms with give-and-take, and thanking Marinette. Maybe even apologizing. This would either be in time-crunch or carried over into S6 as a background arc.

This puts Marinette ascendent, demonstrates her core strengths of intelligence,planning, and compassion, has her interact with her former nemesis from a position of power while showing the difference between them (Does not abuse this position or revel in the disparity) shows boundaries being set, and keeps with the overall theming of hope over despair.

So is that 'salty'?

Okay so, as far as "salt" goes, it tends to have two different meanings that stem from the same root: the first being intense criticism of something, being unhappy with it in some way. And then a more specific meaning, referring to the way "salt" and "salter" were being used when the term first started being used a lot in the ML fandom, to describe the group and conceit that Miraculous as a show was being horribly unfair to Marinette, that her friends and family were all awful to her and needed to be punished while Marinette got what she "deserved" in terms of getting a new group of friends and family who would happily hurt anyone she was mad at and follow her orders without question.

For the record, when I rail at how terrible salters are, I'm talking about the latter category. I don't necessarily want to hear people go on too much about Miraculous being bad or the characters being bad, but some criticism is fair and reasonable. It's just the criticism that spawned out of Chameleon and the entire saltdom that formed around it that tended to not be.

Technically, what you're describing, being unhappy with the way the show did something and wanting to change it, would be classified as salt. HOWEVER, it's not the kind that people generally have a problem with - there's a reason there's been discussion of trying to get "critique" to catch on for that instead, when it's a saner form of criticism that isn't just hell-bent on revenge.

Also something to note with what you're laying out - you're looking to make the situation better, to address it and doing more of a fix-it, rather than just trying to burn the characters at the stake. I think that's what got the people who initially joined up with the salt in the early days, when it was billing itself as being "progressive" and "the right thing to do to address the problems of the show" to slowly bow out of the saltdom. Some people joined up with it because it criticized problematic elements of the show and they wanted to join with what appeared to be the more morally, socially conscious side of the fandom. Some ventfics made sense in that context, but once it became clear that was all it was, and that it wasn't actually setting out to fix things, to make things better, a lot of that group left the saltdom.

I don't personally have a problem with Marinette being upset that Chloe didn't get a greater punishment - she's got plenty of reason to be upset with Chloe and to believe that Chloe will wriggle her way out of any punishment she's given anyway - but your points are fair and I like the way you're seeking to address and fix the situation, laying out an alternative scenario, rather than, you know, burning everything to the ground.

That's a key difference - you seem to be aiming for a fix-it scenario rather than a bashing one.

[generalluxun]

I still say S1-2 at least disguised this? I am running on inference and circumstantial evidence but I think S1-2 were 'Maximum studio/production/investor interference' era. And for once the Execs might not have actually known how to write an original and compelling story, but they could point to other shows in the genre and say 'It should be like that! Make it more like that!' and it helped!

Up to and including the S2 finale like or hate specific moments, the overall arc was towards a very solid sentai show with an interesting cast of characters(you may like or dislike any particular one, but there were choices and they were fun)

Then S3 served, S4 hit the guard rail, and S5 went over a cliff.

Still don’t have the brain juice for a more comprehensive post but the show’s unfailing pattern of having the heroes take one step forward and two steps back during every part of their “journey” feels like it was meant to mock them and the ideals they stood for especially in retrospect when you know how the arc ends

[3deinc]

3D bioprinting of collagen to rebuild components of the human heart

For biofabrication, the goal is to engineer tissue scaffolds to treat diseases for which there are limited options, such as end-stage organ failure. Three-dimensional (3D) bioprinting has achieved important milestones including microphysiological devices (1), patterned tissues (2), perfusable vascular-like networks (3–5), and implantable scaffolds (6). However, direct printing of living cells and soft biomaterials such as extracellular matrix (ECM) proteins has proved difficult (7). A key obstacle is the problem of supporting these soft and dynamic biological materials during the printing process to achieve the resolution and fidelity required to recreate complex 3D structure and function. Recently, Dvir and colleagues 3D-printed a decellularized ECM hydrogel into a heart-like model and showed that human cardiomyocytes and endothelial cells could be integrated into the print and were present as spherical nonaligned cells after 1 day in culture (8). However, no further structural or functional analysis was performed.

We report the ability to directly 3D-bioprint collagen with precise control of composition and microstructure to engineer tissue components of the human heart at multiple length scales. Collagen is an ideal material for biofabrication because of its critical role in the ECM, where it provides mechanical strength, enables structural organization of cell and tissue compartments, and serves as a depot for cell adhesion and signaling molecules (9). However, it is difficult to 3D-bioprint complex scaffolds using collagen in its native unmodified form because gelation is typically achieved using thermally driven self-assembly, which is difficult to control. Researchers have used approaches including chemically modifying collagen into an ultraviolet (UV)–cross-linkable form (10), adjusting pH, temperature, and collagen concentration to control gelation and print fidelity (11, 12), and/or denaturing it into gelatin (13) to make it thermoreversible. However, these hydrogels are typically soft and tend to sag, and they are difficult to print with high fidelity beyond a few layers in height. Instead, we developed an approach that uses rapid pH change to drive collagen self-assembly within a buffered support material, enabling us to (i) use chemically unmodified collagen as a bio-ink, (ii) enhance mechanical properties by using high collagen concentrations of 12 to 24 mg/ml, and (iii) create complex structural and functional tissue architectures. To accomplish this, we developed a substantially improved second generation of the freeform reversible embedding of suspended hydrogels (FRESH v2.0) 3D-bioprinting technique used in combination with our custom-designed open-source hardware platforms (fig. S1) (14, 15). FRESH works by extruding bio-inks within a thermoreversible support bath composed of a gelatin microparticle slurry that provides support during printing and is subsequently melted away at 37°C (Fig. 1, A and B, and movie S1) (16).

The original version of the FRESH support bath, termed FRESH v1.0, consisted of irregularly shaped microparticles with a mean diameter of ~65 μm created by mechanical blending of a large gelatin block (Fig. 1C) (16). In FRESH v2.0, we developed a coacervation approach to generate gelatin microparticles with (i) uniform spherical morphology (Fig. 1D), (ii) reduced polydispersity (Fig. 1E), (iii) decreased particle diameter of ~25 μm (Fig. 1F), and (iv) tunable storage modulus and yield stress (Fig. 1G and fig. S2). FRESH v2.0 improves resolution with the ability to precisely generate collagen filaments and accurately reproduce complex G-code, as shown with a window-frame calibration print (Fig. 1H). Using FRESH v1.0, the smallest collagen filament reliably printed was ~250 μm in mean diameter, with highly variable morphology due to the relatively large and polydisperse gelatin microparticles (Fig. 1I). In contrast, FRESH v2.0 improves the resolution by an order of magnitude, with collagen filaments reliably printed from 200 μm down to 20 μm in diameter (Fig. 1, I and J). Filament morphology from solid-like to highly porous was controlled by tuning the collagen gelation rate using salt concentration and buffering capacity of the gelatin support bath (fig. S3). A pH 7.4 support bath with 50 mM HEPES was the optimal balance between individual strand resolution and strand-to-strand adhesion and was versatile, enabling FRESH printing of multiple bio-inks with orthogonal gelation mechanisms including collagen-based inks, alginate, fibrinogen, and methacrylated hyaluronic acid in the same print by adding CaCl2, thrombin, and UV light exposure (fig. S4) (15).

We first focused on FRESH-printing a simplified model of a small coronary artery–scale linear tube from collagen type I for perfusion with a custom-designed pulsatile perfusion system (Fig. 2A and fig. S5). The linear tube had an inner diameter of 1.4 mm (fig. S6A) and a wall thickness of ~300 μm (fig. S6B), and was patent and manifold as determined by dextran perfusion (fig. S6, C to E, and movie S2) (15). C2C12 cells within a collagen gel were cast around the printed collagen tube to evaluate the ability to support a volumetric tissue. The static nonperfused controls showed minimal compaction over 5 days (Fig. 2B), and a cross section revealed dead cells throughout the interior volume with a layer of viable cells only at the surface (Fig. 2C). In contrast, after active perfusion for 5 days, C2C12 cells compacted the collagen gel around the collagen tube (Fig. 2D), demonstrating viability and active remodeling of the gel through cell-driven compaction. The cross section showed cells alive throughout the entire volume (Fig. 2E), and quantitative analysis using LIVE/DEAD staining confirmed high viability within the perfused vascular construct (Fig. 2F). Others have 3D-bioprinted vasculature by casting cell-laden hydrogels around fugitive filaments, which become the vessel lumens (4, 5). In comparison, we directly print collagen to form the walls of a functional vascular channel, serving as the foundation for engineering more complex architectures.

Engineering smaller-scale vasculature, especially on the order of capillaries (5 to 10 μm in diameter), has been a challenge for extrusion-based 3D bioprinting because this is far below common needle diameters. However, at this length scale, endothelial and perivascular cells can self-assemble vascular networks through angiogenesis (17). We reasoned that the gelatin microparticles in the FRESH v2.0 support bath could be incorporated into the 3D-bioprinted collagen to create a porous microstructure, specifically because pores on the order of 30 μm in diameter have been shown to promote cell infiltration and microvascularization (18). FRESH v2.0–printed constructs contained micropores ~25 μm in diameter resulting from the melting and removal of the gelatin microparticles purposely entrapped during the printing process (Fig. 2G and movie S3). Collagen disks 5 mm thick and 10 mm in diameter were cast in a mold or printed and implanted in an in vivo murine subcutaneous vascularization model (Fig. 2, H and I, and fig. S7, A and B) to observe cellular infiltration. After implantation for 3 and 7 days, collagen disks were extracted and assessed for gross morphology, cellularization, and collagen structure (fig. S7, C to E). The solid-cast collagen showed minimal cell infiltration (Fig. 2J), whereas the printed collagen had extensive cell infiltration and collagen remodeling (Fig. 2K). Quantitative analysis revealed that cells infiltrated throughout the printed collagen disk within 3 days (Fig. 2L and fig. S8) and that the number of cells in the constructs was significantly greater for the printed collagen at 3 and 7 days compared to cast control [N = 6, P < 0.0001, two-way analysis of variance (ANOVA)] (15).

To promote vascularization, we incorporated fibronectin and the proangiogenic molecule recombinant vascular endothelial growth factor (VEGF) into our collagen bio-ink (19). Collagen disks that were FRESH-printed with VEGF and extracted after 10 days in vivo showed enhanced vascularization relative to cast controls (Fig. 2, M and N). By histology, the addition of VEGF to the cast collagen increased cell infiltration without promoting microvascularization (Fig. 2O and fig. S9). In contrast, the addition of VEGF to the printed collagen resulted in widespread vascularization, with CD31-positive vessels and red blood cells visible within the lumens (Fig. 2P). Tail vein injection of fluorescent lectin confirmed an extensive host-derived vascular network with vessels ranging from 8 to 50 μm in diameter throughout the printed collagen disk (Fig. 2Q, fig. S10, and movie S4). Multiphoton microscopy enabled deeper imaging into the printed constructs and showed vessels containing red blood cells at depths of at least 200 μm (Fig. 2R and movie S5).

We next FRESH-printed a model of the left ventricle of the heart using human stem cell–derived cardiomyocytes. We used a dual-material printing strategy with collagen bio-ink as the structural component in combination with a high-density cell bio-ink (Fig. 3A) (15). A test print design (fig. S11A) verified that the collagen pH was neutralized quickly enough to maintain ~96% post-printing viability by LIVE/DEAD staining (fig. S11B). The ventricle was designed as an ellipsoidal shell (Fig. 3B) with inner and outer walls of collagen and a central core region containing human embryonic stem cell–derived cardiomyocytes (hESC-CMs) and 2% cardiac fibroblasts (fig. S11, C to H). Ventricles were printed and cultured for up to 28 days, during which the collagen inner and outer walls provided sufficient structural integrity to maintain their intended geometry (Fig. 3C). After 4 days, the ventricles visibly contracted, and after 7 days they became synchronous with a dense layer of interconnected and striated hESC-CMs, as confirmed by immunofluorescent staining of sarcomeric α-actinin–positive myofibrils (fig. S11, I to K). Calcium imaging revealed contracting hESC-CMs throughout the entire printed ventricles, with directional wave propagation in the direction of the printed hESC-CMs observed from the side (Fig. 3, D and E) and top (Fig. 3, F and G) during spontaneous contractions in multiple ventricles (N = 3) (movie S6). Point stimulation enabled visualization of anisotropic calcium wave propagation with longitudinal conduction velocity of ~2 cm/s and a longitudinal-to-transverse anisotropy ratio of ~1.5 (Fig. 3, H and I). The ventricles had a baseline spontaneous beat rate of ~0.5 Hz and could be captured and paced at 1 and 2 Hz by means of field stimulation (Fig. 3J). We imaged the ventricles top-down to quantify motion of the inner and outer walls (Fig. 3K). Wall thickening is a hallmark of normal ventricular contraction. The printed ventricle expanded both inward and outward during a contraction, as determined by particle tracking to map the deformation field (Fig. 3L). The decrease in cross-sectional area of the interior chamber during peak systole showed a maximum of ~5% at 1-Hz pacing (N = 4) (Fig. 3M and movie S6). We also observed electrophysiologic behavior associated with arrhythmogenic disease states, including multiple propagating waves (fig. S12, A and B) and pinned rotors (fig. S12, C and D).

To demonstrate the mechanical integrity and function of collagen constructs at adult human scale, we printed a tri-leaflet heart valve 28 mm in diameter (Fig. 4A). We first prototyped the valve using alginate, a material previously used to build valve models (20), and then printed a collagen valve and improved the mechanical properties by adapting published fixation protocols for decellularized porcine heart valves (fig. S13A) (15, 21). The collagen valve had well-separated leaflets, was robust enough to be handled in air (Fig. 4, B and C, and movie S7), and was imaged by micro–computed tomography (μCT) (Fig. 4, D and E, and movie S8). Print fidelity was quantified using gauging to overlay the μCT data on the 3D model (fig. S13B), showing average overprinting of +0.55 mm and underprinting of –0.80 mm (Fig. 4F and fig. S13, C and D). Mechanical function was demonstrated by mounting the valve in a flow system with a pulsatile pump to simulate physiologic pressures, and we observed cyclical opening and closing of the valve leaflets (Fig. 4G and movie S7). We quantified flow through the valves (Fig. 4H) and demonstrated <15% regurgitation (Fig. 4I) with a maximum area opening of 19.5% (Fig. 4G). Additionally, the maximum transvalvular pressure was greater than 40 mmHg for the collagen and alginate valves (Fig. 4J), exceeding standard physiologic pressures for the tricuspid and pulmonary valves but less than the aortic and mitral valves (22). Further, human umbilical vein endothelial cells (HUVECs) cultured on unfixed collagen leaflets formed a confluent monolayer (fig. S13E).

A magnetic resonance imaging (MRI)–derived computer-aided design (CAD) model of an adult human heart was created, complete with internal structures such as valves, trabeculae, large veins, and arteries, but lacking smaller-scale vessels. To address this, we developed a computational method that uses the coronary arteries as the template to generate multiscale vasculature (fig. S14 and movie S9). We created a space-filling branching network based on a 3D Voronoi lattice, where vessels further from the left coronary arteries (red to blue) have a denser network and smaller diameters, down to ~100 μm (Fig. 4K). A subregion of the generated vasculature containing the left anterior descending artery (LAD) was selected, rendered, and printed from collagen at adult human scale (Fig. 4, L to N). Patency of large vessels was demonstrated by perfusing the multiscale vasculature through the root of the LAD (Fig. 4O). We confirmed the patency of vessels ~100 μm in diameter by optically clearing and reperfusing the multiscale vasculature (Fig. 4P, fig. S14, N to P, and movie S9).

Finally, to demonstrate organ-scale FRESH v2.0 printing capabilities and the potential to engineer larger scaffolds, we printed a neonatal-scale human heart from collagen (Fig. 4, Q and R, and fig. S15, A to C). To highlight the microscale internal structure, we printed half the heart (Fig. 4S). Structures such as trabeculae were printed from collagen with the same architecture as defined in the G-code file (Fig. 4, T and U). The square-lattice infill pattern within the ventricular walls was similarly well defined (Fig. 4, V and W). We used μCT imaging to confirm reproduction of all the anatomical structures contained within the 3D model of the heart, including the atrial and ventricular chambers, trabeculae, and pulmonary and aortic valves (fig. S15, D to I, and movie S10).

We have used the human heart for proof of concept; however, FRESH v2.0 printing of collagen is a platform that can build advanced tissue scaffolds for a wide range of organ systems. There are still many challenges to overcome, such as generating the billions of cells required to 3D-bioprint large tissues, achieving manufacturing scale, and creating a regulatory process for clinical translation (23). Although the 3D bioprinting of a fully functional organ is yet to be achieved, we now have the ability to build constructs that start to recapitulate the structural, mechanical, and biological properties of native tissues.

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