Next Article in Journal
Ureteral Orifice Detection in Ureteroscopic Images Based on Large-Kernel Convolutional Neural Networks and Attention-Based Feature Fusion
Previous Article in Journal
Physics-Guided Deep Learning for Interpretable Biomedical Image Reconstruction and Pattern Recognition in Diagnostic Frameworks
Previous Article in Special Issue
Design and Validation of a Multi-Modal Bioreactor System: Assessing the Effects of Perfusion and Cyclic Tensile Stimulation on Mechanical and Biological Properties of 3D-Printed Missing-Rib Auxetic Scaffolds
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Bioengineering
Volume 13
Issue 4
10.3390/bioengineering13040458
bioengineering-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Opinion

Prospects and Limitations of Bioprinting in Studying Human Cells’ Responses to Extreme Environments

by
Taieba Tuba Rahman
1,*,
Zhijian Pei
1,
Hongmin Qin
2 and
Hamid R. Parsaei
1
1
Department of Industrial & Systems Engineering, Texas A&M University, College Station, TX 77843, USA
2
Department of Biology, Texas A&M University, College Station, TX 77843, USA
*
Author to whom correspondence should be addressed.
Bioengineering 2026, 13(4), 458; https://doi.org/10.3390/bioengineering13040458
Submission received: 31 January 2026 / Revised: 31 March 2026 / Accepted: 11 April 2026 / Published: 14 April 2026
(This article belongs to the Special Issue Biofabrication of Biologically Realistic Constructs: Needs, Challenges, and Opportunities)
Versions Notes

Abstract

Understanding human’s responses to extreme environments holds significant importance for space exploration, deep-sea research, and environmental adaptation. Traditionally, human subjects were used to study humans’ responses to extreme environments. The main limitations of this approach include the inability to independently investigate specific cellular mechanisms, ethical and safety constraints, limited experimental controllability, and inter-individual variability that complicates mechanistic interpretation. Another approach is to study humans’ responses at the cellular level using 2D culture. This approach often exhibits limited reproducibility due to its inability to recapitulate physiologically relevant microenvironments. Bioprinting can enable studies on human’s responses at the cellular level and within 3D environments. One way is to study human cells’ responses to localized and transient extreme environments created during printing. Another way is to expose 3D printed samples (embedded with human cells) to extreme environments. However, the literature does not contain comprehensive review papers to discuss the prospects and limitations of bioprinting for investigating human cells’ responses to extreme environments. This review paper aims to fill this gap in the literature. It begins with a brief description of the effects of extreme environments on human health and summarizes reported studies on cells’ responses to extreme environments. Afterward, it discusses the prospects and limitations of the two ways of using bioprinting to investigate cells’ responses to extreme environments. Finally, it concludes with identifying knowledge gaps and proposing research directions in the application of bioprinting to study human cells’ responses to extreme environments.

1. Introduction

Exposure to extreme environments (such as high or low temperature, high pressure, and microgravity) can lead to a variety of adverse effects on human health depending on the nature and exposure duration of the extreme environment. Extreme heat exposure may result in dehydration, heat exhaustion, and potentially life-threatening heat stroke [1,2,3], whereas prolonged cold exposure can impair neurological function, reduce metabolic activity, and progress to hypothermia as thermoregulatory mechanisms fail [4]. Hypobaric environments occurring at high altitude reduce oxygen availability and may cause hypoxia, acute mountain sickness, and severe complications such as high-altitude pulmonary edema and cerebral edema [5,6,7,8,9,10,11,12]. Hyperbaric exposure during underwater or pressurized operations increases the risk of barotrauma, nitrogen narcosis, and decompression injury associated with rapid pressure changes [13,14,15,16,17]. Additionally, exposure to microgravity during space missions contributes to musculoskeletal deconditioning, bone density loss, and cardiovascular adaptation [18]. Table 1 summarizes several reported studies on human body’s responses to extreme environments. In these reported studies, human subjects were used to provide critical insight into humans’ responses at the organ or whole-body levels. The main limitation of this approach is the inability to independently investigate specific cellular mechanisms.
Since humans’ responses to extreme environments ultimately originate from the cellular level, it is important to understand how human cells respond to extreme environments. Another approach is to study humans’ responses at the cellular level using 2D culture. Table 2 summarizes several reported studies on cells’ responses to extreme environments using 2D cell culture [21,22,23]. Conventional 2D cell culture systems lack native tissue architecture, extracellular matrix interactions, and three-dimensional mechanical constraints that critically influence cellular behavior under stress [24]. As a result, cells’ responses observed in 2D cultures may not accurately reflect those occurring within tissues that preserve native architecture.
Several review papers have summarized the effects of individual extreme environments (such as heat stress [26,27,28], hypoxia [29], high-pressure environment [30], and microgravity [30,31,32]) on cellular behavior and human health. However, the existing review papers primarily cover reported studies based on conventional cell culture and animal models. None of them was focused on the prospects and limitations of using 3D printing to study human cells’ responses to extreme environments. This paper is intended to fill this gap in the literature.
In this review paper, Section 2 discusses the prospects and limitations of using 3D printing to investigate human cells’ responses to localized and transient extreme environments created during printing. Section 3 discusses the prospects and limitations of using 3D printing to investigate human cells’ responses to extreme environments with 3D printed samples containing human cells. Finally, Section 4 provides concluding remarks.

2. Investigation of Cells’ Responses to Localized and Transient Extreme Environments Created During Printing

2.1. Creation of Localized and Transient Extreme Environments During Printing

Bioprinting techniques can create localized and transient extreme environments during the printing process. In these localized and transient extreme environments, cells embedded in the bioink (mostly hydrogel) experience localized and transient mechanical, thermal, and radiative stresses. The type and magnitude of stresses experienced by cells depend on the printing technique and can be controlled through changes in printing parameters. Table 3 shows several types of localized and transient environmental stress created during printing.
In extrusion-based 3D printing, cells embedded in bioinks primarily experience shear stress during material extrusion. In this technique, cell-laden bioink is continuously extruded through a nozzle, and cells experience localized mechanical stresses, mainly shear stress [33]. Beyond mechanical stress, extrusion-based 3D printing can also introduce thermal and radiative stresses depending on the bioink type and crosslinking approach. Extrusion bioprinting commonly relies on hydrogel-based bioinks to maintain structural stability, which requires a crosslinking mechanism. Depending on the bioink type, crosslinking may occur through photocrosslinking, using ultraviolet or visible light, or through thermal crosslinking, involving temperature-induced gelation [34]. When temperature-sensitive bioinks such as collagen are used, thermal stress arises from temperature changes required to control their phase behavior during printing and gelation. Collagen-based bioinks are typically maintained at low temperatures (≈4–10 °C) to remain in a liquid state prior to printing, and after printing, are exposed to physiological temperatures (≈37 °C) to induce gelation [35]. This temperature transition creates a transient thermal gradient around the encapsulated cells, which can induce thermal stress [36]. In addition, when photocrosslinkable bioinks are employed, UV exposure, used for crosslinking, may generate radiative stress on cells.
In inkjet-based 3D printing, cells experience mechanical stress during droplet formation and ejection, with thermal inkjet systems potentially causing brief temperature fluctuations [37].
In stereolithography-based 3D printing, cells are exposed to ultraviolet or visible light during layer-by-layer photopolymerization, which primarily induces radiative stress [38].
In laser-assisted bioprinting, bioink droplets are transferred onto a substrate using focused laser pulses without nozzle contact. In this technique, cells are exposed to localized energy input during the printing process, which can introduce radiative and thermal stresses [39].
Table 3. Types of localized and transient environmental stress created during printing.
Table 3. Types of localized and transient environmental stress created during printing.
Bioprinting TechniqueType of StressControlling Printing ParameterReference
Extrusion-based Shear stressNozzle shape[40,41]
Nozzle diameter[41,42]
Nozzle length 
Extrusion pressure[41,42,43,44]
Thermal stressPrinting temperature[41]
Thermal and radiative stressUV exposure duration, UV intensity[41]
Inkjet-based Shear stressApplied voltage, frequency, pulse duration[45]
Thermal stressNozzle temperature[37]
Laser-assisted Shear stressJetting speed due to bubble expansion[46]
Thermal and radiative stressLaser exposure, laser intensity[39,47]
StereolithographyRadiative stressUV exposure duration, UV intensity,[38]
 Photoinitiator 

2.2. Variaion in the Intensity of Localized and Transient Extreme Emvironments by Changing Process Parameters

By changing printing process parameters, the intensity of the microenvironmental stress experienced by cells can be varied, which enables the systematic investigation of cells’ stress responses. For example, Rahman et al. investigated the effects of extrusion pressure on human bronchial epithelial (HBE) cells encapsulated in 3D printed samples [48]. Extrusion pressures up to 100 kPa were applied to evaluate how acute mechanical loading during printing influenced cell viability. These pressure levels exceed those typically experienced during normal breathing (≤2.9 kPa) [49,50] or coughing (~19.6 kPa) [51], and therefore serve as controlled mechanical challenges rather than representations of sustained environmental pressure exposure. Representative studies on cells’ responses to localized and transient extreme environments created during 3D printing are summarized in Table 4.
Reported studies have shown that in extrusion-based 3D printing, reducing nozzle diameter below 400 µm increases shear stress during extrusion, which can lead to reduced cell viability [41,42]. Increasing nozzle length above 8.9 mm increases the duration of shear exposure, causing cells to experience shear stress for a longer period during extrusion, which also contributes to reduced cell viability [44]. Besides nozzle geometry, increasing extrusion pressure above 0.5 bar intensifies shear-induced cellular stress, which causes membrane damage and reduced cell viability [42,43]. Shear stress can modulate signaling pathways associated with proliferation, migration, and the metabolism of cells, highlighting its broader impact on cellular function [54]. In addition, shear stress has been associated with increased reactive oxygen species (ROS) generation, which can cause oxidative damage to lipids, proteins, and DNA, ultimately leading to apoptosis or necrosis [33,55]. It is worth noting that at physiological levels of shear stress, apoptosis can be inhibited in human endothelial cells, indicating that cellular responses depend on the magnitude and duration of the applied stress [56].
When temperature-sensitive bioinks are used, thermal stress can induce protein unfolding and the accumulation of non-native proteins, which activate heat shock responses and related signaling pathways. Even under moderate temperature changes, these responses reflect cellular adaptation to thermal perturbations and may influence cellular function and survival [36,57]. In addition, when photocrosslinkable bioinks are employed, UV-induced damage can result in DNA strand breaks and genomic instability, which may activate DNA damage responses and apoptosis pathways, thereby affecting cellular function and survival [37]. For example, UV exposure during hydrogel photocrosslinking has been reported to reduce the viability of HepG2 cells (human hepatocellular carcinoma–derived liver cell line) at UV irradiation doses above 1350 mJ cm−2 [39]. The magnitude of mechanical, thermal, and radiative stresses can be controlled through changes in parameters such as nozzle geometry, extrusion pressure, printing temperature, and UV light exposure [41,42,43,44].
In stereolithography-based 3D printing, UV or visible light exposure can induce cellular damage through photochemical effects, including reactive oxygen species (ROS) generation and DNA damage [38,58,59]. Such damage may activate intracellular signaling pathways associated with stress responses and apoptosis, ultimately affecting cell viability and function [37]. For example, UV doses above 0.5 kJ m−2 affect the cell viability of fibroblasts and mesenchymal stem cells [53]. Stereolithography-based 3D printing can be used to study cells’ responses to localized and transient radiation-driven extreme environments. The magnitude of radiative stress can be controlled by changing parameters such as light intensity (UV or visible), exposure duration, irradiation energy, and photoinitiator properties [38].
It has been reported that in laser-assisted bioprinting, laser exposure at wavelengths such as 355 nm and 532 nm, with fluence in the range of approximately 330–850 mJ cm−2, can affect cell viability due to combined photochemical and photothermal effects [60]. Such exposure can generate reactive oxygen species (ROS) and induce DNA damage, leading to alterations in cellular function and survival [37,60].

2.3. Limitations of the Approach

There are several limitations of this approach. During printing, cells are exposed to mechanical, thermal, or radiative stresses that are localized and transient, occurring primarily within confined regions such as the nozzle or light exposure zone and lasting for short durations. As a result, these conditions mainly reflect the acute stresses that cells experience. Cells’ responses include immediate viability loss or oxidative stress [34,61]. The extreme environments human bodies experience typically involve prolonged exposure that enables physiological adaptation through coordinated biochemical and tissue-level regulation. Due to the lack of sustained environmental exposure, this approach is unable to evaluate long-term environmental adaptation mechanisms. Although having these limitations, studies using this approach can result in new insights into human cells’ responses to extreme environments where the duration is short (transient) and the stress is localized.

3. Investigation of Cells’ Responses to Extreme Environments Using 3D Printed Samples

3.1. Fabrication of 3D Constructs (Embedded with Cells) by 3D Printing

Bioprinting enables cells to be encapsulated within 3D microenvironments that more closely resemble those in native tissues [62,63]. Although bioprinted constructs do not replicate full organ-level or systemic physiology, they enable controlled investigation of cellular-level responses to specific aspects of extreme environments by changing construct design parameters—such as bioink composition, construct geometry, and concentration of cells in printed constructs.
The type of bioink determines the baseline biochemical and structural environment surrounding the cells embedded in the bioink. Different bioinks, such as alginate, PEGDA, hyaluronic acid, chitosan, fibrin, silk fibroin, gelatin, agarose, methylcellulose, and collagen, have distinct properties that influence cell–matrix interactions, adhesion, and signaling [64]. Increasing polymer concentration creates a stiffer matrix [65], which can be used to model mechanical stress environments, where cells experience higher resistance and force transmission. In contrast, using softer or more porous bioinks improves permeability and mass transport [61], enabling the study of diffusion-related conditions, such as localized hypoxia or nutrient limitation.
The geometry of printed constructs directly influences diffusion gradients, leading to spatial variations in oxygen and nutrient availability for the cells embedded in printed constructs. Thick or densely printed constructs often develop hypoxic cores, which can alter cell viability, metabolic activity, and stress responses [66,67]. Reported studies have shown that such gradients can be intentionally leveraged to model oxygen-limited environments and investigate hypoxia-induced cellular behavior [68]. Additionally, constructs can be printed in a way that different portions of the constructs can have different oxygen levels. Exposing such constructs to externally applied extreme environments can reveal cellular responses in these different portions of the constructs. In several reported studies, mammalian cells have been co-printed with photosynthetic microalgae that produce oxygen within the constructs and help regulate local oxygen availability [69,70].

3.2. Exposure of Printed Constructs to Different Environmental Stressors by Using Different Test Chambers

Bioprinting can be used to study cells’ responses to extreme environments by placing bioprinted 3D constructs into environmental testing chambers. Following printing, customized 3D constructs can be transferred into temperature-controlled systems, hypoxia workstations, pressure chambers, or other defined environmental platforms. This approach allows controlled exposure to specific environmental stress condition. For instance, Rahman et al. investigated the effects of thermal stress (37 °C, 45 °C, and 55 °C for 10 min) on HBE cells embedded in 3D printed constructs [71]. In this case, the extrusion-based platform provided a reproducible three-dimensional tissue model, while the thermal stress was applied post-printing as a separate environmental variable using a heat block.

3.3. Limitations of the Approach

Despite the advantages of 3D printed constructs for studying cells’ responses under defined post-printing environmental conditions, these printed constructs are subject to fundamental limitations related to mass transport within hydrogel matrices. In most printed constructs, oxygen, nutrients, and metabolic waste are transported primarily through passive diffusion. Because bioprinted hydrogels typically lack perfusable vasculature, oxygen supply to cells depends largely on diffusion through the surrounding matrix. The extent of diffusion limitation therefore depends on several experimental parameters, including construct thickness, cell density, and hydrogel properties such as porosity, polymer concentration, crosslinking density, and water content [66]. For example, as construct thickness or cell density increases, cellular oxygen consumption may exceed the rate at which oxygen diffuses through the matrix. In hydrogel-based constructs, oxygen diffusion is generally limited to approximately 100–200 µm from the construct surface, beyond which oxygen delivery may become insufficient to sustain cellular metabolism [72]. Consequently, cells located deep inside of thick constructs may experience hypoxia even under incubator conditions with normal oxygen concentration.
These diffusion-induced gradients of oxygen level can complicate the interpretation of results from experiments designed to study cells’ responses to externally applied extreme environments. For example, when studying hypobaric or hypoxic conditions, oxygen deprivation may arise not only from the imposed environmental conditions but also from intrinsic diffusion limitations within the construct. It may therefore be difficult to distinguish between environmentally induced hypoxia and diffusion-driven oxygen depletion. Moreover, due to diffusion constraints, the absence of functional vascular networks, and the lack of systemic physiological regulation, this approach remains limited in their ability to reproduce complex tissue-level and organism-level responses to extreme environments.
Several strategies have been reported to mitigate diffusion-induced gradients of oxygen and nutrient level in three-dimensional constructs. One strategy is the incorporation of vascular channels or perfusable microvascular networks, which enable convective transport of oxygen and nutrients within the construct [73]. This is critical because oxygen and nutrient delivery in bioprinted tissues rely largely on diffusion in the absence of a functional vascular network. Another strategy is dynamic culture systems, such as perfusion bioreactors or microfluidic platforms, which can further improve mass transport by introducing controlled fluid flow [74,75]. An additional strategy is limiting construct thickness as thin constructs (≤200–300 µm) that are less susceptible to diffusion limitations, whereas thicker constructs (>500 µm) are more prone to central hypoxia unless active transport mechanisms are introduced [76]. Finally, proper hydrogel design, including increased porosity or optimized crosslinking density, can improve molecular diffusion within the matrix [73].

4. Concluding Remarks

Bioprinting can be used for studying human cells’ responses to extreme environments. First, localized and transient extreme environments can be intentionally created during the printing process. The intensity of the extreme environments can be varied by adjusting printing parameters (such as nozzle geometry, extrusion pressure, printing speed, temperature, or light exposure). This approach enables the investigation of cells’ responses to localized and transient mechanical, thermal, or radiative stresses. However, because these stresses are localized and transient, they do not represent long-duration environmental exposure. This paper summarizes, for the first time in the literature, publicly available information in the use of 3D printing to create extreme environments during 3D printing to study cells’ responses to localized and transient extreme environments. In some relevant papers, this use of 3D printing might be implied but is not explicitly stated.
Second, bioprinting enables fabrication of 3D cell-laden constructs that better mimic native tissue architecture. These 3D constructs can be exposed to extreme environments under controlled conditions and can contain integrated sensors that allow real-time monitoring of cellular responses. However, because bioprinted constructs lack full vascularization, perfusion, and systemic physiological interactions, the results obtained using these constructs might not be directly translatable to whole-body human responses. This paper summarizes, for the first time in the literature, publicly available information on the use of 3D printed constructs to investigate the responses of cells (embedded in the printed constructs) to extreme environments. In some relevant papers, this use of 3D printing might be implied but is not explicitly stated.

Author Contributions

Writing—original draft preparation, T.T.R.; writing—review and editing, Z.P., H.Q., H.R.P. and T.T.R.; supervision, Z.P. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Air Force Office of Scientific Research, grant number FA9550-23-1-0599.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The authors confirm that the data to support the findings of this study are available within the article or upon request to the corresponding author.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Leon, L.R.; Kenefick, R. Pathophysiology of Heat-Related Illnesses; Defense Technical Information Center: Fort Belvoir, VA, USA, 2012. [Google Scholar]
  2. Gomez, C.R. Disorders of body temperature. In Handbook of Clinical Neurology; Elsevier: Amsterdam, The Netherlands, 2014; Volume 120, pp. 947–957. [Google Scholar]
  3. Shrimanker, I.; Bhattarai, S. Electrolytes. In StatPearls [Internet]; StatPearls Publishing: Treasure Island, FL, USA, 2019. [Google Scholar]
  4. Beker, B.M.; Cervellera, C.; De Vito, A.; Musso, C.G. Human physiology in extreme heat and cold. Int. Arch. Clin. Physiol 2018, 1, 1–8. [Google Scholar]
  5. Hackett, P. High-altitude medicine. In Wilderness Medicine; Mosby Inc.: St Louis, MO, USA, 2001; pp. 2–43. [Google Scholar]
  6. Barker, K.R.; Conroy, A.L.; Hawkes, M.; Murphy, H.; Pandey, P.; Kain, K.C. Biomarkers of hypoxia, endothelial and circulatory dysfunction among climbers in Nepal with AMS and HAPE: A prospective case-control study. J. Travel Med. 2016, 23, taw005. [Google Scholar] [CrossRef][Green Version]
  7. Mehta, S.; Chawla, A.; Kashyap, A. Acute mountain sickness, high altitude cerebral oedema, high altitude pulmonary oedema: The current concepts. Med. J. Armed Forces India 2008, 64, 149–153. [Google Scholar] [CrossRef]
  8. Auerbach, P.S.; Cushing, T.A.; Harris, N.S. Auerbach’s Wilderness Medicine e-Book; Elsevier Health Sciences: Amsterdam, The Netherlands, 2016. [Google Scholar]
  9. Ward, M.; Milledge, J.; West, J. Human and medical geography of mountain regions. In Ward, Milledge and West, Editors. High Altitude Medicine and Physiology; Chapman and Hall Medical: London, UK, 1989; pp. 45–62. [Google Scholar]
  10. Jensen, J.D.; Vincent, A.L. High altitude pulmonary edema. In StatPearls [Internet]; StatPearls Publishing: Treasure Island, FL, USA, 2022. [Google Scholar]
  11. Jensen, J.D.; Vincent, A.L. High altitude cerebral edema. In StatPearls [Internet]; StatPearls Publishing: Treasure Island, FL, USA, 2022. [Google Scholar]
  12. Sutton, J.R.; Coates, G.; Houston, C.S. Hypoxia and mountain medicine. In Proceedings of the 7th International Hypoxia Symposium, Lake Louise, AB, Canada, 26 February–2 March 1991. [Google Scholar]
  13. Rupp, T.; Maufrais, C.; Walther, G.; Esteve, F.; Macdonald, J.H.; Bouzat, P.; Verges, S. MEDEX 2015: Prophylactic Effects of Positive Expiratory Pressure in Trekkers at Very High Altitude. Front. Physiol. 2021, 12, 710622. [Google Scholar] [CrossRef]
  14. Battisti, A.S.; Haftel, A.; Murphy-Lavoie, H.M. Barotrauma. In StatPearls [Internet]; StatPearls Publishing: Treasure Island, FL, USA, 2021. [Google Scholar]
  15. Wienke, B.R. Basic Decompression: Theory and Application; Best Publishing Company: North Palm Beach, FL, USA, 2008. [Google Scholar]
  16. Hunger, W., Jr.; Bennett, P. The causes, mechanisms and prevention of the high pressure nervous syndrome. Undersea Biomed. Res. 1974, 1, 1–28. [Google Scholar]
  17. Halsey, M. Effects of high pressure on the central nervous system. Physiol. Rev. 1982, 62, 1341–1377. [Google Scholar] [CrossRef]
  18. Horneck, G.; Facius, R.; Reichert, M.; Rettberg, P.; Seboldt, W.; Manzey, D.; Comet, B.; Maillet, A.; Preiss, H.; Schauer, L. HUMEX, a study on the survivability and adaptation of humans to long-duration exploratory missions, part II: Missions to Mars. Adv. Space Res. 2006, 38, 752–759. [Google Scholar] [CrossRef]
  19. Monnoyer, R.; Eftedal, I.; Hjelde, A.; Deb, S.; Haugum, K.; Lautridou, J. Functional Profiling Reveals Altered Metabolic Activity in Divers’ Oral Microbiota During Commercial Heliox Saturation Diving. Front. Physiol. 2021, 12, 702634. [Google Scholar] [CrossRef] [PubMed]
  20. Bao, X.C.; Shen, Q.; Fang, Y.Q.; Wu, J.G. Human Physiological Responses to a Single Deep Helium-Oxygen Diving. Front. Physiol. 2021, 12, 735986. [Google Scholar] [CrossRef] [PubMed]
  21. Omori, H.; Otsu, M.; Nogami, H.; Shibata, M. Heat shock response enhanced by cell culture treatment in mouse embryonic stem cell-derived proliferating neural stem cells. PLoS ONE 2021, 16, e0249954. [Google Scholar] [CrossRef]
  22. Kouznetsov, N.V. Cell Responses to Simulated Microgravity and Hydrodynamic Stress Can Be Distinguished by Comparative Transcriptomics. Int. J. Transl. Med. 2022, 2, 364–386. [Google Scholar] [CrossRef]
  23. Aissiou, A.K.; Jha, S.; Dhunnoo, K.; Ma, Z.; Li, D.X.; Ravin, R.; Kunze, M.; Wong, K.; Adesida, A.B. Transcriptomic response of bioengineered human cartilage to parabolic flight microgravity is sex-dependent. npj Microgravity 2023, 9, 5. [Google Scholar] [CrossRef]
  24. Duval, K.; Grover, H.; Han, L.-H.; Mou, Y.; Pegoraro, A.F.; Fredberg, J.; Chen, Z. Modeling physiological events in 2D vs. 3D cell culture. Physiology 2017, 32, 266–277. [Google Scholar] [CrossRef]
  25. Reissis, Y.; García-Gareta, E.; Korda, M.; Blunn, G.W.; Hua, J. The effect of temperature on the viability of human mesenchymal stem cells. Stem Cell Res. Ther. 2013, 4, 139. [Google Scholar] [CrossRef] [PubMed]
  26. Kregel, K.C. Invited review: Heat shock proteins: Modifying factors in physiological stress responses and acquired thermotolerance. J. Appl. Physiol. 2002, 92, 2177–2186. [Google Scholar] [CrossRef]
  27. Richter, K.; Haslbeck, M.; Buchner, J. The heat shock response: Life on the verge of death. Mol. Cell 2010, 40, 253–266. [Google Scholar] [CrossRef] [PubMed]
  28. Lepock, J.R. How do cells respond to their thermal environment? Int. J. Hyperth. 2005, 21, 681–687. [Google Scholar] [CrossRef] [PubMed]
  29. Semenza, G.L. Hypoxia-inducible factors in physiology and medicine. Cell 2012, 148, 399–408. [Google Scholar] [CrossRef] [PubMed]
  30. Macdonald, A. Adaptation to High Pressure in the Laboratory. In Life at High Pressure: In the Deep Sea and Other Environments; Springer: Berlin/Heidelberg, Germany, 2021; pp. 327–352. [Google Scholar]
  31. Marchal, S.; Choukér, A.; Bereiter-Hahn, J.; Kraus, A.; Grimm, D.; Krüger, M. Challenges for the human immune system after leaving Earth. npj Microgravity 2024, 10, 106. [Google Scholar] [CrossRef]
  32. Pan, C.; Zhang, Y.; Yan, J.; Zhou, Y.; Wang, S.; Liu, X.; Zhang, P.; Yang, H. Extreme environments and human health: From the immune microenvironments to immune cells. Environ. Res. 2023, 236, 116800. [Google Scholar] [CrossRef] [PubMed]
  33. Blaeser, A.; Duarte Campos, D.F.; Puster, U.; Richtering, W.; Stevens, M.M.; Fischer, H. Controlling shear stress in 3D bioprinting is a key factor to balance printing resolution and stem cell integrity. Adv. Healthc. Mater. 2016, 5, 326–333. [Google Scholar] [CrossRef] [PubMed]
  34. Ozbolat, I.T.; Hospodiuk, M. Current advances and future perspectives in extrusion-based bioprinting. Biomaterials 2016, 76, 321–343. [Google Scholar] [CrossRef]
  35. Stepanovska, J.; Supova, M.; Hanzalek, K.; Broz, A.; Matejka, R. Collagen bioinks for bioprinting: A systematic review of hydrogel properties, bioprinting parameters, protocols, and bioprinted structure characteristics. Biomedicines 2021, 9, 1137. [Google Scholar] [CrossRef] [PubMed]
  36. Somero, G.N. The cellular stress response and temperature: Function, regulation, and evolution. J. Exp. Zool. Part A Ecol. Integr. Physiol. 2020, 333, 379–397. [Google Scholar] [CrossRef] [PubMed]
  37. Xu, H.-Q.; Liu, J.-C.; Zhang, Z.-Y.; Xu, C.-X. A review on cell damage, viability, and functionality during 3D bioprinting. Mil. Med. Res. 2022, 9, 70. [Google Scholar] [CrossRef]
  38. Lin, H.; Zhang, D.; Alexander, P.G.; Yang, G.; Tan, J.; Cheng, A.W.-M.; Tuan, R.S. Application of visible light-based projection stereolithography for live cell-scaffold fabrication with designed architecture. Biomaterials 2013, 34, 331–339. [Google Scholar] [CrossRef]
  39. Gudapati, H.; Yan, J.; Huang, Y.; Chrisey, D.B. Alginate gelation-induced cell death during laser-assisted cell printing. Biofabrication 2014, 6, 035022. [Google Scholar] [CrossRef]
  40. Willson, K.; Ke, D.; Kengla, C.; Atala, A.; Murphy, S.V. Extrusion-based bioprinting: Current standards and relevancy for human-sized tissue fabrication. In 3D Bioprinting: Principles and Protocols; Springer: New York, NY, USA, 2020; pp. 65–92. [Google Scholar]
  41. Billiet, T.; Gevaert, E.; De Schryver, T.; Cornelissen, M.; Dubruel, P. The 3D printing of gelatin methacrylamide cell-laden tissue-engineered constructs with high cell viability. Biomaterials 2014, 35, 49–62. [Google Scholar] [CrossRef]
  42. Chang, R.; Nam, J.; Sun, W. Effects of dispensing pressure and nozzle diameter on cell survival from solid freeform fabrication-based direct cell writing. Tissue Eng. Part A 2008, 14, 41–48. [Google Scholar] [CrossRef] [PubMed]
  43. Fakhruddin, K.; Hamzah, M.S.A.; Razak, S.I.A. Effects of extrusion pressure and printing speed of 3D bioprinted construct on the fibroblast cells viability. In Proceedings of the IOP Conference Series: Materials Science and Engineering; IOP Publishing: Bristol, UK, 2018; p. 012042. [Google Scholar]
  44. Faulkner-Jones, A.; Fyfe, C.; Cornelissen, D.-J.; Gardner, J.; King, J.; Courtney, A.; Shu, W. Bioprinting of human pluripotent stem cells and their directed differentiation into hepatocyte-like cells for the generation of mini-livers in 3D. Biofabrication 2015, 7, 044102. [Google Scholar] [CrossRef]
  45. Adhikari, J.; Roy, A.; Das, A.; Ghosh, M.; Thomas, S.; Sinha, A.; Kim, J.; Saha, P. Effects of processing parameters of 3D bioprinting on the cellular activity of bioinks. Macromol. Biosci. 2021, 21, 2000179. [Google Scholar] [CrossRef]
  46. Wang, W.; Li, G.; Huang, Y. Modeling of bubble expansion-induced cell mechanical profile in laser-assisted cell direct writing. J. Manuf. Sci. Eng. 2009, 131, 051013. [Google Scholar] [CrossRef]
  47. Akerfelt, M.; Morimoto, R.I.; Sistonen, L. Heat shock factors: Integrators of cell stress, development and lifespan. Nat. Rev. Mol. Cell Biol. 2010, 11, 545–555. [Google Scholar] [CrossRef] [PubMed]
  48. Rahman, T.T.; Wood, N.; Pei, Z.; Qin, H.; Mohan, P. Effects of Extrusion Pressure During 3D Printing on Viability of Human Bronchial Epithelial Cells in 3D Printed Samples. Biomimetics 2025, 10, 297. [Google Scholar] [CrossRef]
  49. Major, V.J.; Chiew, Y.S.; Shaw, G.M.; Chase, J.G. Biomedical engineer’s guide to the clinical aspects of intensive care mechanical ventilation. Biomed. Eng. Online 2018, 17, 169. [Google Scholar] [CrossRef] [PubMed]
  50. Pleil, J.D.; Ariel Geer Wallace, M.; Davis, M.D.; Matty, C.M. The physics of human breathing: Flow, timing, volume, and pressure parameters for normal, on-demand, and ventilator respiration. J. Breath Res. 2021, 15, 042002. [Google Scholar] [CrossRef]
  51. Dickey, B.F. What it takes for a cough to expel mucus from the airway. Proc. Natl. Acad. Sci. USA 2018, 115, 12340–12342. [Google Scholar] [CrossRef]
  52. Saunders, R.E.; Gough, J.E.; Derby, B. Delivery of human fibroblast cells by piezoelectric drop-on-demand inkjet printing. Biomaterials 2008, 29, 193–203. [Google Scholar] [CrossRef]
  53. Godar, D.E.; Gurunathan, C.; Ilev, I. 3D Bioprinting with UVA1 Radiation and Photoinitiator Irgacure 2959: Can the ASTM Standard L929 Cells Predict Human Stem Cell Cytotoxicity? Photochem. Photobiol. 2019, 95, 581–586. [Google Scholar] [CrossRef]
  54. Zhang, Y.; O’Mahony, A.; He, Y.; Barber, T. Hydrodynamic shear stress’ impact on mammalian cell properties and its applications in 3D bioprinting. Biofabrication 2024, 16, 022003. [Google Scholar] [CrossRef]
  55. Espina, J.A.; Cordeiro, M.H.; Milivojevic, M.; Pajić-Lijaković, I.; Barriga, E.H. Response of cells and tissues to shear stress. J. Cell Sci. 2023, 136, jcs260985. [Google Scholar] [CrossRef]
  56. Dimmeler, S.; Haendeler, J.; Rippmann, V.; Nehls, M.; Zeiher, A.M. Shear stress inhibits apoptosis of human endothelial cells. FEBS Lett. 1996, 399, 71–74. [Google Scholar] [CrossRef]
  57. Park, H.; Han, S.; Oh, S.; Kang, H. Cellular responses to mild heat stress. Cell. Mol. Life Sci. 2005, 62, 10–23. [Google Scholar] [CrossRef] [PubMed]
  58. de Gruijl, F.R.; van Kranen, H.J.; Mullenders, L.H. UV-induced DNA damage, repair, mutations and oncogenic pathways in skin cancer. J. Photochem. Photobiol. B Biol. 2001, 63, 19–27. [Google Scholar] [CrossRef]
  59. Kulms, D.; Zeise, E.; Pöppelmann, B.; Schwarz, T. DNA damage, death receptor activation and reactive oxygen species contribute to ultraviolet radiation-induced apoptosis in an essential and independent way. Oncogene 2002, 21, 5844–5851. [Google Scholar] [CrossRef]
  60. Karakaidos, P.; Kryou, C.; Simigdala, N.; Klinakis, A.; Zergioti, I. Laser bioprinting of cells using UV and visible wavelengths: A comparative DNA damage study. Bioengineering 2022, 9, 378. [Google Scholar] [CrossRef] [PubMed]
  61. Murphy, S.V.; Atala, A. 3D bioprinting of tissues and organs. Nat. Biotechnol. 2014, 32, 773–785. [Google Scholar] [CrossRef] [PubMed]
  62. Libretti, S.; Puckett, Y. Physiology, homeostasis. In StatPearls [Internet]; StatPearls Publishing: Treasure Island, FL, USA, 2020. [Google Scholar]
  63. Tsibulnikov, S.; Maslov, L.; Voronkov, N.; Oeltgen, P. Thyroid hormones and the mechanisms of adaptation to cold. Hormones 2020, 19, 329–339. [Google Scholar] [CrossRef]
  64. Chimene, D.; Lennox, K.K.; Kaunas, R.R.; Gaharwar, A.K. Advanced bioinks for 3D printing: A materials science perspective. Ann. Biomed. Eng. 2016, 44, 2090–2102. [Google Scholar] [CrossRef]
  65. Rahman, T.T.; Wood, N.; Akib, Y.M.; Qin, H.; Pei, Z. Experimental Study on Compatibility of Human Bronchial Epithelial Cells in Collagen–Alginate Bioink for 3D Printing. Bioengineering 2024, 11, 862. [Google Scholar] [CrossRef]
  66. Malda, J.; Rouwkema, J.; Martens, D.E.; le Comte, E.P.; Kooy, F.; Tramper, J.; van Blitterswijk, C.A.; Riesle, J. Oxygen gradients in tissue-engineered PEGT/PBT cartilaginous constructs: Measurement and modeling. Biotechnol. Bioeng. 2004, 86, 9–18. [Google Scholar] [CrossRef] [PubMed]
  67. Rouwkema, J.; Rivron, N.C.; van Blitterswijk, C.A. Vascularization in tissue engineering. Trends Biotechnol. 2008, 26, 434–441. [Google Scholar] [CrossRef] [PubMed]
  68. Caliari, S.R.; Burdick, J.A. A practical guide to hydrogels for cell culture. Nat. Methods 2016, 13, 405–414. [Google Scholar] [CrossRef] [PubMed]
  69. Maharjan, S.; Alva, J.; Cámara, C.; Rubio, A.G.; Hernández, D.; Delavaux, C.; Correa, E.; Romo, M.D.; Bonilla, D.; Santiago, M.L. Symbiotic photosynthetic oxygenation within 3D-bioprinted vascularized tissues. Matter 2021, 4, 217–240. [Google Scholar] [CrossRef]
  70. Dani, F.; Windisch, J.; Valencia Guerrero, X.M.; Bernhardt, A.; Gelinsky, M.; Krujatz, F.; Lode, A. Selection of a suitable photosynthetically active microalgae strain for the co-cultivation with mammalian cells. Front. Bioeng. Biotechnol. 2022, 10, 994134. [Google Scholar] [CrossRef]
  71. Rahman, T.T.; Wood, N.; Pei, Z.; Qin, H. Effects of Extreme Temperature on Human Bronchial Epithelial Cells in 3D Printed Samples. Bioengineering 2024, 11, 1201. [Google Scholar] [CrossRef]
  72. Lovett, M.; Lee, K.; Edwards, A.; Kaplan, D.L. Vascularization strategies for tissue engineering. Tissue Eng. Part B Rev. 2009, 15, 353–370. [Google Scholar] [CrossRef]
  73. Ke, D.; Murphy, S.V. Current challenges of bioprinted tissues toward clinical translation. Tissue Eng. Part B Rev. 2019, 25, 1–13. [Google Scholar] [CrossRef]
  74. Figueiredo, L.; Le Visage, C.; Weiss, P.; Yang, J. Quantifying oxygen levels in 3D bioprinted cell-laden thick constructs with perfusable microchannel networks. Polymers 2020, 12, 1260. [Google Scholar] [CrossRef]
  75. Grebenyuk, S.; Abdel Fattah, A.R.; Kumar, M.; Toprakhisar, B.; Rustandi, G.; Vananroye, A.; Salmon, I.; Verfaillie, C.; Grillo, M.; Ranga, A. Large-scale perfused tissues via synthetic 3D soft microfluidics. Nat. Commun. 2023, 14, 193. [Google Scholar] [CrossRef] [PubMed]
  76. Hudson, A.R.; Shiwarski, D.J.; Kramer, A.J.; Feinberg, A.W. Enhancing viability in static and perfused 3D tissue constructs using sacrificial gelatin microparticles. ACS Biomater. Sci. Eng. 2025, 11, 2888–2897. [Google Scholar] [CrossRef] [PubMed]
Table 1. Several reported studies on human body’s responses to extreme environments.
Table 1. Several reported studies on human body’s responses to extreme environments.
Extreme EnvironmentTest ModelResponseReference
Heat Human,
male and female		Transcriptomic response of peripheral blood mononuclear cell[18]
HypobaricHuman, male trekkerAcute mountain sickness evaluation, electrocardiograph, extravascular lung water accumulation by thoracic ultrasound, middle cerebral artery blood flow velocity, muscle and cerebral oxygenation[13]
HyperbaricHuman, male diversOral bacterial metabolism, bacterial oxidative stress response[19]
HyperbaricHuman, male diversBlood cell counts, cardiac damage, oxidative stress, vascular endothelial activation, and hormonal biomarkers[20]
Table 2. Several reported studies on cells’ responses to extreme environments.
Table 2. Several reported studies on cells’ responses to extreme environments.
Extreme EnvironmentTest ModelResponseReference
Heat Mouse neural stem cells The number of adherent cells, expression ratios of HS protein (Hsp)40and Hsp70genes[21]
HeatHuman mesenchymal stem cellsMetabolic activity and viability[25]
Simulated microgravityMouse primary T cells, Human T lymphocytes cellsT-cell transcriptome analysis using RNA sequencing[22]
Spaceflight microgravityHuman bone marrow mesenchymal stem cellsTranscriptomic response via RNA sequencing[23]
Table 4. Reported studies on cells’ responses to localized and transient extreme environments created during 3D printing.
Table 4. Reported studies on cells’ responses to localized and transient extreme environments created during 3D printing.
Bioprinting TechniquePrinting ParameterRangeCell TypeResultReference
Extrusion-basedNozzle diameter150–400 µmHepG2 cellsDecreased cell viability as nozzle diameter decreased[41,42]
 Extrusion pressure0.5–5 barHepG2 cells, hiPSCS cells, human skin fibroblast cells Decreased cell viability and increased membrane damage as extrusion pressure increased[41,42,43,44]
 Nozzle length variation8.9–24.4 mm.Human pluripotent stem cells (hPSCs)Decreased cell viability as nozzle length increase[44]
 UV-A irradiation dose1350–5400 mJ cm−2HepG2 cellsReduced viability with prolonged exposure (phototoxicity)[41]
Inkjet-basedPulse amplitude40–80 VHuman fibroblasts cellsDecreased cell viability as pulse amplitude increased[52]
Laser-assistedLaser fluence800–1600 mJ cm−2 NIH 3T3 mouse fibroblast cellsDecreased cell viability
as the laser fluence increases		[39]
Light-based bioprintingUV dose0.5–20 kg/m2L929 mouse fibroblasts, human mesenchymal stem cellsInduced DNA damage and increased apoptosis under higher UV intensity/exposure conditions[53]
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Rahman, T.T.; Pei, Z.; Qin, H.; Parsaei, H.R. Prospects and Limitations of Bioprinting in Studying Human Cells’ Responses to Extreme Environments. Bioengineering 2026, 13, 458. https://doi.org/10.3390/bioengineering13040458

AMA Style

Rahman TT, Pei Z, Qin H, Parsaei HR. Prospects and Limitations of Bioprinting in Studying Human Cells’ Responses to Extreme Environments. Bioengineering. 2026; 13(4):458. https://doi.org/10.3390/bioengineering13040458

Chicago/Turabian Style

Rahman, Taieba Tuba, Zhijian Pei, Hongmin Qin, and Hamid R. Parsaei. 2026. "Prospects and Limitations of Bioprinting in Studying Human Cells’ Responses to Extreme Environments" Bioengineering 13, no. 4: 458. https://doi.org/10.3390/bioengineering13040458

APA Style

Rahman, T. T., Pei, Z., Qin, H., & Parsaei, H. R. (2026). Prospects and Limitations of Bioprinting in Studying Human Cells’ Responses to Extreme Environments. Bioengineering, 13(4), 458. https://doi.org/10.3390/bioengineering13040458

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop