bims-biprem 2024-07-21 papers

bims-biprem

Biomed News

on Bioprinting for regenerative medicine

Issue of 2024–07–21
eleven papers selected by
Seerat Maqsood, University of Teramo

Application of 4D printing in dentistry: A narrative review.
Fabrication of chitin-fibrin hydrogels to construct the 3D artificial extracellular matrix scaffold for vascular regeneration and cardiac tissue engineering.
Fabrication, characterization and evaluating properties of 3D printed PLA-Mn scaffolds.
Bioengineered Silk Protein-Based 3D In Vitro Models for Tissue Engineering and Drug Development: From Silk Matrix Properties to Biomedical Applications.
Fabrication of 3D-Printed Scaffolds with Multiscale Porosity.
Pneumatic extrusion bioprinting-based high throughput fabrication of a melanoma 3D cell culture model for anti-cancer drug screening.
Direct 3D printing of freeform anisotropic bioactive structure based on shear-oriented ink system.
A practical machine learning approach for predicting the quality of 3D (bio)printed scaffolds.
3D Printed O2-Generating Scaffolds Enhance Osteoprogenitor- and Type H Vessel Recruitment During Bone Healing.
Viscoelasticity in 3D Cell Culture and Regenerative Medicine: Measurement Techniques and Biological Relevance.
Extrusion-based 3D Bioprinting of Bioactive and Piezoelectric Scaffolds for Treating Critical Soft Tissue Wounds.

J Adv Periodontol Implant Dent. 2024 ;16(1): 55-63

Application of 4D printing in dentistry: A narrative review.

Yeshwanth Perambudhuru, Lata Goyal, Meghna Dewan, Ajay Mahajan, Prabhat Kumar Chaudhari.

  4D printing is an innovative digital manufacturing technology that originated by adding a fourth dimension, i.e., time, to pre-existing 3D technology or additive manufacturing (AM). AM is a fast-growing technology used in many fields, which develops accurate 3D objects based on models designed by computers. Dentistry is one such field in which 3D technology is used for manufacturing objects in periodontics (scaffolds, local drug-delivering agents, augmentation of ridges), implants, prosthodontics (partial and complete dentures, obturators), oral surgery for reconstructing jaw, and orthodontics. Dynamism is a vital property needed for the survival of materials used in the oral cavity since the oral cavity is constantly subjected to various insults. 4D printing technology has overcome the disadvantages of 3D printing technology, i.e., it cannot create dynamic objects. Therefore, constant knowledge of 4D technology is required. 3D printing technology has shortcomings, which are discussed in this review. This review summaries various printing technologies, materials used, stimuli, and potential applications of 4D technology in dentistry.

Keywords:  3D printing; Computers; Dentistry; Dynamic; Printing; Smart material; Technology; Three-dimensional; time

DOI:  https://doi.org/10.34172/japid.2024.003
J Biomed Mater Res A. 2024 Jul 15.

Fabrication of chitin-fibrin hydrogels to construct the 3D artificial extracellular matrix scaffold for vascular regeneration and cardiac tissue engineering.

Pengcheng Yang, Fang Xie, Lihang Zhu, Jonathan Nimal Selvaraj, Donghui Zhang, Jie Cai.

  As the cornerstone of tissue engineering and regeneration medicine research, developing a cost-effective and bionic extracellular matrix (ECM) that can precisely modulate cellular behavior and form functional tissue remains challenging. An artificial ECM combining polysaccharides and fibrillar proteins to mimic the structure and composition of natural ECM provides a promising solution for cardiac tissue regeneration. In this study, we developed a bionic hydrogel scaffold by combining a quaternized β-chitin derivative (QC) and fibrin-matrigel (FM) in different ratios to mimic a natural ECM. We evaluated the stiffness of those composite hydrogels with different mixing ratios and their effects on the growth of human umbilical vein endothelial cells (HUVECs). The optimal hydrogels, QCFM1 hydrogels were further applied to load HUVECs into nude mice for in vivo angiogenesis. Besides, we encapsulated human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) into QCFM hydrogels and employed 3D bioprinting to achieve batch fabrication of human-engineered heart tissue (hEHT). Finally, the myocardial structure and electrophysiological function of hEHT were evaluated by immunofluorescence and optical mapping. Designed artificial ECM has a tunable modulus (220-1380 Pa), which determines the different cellular behavior of HUVECs when encapsulated in these. QCFM1 composite hydrogels with optimal stiffness (800 Pa) and porous architecture were finally identified, which could adapt for in vitro cell spreading and in vivo angiogenesis of HUVECs. Moreover, QCFM1 hydrogels were applied in 3D bioprinting successfully to achieve batch fabrication of both ring-shaped and patch-shaped hEHT. These QCFM1 hydrogels-based hEHTs possess organized sarcomeres and advanced function characteristics comparable to reported hEHTs. The chitin-derived hydrogels are first used for cardiac tissue engineering and achieve the batch fabrication of functionalized artificial myocardium. Specifically, these novel QCFM1 hydrogels provided a reliable and economical choice serving as ideal ECM for application in tissue engineering and regeneration medicine.

Keywords:  3D bioprinting; angiogenesis; artificial extracellular matrix; cardiac tissue engineering; chitin hydrogels; fibrin hydrogels

DOI:  https://doi.org/10.1002/jbm.a.37774
Sci Rep. 2024 Jul 18. 14(1): 16592

Fabrication, characterization and evaluating properties of 3D printed PLA-Mn scaffolds.

Sina Dehghan-Toranposhti, Rasoul Bakhshi, Reza Alizadeh, Mahboubeh Bohlouli.

  Polylactic acid (PLA) based scaffolds have attained considerable attention in recent years for being used as biodegradable implants in bone tissue engineering (BTE), owing to their suitable biocompatibility and processability. Nevertheless, the mechanical properties, bioactivity and biodegradation rate of PLA need to be improved for practical application. In this investigation, PLA-xMn composite filaments (x = 0, 1, 3, 5 and 7 wt%) were fabricated, characterized, and used for 3D printing of scaffolds by the fused deposition modeling process. The effect of Mn addition on the thermal, physical, mechanical, and structural properties, as well as the degradability and cell viability of 3D printed scaffolds were investigated in details. The obtained results indicate that the PLA-Mn composite filaments exhibit higher chain mobility and melt flow index values, with lower cold crystallization temperature and a higher degree of crystallinity. This higher flowability led to lower dimensional accuracy of 3D printed scaffolds, but resulted in higher interlayer adhesion. It was found that the mechanical properties of composite scaffolds were remarkably enhanced with the addition of Mn particles. The incorporation of Mn particles also caused higher surface roughness and hydrophilicity, a superior biodegradation rate of the scaffolds as well as better biocompatibility, indicating a promising candidate for (BTE) applications.

Keywords:  3D printing; Biodegradation; Bone tissue scaffold; FDM; Manganese; PLA

DOI:  https://doi.org/10.1038/s41598-024-67478-9
Adv Healthc Mater. 2024 Jul 15. e2401458

Bioengineered Silk Protein-Based 3D In Vitro Models for Tissue Engineering and Drug Development: From Silk Matrix Properties to Biomedical Applications.

Yajun Shuai, Meidan Zheng, Subhas C Kundu, Chuanbin Mao, Mingying Yang.

  3D in vitro model has emerged as a valuable tool for studying tissue development, drug screening, and disease modeling. 3D systems can accurately replicate tissue microstructures and physiological features, mirroring the in vivo microenvironment departing from conventional 2D cell cultures. Various 3D in vitro models utilizing biomacromolecules like collagen and synthetic polymers have been developed to meet diverse research needs and address the complex challenges of contemporary research. Silk proteins, bearing structural and functional similarities to collagen, have been increasingly employed to construct advanced 3D in vitro systems, surpassing the limitations of 2D cultures. This review examines silk proteins' composition, structure, properties, and functions, elucidating their role in 3D in vitro models. Furthermore, recent advances in biomedical applications involving silk-based organoid models are discussed. In particular, the unique physiological attributes of silk matrix constituents in in vitro tissue constructs are highlighted, providing a meticulous evaluation of their importance. Additionally, it outlines the current research hurdles and complexities while contemplating future avenues, thereby paving the way for developing complex and biomimetic silk protein-based microtissues.

Keywords:  3D printing; hydrogels; organoids; silk fibroin; tissue regeneration

DOI:  https://doi.org/10.1002/adhm.202401458
ACS Omega. 2024 Jul 09. 9(27): 29186-29204

Fabrication of 3D-Printed Scaffolds with Multiscale Porosity.

Rafał Podgórski, Michał Wojasiński, Artur Małolepszy, Jakub Jaroszewicz, Tomasz Ciach.

3D printing is a promising technique for producing bone implants, but there is still a need to adjust efficiency, facilitate production, and improve biocompatibility. Porous materials have a proven positive effect on the regeneration of bone tissue, but their production is associated with numerous limitations. In this work, we described a simple method of producing polymer or polymer-ceramic filaments for 3D-printing scaffolds by adding micrometer-scale porous structures on scaffold surfaces. Scaffolds included polycaprolactone (PCL) as the primary polymer, β-tricalcium phosphate (β-TCP) as the ceramic filler, and poly(ethylene glycol) (PEG) as a porogen. The pressurized filament extrusion gave flexible filaments composed of PCL, β-TCP, and PEG, which are ready to use in fused filament fabrication (FFF) 3D printers. Washing of 3D-printed scaffolds in ethanol solution removed PEG and revealed a microporous structure and ceramic particles on the scaffold's surfaces. Furthermore, 3D-printed materials exhibit good printing precision, no cytotoxic properties, and highly impact MG63 cell alignment. Although combining PCL, PEG, and β-TCP is quite popular, the presented method allows the production of porous scaffolds with a well-organized structure without advanced equipment, and the produced filaments can be used to 3D print scaffolds on a simple commercially available 3D printer.

DOI: https://doi.org/10.1021/acsomega.3c09035
Biomed Mater. 2024 Jul 18.

Pneumatic extrusion bioprinting-based high throughput fabrication of a melanoma 3D cell culture model for anti-cancer drug screening.

Maryke de Villiers, Awie F Kotzé, Lissinda H du Plessis.

  The high incidence of malignant melanoma highlights the need for in vitro models that accurately represent the tumour microenvironment, enabling developments in melanoma therapy and drug screening. Despite several advancements in 3D cell culture models, appropriate melanoma models for evaluating drug efficacy are still in high demand. The 3D pneumatic extrusion-based bioprinting technology offers numerous benefits, including the ability to achieve high-throughput capabilities. However, there is a lack of research that combines pneumatic extrusion-based bioprinting with analytical assays to enable efficient drug screening in 3D melanoma models. To address this gap, this study developed a simple and highly reproducible approach to fabricate a 3D A375 melanoma cell culture model using the pneumatic extrusion-based bioprinting technology. To optimise this method, the bioprinting parameters for producing 3D cell cultures in a 96-well plate were adjusted to improve reproducibility while maintaining the desired droplet size and a cell viability of 92.13± 6.02%. The cross-linking method was optimised by evaluating cell viability and proliferation of the 3D bioprinted cells in three different concentrations of calcium chloride. The lower concentration of 50 mM resulted in higher cell viability and increased cell proliferation after 9 days of incubation. The A375 cells exhibited a steadier proliferation rate in the 3D bioprinted cell cultures, and tended to aggregate into spheroids, whereas the 2D cell cultures generally formed monolayered cell sheets. In addition, we evaluated the drug responses of four different anti-cancer drugs on the A375 cells in both the 2D and 3D cell cultures. The 3D cell cultures exhibited higher levels of drug resistance in all four tested anti-cancer drugs. This method presents a simple and cost-effective method of producing and analysing 3D cell culture models that do not add additional complexity to current assays and shows considerable potential for advancing 3D cell culture models' drug efficacy evaluations.

Keywords:  3D cell culture models; droplet-based extrusion bioprinting; extrusion-based bioprinting; high-throughput bioprinting; high-throughput drug screening

DOI:  https://doi.org/10.1088/1748-605X/ad651f
Biofabrication. 2024 Jul 15.

Direct 3D printing of freeform anisotropic bioactive structure based on shear-oriented ink system.

Chenhui Yuan, Jinhong Jiang, Xinyu Zhang, Lin Gu, Xueping Wang, Lei Shao.

  Various anisotropic tissue structures exist in organisms, including muscle tissue, skin tissue, and nerve tissue. Replicating anisotropic tissue structures in vitro has posed a significant challenge. 3D printing technology is often used to fabricate biomimetic structures due to its advantages in manufacturing principle. However, direct 3D printing of freeform anisotropic bioactive structures has not been reported. To tackle this challenge, we developed a ternary F/G/P ink system that integrates the printability of Pluronic F127 (F), the robust bioactivity and photocrosslinking properties of GelMA (G), and the shear-induced alignment functionality of high-molecular-weight PEG (P). And through this strategic ternary system combination, freeform anisotropic tissue structures can be 3D printed directly. Moreover, these anisotropic structures exhibit excellent bioactivity, and promote orientational growth of different cells. This advancement holds promise for the repair and replacement of anisotropic tissues within the human body.

Keywords:  3D printing; Anisotropic structure; Cell orientation; High printability; Shear orientation

DOI:  https://doi.org/10.1088/1758-5090/ad6375
Biofabrication. 2024 Jul 15.

A practical machine learning approach for predicting the quality of 3D (bio)printed scaffolds.

Saeed Rafieyan, Elham Ansari, Ebrahim Vasheghani-Farahani.

  3D (Bio)printing is a highly effective method for fabricating tissue engineering scaffolds, renowned for their exceptional precision and control. Artificial intelligence (AI) has become a crucial technology in this field, capable of learning and replicating complex patterns that surpass human capabilities. However, the integration of AI in tissue engineering is often hampered by the lack of comprehensive and reliable data. This study addresses these challenges by providing one of the most extensive datasets on 3D-printed scaffolds. It provides the most comprehensive open-source dataset and employs various AI techniques, from unsupervised to supervised learning. This dataset includes detailed information on 1,171 scaffolds, featuring a variety of biomaterials and concentrations-including 60 biomaterials such as natural and synthesized biomaterials, crosslinkers, enzymes, etc.-along with 49 cell lines, cell densities, and different printing conditions. We used over 40 machine learning and deep learning algorithms, tuning their hyperparameters to reveal hidden patterns and predict cell response, printability, and scaffold quality. The clustering analysis using KMeans identified five distinct ones. In classification tasks, algorithms such as XGBoost, Gradient Boosting, Extra Trees Classifier, Random Forest Classifier, and LightGBM demonstrated superior performance, achieving higher accuracy and F1 scores. A fully connected neural network with six hidden layers from scratch was developed, precisely tuning its hyperparameters for accurate predictions. To promote future research, the developed dataset and the associated code are publicly available on www.github.com/saeedrafieyan/MLATE.&#xD.

Keywords:  3D printing; Clustering; Deep learning; Machine learning; bioprinting; tissue engineering

DOI:  https://doi.org/10.1088/1758-5090/ad6374
Acta Biomater. 2024 Jul 13. pii: S1742-7061(24)00382-9. [Epub ahead of print]

3D Printed O2-Generating Scaffolds Enhance Osteoprogenitor- and Type H Vessel Recruitment During Bone Healing.

Naboneeta Sarkar, Jingtong Zhao, Nicholas Y Zhang, Allison L Horenberg, Warren L Grayson.

  Oxygen (O2)-delivering tissue substitutes have shown tremendous potential for enhancing tissue regeneration, maturation, and healing. As O2 is both a metabolite and powerful signaling molecule, providing controlled delivery is crucial for optimizing its beneficial effects in the treatment of critical-sized injuries. Here, we report the design and fabrication of 3D-printed, biodegradable, O2-generating bone scaffold comprising calcium peroxide (CPO) that once hydrolytically activated, provides long-term generation of oxygen at a controlled, concentration-dependent manner, and polycaprolactone (PCL), a hydrophobic polymer that regulate the interaction of CPO with water, preventing burst release of O2 at early time points. When anoxic conditions were simulated in vitro, CPO-PCL scaffolds maintained the survival and proliferation of human adipose-derived stem/stromal cells (hASCs) relative to PCL-only controls. We assessed the in vivo osteogenic efficacy of hASC-seeded CPO-PCL scaffolds implanted in a non-healing critical-sized 4-mm calvarial defects in nude mice for 8 weeks. Even without exogenous osteoinductive factors, CPO-PCL scaffolds demonstrated increased new bone volume compared to PCL-only scaffolds as verified by both microcomputed tomography analysis and histological assessments. Lastly, we employed a quantitative 3D lightsheet microscopy platform to determine that O2-generating scaffolds had similar vascular volumes with slightly higher presence of CD31hiEmcnhi pro-osteogenic, type H vessels and increased number of Osterix+ skeletal progenitor cells relative to PCL-only scaffolds. In summary, 3D-printed O2 generating CPO-PCL scaffolds with tunable O2 release rates provide a facile, customizable strategy for effectively treating, craniofacial bone defects. STATEMENT OF SIGNIFICANCE: Oxygen(O2)-delivering bone substitutes show promise in defect repair applications by supplying O2 to the cells within or around the graft, improving cell survivability and enhancing bone matrix mineralization. A novel O2-generating bone scaffold has been 3D printed for the first-time which ensures patient and defect specificity. 3D printed calcium peroxide-polycaprolactone (CPO-PCL) bone scaffold provides uninterrupted O2 supply for 22 days allowing cell survival in deprived O2 and nutrient conditions. For the first time, O2-driven bone regenerative environment in mice calvaria has been captured by light-sheet imaging which illuminates abundance of Osterix+ cells, angiogenesis at a single cell resolution indicating active site of bone remodeling and growth in the presence of O2.

Keywords:  Angiogenesis; Bone regeneration; Bone tissue engineering; Calcium peroxide; Oxygen

DOI:  https://doi.org/10.1016/j.actbio.2024.07.011
ACS Mater Au. 2024 Jul 10. 4(4): 354-384

Viscoelasticity in 3D Cell Culture and Regenerative Medicine: Measurement Techniques and Biological Relevance.

Payam Eliahoo, Hesam Setayesh, Tyler Hoffman, Yifan Wu, Song Li, Jennifer B Treweek.

The field of mechanobiology is gaining prominence due to recent findings that show cells sense and respond to the mechanical properties of their environment through a process called mechanotransduction. The mechanical properties of cells, cell organelles, and the extracellular matrix are understood to be viscoelastic. Various technologies have been researched and developed for measuring the viscoelasticity of biological materials, which may provide insight into both the cellular mechanisms and the biological functions of mechanotransduction. Here, we explain the concept of viscoelasticity and introduce the major techniques that have been used to measure the viscoelasticity of various soft materials in different length- and timescale frames. The topology of the material undergoing testing, the geometry of the probe, the magnitude of the exerted stress, and the resulting deformation should be carefully considered to choose a proper technique for each application. Lastly, we discuss several applications of viscoelasticity in 3D cell culture and tissue models for regenerative medicine, including organoids, organ-on-a-chip systems, engineered tissue constructs, and tunable viscoelastic hydrogels for 3D bioprinting and cell-based therapies.

DOI: https://doi.org/10.1021/acsmaterialsau.3c00038
Adv Wound Care (New Rochelle). 2024 Jul 13.

Extrusion-based 3D Bioprinting of Bioactive and Piezoelectric Scaffolds for Treating Critical Soft Tissue Wounds.

Surendrasingh Y Sonaye, Smriti Bohara, Breanne Welsh, Elif G Ertugral, Chandrasekhar R Kothapalli, Hanz Richter, Koyal Garg, Prabaha Sikder.

OBJECTIVE: This study focuses on developing bioactive piezoelectric scaffolds that could deliver bioelectrical cues to potentially treat injuries to soft tissues such as skeletal muscles and promote muscle regeneration.
APPROACH: To address the underexplored aspect of bioelectrical cues in skeletal muscle tissue engineering (SMTE), we developed piezoelectric bioinks based on natural bioactive materials such as alginate, gelatin, and chitosan. Extrusion-based 3D bioprinting was utilized to develop scaffolds that mimic muscle stiffness and generate electrical stimulation when subjected to forces. The biocompatibility of these scaffolds was tested with C2C12 muscle cell line.
RESULTS: The bioinks demonstrated suitable rheological properties for 3D bioprinting, resulting in high-resolution composite alginate-gelatin-chitosan scaffolds with good structural fidelity. The scaffolds exhibited a 42-60 kPa stiffness, similar to muscles. When a controlled force of 5 N was applied to the scaffolds at a constant frequency of 4 Hz, they generated electrical fields and impulses (charge), indicating their suitability as a standalone scaffold to generate electrical stimulation and instill bioelectrical cues in the wound region. The cell viability and proliferation test results confirm the scaffold's biocompatibility with C2C12s and the benefit of piezoelectricity in promoting muscle cell growth kinetics. Our study indicates that our piezoelectric bioinks and scaffolds offer promise as autonomous electrical stimulation-generating regenerative therapy for SMTE.
INNOVATION: A novel approach for treating skeletal muscle wounds was introduced by developing a bioactive electroactive scaffold capable of autonomously generating electrical stimulation without stimulators and electrodes. This scaffold offers a unique approach to enhancing skeletal muscle regeneration through bioelectric cues, addressing a major gap in the SMTE, i.e., fibrotic tissue formation due to delayed muscle regeneration.
CONCLUSION: A piezoelectric scaffold was developed, providing a promising solution for promoting skeletal muscle regeneration. This development can potentially address skeletal muscle injuries and offer a unique approach to facilitating skeletal muscle wound healing.

DOI: https://doi.org/10.1089/wound.2024.0073