Three-dimensional bioprinted GelMA/GO composite hydrogel for stem cell osteogenic differentiation both in vitro and in vivo.

In this study, we evaluated the use of graphene oxide (GO) mixed with methyl methacrylate gelatin (GelMA) for the construction of a microenvironmental implant to repair bone defects in orthopedic surgery. A scaffold containing a GelMA/GO composite with mesenchymal stem cells (MSCs) was constructed using three-dimensional bioprinting. The survival and osteogenic capacity of MSCs in the composite bioink were evaluated using cell viability and proliferation assays, osteogenesis-related gene expression analysis, and implantation under the skin of nude mice. The printing process had little effect on cell viability. We found that GO enhanced cell proliferation but had no significant effect on cell viability. In vitro experiments suggested that GO promoted material-cell interactions and the expression of osteogenesis-related genes. In vivo experiments showed that GO decreased the degradation time of the material and increased calcium nodule deposition. In contrast to pure GelMA, the addition of GO created a suitable microenvironment to promote the differentiation of loaded exogenous MSCs in vitro and in vivo, providing a basis for the repair of bone defects.

Iodine-Doped Hollow Carbon Nanocages without Templates Strategy for Boosting Zinc-Ion Storage by Nucleophilicity.

Zn-ion hybrid supercapacitors (ZHCs) combining merits of battery-type and capacitive electrodes are considered to be a prospective candidate in energy storage systems. Tailor-made carbon cathodes with high zincophilicity and abundant physi/chemisorption sites are critical but it remains a great challenge to achieve both features by a sustainable means. Herein, a hydrogen-bonding interaction-guided self-assembly strategy is presented to prepare iodine-doped carbon nanocages without templates for boosting zinc-ion storage by nucleophilicity. The biomass ellagic acid contains extensional hydroxy and acyloxy groups with electron-donating ability, which interact with melamine and ammonium iodide to form organic supermolecules. The organic supermolecules further self-assemble into a nanocage-like structure with cavities under hydrothermal processes via hydrogen-bonding and π-π stacking. The carbon nanocages as ZHCs cathodes enable the high approachability of zincophilic sites and low ion migration resistance resulting from the interconnected conductive network and nanoscale architecture. The experimental analyses and theoretical simulations reveal the pivotal role of iodine dopants. The I5-/I3- doping anions in carbon cathodes have a nucleophilicity to preferentially adsorb the Zn2+ cation by the formation of C+-I5--Zn2+ and C+-I3--Zn2+. Of these, the C+-I3- shows stronger bonding with Zn2+ than C+-I5-. As a result, the iodine-doped carbon nanocages produced via this template-free strategy deliver a high capacity of 134.2 mAh/g at 1 A/g and a maximum energy and power density of 114.1 Wh/kg and 42.5 kW/kg.

Metabolomics analysis of the potential mechanism of Yi-Guan-Jian decoction to reverse bone loss in glucocorticoid-induced osteoporosis.

BACKGROUND: Glucocorticoid-induced osteoporosis (GIOP) is a disease in which long-term use of glucocorticoid causes bone loss, deterioration of bone microstructure and fracture. Currently, clinical drugs targeting this disease have certain side effects. There is still a need to find effective drugs with fewer side effects. The theory of traditional Chinese medicine suggests that YGJ has therapeutic effect on GIOP, but it has not been explained. Therefore, this study aims to explore the protective effect of YGJ on GIOP mouse models and elucidate the underlying mechanism through LC-MS-based metabolomics analysis. METHODS: The general condition of 8 week age male C57BL/6J mice was recorded after 8 weeks of treatment with dexamethasone (DEX) and YGJ. Bone-related parameters and bone morphology were determined by Micro-CT. HE staining was used to observe the pathological changes of bone tissue. Serum levels of bone metabolism markers were detected by ELISA. Liver metabolomics analysis was conducted to search for the significant markers of anti-GIOP of YGJ and the metabolic pathway affecting it. RESULTS: After treatment, YGJ significantly reversed the weight loss caused by DEX; increase the number of bone trabecular in ROI region, significantly improve the bone-related parameters of GIOP mice, and increase the levels of alkaline phosphatase and osteocalcin. In the study of metabolic mechanism, YGJ reversed 24 potential markers in GIOP mice. These included cortisol, 3-hydroxybutyric acid, taurine, esculin and uric acid, which are closely associated with osteoporosis. Topological analysis results showed that YGJ had the most significant effect on taurine and hypotaurine metabolism, with - log10 (P) > 2.0 and Impact > 0.4. CONCLUSIONS: Yi-Guan-Jian decoction can increase bone density and improve bone microstructure by regulating the levels of alkaline phosphatase and osteocalcin and reverse bone loss in GIOP mouse model. The underlying metabolic mechanism may be related to taurine and hypotaurine metabolic pathway.

3D bioprinted autologous bone particle scaffolds for cranioplasty promote bone regeneration with both implanted and native BMSCs.

Although autologous bone (AB) grafting is considered to be the gold standard for cranioplasty, unresolved problems remain, such as surgical-site infections and bone flap absorption. In this study, an AB scaffold was constructed via three-dimensional (3D) bedside-bioprinting technology and used for cranioplasty. To simulate the skull structure, a polycaprolactone shell was designed as an external lamina, and 3D-printed AB and a bone marrow-derived mesenchymal stem cell (BMSC) hydrogel was used to mimic cancellous bone for bone regeneration. Ourin vitroresults showed that the scaffold exhibited excellent cellular affinity and promoted osteogenic differentiation of BMSCs in both two-dimensional and 3D culture systems. The scaffold was implanted in beagle dog cranial defects for up to 9 months, and the scaffold promoted new bone and osteoid formation. Furtherin vivostudies indicated that transplanted BMSCs differentiated into vascular endothelium, cartilage, and bone tissues, whereas native BMSCs were recruited into the defect. The results of this study provide a method for bedside bioprinting of a cranioplasty scaffold for bone regeneration, which opens up another window for clinical applications of 3D printing in the future.

Multiscale Physics-Informed Neural Networks for Stiff Chemical Kinetics.

In this paper, a multiscale physics-informed neural network (MPINN) approach is proposed based on the regular physics-informed neural network (PINN) for solving stiff chemical kinetic problems with governing equations of stiff ordinary differential equations (ODEs). In MPINNs, chemical species with different time scales are grouped and trained by multiple corresponding neural networks with the same structure. The adaptive weight based on a key performance indicator is assigned to each loss term when calculating the summation of loss residues. With this structure, MPINNs provide a framework to solve challenging stiff chemical kinetic problems without any stiffness-removal artifacts before training. In addition, by introducing a small number of ground truth data (GTD) points (less than 10% of the number required for residual loss calculation) and adding data loss terms into loss functions, MPINNs show superior ability to represent stiff ODE solutions at any desired time. The accuracy of MPINNs is tested with classical chemical kinetic problems, by comparing with the regular PINN and other state-of-the-art methods with special consideration for solving stiff chemical kinetic problems with PINNs. The validation results show that MPINNs can effectively avoid the influence of stiffness on neural network optimization. Compared with the traditional deep neural network only trained by GTD, MPINNs can use no data or a relatively small amount of data to achieve high-precision prediction of stiff chemical ODEs. The proposed approach is very promising for solving stiff chemical kinetics, opening up possibilities of MPINN application in different fields involving stiff chemical dynamics.

Mechanical engineering of hair follicle regeneration by in situ bioprinting.

Hair loss caused by various factors such as trauma, stress, and diseases hurts patient psychology and seriously affects patients' quality of life, but there is no effective method to control it. In situ bioprinting is a method for printing bioinks directly into defective sites according to the shape and characteristics of the defective tissue or organ to promote tissue or organ repair. In this study, we applied a 3D bioprinting machine in situ bioprinting of epidermal stem cells (Epi-SCs), skin-derived precursors (SKPs), and Matrigel into the wounds of nude mice to promote hair follicle regeneration based on their native microenvironment. The results showed successful regeneration of hair follicles and other skin appendages at 4 weeks after in situ bioprinting. Moreover, we confirmed that bioprinting only slightly decreased stem cell viability and maintained the stemness of the stem cells. These findings demonstrated a mechanical engineering method for hair follicle regeneration by in situ bioprinting which has potential in the clinic.

Adaptive multi-degree-of-freedom in situ bioprinting robot for hair-follicle-inclusive skin repair: A preliminary study conducted in mice.

Skin acts as an essential barrier, protecting organisms from their environment. For skin trauma caused by accidental injuries, rapid healing, personalization, and functionality are vital requirements in clinical, which are the bottlenecks hindering the translation of skin repair from benchside to bedside. Herein, we described a novel design and a proof-of-concept demonstration of an adaptive bioprinting robot to proceed rapid in situ bioprinting on a full-thickness excisional wound in mice. The three-dimensional (3D) scanning and closed-loop visual system integrated in the robot and the multi-degree-of-freedom mechanism provide immediate, precise, and complete wound coverage through stereotactic bioprinting, which hits the key requirements of rapid-healing and personalization in skin repair. Combined with the robot, epidermal stem cells and skin-derived precursors isolated from neonatal mice mixed with Matrigel were directly printed into the injured area to replicate the skin structure. Excisional wounds after bioprinting showed complete wound healing and functional skin tissue regeneration that closely resembling native skin, including epidermis, dermis, blood vessels, hair follicles and sebaceous glands etc. This study provides an effective strategy for skin repair through the combination of the novel robot and a bioactive bioink, and has a promising clinical translational potential for further applications.

3D bioprinted GelMA/GO composite induces osteoblastic differentiation.

Graft substitute is a mature treatment tool in craniofacial bone repair. However, stress shielding and immutability of structure limit its use in patients with congenital defects. Therefore, a regenerative graft would be best suited for repair. Mesenchymal stem cells (MSCs) have been shown to be feasible in regenerative medicine and the clinical treatment of bone repair. The aim of this study was to propose a strategy that would directly blend graphene oxide (GO) and MSCs with gelatin methacrylate anhydride (GelMA), as bioink, to generate the scaffold for bone regenerative repair. The survival and osteogenic capacity of MSCs in the composite bioink were assessed by cell viability and proliferation assays, along with expression analysis of osteogenesis-related genes and proteins, and targeted immunofluorescence. The introduction of GO to the printing process had no influence on cell printing, viability, or printability of GelMa. However, the GO-involved structure exhibited a positive influence on MSC proliferation, without significantly affecting cell viability. Alkaline phosphatase was expressed more in cells cultured with GO than in those with pure GelMA. In addition, GO promoted the expression of osteogenesis-related genes and proteins, such as osteopontin, osteocalcin, and RUNX2. Collectively, the composite bioink enhanced cell proliferation and adhesion, as well as osteogenic differentiation properties, compared with pure GelMA.

A scalable coaxial bioprinting technology for mesenchymal stem cell microfiber fabrication and high extracellular vesicle yield.

Mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) are promising candidates for regenerative medicine; however, the lack of scalable methods for high quantity EV production limits their application. In addition, signature EV-derived proteins shared in 3D environments and 2D surfaces, remain mostly unknown. Herein, we present a platform combining MSC microfiber culture with ultracentrifugation purification for high EV yield. Within this platform, a high quantity MSC solution (∼3 × 108total cells) is encapsulated in a meter-long hollow hydrogel-microfiber via coaxial bioprinting technology. In this 3D core-shell microfiber environment, MSCs express higher levels of stemness markers (Oct4, Nanog, Sox2) than in 2D culture, and maintain their differentiation capacity. Moreover, this platform enriches particles by ∼1009-fold compared to conventional 2D culture, while preserving their pro-angiogenic properties. Liquid chromatography-mass spectrometry characterization results demonstrate that EVs derived from our platform and conventional 2D culturing have unique protein profiles with 3D-EVs having a greater variety of proteins (1023 vs 605), however, they also share certain proteins (536) and signature MSC-EV proteins (10). This platform, therefore, provides a new tool for EV production using microfibers in one culture dish, thereby reducing space, labor, time, and cost.

A facile, versatile hydrogel bioink for 3D bioprinting benefits long-term subaqueous fidelity, cell viability and proliferation.

Both of the long-term fidelity and cell viability of three-dimensional (3D)-bioprinted constructs are essential to precise soft tissue repair. However, the shrinking/swelling behavior of hydrogels brings about inadequate long-term fidelity of constructs, and bioinks containing excessive polymer are detrimental to cell viability. Here, we obtained a facile hydrogel by introducing 1% aldehyde hyaluronic acid (AHA) and 0.375% N-carboxymethyl chitosan (CMC), two polysaccharides with strong water absorption and water retention capacity, into classic gelatin (GEL, 5%)-alginate (ALG, 1%) ink. This GEL-ALG/CMC/AHA bioink possesses weak temperature dependence due to the Schiff base linkage of CMC/AHA and electrostatic interaction of CMC/ALG. We fabricated integrated constructs through traditional printing at room temperature and in vivo simulation printing at 37°C. The printed cell-laden constructs can maintain subaqueous fidelity for 30 days after being reinforced by 3% calcium chloride for only 20 s. Flow cytometry results showed that the cell viability was 91.38 ± 1.55% on day 29, and the cells in the proliferation plateau at this time still maintained their dynamic renewal with a DNA replication rate of 6.06 ± 1.24%. This work provides a convenient and practical bioink option for 3D bioprinting in precise soft tissue repair.

Bioprinting of Human Cord Blood-Derived CD34+ Cells and Exploration of the Multilineage Differentiation Ability in Vitro.

The three-dimensional (3D) marrow microenvironment plays an essential role in regulating human cord blood-derived CD34+ cells (hCB-CD34+) migration, proliferation, and differentiation. Extensive in vitro and in vivo studies have aimed to recapitulate the main components of the bone marrow (BM) niche. Nonetheless, the models are limited by a lack of heterogeneity and compound structure. Here, we fabricated coaxial extruded core-shell tubular scaffolds and extrusion-based bioprinted cell-laden mesh scaffolds to mimic the functional niche in vitro. A multicellular mesh scaffold and two different core-shell tubular scaffolds were developed with human bone marrow-derived mesenchymal stromal cells (BMSCs) in comparison with a conventional 2D coculture system. A clear cell-cell connection was established in all three bioprinted constructs. Cell distribution and morphology were observed in different systems with scanning electron microscopy (SEM). Collected hCB-CD34+ cells were characterized by various stem cell-specific and lineage-specific phenotypic parameters. The results showed that compared with hCB-CD34+ cells cocultured with BMSCs in Petri dishes, the self-renewal potential of hCB-CD34+ cells was stronger in the tubular scaffolds after 14 days. Besides, cells in these core-shell constructs tended to obtain stronger differentiation potential of lymphoid and megakaryocytes, while cells encapsulated in mesh scaffolds obtained stronger differentiation tendency into erythroid cells. Consequently, 3D bioprinting technology could partially simulate the niche of human hematopoietic stem cells. The three models have their potential in stemness maintenance and multilineage differentiation. This study can provide initial effective guidance in the directed differentiation research and related screening of drug models for hematological diseases.

A structure-supporting, self-healing, and high permeating hydrogel bioink for establishment of diverse homogeneous tissue-like constructs.

The ready-to-use, structure-supporting hydrogel bioink can shorten the time for ink preparation, ensure cell dispersion, and maintain the preset shape/microstructure without additional assistance during printing. Meanwhile, ink with high permeability might facilitate uniform cell growth in biological constructs, which is beneficial to homogeneous tissue repair. Unfortunately, current bioinks are hard to meet these requirements simultaneously in a simple way. Here, based on the fast dynamic crosslinking of aldehyde hyaluronic acid (AHA)/N-carboxymethyl chitosan (CMC) and the slow stable crosslinking of gelatin (GEL)/4-arm poly(ethylene glycol) succinimidyl glutarate (PEG-SG), we present a time-sharing structure-supporting (TSHSP) hydrogel bioink with high permeability, containing 1% AHA, 0.75% CMC, 1% GEL and 0.5% PEG-SG. The TSHSP hydrogel can facilitate printing with proper viscoelastic property and self-healing behavior. By crosslinking with 4% PEG-SG for only 3 min, the integrity of the cell-laden construct can last for 21 days due to the stable internal and external GEL/PEG-SG networks, and cells manifested long-term viability and spreading morphology. Nerve-like, muscle-like, and cartilage-like in vitro constructs exhibited homogeneous cell growth and remarkable biological specificities. This work provides not only a convenient and practical bioink for tissue engineering, targeted cell therapy, but also a new direction for hydrogel bioink development.

Risk assessment of airborne transmission of COVID-19 by asymptomatic individuals under different practical settings.

The lack of quantitative risk assessment of airborne transmission of COVID-19 under practical settings leads to large uncertainties and inconsistencies in our preventive measures. Combining in situ measurements and computational fluid dynamics simulations, we quantify the exhaled particles from normal respiratory behaviors and their transport under elevator, small classroom, and supermarket settings to evaluate the risk of inhaling potentially virus-containing particles. Our results show that the design of ventilation is critical for reducing the risk of particle encounters. Inappropriate design can significantly limit the efficiency of particle removal, create local hot spots with orders of magnitude higher risks, and enhance particle deposition causing surface contamination. Additionally, our measurements reveal the presence of a substantial fraction of faceted particles from normal breathing and its strong correlation with breathing depth.

Inkjet Bioprinting of Biomaterials.

The inkjet technique has the capability of generating droplets in the picoliter volume range, firing thousands of times in a few seconds and printing in the noncontact manner. Since its emergence, inkjet technology has been widely utilized in the publishing industry for printing of text and pictures. As the technology developed, its applications have been expanded from two-dimensional (2D) to three-dimensional (3D) and even used to fabricate components of electronic devices. At the end of the twentieth century, researchers were aware of the potential value of this technology in life sciences and tissue engineering because its picoliter-level printing unit is suitable for depositing biological components. Currently inkjet technology has been becoming a practical tool in modern medicine serving for drug development, scaffold building, and cell depositing. In this article, we first review the history, principles and different methods of developing this technology. Next, we focus on the recent achievements of inkjet printing in the biological field. Inkjet bioprinting of generic biomaterials, biomacromolecules, DNAs, and cells and their major applications are introduced in order of increasing complexity. The current limitations/challenges and corresponding solutions of this technology are also discussed. A new concept, biopixels, is put forward with a combination of the key characteristics of inkjet printing and basic biological units to bring a comprehensive view on inkjet-based bioprinting. Finally, a roadmap of the entire 3D bioprinting is depicted at the end of this review article, clearly demonstrating the past, present, and future of 3D bioprinting and our current progress in this field.

A 3D engineered scaffold for hematopoietic progenitor/stem cell co-culture in vitro.

Proliferation of HPSCs in vitro can promote its broad clinical therapeutic use. For in vitro co-culture, interaction between the stem cell and feeder cell as well as their spatial position are essential. To imitate the natural microenvironment, a 3D engineered scaffold for CD34+ cells co-culture was established via 3D bioprinting. Herein, the concentration of hydrogel and the ratio of two kinds of cells were optimized. Flow cytometry, real time PCR and RNA-seq technology were applied to analyze the effect of the engineered scaffold on expanded cells. After 10 days co-culture with the engineered scaffold, the expansion of CD34+CD38- cells can reach 33.57-folds and the expansion of CD34+CD184+ cells can reach 16.66-folds. Result of PCR and RNA-seq indicates that the CD34+ cells in 3D group exhibited a tendency of interaction with the engineered scaffold. Compared to 2D co-culture, this customizable 3D engineered scaffold can provide an original and integrated environment for HPSCs growth. Additionally, this scaffold can be modified for different cell co-culture or cell behavior study.

Risk assessment of airborne transmission of COVID-19 by asymptomatic individuals under different practical settings.

The lack of quantitative risk assessment of airborne transmission of COVID-19 under practical settings leads to large uncertainties and inconsistencies in our preventive measures. Combining in situ measurements and numerical simulations, we quantify the exhaled particles from normal respiratory behaviors and their transport under elevator, small classroom and supermarket settings to evaluate the risk of inhaling potentially virus-containing particles. Our results show that the design of ventilation is critical for reducing the risk of particle encounters. Inappropriate design can significantly limit the efficiency of particle removal, create local hot spots with orders of magnitude higher risks, and enhance particle deposition causing surface contamination. Additionally, our measurements reveal the presence of substantial fraction of crystalline particles from normal breathing and its strong correlation with breathing depth.

A coaxially extruded heterogeneous core-shell fiber with Schwann cells and neural stem cells.

Cellular therapies play a critical role in the treatment of spinal cord injury (SCI). Compared with cell-seeded conduits, fully cellular grafts have more similarities with autografts, and thus might result in better regeneration effects. In this study, we fabricated Schwann cell (SC)-neural stem cell (NSC) core-shell alginate hydrogel fibers in a coaxial extrusion manner. The rat SC line RSC96 and mouse NSC line NE-4C were used in this experiment. Fully cellular components were achieved in the core portion and the relative spatial positions of these two cells partially mimic the construction of nerve fibers in vivo. SCs were demonstrated to express more genes of neurotrophic factors in alginate shell. Enhanced proliferation and differentiation tendency of NSCs was observed when they were co-cultured with SCs. This model has strong potential for application in SCI repair.

Biomaterials Based on Marine Resources for 3D Bioprinting Applications.

Three-dimensional (3D) bioprinting has become a flexible tool in regenerative medicine with potential for various applications. Further development of the new 3D bioprinting field lies in suitable bioink materials with satisfied printability, mechanical integrity, and biocompatibility. Natural polymers from marine resources have been attracting increasing attention in recent years, as they are biologically active and abundant when comparing to polymers from other resources. This review focuses on research and applications of marine biomaterials for 3D bioprinting. Special attention is paid to the mechanisms, material requirements, and applications of commonly used 3D bioprinting technologies based on marine-derived resources. Commonly used marine materials for 3D bioprinting including alginate, carrageenan, chitosan, hyaluronic acid, collagen, and gelatin are also discussed, especially in regards to their advantages and applications.

Effects of straw amendment on selenium aging in soils: Mechanism and influential factors.

Soil dissolved organic matter (DOM) alters heavy metal availability, but whether straw amendment can manipulate soil selenium (Se) speciation and availability through DOM mineralization remains unclear. In this study, allochthonous maize straw and selenate were incubated together in four different soils for 1 y. The transformation and availability of DOM associated Se (DOM-Se) was investigated during aging. Results indicated that soil solution and soil particle surfaces were dominated by hexavalent hydrophilic acid-bound Se (Hy-Se). The amount of fulvic acid bound Se in soil solution (SOL-FA-Se) was higher than humic acid bound Se in soil solution (SOL-HA-Se), except in krasnozems, and mainly existed as hexavalent Se (Se(VI)). Tetravalent Se (Se(IV)) was the main valence state of FA-Se adsorbed on soil particle surfaces (EX-FA-Se) after 5 w of aging. The proportion of soil-available Se (SOL + EX-Se) decreased with increasing straw rate. However, under an application rate of 7500 kg·hm-2, soluble Se fraction (SOL-Se) reduction was minimal in acidic soils (18.7%-34.7%), and the organic bound Se fraction (OM-Se) was maximally promoted in alkaline soils (18.2%-39.1%). FA and HON could enhance the availability of Se in the soil solution and on particle surfaces of acidic soil with high organic matter content. While Se incorporation with HA could accelerate the fixation of Se into the solid phase of soil. Three mechanisms were involved in DOM-Se aging: (1) Reduction, ligand adsorption, and inner/outer-sphere complexation associated with the functional groups of straw-derived DOM, including hydroxyls, carboxyl, methyl, and aromatic phenolic compounds; (2) interconnection of EX-FA-Se between non-residual and residual Se pools; and (3) promotion by soil electrical conductivity (EC), clay, OM, and straw application. The dual effect of DOM on Se aging was highly reliant on the characteristics of the materials and soil properties. In conclusion, straw amendment could return selenium in soil and reduce soluble Se loss.

Human embryonic lung fibroblasts treated with artesunate exhibit reduced rates of proliferation and human cytomegalovirus infection in vitro.

BACKGROUND: Cytomegalovirus (CMV) pneumonia is a major cause of death in immunosuppressed patients. Despite the effective treatment with ganciclovir (GCV) and other antiviral agents, the mortality rate remains between 30% to 50%. Recently, the anti-malarial drug artesunate (ART) wasfound to exhibit significant anti-viral activity. Here, we examined the effects of ART on human cytomegalovirus (HCMV) infection and human embryonic lung fibroblast (HELF) proliferation in vitro. METHODS: HELFs infected with the GFP-expressing Towne-BAC strain of HCMV were divided into three treatment groups: Group I, cells treated with ART for 1.5 h before HCMV inoculation; Group II, cells infected with HCMV that was pre-treated with ART for 1.5 h before HCMV inoculation; Group III, cells that were treated with ART at 1.5 h post-HCMV inoculation. GFP expression was observed daily by fluorescence microscopy, and the number of GFP-positive cells in each experimental group was recorded at 4-5 days post-infection. At 10 days post-infection, the viability of cells in each group was recorded. GCV treatment was used as a control. RESULTS: While no significant effects on cytotoxicity, cell viability, viral infection rates, or antiviral activity were observed upon treatment of Group I or II cells with GCV or low levels of ART, the ART-treated Group III population exhibited significantly reduced rates of infection at drug concentrations higher than 12.5 µM. Similarly, we observed a GCV concentration-dependent reduction in the viral infection rate in Group III cells. Notably, ART-treated, but not GCV-treated, cells also exhibited decreased proliferation. The 50% cytostatic concentrations (CC50) and the half maximal inhibitory concentrations (IC50) of ART and GCV were 54.382 µM and 12.679 µM, and 3.76 M and 14.479 µM, respectively. CONCLUSIONS: In addition to its robust antiviral activity, ART inhibits proliferation of HCMV-infected lung fibroblasts, making it a potential next-generation drug for CMV pneumonia treatment and for reducing fibroproliferation and fibrosis in these patients.