Shivesh Anand

All Publications

• Human iPSC-Based in Vitro Cardiovascular Tissue Models for Drug Screening Applications. Current cardiology reports Anand, S., Chen, G., Khanna, A., Huang, N. F. 2025; 27 (1): 145

  Abstract

  To provide an overview of human induced pluripotent stem cell (hiPSC)-derived cardiovascular lineages and describe their impact on drug testing in vitro.hiPSCs have garnered tremendous interest over the last decade due to their potential for unlimited proliferation and differentiation into cardiovascular lineages. Technologies using tissue engineering, 3D bioprinting, and organ-on-a-chip platforms composed of hiPSC derivatives can produce cardiovascular tissue mimetics that enhance drug screening applications. hiPSC-derived cardiovascular lineages advance drug screening efforts by using autologous cells that are more therapeutically relevant. Established approaches to reproducibly generate hiPSC-derived cardiovascular lineages and their subsequent organization into 3D constructs more accurately mimic the physiological organization of cardiac tissue, leading to improved identification of potential drug targets for therapeutic testing.

  View details for DOI 10.1007/s11886-025-02284-x

  View details for PubMedID 41128948

  View details for PubMedCentralID 10922734

• Acoustically Responsive Nanofibrous Scaffolds with 3D Hierarchy for Tympanic Membrane Regeneration SMALL STRUCTURES Anand, S., Del Toro Runzer, C., Rosado Balmayor, E., van Griensven, M., Moroni, L., Mota, C. 2025

  View details for DOI 10.1002/sstr.202500155

  View details for Web of Science ID 001531650200001

• Tunable ciprofloxacin delivery through personalized electrospun patches for tympanic membrane perforations BIOACTIVE MATERIALS Anand, S., Fusco, A., Guenday, C., Guenday-Tuereli, N., Donnarumma, G., Danti, S., Moroni, L., Mota, C. 2024; 38: 109-123

  Abstract

  Approximately 740 million symptomatic patients are affected by otitis media every year. Being an inflammatory disease affecting the middle ear, it is one of the primary causes of tympanic membrane (TM) perforations, often resulting in impaired hearing abilities. Antibiotic therapy using broad-spectrum fluoroquinolones, such as ciprofloxacin (CIP), is frequently employed and considered the optimal route to treat otitis media. However, patients often get exposed to high dosages to compensate for the low drug concentration reaching the affected site. Therefore, this study aims to integrate tissue engineering with drug delivery strategies to create biomimetic scaffolds promoting TM regeneration while facilitating a localized release of CIP. Distinct electrospinning (ES) modalities were designed in this regard either by blending CIP into the polymer ES solution or by incorporating nanoparticles-based co-ES/electrospraying. The combination of these modalities was investigated as well. A broad range of release kinetic profiles was achieved from the fabricated scaffolds, thereby offering a wide spectrum of antibiotic concentrations that could serve patients with diverse therapeutic needs. Furthermore, the incorporation of CIP into the TM patches demonstrated a favorable influence on their resultant mechanical properties. Biological studies performed with human mesenchymal stromal cells confirmed the absence of any cytotoxic or anti-proliferative effects from the released antibiotic. Finally, antibacterial assays validated the efficacy of CIP-loaded scaffolds in suppressing bacterial infections, highlighting their promising relevance for TM applications.

  View details for DOI 10.1016/j.bioactmat.2024.04.001

  View details for Web of Science ID 001228871400001

  View details for PubMedID 38699239

  View details for PubMedCentralID PMC11063525

• Embracing Remote Fields as the Fourth Dimension of Tissue Biofabrication ADVANCED FUNCTIONAL MATERIALS Anand, S., Mueller, C., Jensen, B., Chen, M. 2024; 34 (32)

  View details for DOI 10.1002/adfm.202401654

  View details for Web of Science ID 001204667200001

• Chitin nanofibrils modulate mechanical response in tympanic membrane replacements. Carbohydrate polymers Anand, S., Azimi, B., Lucena, M., Ricci, C., Candito, M., Zavagna, L., Astolfi, L., Coltelli, M. B., Lazzeri, A., Berrettini, S., Moroni, L., Danti, S., Mota, C. 2023; 310: 120732

  Abstract

  The tympanic membrane (TM), is a thin tissue lying at the intersection of the outer and the middle ear. TM perforations caused by traumas and infections often result in a conductive hearing loss. Tissue engineering has emerged as a promising approach for reconstructing the damaged TM by replicating the native material characteristics. In this regard, chitin nanofibrils (CN), a polysaccharide-derived nanomaterial, is known to exhibit excellent biocompatibility, immunomodulation and antimicrobial activity, thereby imparting essential qualities for an optimal TM regeneration. This work investigates the application of CN as a nanofiller for poly(ethylene oxide terephthalate)/poly(butylene terephthalate) (PEOT/PBT) copolymer to manufacture clinically suitable TM scaffolds using electrospinning and fused deposition modelling. The inclusion of CN within the PEOT/PBT matrix showed a three-fold reduction in the corresponding electrospun fiber diameters and demonstrated a significant improvement in the mechanical properties required for TM repair. Furthermore, in vitro biodegradation assay highlighted a favorable influence of CN in accelerating the scaffold degradation over a period of one year. Finally, the oto- and cytocompatibility response of the nanocomposite substrates corroborated their biological relevance for the reconstruction of perforated eardrums.

  View details for DOI 10.1016/j.carbpol.2023.120732

  View details for PubMedID 36925264

• Cellular uptake of modified mRNA occurs via caveolae-mediated endocytosis, yielding high protein expression in slow-dividing cells. Molecular therapy. Nucleic acids Del Toro Runzer, C., Anand, S., Mota, C., Moroni, L., Plank, C., van Griensven, M., Balmayor, E. R. 2023; 32: 960-979

  Abstract

  Nucleic acids have clear clinical potential for gene therapy. Plasmid DNA (pDNA) was the first nucleic acid to be pursued as a therapeutic molecule. Recently, mRNA came into play as it offers improved safety and affordability. In this study, we investigated the uptake mechanisms and efficiencies of genetic material by cells. We focused on three main variables (1) the nucleic acid (pDNA, or chemically modified mRNA), (2) the delivery vector (Lipofectamine 3000 or 3DFect), and (3) human primary cells (mesenchymal stem cells, dermal fibroblasts, and osteoblasts). In addition, transfections were studied in a 3D environment using electrospun scaffolds. Cellular internalization and intracellular trafficking were assessed by using enhancers or inhibitors of endocytosis and endosomal escape. The polymeric vector TransIT-X2 was included for comparison purposes. While lipoplexes utilized several entry routes, uptake via caveolae served as the main route for gene delivery. pDNA yielded higher expression levels in fast-dividing fibroblasts, whereas, in slow-dividing osteoblasts, cmRNA was responsible for high protein production. In the case of mesenchymal stem cells, which presented an intermediate doubling time, the combination vector/nucleic acid seemed more relevant than the nucleic acid per se. In all cases, protein expression was higher when the cells were seeded on 3D scaffolds.

  View details for DOI 10.1016/j.omtn.2023.05.019

  View details for PubMedID 37305166

  View details for PubMedCentralID PMC10250585

• DRUG RELEASING HIERARCHICAL SCAFFOLDS FOR HUMAN EARDRUM RECONSTRUCTION Anand, S., Gunday, C., Munafo, S., Tureli, N., Danti, S., Moroni, L., Mota, C. MARY ANN LIEBERT, INC. 2022: S410

  View details for Web of Science ID 000821187301595

• TYMPANIC MEMBRANE SCAFFOLDS AIDED BY CHITIN NANOFIBRILS TO MODULATE INFLAMMATORY AND IMMUNE RESPONSE Azimi, B., Zavagna, L., Ricci, C., Fusco, A., Milazzo, M., Anand, S., Donnarumma, G., Mota, C., Danti, S. MARY ANN LIEBERT, INC. 2022: S651-S652

  View details for Web of Science ID 000821187303399

• MIMICKING HUMAN TYMPANIC MEMBRANE: THE SIGNIFICANCE OF GEOMETRY Anand, S., Stoppe, T., Lucena, M., Neudert, M., Danti, S., Moroni, L., Mota, C. MARY ANN LIEBERT, INC. 2022: S406-S407

  View details for Web of Science ID 000821187301584

• Regenerative therapies for tympanic membrane PROGRESS IN MATERIALS SCIENCE Anand, S., Danti, S., Moroni, L., Mota, C. 2022; 127

  View details for DOI 10.1016/j.pmatsci.2022.100942

  View details for Web of Science ID 000793420700001

• Shaping and properties of thermoplastic scaffolds in tissue regeneration: The effect of thermal history on polymer crystallization, surface characteristics and cell fate JOURNAL OF MATERIALS RESEARCH Calore, A., Srinivas, V., Anand, S., Albillos-Sanchez, A., Looijmans, S. P., van Breemen, L. C. A., Mota, C., Bernaerts, K., Harings, J. A. W., Moroni, L. 2021; 36 (19): 3914-3935

  View details for DOI 10.1557/s43578-021-00403-2

  View details for Web of Science ID 000709661900002

• Chitin Nanofibril Application in Tympanic Membrane Scaffolds to Modulate Inflammatory and Immune Response. Pharmaceutics Danti, S., Anand, S., Azimi, B., Milazzo, M., Fusco, A., Ricci, C., Zavagna, L., Linari, S., Donnarumma, G., Lazzeri, A., Moroni, L., Mota, C., Berrettini, S. 2021; 13 (9)

  Abstract

  Chitin nanofibrils (CNs) are an emerging bio-based nanomaterial. Due to nanometric size and high crystallinity, CNs lose the allergenic features of chitin and interestingly acquire anti-inflammatory activity. Here we investigate the possible advantageous use of CNs in tympanic membrane (TM) scaffolds, as they are usually implanted inside highly inflamed tissue environment due to underlying infectious pathologies. In this study, the applications of CNs in TM scaffolds were twofold. A nanocomposite was used, consisting of poly(ethylene oxide terephthalate)/poly(butylene terephthalate) (PEOT/PBT) copolymer loaded with CN/polyethylene glycol (PEG) pre-composite at 50/50 (w/w %) weight ratio, and electrospun into fiber scaffolds, which were coated by CNs from crustacean or fungal sources via electrospray. The degradation behavior of the scaffolds was investigated during 4 months at 37 °C in an otitis-simulating fluid. In vitro tests were performed using cell types to mimic the eardrum, i.e., human mesenchymal stem cells (hMSCs) for connective, and human dermal keratinocytes (HaCaT cells) for epithelial tissues. HMSCs were able to colonize the scaffolds and produce collagen type I. The inflammatory response of HaCaT cells in contact with the CN-coated scaffolds was investigated, revealing a marked downregulation of the pro-inflammatory cytokines. CN-coated PEOT/PBT/(CN/PEG 50:50) scaffolds showed a significant indirect antimicrobial activity.

  View details for DOI 10.3390/pharmaceutics13091440

  View details for PubMedID 34575515

  View details for PubMedCentralID PMC8468799

• Mimicking the Human Tympanic Membrane: The Significance of Scaffold Geometry. Advanced healthcare materials Anand, S., Stoppe, T., Lucena, M., Rademakers, T., Neudert, M., Danti, S., Moroni, L., Mota, C. 2021; 10 (11): e2002082

  Abstract

  The human tympanic membrane (TM) captures sound waves from the environment and transforms them into mechanical motion. The successful transmission of these acoustic vibrations is attributed to the unique architecture of the TM. However, a limited knowledge is available on the contribution of its discrete anatomical features, which is important for fabricating functional TM replacements. This work synergizes theoretical and experimental approaches toward understanding the significance of geometry in tissue-engineered TM scaffolds. Three test designs along with a plain control are chosen to decouple some of the dominant structural elements, such as the radial and circumferential alignment of the collagen fibrils. In silico models suggest a geometrical dependency of their mechanical and acoustical responses, where the presence of radially aligned fibers is observed to have a more prominent effect compared to their circumferential counterparts. Following which, a hybrid fabrication strategy combining electrospinning and additive manufacturing has been optimized to manufacture biomimetic scaffolds within the dimensions of the native TM. The experimental characterizations conducted using macroindentation and laser Doppler vibrometry corroborate the computational findings. Finally, biological studies with human dermal fibroblasts and human mesenchymal stromal cells reveal a favorable influence of scaffold hierarchy on cellular alignment and subsequent collagen deposition.

  View details for DOI 10.1002/adhm.202002082

  View details for PubMedID 33945239

  View details for PubMedCentralID PMC11469228

• Ciprofloxacin-loaded polymeric nanoparticles incorporated electrospun fibers for drug delivery in tissue engineering applications. Drug delivery and translational research Günday, C., Anand, S., Gencer, H. B., Munafò, S., Moroni, L., Fusco, A., Donnarumma, G., Ricci, C., Hatir, P. C., Türeli, N. G., Türeli, A. E., Mota, C., Danti, S. 2020; 10 (3): 706-720

  Abstract

  Presented work focuses on the development of biodegradable polymer nanoparticles loaded with antibiotics as drug delivery systems deposited on electrospun scaffolds for tissue engineering. The innovative ciprofloxacin-loaded poly(DL-lactide-co-glycolide) NPs ensure a continuous slow release and high local concentration at the site of action for an optimal therapy. The local delivery of antibiotics as an integrated part of electrospun scaffolds offers an effective, safe, and smart enhancement supporting tissue regeneration. Presented data provides solid scientific evidence for fulfilling the requirements of local nano antibiotic delivery systems with biodegradability and biocompatibility for a wide range of tissue engineering applications, including middle ear tissues (e.g., tympanic membranes) which are subject to bacterial infections. Further characterization of such systems, including in vivo studies, is required to ensure successful transfer from lab to clinical applications. Graphical abstract .

  View details for DOI 10.1007/s13346-020-00736-1

  View details for PubMedID 32100267

• Dimensionality changes actin network through lamin A/C and zyxin. Biomaterials Zonderland, J., Moldero, I. L., Anand, S., Mota, C., Moroni, L. 2020; 240: 119854

  Abstract

  Mechanosensing proteins have mainly been investigated in 2D culture platforms, while understanding their regulation in 3D enviroments is critical for tissue engineering. Among mechanosensing proteins, the actin cytoskeleton plays a key role in human mesenchymal stromal cells (hMSCs) activity, but its regulation in 3D tissue engineered scaffolds remains poorly studied. Here, we show that human mesenchymal stromal cells (hMSCs) cultured on 3D electrospun scaffolds made of a stiff material do not form actin stress fibers, contrary to hMSCs on 2D films of the same material. On 3D electrospun and additive manufactured scaffolds, hMSCs also displayed fewer focal adhesions, lower lamin A and C expression and less YAP1 nuclear localization and myosin light chain phosphorylation. Together, this strongly suggests that dimensionality prevents the build-up of cellular tension, even on stiff materials. Knock down of either lamin A and C or zyxin resulted in fewer stress fibers in the cell center. Zyxin knock down reduced lamin A and C expression, but not vice versa, showing that this signal chain starts from the outside of the cell. Lineage commitment was not affected by the lack of these important osteogenic proteins in 3D, as all cells committed to osteogenesis in bi-potential medium. Our study demonstrates that dimensionality changes the actin cytoskeleton through lamin A and C and zyxin, and highlights the difference in the regulation of lineage commitment in 3D enviroments. Together, these results can have important implications for future scaffold design for both stiff- and soft tissue engineering constructs.

  View details for DOI 10.1016/j.biomaterials.2020.119854

  View details for PubMedID 32087459

• Force Modulation and Adaptability of 3D-Bioprinted Biological Actuators Based on Skeletal Muscle Tissue ADVANCED MATERIALS TECHNOLOGIES Mestre, R., Patino, T., Barcelo, X., Anand, S., Perez-Jimenez, A., Sanchez, S. 2019; 4 (2)

  View details for DOI 10.1002/admt.201800631

  View details for Web of Science ID 000459632800059

• Reduction of efficiency droop in GaN/InGaN based multiple quantum well light emitting diode by varying Si-doping and thickness in barrier layers OPTIK Anand, S., Mahala, P., Singh, S., Pal, S. 2019; 178: 645-649

  View details for DOI 10.1016/j.ijleo.2018.09.151

  View details for Web of Science ID 000454472000081

• Development of a Water-Dispersible SBA-15-Benzothiazole-Derived Fluorescence Nanosensor by Physisorption and Its Use in Organic-Solvent-Free Detection of Perborate and Hydrazine CHEMISTRYSELECT Gawas, R. U., Anand, S., Ghosh, B. K., Shivbhagwan, P., Choudhary, K., Ghosh, N., Banerjee, M., Chatterjee, A. 2018; 3 (38): 10585-10592

  View details for DOI 10.1002/slct.201802328

  View details for Web of Science ID 000447546900007

• The arrival of commercial bioprinters - Towards 3D bioprinting revolution! International journal of bioprinting Choudhury, D., Anand, S., Naing, M. W. 2018; 4 (2): 139

  Abstract

  The dawn of commercial bioprinting is rapidly advancing the tissue engineering field. In the past few years, new bioprinting approaches as well as novel bioinks formulations have emerged, enabling biological research groups to demonstrate the use of such technology to fabricate functional and relevant tissue models. In recent years, several companies have launched bioprinters pushing for early adoption and democratisation of bioprinting. This article reviews the progress in commercial bioprinting since the inception, with a particular focus on the comparison of different available printing technologies and important features of the individual technologies as well as various existing applications. Various challenges and potential design considerations for next generations of bioprinters are also discussed.

  View details for DOI 10.18063/IJB.v4i2.139

  View details for PubMedID 33102917

  View details for PubMedCentralID PMC7582003

• Bioprinting for Neural Tissue Engineering. Trends in neurosciences Knowlton, S., Anand, S., Shah, T., Tasoglu, S. 2018; 41 (1): 31-46

  Abstract

  Bioprinting is a method by which a cell-encapsulating bioink is patterned to create complex tissue architectures. Given the potential impact of this technology on neural research, we review the current state-of-the-art approaches for bioprinting neural tissues. While 2D neural cultures are ubiquitous for studying neural cells, 3D cultures can more accurately replicate the microenvironment of neural tissues. By bioprinting neuronal constructs, one can precisely control the microenvironment by specifically formulating the bioink for neural tissues, and by spatially patterning cell types and scaffold properties in three dimensions. We review a range of bioprinted neural tissue models and discuss how they can be used to observe how neurons behave, understand disease processes, develop new therapies and, ultimately, design replacement tissues.

  View details for DOI 10.1016/j.tins.2017.11.001

  View details for PubMedID 29223312