E2-Loaded Microcapsules and Bone Marrow–Derived Mesenchymal Stem Cells with Injectable Scaffolds for Endometrial Regeneration Application
Tissue Engineering Part AYuelin Wu, Shengyi Gu, Jonathan M. Cobb, Griffin H. Dunn, Taylor A. Muth, Chloe J. Simchick, Baoguo Li*, Wujie Zhang*, and Xiaolin Hua*
2024
Abstract
Bone marrow–derived mesenchymal stem cells (BMSCs) have been recognized as new candidates for the treatment of serious endometrial injuries. However, owing to the local microenvironment of damaged endometrium, transplantation of BMSCs yielded disappointing results. In this study, Pectin-Pluronic® F-127 hydrogel as scaffolds were fabricated to provide three-dimensional architecture for the attachment, growth, and migration of BMSCs. E2 was encapsulated into the W/O/W microspheres to construct pectin-based E2-loaded microcapsules (E2 MPs), which has the potential to serve as a long-term reliable source of E2 for endometrial regeneration. Then, the BMSCs/E2 MPs/scaffolds system was injected into the uterine cavity of mouse endometrial injury model for treatment. At 4 weeks after transplantation, the system increased proliferative abilities of uterine endometrial cells, facilitated microvasculature regeneration, and restored the ability of endometrium to receive an embryo, suggesting that the BMSCs/E2 MPs/scaffolds system is a promising treatment option for endometrial regeneration. Furthermore, the mechanism of E2 in promoting the repair of endometrial injury was also investigated. Exosomes are critical paracrine mediators that act as biochemical cues to direct stem cell differentiation. In this study, it was found that the expression of endometrial epithelial cell (EEC) markers was upregulated in BMSCs treated by exosomes secreted from endometrial stromal cells (ESCs-Exos). Exosomes derived from E2-stimulated ESCs further promoted the expression level of EECs markers in BMSCs, suggesting exosomes released from ESCs by E2 stimulation could enhance the differentiation efficiency of BMSCs. Therefore, exosomes derived from ESCs play paracrine roles in endometrial regeneration stimulated by E2 and provide optimal estrogenic response.
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Development of Gelatin-Coated Hydrogel Microspheres for Novel Bioink Design: A Crosslinker Study
Invited Feature Paper: PharmaceuticsJoshua Zieman, Megan Cohan, Yang Wang, Alexa De La Sancha, Muskan Kanungo, Ryan Azzouz, Rebekah Smith, Keagan Schmidt, Subha Kumpaty, Junhong Chen, Wujie Zhang*
2023
Design of Pectin-Based Hydrogel Microspheres for Targeted Pulmonary Delivery
Invited Feature Paper: GelsAndy Chai, Keagan Schmidt, Gregory Brewster, Lu Shi Peng Xiong, Benjamin Church, Timothy Wahl, Hamed Sadabadi, Subha Kumpaty, and Wujie Zhang*
2023
Abstract
Pulmonary drug delivery via microspheres has gained growing interest as a noninvasive method for therapy. However, drug delivery through the lungs via inhalation faces great challenges due to the natural defense mechanisms of the respiratory tract, such as the removal or deactivation of drugs. This study aims to develop a natural polymer-based microsphere system with a diameter of around 3 μm for encapsulating pulmonary drugs and facilitating their delivery to the deep lungs. Pectin was chosen as the foundational material due to its biocompatibility and degradability in physiological environments. Electrospray was used to produce the pectin-based hydrogel microspheres, and Design-Expert software was used to optimize the production process for microsphere size and uniformity. The optimized conditions were determined to be as follows: pectin/PEO ratio of 3:1, voltage of 14.4 kV, distance of 18.2 cm, and flow rate of 0.95 mL/h. The stability and responsiveness of the pectin-based hydrogel microspheres can be altered through coatings such as gelatin. Furthermore, the potential of the microspheres for pulmonary drug delivery (i.e., their responsiveness to the deep lung environment) was investigated. Successfully coated microspheres with 0.75% gelatin in 0.3 M mannitol exhibited improved stability while retaining high responsiveness in the simulated lung fluid (Gamble's solution). A gelatin-coated pectin-based microsphere system was developed, which could potentially be used for targeted drug delivery to reach the deep lungs and rapid release of the drug.
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Recent Progress in Bioprinting: From Bioink Design to Applications
BioengineeringWujie Zhang
2022
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One-pot synthesis and enzyme-responsiveness of amphiphilic doxorubicin prodrug nanomicelles for cancer therapeutics
RSC AdvancesLing-Na Han, Kai-Qiang Wang, Zi-Ning Ren, Xue Yang, Xiao Duan, Sasirekha Krishnan, Abinaya Jaisankar, Jeong-Hui Park, Khandmaa Dashnyam, Wujie Zhang, José Luis Pedraz, Seeram Ramakrishna, Hae-Won Kim, Chang-Feng Li, Li-Hua Song, and Murugan Ramalingam
2022
Abstract
In this study, we report a one-pot synthesis and enzyme-responsiveness of polyethylene glycol (PEG) and glutamic acid (Glu)-based amphiphilic doxorubicin (DOX) prodrug nanomicelles for cancer therapeutics. The nanomicelles were accomplished by esterification and amidation reactions. The nuclear magnetic resonance (NMR) and Fourier transform infrared (FTIR) data confirmed the structure of nanomicelles. The DOX-loaded nanomicelles showed a DLS-measured average size of 107 nm and excellent stability in phosphate-buffered saline (PBS) for 7 days. The drug loading and cumulative release rates were measured by ultraviolet-visible (UV-vis) spectrophotometry at 481 nm. The cumulative release rate could reach 100% in an enzyme-rich environment. Further, the therapeutic efficiency of nanomicelles to cancer cells was determined by cell viability and cellular uptake and distribution using HeLa cells. The cell viability study showed that the DOX-loaded nanomicelles could effectively inhibit the HeLa cell proliferation. The cellular uptake study confirmed that the nanomicelles could be effectively ingested by HeLa cells and distributed into cell nuclei. Based on the collective experimental data, this study demonstrated that the synthesized nanomicellar prodrug of DOX is a potential candidate for cancer therapeutics.
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Green Synthesis of Gold Nanoparticles Using Upland Cress and Their Biochemical Characterization and Assessment
NanomaterialsNoah Hutchinson, Yuelin Wu, Yale Wang, Muskan Kanungo, Anna DeBruine, Emma Kroll, De’Jorra Gilmore, Zachary Eckrose, Stephanie Gaston, Phoebe Matel, Matey Kaltchev, Anne-Marie Nickel, Subha Kumpaty, Xiaolin Hua*, and Wujie Zhang*
2022 Abstract
This research focuses on the plant-mediated green synthesis process to produce gold nanoparticles (Au NPs) using upland cress (Barbarea verna), as various biomolecules within the upland cress act as both reducing and capping agents. The synthesized gold nanoparticles were thoroughly characterized using UV-vis spectroscopy, surface charge (zeta potential) analysis, scanning electron microscopy-energy-dispersive X-ray spectroscopy (SEM-EDX), atomic force microscopy (AFM), attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), and X-ray diffraction (XRD). The results indicated the synthesized Au NPs are spherical and well-dispersed with an average diameter ~11 nm and a characteristic absorbance peak at ~529 nm. EDX results showed an 11.13% gold content. Colloidal Au NP stability was confirmed with a zeta potential (ζ) value of −36.8 mV. X-ray diffraction analysis verified the production of crystalline face-centered cubic gold. Moreover, the antimicrobial activity of the Au NPs was evaluated using Gram-negative Escherichia coli and Gram-positive Bacillus megaterium. Results demonstrated concentration-dependent antimicrobial properties. Lastly, applications of the Au NPs in catalysis and biomedicine were evaluated. The catalytic activity of Au NPs was demonstrated through the conversion of 4-nitrophenol to 4-aminophenol which followed first-order kinetics. Cellular uptake and cytotoxicity were evaluated using both BMSCs (stem) and HeLa (cancer) cells and the results were cell type dependent. The synthesized Au NPs show great potential for various applications such as catalysis, pharmaceutics, and biomedicine.
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RSC Advances
One-pot synthesis and enzyme-responsiveness of amphiphilic doxorubicin prodrug nanomicelles for cancer therapeuticsLing-Na Han, Kai-Qiang Wang, Zi-Ning Ren, Xue Yang, Xiao Duan, Sasirekha Krishnan, Abinaya Jaisankar, Jeong-Hui Park, Khandmaa Dashnyam, Wujie Zhang, José Luis Pedraz, Seeram Ramakrishna, Hae-Won Kim, Chang-Feng Li, Li-Hua Song, and Murugan Ramalingam
2022
Abstract
In this study, we report a one-pot synthesis and enzyme-responsiveness of polyethylene glycol (PEG) and glutamic acid (Glu)-based amphiphilic doxorubicin (DOX) prodrug nanomicelles for cancer therapeutics. The nanomicelles were accomplished by esterification and amidation reactions. The nuclear magnetic resonance (NMR) and Fourier transform infrared (FTIR) data confirmed the structure of nanomicelles. The DOX-loaded nanomicelles showed a DLS-measured average size of 107 nm and excellent stability in phosphate-buffered saline (PBS) for 7 days. The drug loading and cumulative release rates were measured by ultraviolet-visible (UV-vis) spectrophotometry at 481 nm. The cumulative release rate could reach 100% in an enzyme-rich environment. Further, the therapeutic efficiency of nanomicelles to cancer cells was determined by cell viability and cellular uptake and distribution using HeLa cells. The cell viability study showed that the DOX-loaded nanomicelles could effectively inhibit the HeLa cell proliferation. The cellular uptake study confirmed that the nanomicelles could be effectively ingested by HeLa cells and distributed into cell nuclei. Based on the collective experimental data, this study demonstrated that the synthesized nanomicellar prodrug of DOX is a potential candidate for cancer therapeutics.
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Investigating the stabilisation of INFα2a by replica-exchange molecular dynamic simulation
Journal of Molecular ModelingDaixi Li, Peiqin Chen, Qingli Dong, Baolin Liu, Wujie Zhang, Dongqing Wei, and Baisong Guo
2022
Abstract
Current biopharmaceutical drugs are mainly a class of peptides or proteins that play an essential role in the treatment of many diseases. Such peptides/proteins are usually thermally unstable and may lose their bioactivity when exposed to ambient conditions. Therefore, they are not suitable for long-term storage. Lyophilisation is the most common method to prolong shelf life of solid peptide/protein drugs; however, the freeze-drying process can lead to irreversible damage. In the present study, human interferon-alpha 2a (IFN-α2a) was selected as a model protein drug; four disaccharides (β-lactose, β-maltose, sucrose, and trehalose) were selected as bioactive protectants. We investigated the effects of different protectants on IFN-α2a under various ambient conditions (vacuum, dry state, and aqueous solution) using replica exchange molecular dynamics simulation. The protective effect of β-maltose on IFN-α2a was the highest in aqueous solution and dry state, β-lactose showed a poor protective effect in all three conditions, the performance of sucrose was good in all conditions, and trehalose showed a better protective effect under vacuum conditions and in aqueous solution. Disaccharides form H-bonds with water, thereby preventing water from the tertiary structure of proteins. Trehalose forms strong H-bonds with water which explains its extraordinary stability.
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Validation of Solar Dehydrator for Food Drying Applications: A Granny Smith Apple Study
Journal of Chemical Engineering Research UpdatesJude Ingham, Muskan Kanungo, Brandon Beauchamp, Michael Korbut, Michael Swedish, Michael Navin, Wujie Zhang*
2022
Abstract
Food loss is a global issue that may be alleviated with effective dehydration strategies. Solar dehydration, rather than traditional sun-drying, is one method that could allow for the safe, efficient preservation of food materials. In this study, passive solar dehydration was achieved using a psychrometric chamber to model the environment of sub-Saharan Africa, where the temperature was the major focus (24.3 °C to 29.4 °C). A mass decrease of 88.56% was achieved within 9 hours. Microbial testing (total aerobic bacteria, Gram-negative bacteria, and total yeasts and molds) demonstrated no difference (all negative) between food stored at 4 °C and dehydrated food, indicating that the dehydrator introduced no new contamination. A 16.0% decrease in vitamin C (VC) concentration was observed due to the lability of VC. Insight into the visual appeal of the food samples was provided by measuring browning values, where it was found that dehydrated green apples are significantly less brown than the sample exposed to air for the same length of time. Passive solar dehydrators could provide a simple method to reduce food waste and maintain nutritional content and visual appeal.
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Development of Gelatin-Coated Microspheres for Novel Bioink Design
PolymersMuskan Kanungo, Yale Wang, Noah Hutchinson, Emma Kroll, Anna DeBruine, Subha Kumpaty, Lixia Ren, Yuelin Wu, Xiaolin Hua*, and Wujie Zhang*
2021 Abstract
A major challenge in tissue engineering is the formation of vasculature in tissue and organs. Recent studies have shown that positively charged microspheres promote vascularization, while also supporting the controlled release of bioactive molecules. This study investigated the development of gelatin-coated pectin microspheres for incorporation into a novel bioink. Electrospray was used to produce the microspheres. The process was optimized using Design-Expert® software. Microspheres underwent gelatin coating and EDC catalysis modifications. The results showed that the concentration of pectin solution impacted roundness and uniformity primarily, while flow rate affected size most significantly. The optimal gelatin concentration for microsphere coating was determined to be 0.75%, and gelatin coating led to a positively charged surface. When incorporated into bioink, the microspheres did not significantly alter viscosity, and they distributed evenly in bioink. These microspheres show great promise for incorporation into bioink for tissue engineering applications.
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Design and Stability Improvement of Pectin-based Red Blood Cell-mimicking Microcapsules for Oxygen Therapeutics
Journal of Biomedical NanotechnologyDamien Phakousonh, Yale Wang, Sabrina Schlicht, Sam Wiskirchen, Trevor Bos, Lixia Ren, Junhong Chen*, Xiaolin Hua, Jung Lee, Sasirekha Krishnan, Shoma Suresh K, Abinaya Jaisankar, Murugan Ramalingam*, and Wujie Zhang*
2021 Abstract
A pectin-oligochitosan microcapsule system has recently been developed for novel oxygen therapeutic design. To improve the stability of the pectin-oligochitosan microcapsules in physiological conditions, both covalent (glutaraldehyde) and noncovalent (Mn2+ and Ca2+) cross-linkers were tested. The chemistry and morphology of the microcapsules were studied using FTIR and SEM, respectively. Results showed that glutaraldehyde is an effective cross-linker, even at low concentrations and short incubation times, and the glutaraldehyde cross-linking does not negatively impact the morphology of the microcapsules. Moreover, it was confirmed that the hemoglobin could be retained within the microcapsules with a minimal release.
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Biosynthesis of Silver Nanoparticles using Upland Cress: Purification, Characterisation, and Antimicrobial Activity
Micro & Nano LettersDavid Johnson, Yale Wang, Samuel Stealey, Anne Alexander, Matey Kaltchev, Junhong Chen, and Wujie Zhang
2020
Silver nanoparticles have traditionally been synthesised using physical and chemical methods, often requiring expensive equipment and reagents that pose risks to the environment. This work provides a green method for the biosynthesis of silver nanoparticles using leaf extracts from upland cress: Barbarea verna. Natural reducing agents within the leaf extracts of upland cress reduce silver ions from silver nitrates, resulting in the formation of silver nanoparticles. The silver nanoparticles were purified using centrifugation and extraction using Triton X-114. The resulting nanoparticles were characterised using UV–Vis spectroscopy, dynamic light scattering, atomic force microscopy, and scanning electron microscopy. Silver nanoparticles were shown to have a diameter of 30–40 nm with a characteristic UV–Vis absorption peak at 420 nm. Antimicrobial properties of the synthesised silver nanoparticles were also confirmed using S. epidermis and E. coli bacteria.
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Calcium-Oligochitosan-Pectin Microcarrier for Colonic Drug Delivery
Pharmaceutical Development and TechnologySamuel Stealey, Xiaoru Guo, Rebecca Majewski, Alexander Dyble, Kendra Lehman, Michael Wedemeyer, Douglas A. Steeber, Matey G. Kaltchev, Junhong Chen, and Wujie Zhang
2020
Pectin-based hydrogel microcarriers have shown promise for drug delivery to the colonic region. Microcarriers must remain stable throughout the upper gastrointestinal tract for effective colonic delivery, an issue that traditional pectin-based microcarriers have faced. The positively-charged natural biopolymer oligochitosan and divalent cation Ca2+ were used to dually cross-link pectin-based hydrogel microcarriers to improve carrier stability through simulated gastric and intestinal environments. Microcarriers were characterized with Scanning Electron Microscope and Fourier-Transform Infrared analysis. An optical microscope was used to observe the change of microcarrier size and morphology over time in the simulated gastrointestinal environments. Fluorescently-labeled Dextran was used as a model drug for this system. Calcium-Oligochitosan-Pectin microcarriers exhibited relatively small drug release in the upper gastrointestinal regions and were responsive to the high pH and enzymatic activity of simulated col
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Nanotechnology for Bioengineers
Part of the book series: Synthesis Lectures on Biomedical Engineering (SLBE)Wujie Zhang
2020
Nanotechnology is an interdisciplinary field that is rapidly evolving and expanding. Significant advancements have been made in nanotechnology-related disciplines in the past few decades and continued growth and progression in the field are anticipated. Moreover, nanotechnology, omnipresent in innovation, has been applied to resolve critical challenges in nearly every field, especially those related to biological technologies and processes. This book, used as either a textbook for a short course or a reference book, provides state-of-the-art analysis of essential topics in nanotechnology for bioengineers studying and working in biotechnology, chemical/biochemical, pharmaceutical, biomedical, and other related fields. The book topics range from introduction to nanotechnology and nanofabrication to applications of nanotechnology in various biological fields. This book not only intends to introduce bioengineers to the amazing world of nanotechnology, but also inspires them to use nanotechnology to address some of the world's biggest challenges.
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Stability improvement and characterization of bioprinted pectin-based scaffold
Journal of Applied Biomaterials & Functional MaterialsStealey, S., Guo, X., Ren, L., Bryant, E., Kaltchev, M., Chen, J., Kumpaty, S., Hua, X., Zhang, W.
2019
Bioprinting is an alternative method for constructing tissues/organs for transplantation. This study investigated the cross-linker influence and post-printing modification using oligochitosan and chitosan for stability improvement.
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Design of A Novel Oxygen Therapeutic Using Polymeric Hydrogel Microcapsules Mimicking Red Blood Cells
PharmaceuticsAmanda Cherwin, Shelby Namen, Justyna Rapacz, Grace Kusik, Alexa Anderson, Yale Wang, Matey Kaltchev, Rebecca Schroeder, Kellen O'Connell, Sydney Stephens, Junhong Chen, and Wujie Zhang
2019
The goal of this research was to develop a novel oxygen therapeutic made from a pectin-based hydrogel microcapsule carrier mimicking red blood cells. The study focused on three main criteria for developing the oxygen therapeutic to mimic red blood cells: size (5–10 μm), morphology (biconcave shape), and functionality (encapsulation of oxygen carriers; e.g., hemoglobin (Hb)). The hydrogel carriers were generated via the electrospraying of the pectin-based solution into an oligochitosan crosslinking solution using an electrospinning setup. The pectin-based solution was investigated first to develop the simplest possible formulation for electrospray. Then, Design-Expert® software was used to optimize the production process of the hydrogel microcapsules. The optimal parameters were obtained through the analysis of a total of 17 trials and the microcapsule with the desired morphology and size was successfully prepared under the optimized condition. Fourier transform infrared spectroscopy (FTIR) was used to ana
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Design of pectin-based bioink containing bioactive agent-loaded microspheres for bioprinting
Biomedical Physics & Engineering ExpressDavid L. Johnson, Rachel M. Ziemba, Johnathan H. Shebesta, Jesseka C. Lipscomb, Yale Wang, Kellen D. O’Connell, Matey G. Kaltchev, Anthony van Groningen, Junhong Chen, Xiaolin Hua, and Wujie Zhang
2019
Bioprinting offers an alternative approach for tissue engineering and exhibits the great potential to play a key role in personalized medicine. One of the major advantages of bioprinting is its capability of achieving the homogeneous distribution of cells within large tissue scaffolds. Microspheres have been used for controlled release of bioactive molecules in tissue engineering. Recently studies show that microspheres, especially positively charged, could promote the vascularized tissue formation. This study aims to develop a bioprinted scaffold containing microspheres. The double emulsion system, water in oil in water (W/O/W), was used to produce the microspheres considering its advantage for controlled release of small molecules. Design Expert® Software was used to optimize the microsphere production process and chitosan coating was performed to provide the positively charged surface of the microspheres. Fourier-transform infrared spectroscopy (FTIR) and confocal microscopic analysis confirmed the suc
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Electrospinning pectin-based nanofibers: a parametric and cross-linker study
Applied NanoscienceMcCune, D., Guo, X., Shi, T., Stealey, S., Antrobus, R., Kaltchev, M., Chen, J., Kumpaty, S., Hua, X., Ren, W., Zhang, W.
2018
Pectin, a natural biopolymer mainly derived from citrus fruits and apple peels, shows excellent biodegradable and biocompatible properties. This study investigated the electrospinning of pectin-based nanofibers. The parameters, pectin:PEO (polyethylene oxide) ratio, surfactant concentration, voltage, and flow rate, were studied to optimize the electrospinning process for generating the pectin-based nanofibers. Oligochitosan, as a novel and nonionic cross-liker of pectin, was also researched. Nanofibers were characterized by using AFM, SEM, and FTIR spectroscopy. The results showed that oligochitosan was preferred over Ca2+ because it cross-linked pectin molecules without negatively affecting the nanofiber morphology. Moreover, oligochitosan treatment produced a positive surface charge of nanofibers, determined by zeta potential measurement, which is desired for tissue engineering applications.
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Novel bioprinting method using a pectin based bioink
Technology and Health Care,Banks, A., Guo, X., Chen, J., Kumpaty, S., Zhang, W.
2017
One major challenge of bioprinting is to develop a viable bioink to act as an extracellular matrix. This study investigated a novel method for bioprinting using a pectin based bioink. Besides pectin, Pluronic® F-127 was incorporated into the bioink to obtain the desired shape during the initial bioprinting process at 37∘C. Once an object was printed it was treated with Ca2+ (pectin cross-linker) to create the final tissue/organ structure. The results indicated that pectin/Pluronic® F-127 is a potential bioink. Moreover, this methodology provides a novel and fast approach for bioprinting.
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Engineering microvascularized 3D tissue using alginate-chitosan microcapsules
Journal of Biomaterials and Tissue EngineeringZhang, W., Choi, J.K. and He, X
2017
Construction of vascularized tissues is one of the major challenges of tissue engineering. The goal of this study was to engineer 3D microvascularized tissues by incorporating the HUVEC-CS cells with a collagen/alginate-chitosan (AC) microcapsule scaffold. In the presence of AC microcapsules, a 3D vascular-like network was clearly observable. The results indicate the importance of AC microcapsules in engineering microvascularized tissues—providing support and guiding alignment of HUVEC-CS cells. This provides a promising strategy for constructing vascularized tissues.
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Hydrogels: A novel red‐blood‐cell‐shaped pectin‐oligochitosan hydrogels system
Particle & Particle Systems CharacterizationHarvestine, J.N., Mikulski, B.A., Mahuta, K.M., Crouse, J.Z., Guo, X., Lee, J.C., Midelfort, K.S., Chen, J., Zhang, W.
2014
A novel pectin‐oligochitosan hydrogel microcapsule system is reported by W. Zhang and co‐workers on page 955. This kind of microcapsule shows red‐blood‐cell‐like (biconcave) morphology. The biological applications of this system are drug encapsulation/delivery, cell encapsulation, and creating artificial red blood cells.
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