Designing a new sustainable Test Kit supply chain network utilizing Internet of Things.

The advent of COVID-19 put much economic pressure on countries worldwide, especially low-income countries. Providing test kits for Covid-19 posed a huge challenge at the beginning of the pandemic. Especially the low-income and less developed countries that did not have the technology to produce this kit and had to import it into the country, which itself cost a lot to buy and distribute these kits. This paper proposes a sustainable COVID-19 test kits supply chain network (STKSCN) for the first time to fill this gap. Distribution and transportation of test kits, location of distribution centers, and management of used test kits are considered in this network. A mixed integer linear programming Multi-Objective (MO), multi-period, multi-resource mathematical model is extended for the proposed supply chain. Another contribution is designing a platform based on the Internet of Things (IoT) to increase the speed, accuracy and security of the network. In this way, patients set their appointment online by registering their personal details and clinical symptoms. An augmented ɛ-constraint2 (AUGMECON2) is proposed for solving small and medium size of problem. Also, two meta-heuristic algorithms, namely NSGA-II and PESA-II are presented to solve the small, medium and large size of the problem. Taguchi method is utilized to control the parameters, and for comparison between meta-heuristic, five performance metrics are suggested. In addition, a case study in Iran is presented to validate the proposed model. Finally, the results show that PESA-II is more efficient and has better performance than the others based on assessment metrics and computational time.

Enabling Allogeneic T Cell-Based Therapies: Scalable Stirred-Tank Bioreactor Mediated Manufacturing.

Allogeneic T cells are key immune therapeutic cells to fight cancer and other clinical indications. High T cell dose per patient and increasing patient numbers result in clinical demand for a large number of allogeneic T cells. This necessitates a manufacturing platform that can be scaled up while retaining cell quality. Here we present a closed and scalable platform for T cell manufacturing to meet clinical demand. Upstream manufacturing steps of T cell activation and expansion are done in-vessel, in a stirred-tank bioreactor. T cell selection, which is necessary for CAR-T-based therapy, is done in the bioreactor itself, thus maintaining optimal culture conditions through the selection step. Platform's attributes of automation and performing the steps of T cell activation, expansion, and selection in-vessel, greatly contribute to enhancing process control, cell quality, and to the reduction of manual labor and contamination risk. In addition, the viability of integrating a closed, automated, downstream process of cell concentration, is demonstrated. The presented T cell manufacturing platform has scale-up capabilities while preserving key factors of cell quality and process control.

Designing an integrated responsive-green-cold vaccine supply chain network using Internet-of-Things: artificial intelligence-based solutions.

In this paper, a new responsive-green-cold vaccine supply chain network during the COVID-19 pandemic is developed for the first time. According to the proposed network, a new multi-objective, multi-period, multi-echelon mathematical model for the distribution-allocation-location problem is designed. Another important novelty in this paper is that it considers an Internet-of-Things application in the COVID-19 condition in the suggested model to enhance the accuracy, speed, and justice of vaccine injection with existing priorities. Waste management, environmental effects, coverage demand, and delivery time of COVID-19 vaccine simultaneously are therefore considered for the first time. The LP-metric method and meta-heuristic algorithms called Gray Wolf Optimization (GWO), and Variable Neighborhood Search (VNS) algorithms are then used to solve the developed model. The other significant contribution, based on two presented meta-heuristic algorithms, is a new heuristic method called modified GWO (MGWO), and is developed for the first time to solve the model. Therefore, a set of test problems in different sizes is provided. Hence, to evaluate the proposed algorithms, assessment metrics including (1) percentage of domination, (2) the number of Pareto solutions, (3) data envelopment analysis, and (4) diversification metrics and the performance of the convergence are considered. Moreover, the Taguchi method is used to tune the algorithm's parameters. Accordingly, to illustrate the efficiency of the model developed, a real case study in Iran is suggested. Finally, the results of this research show MGO offers higher quality and better performance than other proposed algorithms based on assessment metrics, computational time, and convergence.

Relationship Between BMI and Disease Severity in COVID-19 Patients.

Background: Obesity and increased body mass index (BMI) are associated with coronavirus disease 2019 (COVID-19)-related complications and severity. They can exacerbate the cytokine storm and lead to severe symptoms or death in obese patients. Objectives: This cross-sectional descriptive study included patients with COVID-19 admitted to the Razi Hospital in Ahvaz, Iran, from January 2019 to December 2020. Methods: We evaluated the effect of BMI of patients admitted to the general ward and invasive unit care (ICU) on the length of hospitalization. Results: We included a total of 466 patients (male: 281 or 60.3% vs. female: 185 or 39.7%) with a mean age of 59.49 ± 14.5 years in the study. Also, 47 (10.1%) patients were admitted to the ICU, and 418 (89.7%) patients to the general ward. A higher BMI was associated with longer hospitalization (P < 0.001). Patients with BMI in the range of 18.5 - 24.9 experienced a longer hospitalization (10-20 days) (P < 0.001). BMI had no significant effect on ICU hospitalization (P = 0.36). Also, there was no significant difference between the two groups regarding the length of hospitalization (P = 0.49). Furthermore, non-diabetic patients were less likely to be admitted to the ICU (73.3% vs. 26.7%) (P < 0.001). The number of discharged patients was higher in patients admitted to the general ward compared to those admitted to the ICU (93.8% vs. 63.8%) (P < 0.001). Conclusions: According to our results, a higher BMI was a risk factor for COVID-19, especially in the early stage of infection.

The influence of electrically conductive and non-conductive nanocomposite scaffolds on the maturation and excitability of engineered cardiac tissues.

Utilization of electrically conductive nanomaterials for developing nanocomposite scaffolds has been at the center of attention for engineering functional cardiac tissues. The primary motive in the use of conductive nanomaterials has been to develop biomimetic scaffolds to recapitulate the extracellular matrix (ECM) of the native heart and to promote cardiac tissue maturity, excitability and electrical signal propagation. Alternatively, it is well accepted that the inclusion of nanomaterials also alters the stiffness and nano-scale topography of the scaffolds. However, what is missing in the literature is that to what extent the sole presence of nanomaterials within a scaffold, regardless of their conductivity, influences the maturation and excitability of engineered cardiac tissues. To address this knowledge gap, we developed four different classes of gelatin methacrylate (GelMA) hydrogels, with varied concentrations, embedded electrically conductive gold nanorods (GNRs) and non-conductive silica nanomaterials (SNPs), to assess the influence of matrix stiffness and the presence of nanomaterials on cardiac cell adhesion, protein expression (i.e. maturation), and tissue-level excitability. Our results demonstrated that either embedding nanomaterials (i.e. GNRs and SNPs) or increasing the matrix stiffness significantly promoted cellular retention and the expression of cardiac-specific markers, including sarcomeric α-actinin (SAC), cardiac troponin I (cTnI) and connexin43 (Cx43) gap junctions. Notably, excitation voltage thresholds at a high frequency (i.e. 2 Hz and higher), in both coupled and uncoupled gap junctions induced by heptanol, were lower for scaffolds embedded conductive GNRs or non-conductive SNPs, independent of matrix stiffness. Overall, our findings demonstrated that the sole presence of nanomaterials within the scaffolding matrix had a more pronounced influence as compared to the scaffold stiffness on the cell-cell coupling, maturation and excitability of engineered cardiac tissues.

Wireless Passive Stimulation of Engineered Cardiac Tissues.

We present a battery-free radio frequency (RF) microwave activated wireless stimulator, 25 × 42 × 1.6 mm3 on a flexible substrate, featuring high current delivery, up to 60 mA, to stimulate engineered cardiac tissues. An external antenna shines 2.4 GHz microwave, which is modulated by an inverted pulse to directly control the stimulating waveform, to the wireless passive stimulator. The stimulator is equipped with an on-board antenna, multistage diode multipliers, and a control transistor. Rat cardiomyocytes, seeded on electrically conductive gelatin-based hydrogels, demonstrate synchronous contractions and Ca2+ transients immediately upon stimulation. Notably, the stimulator output voltage and current profiles match the tissue contraction frequency within 0.5-2 Hz. Overall, our results indicate the promising potential of the proposed wireless passive stimulator for cardiac stimulation and therapy by induction of precisely controlled and synchronous contractions.

Bioprinting technologies for disease modeling.

There is a great need for the development of biomimetic human tissue models that allow elucidation of the pathophysiological conditions involved in disease initiation and progression. Conventional two-dimensional (2D) in vitro assays and animal models have been unable to fully recapitulate the critical characteristics of human physiology. Alternatively, three-dimensional (3D) tissue models are often developed in a low-throughput manner and lack crucial native-like architecture. The recent emergence of bioprinting technologies has enabled creating 3D tissue models that address the critical challenges of conventional in vitro assays through the development of custom bioinks and patient derived cells coupled with well-defined arrangements of biomaterials. Here, we provide an overview on the technological aspects of 3D bioprinting technique and discuss how the development of bioprinted tissue models have propelled our understanding of diseases' characteristics (i.e. initiation and progression). The future perspectives on the use of bioprinted 3D tissue models for drug discovery application are also highlighted.

Emerging Biofabrication Strategies for Engineering Complex Tissue Constructs.

The demand for organ transplantation and repair, coupled with a shortage of available donors, poses an urgent clinical need for the development of innovative treatment strategies for long-term repair and regeneration of injured or diseased tissues and organs. Bioengineering organs, by growing patient-derived cells in biomaterial scaffolds in the presence of pertinent physicochemical signals, provides a promising solution to meet this demand. However, recapitulating the structural and cytoarchitectural complexities of native tissues in vitro remains a significant challenge to be addressed. Through tremendous efforts over the past decade, several innovative biofabrication strategies have been developed to overcome these challenges. This review highlights recent work on emerging three-dimensional bioprinting and textile techniques, compares the advantages and shortcomings of these approaches, outlines the use of common biomaterials and advanced hybrid scaffolds, and describes several design considerations including the structural, physical, biological, and economical parameters that are crucial for the fabrication of functional, complex, engineered tissues. Finally, the applications of these biofabrication strategies in neural, skin, connective, and muscle tissue engineering are explored.

Gold nanorod-incorporated gelatin-based conductive hydrogels for engineering cardiac tissue constructs.

UNLABELLED: The development of advanced biomaterials is a crucial step to enhance the efficacy of tissue engineering strategies for treatment of myocardial infarction. Specific characteristics of biomaterials including electrical conductivity, mechanical robustness and structural integrity need to be further enhanced to promote the functionalities of cardiac cells. In this work, we fabricated UV-crosslinkable gold nanorod (GNR)-incorporated gelatin methacrylate (GelMA) hybrid hydrogels with enhanced material and biological properties for cardiac tissue engineering. Embedded GNRs promoted electrical conductivity and mechanical stiffness of the hydrogel matrix. Cardiomyocytes seeded on GelMA-GNR hybrid hydrogels exhibited excellent cell retention, viability, and metabolic activity. The increased cell adhesion resulted in abundance of locally organized F-actin fibers, leading to the formation of an integrated tissue layer on the GNR-embedded hydrogels. Immunostained images of integrin ß-1 confirmed improved cell-matrix interaction on the hybrid hydrogels. Notably, homogeneous distribution of cardiac specific markers (sarcomeric α-actinin and connexin 43), were observed on GelMA-GNR hydrogels as a function of GNRs concentration. Furthermore, the GelMA-GNR hybrids supported synchronous tissue-level beating of cardiomyocytes. Similar observations were also noted by, calcium transient assay that demonstrated the rhythmic contraction of the cardiomyocytes on GelMA-GNR hydrogels as compared to pure GelMA. Thus, the findings of this study clearly demonstrated that functional cardiac patches with superior electrical and mechanical properties can be developed using nanoengineered GelMA-GNR hybrid hydrogels. STATEMENT OF SIGNIFICANCE: In this work, we developed gold nanorod (GNR) incorporated gelatin-based hydrogels with suitable electrical conductivity and mechanical stiffness for engineering functional cardiac tissue constructs (e.g. cardiac patches). The synthesized conductive hybrid hydrogels properly accommodated cardiac cells and subsequently resulted in excellent cell retention, spreading, homogeneous distribution of cardiac specific markers, cell-cell coupling as well as robust synchronized (tissue-level) beating behavior.

PNIPAAm-based biohybrid injectable hydrogel for cardiac tissue engineering.

Injectable biomaterials offer a non-invasive approach to deliver cells into the myocardial infarct region to maintain a high level of cell retention and viability and initiate the regeneration process. However, previously developed injectable matrices often suffer from low bioactivity or poor mechanical properties. To address this need, we introduced a biohybrid temperature-responsive poly(N-isopropylacrylamide) PNIPAAm-Gelatin-based injectable hydrogel with excellent bioactivity as well as mechanical robustness for cardiac tissue engineering. A unique feature of our work was that we performed extensive in vitro biological analyses to assess the functionalities of cardiomyocytes (CMs) alone and in co-culture with cardiac fibroblasts (CFs) (2:1 ratio) within the hydrogel matrix. The synthesized hydrogel exhibited viscoelastic behavior (storage modulus: 1260 Pa) and necessary water content (75%) to properly accommodate the cardiac cells. The encapsulated cells demonstrated a high level of cell survival (90% for co-culture condition, day 7) and spreading throughout the hydrogel matrix in both culture conditions. A dense network of stained F-actin fibers (∼ 6 × 10(4) µm(2) area coverage, co-culture condition) illustrated the formation of an intact and three dimensional (3D) cell-embedded matrix. Furthermore, immunostaining and gene expression analyses revealed mature phenotypic characteristics of cardiac cells. Notably, the co-culture group exhibited superior structural organization and cell-cell coupling, as well as beating behavior (average ∼ 45 beats per min, co-culture condition, day 7). The outcome of this study is envisioned to open a new avenue for extensive in vitro characterization of injectable matrices embedded with 3D mono- and co-culture of cardiac cells prior to in vivo experiments. STATEMENT OF SIGNIFICANCE: In this work, we synthesized a new class of biohybrid temperature-responsive poly(N-isopropylacrylamide) PNIPAAm-Gelatin-based injectable hydrogel with suitable bioactivity and mechanical properties for cardiac tissue engineering. A significant aspect of our work was that we performed extensive in vitro biological analyses to assess the functionality of cardiomyocytes alone and in co-culture with cardiac fibroblasts encapsulated within the 3D hydrogel matrix.

3D cardiac microtissues encapsulated with the co-culture of cardiomyocytes and cardiac fibroblasts.

Cardiac tissue engineering has major applications in regenerative medicine, disease modeling and biological studies. Despite the significance, numerous questions still need to be explored to enhance the functionalities of engineered tissue substitutes. In this study, 3D cardiac microtissues are developed through encapsulation of cardiomyocytes and cardiac fibroblasts, as the main cellular constituents of native myocardium. The geometries of the constructs are precisely controlled and assessed for their role on synchronous contraction of the cells. Cardiomyocytes exhibit a native-like phenotype when co-cultured with cardiac fibroblasts as compared to the monoculture condition. Particularly, elongated F-actin fibers with abundance of sarcomeric α-actinin and troponin-I are observed within all layers of the constructs. Higher expressions of connexin-43 and integrin-ß1 indicate improved cell-cell and cell-matrix interactions. Amongst co-culture conditions, 2:1 (cardiomyocytes: cardiac fibroblasts) ratio exhibits enhanced functionalities, whereas decreasing the construct size adversely affects the synchronous contraction of the cells. Overall, the study here indicates that the cell-cell ratio and the construct geometry are crucial parameters, which need to be optimized to enhance the functionalities of the engineered tissue substitutes.

Double-walled microspheres loaded with meglumine antimoniate: preparation, characterization and in vitro release study.

OBJECTIVE: The objective of this study was to fabricate double-walled poly(lactide-co-glycolide) (PLGA) microspheres to increase encapsulation efficiency and avoid rapid release of hydrophilic drugs such as meglumine antimoniate. METHODS: In this study, double-walled and one-layered microspheres of PLGA were prepared using the emulsion solvent evaporation technique to better control the release of a hydrophilic drug, meglumine antimoniate (Glucantime®), which is the first choice treatment of cutaneous leishmaniasis. The effect of hydrophobic coating on microspheres' size, morphology, encapsulation efficiency and drug release characteristics was evaluated. Furthermore, the presence of antimony in meglumine antimoniate made it possible to observe the drug distribution within the microspheres' cross section by means of energy dispersive X-ray spectroscopy. RESULTS: Drug distribution images confirmed accumulation of the drug within the inner core of double-walled microspheres. In addition, these microspheres encapsulated the drug more efficiently up to 87% and demonstrated reduced initial burst and prolonged release compared to one-layered microspheres. These superiorities make double-walled microspheres an optimum candidate for sustained delivery of hydrophilic drugs. CONCLUSION: Double-walled microspheres provide some advantages over traditional microspheres overcoming most of their limitations. Double-walled microspheres were found to be more efficient than their corresponding one-layered microspheres in terms of encapsulation efficiencies and release characteristics.

Development of liposomes loaded with anti-leishmanial drugs for the treatment of cutaneous leishmaniasis.

Cutaneous leishmaniasis is caused by different species of Leishmania parasites and its available treatments have not yet provided a strong consistent result. The weak response of current chemotherapeutics is due to their deficient effects on stealth parasites inside macrophages, rapid clearance from the site of action and systemic side effects in high doses. Liposomal formulation of anti-leishmanial drugs could overcome these problems. In this study, different liposomal formulations of three famous anti-leishmanial drugs: Glucantime®, miltefosine and paromomycin were prepared by a modified freeze-drying double emulsion method. Liposome size, zeta potential and encapsulation efficiency were evaluated, and their imaging was carried out by means of atomic force microscopy. Three formulations were evaluated in vivo by subcutaneous injection into skin lesions caused by Leishmania major in BALB/c mice. Encapsulation efficiency of prepared liposomes was up to 90%; however, they inherited a bimodal size distribution that caused their encapsulation efficiency to decrease to 50% during filtering sterilization. Besides, the effect of surface charge was significant on preparation procedure, size and encapsulation efficiency. All three formulations reduced amastigote counts and lesion size but only miltefosine-loaded formulations had significant therapeutic effects compared with control group (p < 0.05).