Bioengineering Methods Research Articles

Stem Cells: Present Understanding and Prospects for Regenerative Dentistry.

Regenerative medicine in dentistry focuses on repairing damaged oral tissues using advanced tools like stem cells, biomaterials, and tissue engineering (TE). Mesenchymal stem cells (MSCs) from dental sources, such as dental pulp and periodontal ligament, show significant potential for tissue regeneration due to their proliferative and differentiative abilities. This systematic review, following PRISMA guidelines, evaluated fifteen studies and identified effective strategies for improving dental, periodontal, and bone tissue regeneration through scaffolds, secretomes, and bioengineering methods. Key advancements include the use of dental pulp stem cells (DPSCs) and periodontal ligament stem cells (PDLSCs) to boost cell viability and manage inflammation. Additionally, pharmacological agents like matrine and surface modifications on biomaterials improve stem cell adhesion and promote osteogenic differentiation. By integrating these approaches, regenerative medicine and TE can optimize dental therapies and enhance patient outcomes. This review highlights the potential and challenges in this field, providing a critical assessment of current research and future directions.

Formation of a scientific school on genetic resources, selection and genetics of grapes in Russia from N.I. Vavilov to the present day

While discussing the establishment of scientific school of grapevine genetic resources, breeding and genetics in Russia, its formation is of undoubted scientific importance not by one individual only, but generally by studying and analyzing the contribution of generations of scientists in the country, focusing, as a baseline, on fundamental researches and stated theories of N.I. Vavilov. Analyzing the historical and modern stages of grape breeding methodology in conjunction with source material – genetic resources, and those determined by the specificity of pattern genome manifesting and inheriting genetically determined traits, it is necessary to highlight two aspects: the process of grape gene pool formation, the origin of individual varieties in the process of natural evolution, and task-oriented creation of new cultivars in the process of experimental evolution. Understanding of the methodology of grapevine natural morphogenesis in due course generated the methodology of targeted creation of new cultivars. Natural hybridization, natural mutagenesis and natural selection as initial methods of grapevine morphogenesis for generations of Russian scientists made it possible, using descriptive ampelography, to further move to directed hybridization, induced mutagenesis and induced polyploidization in combination with artificial selection. And only recently the transition from breeding at the organismal level to genome editing at the gene level became possible using modern bioengineering methods and various approaches to gene engineering.

Growth factor-loaded ovarian extracellular matrix hydrogels promote in vivo ovarian niche regeneration and enhance fertility in premature ovarian insufficiency preclinical models

Premature ovarian insufficiency (POI) means menopause before 40 years of age affecting about 1 % of women. Approaches based on cell therapy and the paracrine effects of stem cells or bioproducts such as platelet-rich plasma have been proposed, but concerns remain about undesired systemic effects, as well as the need to optimize delivery methods through bioengineering methods. This study explores the efficacy of decellularized bovine ovarian cortex extracellular matrix (OvaECM) hydrogels alone and as a growth factor (GF) carrier (OvaECM+GF) in a chemotherapy-induced POI murine model. In vitro assays showed a gradual release of GF from the OvaECM sustained for two weeks. Chemotherapy drastically reduced follicle numbers, but OvaECM+GF treatment restored pre-antral follicle development. Moreover, this treatment notably regenerated the ovarian microenvironment by increasing cell proliferation and microvessel density while reducing chemotherapy-induced apoptosis and fibrosis. Whole-ovary RNA sequencing and gene set enrichment analysis revealed an upregulation of regeneration-related genes and a downregulation of apoptotic pathways. The OvaECM+GF treatment also yielded significantly better outcomes following ovarian stimulation and in vitro fertilization. After two consecutive crossbreeding cycles, OvaECM+GF-treated mice showed normal reproductive function. This research showcases the biocompatibility and efficacy of OvaECM to reverse POI in mice, setting a foundation to explore innovative bioengineering-based POI therapies. Statement of significancePremature ovarian insufficiency (POI) affects about 1 % of women worldwide, causing early menopause before 40 years old. Current treatments alleviate symptoms but do not restore ovarian function. This study explores an innovative approach using ovarian cortex extracellular matrix hydrogels to deliver growth factors into the murine ovarian niche and reverse POI. In vitro release kinetic assays demonstrated a gradual and sustained release of growth factors. In a POI-induced mouse model, intraovarian injections of the hydrogel encapsulating growth factors restored pre-antral follicle development, increased cell proliferation, reduced apoptosis and fibrosis, and improved ovarian response and in vitro fertilization outcomes. Long-term benefits included larger litter sizes. This innovative technique shows promise in regenerating the ovarian environment and improving reproductive outcomes.

Applications of Exosomal miRNAs from Mesenchymal Stem Cells as Skin Boosters.

The skin is the outer layer of the human body, and it is crucial in defending against injuries and damage. The regenerative capacity of aging and damaged skin caused by exposure to external stimuli is significantly impaired. Currently, the rise in average life expectancy and the modern population's aesthetic standards have sparked a desire for stem-cell-based therapies that can address skin health conditions. In recent years, mesenchymal stem cells (MSCs) as therapeutic agents have provided a promising and effective alternative for managing skin regeneration and rejuvenation, attributing to their healing capacities that can be applied to damaged and aged skin. However, it has been established that the therapeutic effects of MSC may be primarily mediated by paracrine mechanisms, particularly the release of exosomes (Exos). Exosomes are nanoscale extracellular vesicles (EVs) that have lipid bilayer and membrane structures and can be naturally released by different types of cells. They influence the physiological and pathological processes of recipient cells by transferring a variety of bioactive molecules, including lipids, proteins, and nucleic acids such as messenger RNAs (mRNAs) and microRNAs (miRNAs) between cells, thus playing an important role in intercellular communication and activating signaling pathways in target cells. Among them, miRNAs, a type of endogenous regulatory non-coding RNA, are often incorporated into exosomes as important signaling molecules regulating protein biosynthesis. Emerging evidence suggests that exosomal miRNAs from MSC play a key role in skin regeneration and rejuvenation by targeting multiple genes and regulating various biological processes, such as participating in inflammatory responses, cell migration, proliferation, and apoptosis. In this review, we summarize the recent studies and observations on how MSC-derived exosomal miRNAs contribute to the regeneration and rejuvenation of skin tissue, with particular attention to the applications of bioengineering methods for manipulating the miRNA content of exosome cargo to improve their therapeutic potential. This review can provide new clues for the diagnosis and treatment of skin damage and aging, as well as assist investigators in exploring innovative therapeutic strategies for treating a multitude of skin problems with the aim of delaying skin aging, promoting skin regeneration, and maintaining healthy skin.

Development of CNS tropic AAV1-like variants with reduced liver-targeting following systemic administration in mice

Directed evolution of natural AAV9 using peptide display libraries have been widely used in the search for an optimal recombinant AAV (rAAV) for transgene delivery across the blood-brain barrier (BBB) to the CNS following intravenous ( IV) injection. In this study, we used a different approach by creating a shuffled rAAV capsid library based on parental AAV serotypes 1 through 12. Following selection in mice, 3 novel variants closely related to AAV1, AAV-BBB6, AAV-BBB28, and AAV-BBB31, emerged as top candidates. In direct comparisons with AAV9, our novel variants demonstrated an over 270-fold improvement in CNS transduction and exhibited a clear bias toward neuronal cells. Intriguingly, our AAV-BBB variants relied on the LY6A cellular receptor for CNS entry, similar to AAV9 peptide variants AAV-PHP.eB and AAV.CAP-B10, despite the different bioengineering methods used and parental backgrounds. The variants also showed reduced transduction of both mouse liver and human primary hepatocytes invivo. To increase clinical translatability, we enhanced the immune escape properties of our new variants by introducing additional modifications based on rational design. Overall, our study highlights the potential of AAV1-like vectors for efficient CNS transduction with reduced liver tropism, offering promising prospects for CNS gene therapies.

NucleoCraft: The Art of Stimuli-Responsive Precision in DNA and RNA Bioengineering.

Recent advancements in DNA and RNA bioengineering have paved the way for developing stimuli-responsive nanostructures with remarkable potential across various applications. These nanostructures, crafted through sophisticated bioengineering techniques, can dynamically and precisely respond to both physiological and physical stimuli, including nucleic acids (DNA/RNA), adenosine triphosphate, proteins, ions, small molecules, pH, light, and temperature. They offer high sensitivity and specificity, making them ideal for applications such as biomarker detection, gene therapy, and controlled targeted drug delivery. In this review, we summarize the bioengineering methods used to assemble versatile stimuli-responsive DNA/RNA nanostructures and discuss their emerging applications in structural biology and biomedicine, including biosensing, targeted drug delivery, and therapeutics. Finally, we highlight the challenges and opportunities in the rational design of these intelligent bioengineered nanostructures.

The brief introduction of CAR T-cell: From mechanism to application

Chimeric antigen receptor T-cell (CAR-T) gene therapy is a newly developed solution to fight tumor cells. The therapy develops from the combination of the immunity and cell bioengineering, taking advantage of the cell killing function of T-cells, scientists modify them with a bioengineering method to target specific cells. In recent days, this therapy shows great results in several trials especially in hematological malignancies. However, there are still many obstacles, such as toxicities, that need to be overcome. Taking the promising potentials of CAR T-cell gene therapy, it is still a revolutionary step in the improvement of immunotherapy. In this paper, we focus on the structure and target antigens of CAR T-cell, summarize the generations of CAR T-cell gene therapies, as well as list out the FDA approved treatments using CAR. Most importantly, we will discuss the past advancements, recent innovations, and the existing factors that still prevent CAR T-cells from becoming the first line treatment for cancers.

Progress in the generation of spinal cord organoids over the past decade and future perspectives.

Spinal cord organoids are three-dimensional tissues derived from stem cells that recapitulate the primary morphological and functional characteristics of the spinal cord in vivo. As emerging bioengineering methods have led to the optimization of cell culture protocols, spinal cord organoids technology has made remarkable advancements in the past decade. Our literature search found that current spinal cord organoids do not only dynamically simulate neural tube formation but also exhibit diverse cytoarchitecture along the dorsal-ventral and rostral-caudal axes. Moreover, fused organoids that integrate motor neurons and other regionally specific organoids exhibit intricate neural circuits that allows for functional assessment. These qualities make spinal cord organoids valuable tools for disease modeling, drug screening, and tissue regeneration. By utilizing this emergent technology, researchers have made significant progress in investigating the pathogenesis and potential therapeutic targets of spinal cord diseases. However, at present, spinal cord organoid technology remains in its infancy and has not been widely applied in translational medicine. Establishment of the next generation of spinal cord organoids will depend on good manufacturing practice standards and needs to focus on diverse cell phenotypes and electrophysiological functionality evaluation.

Sonogenetics: a mini review

Sonogenetics has emerged as a tool of therapeutic ultrasound which is revolutionizing the ability to non-invasively modulate the activity of neurons and other excitatory cells. This technology utilizes bioengineering methods to confer or amplify ultrasound sensitivity in target cells using engineered or modified protein mediators. The neuromodulation community has shown a growing interest in sonogenetics due to ultrasound’s ability to penetrate the skull and reach deep brain tissue, enabling non-invasive modulation of neurons. Novel methods of sonogenetics aim to enhance cellular control in humans by leveraging mechanosensitive and thermosensitive cellular mechanisms activated by ultrasound to address cellular dysfunction and degeneration. This mini review summarizes the progress of sonogenetic mediators proposed for neuromodulation and looks at new therapeutic applications of sonogenetics for cancer treatment and vision restoration.

Preconditioning Concepts for the Therapeutic Use of Extracellular Vesicles Against Stroke.

Various preclinical stroke models have demonstrated the neuroprotective effects of extracellular vesicles (EVs) obtained from several types of cells, including neurons, astrocytes, microglia, neuronal progenitor cells, bone marrow stem cells, and mesenchymal stem cells. EVs interfere with key mechanisms in stroke pathophysiology such as cell death, neuroinflammation, autophagy, and angiogenesis. The mode of action and efficacy depend on the specific EV content, including miRNAs, proteins, and lipids, which can be modified through (I) bioengineering methods, (II) choice of source cells, and (III) modification of the source cell environment. Indeed, modifying the environment by preconditioning the EV-secreting cells with oxygen-glucose deprivation or medium modification revealed superior neuroprotective effects in stroke models. Although the concept of preconditioned EVs is relatively novel, it holds promise for the future treatment of ischemic stroke. Here, we give a brief overview about the main mechanisms of EV-induced neuroprotection and discuss the current status of preconditioning concepts for EV-treatment of ischemic stroke.

Short-chain polyols from bio-based carboxylic acids for high-performance polyurethane coatings

Emerging bioengineering methods have created an opportunity to introduce a range of novel sustainable raw materials into industrial chemical processes. Among others, a multitude of bio-based acids can become available on the market on a commercial scale. These functional molecules can be useful as reagents for manufacturing sustainable high-performance materials. To this end, we evaluated a series of seven bio-based carboxylic acids as starting materials in the development of cross-linked 2K polyurethane (PU) coatings. By reacting the acids with propylene oxide (PO), seven short-chain polyols were synthesized. The synthesized polyols, consisting of mixtures of polyfunctional hydroxy-terminated products, were cross-linked with two isocyanate trimers (hexamethylene diisocyanate (HDI) trimer and isophorone diisocyanate (IPDI) trimer) to produce polyurethane coatings. The thermal, mechanical, and coating properties of PU films can be varied depending on the polyol and isocyanate used. Thus, coatings formulated using IPDI trimer were hard and brittle, whereas films made with the aliphatic HDI trimer offered a valuable combination of excellent hardness and extraordinary flexibility. Using polyols made from several isomeric carboxylic acids resulted in a significant variation of properties in the PU coatings. The results illustrate the applicability of novel bio-based acids as starting materials in the coatings industry and highlight the necessity to explore their potential application as components of sustainable materials.

Bioengineering Applied to Oral Implantology, a New Protocol: “Digital Guided Surgery”

Rehabilitative dentistry has made enormous progress in recent years, not only due to the advent of new implant-prosthetic methods, but also thanks to new information technologies that support the doctor. This study aims to present a new implant protocol that involves the application of bioengineering methods. With the application of the finite element analysis, it is possible to evaluate the distribution of the forces of a fixture and possible implant rehabilitation on each patient, even before performing the surgery. This protocol provides for the combination of radiographic images and three-dimensional files to obtain predictable results on possible rehabilitation, guiding its planning in the best possible way. Surely, the evolution of machines and computers will enable the surgeon to carry out and maintain these protocols in a chair-side manner, and to carry out safe and predictable rehabilitations.

Photopolymerization in 3D printing of tissue-engineered constructs for regenerative medicine

The progress in the field of tissue engineering is largely driven by the development of 3D laser printing technologies, which allow precise creation of hydrogel scaffolds containing cells (the so-called tissue-engineered constructs), using photoinduced radical reactions of polymerization and crosslinking. The review considers the main mechanisms and features of such reactions, presents the most common materials for photocompositions, including natural and synthetic polymers and precursors, and describes various mechanisms for photoinitiator activation. Advances in the field of photopolymerization enable application of modern laser 3D printing techniques based on extrusion and stereolithography to design tissue-engineered constructs in a wide range of sizes and shapes with a finely organized architecture. The integration of such methods with the methods of bioengineering and cell technology is discussed, including for the creation of tissue-specific and <i>in vivo</i> polymerized constructs. The bibliography includes 225 references.<br>

Exosome engineering in cell therapy and drug delivery.

Cell-derived exosomes have opened new horizons in modern therapy for advanced drug delivery and therapeutic applications, due to their key features such as low immunogenicity, high physicochemical stability, capacity to penetrate into tissues, and the innate capacity to communicate with other cells over long distances. Exosome-based liquid biopsy has been potentially used for the diagnosis and prognosis of a range of disorders. Exosomes deliver therapeutic agents, including immunological modulators, therapeutic drugs, and antisense oligonucleotides to certain targets, and can be used as vaccines, though their clinical application is still far from reality. Producing exosomes on a large-scale is restricted to their low circulation lifetime, weak targeting capacity, and inappropriate controls, which need to be refined before being implemented in practice. Several bioengineering methods have been used for refining therapeutic applications of exosomes and promoting their effectiveness, on the one hand, and addressing the existing challenges, on the other. In the short run, new diagnostic platforms and emerging therapeutic strategies will further develop exosome engineering and therapeutic potential. This requires a thorough analysis of exosome engineering approaches along with their merits and drawbacks, as outlined in this paper. The present study is a comprehensive review of novel techniques for exosome development in terms of circulation time in the body, targeting capacity, and higher drug loading/delivery efficacies.

Sosoalisasi Pemanfaatan Rumput Vetiver (Chrysopogon zizanioides) Dalam Mitigasi Bencana Longsor di Desa Gelangsar, Kabupaten Lombok Barat

Gelangsar Village, West Lombok is one of the villages located in a hilly area which resulted in several points in the village of Gelangsar experiencing landslides. The topography which is hilly and steep, triggers landslides in the village of Gelangsar. In addition, the relatively high rainfall in the last few months has made the soil conditions more flexible. Areas that have the potential to cause landslides can be overcome by planting plants that can bind the soil tightly so that it can overcome the potential for landslides. Therefore, this service aims to socialize the use of vetiver grass as a bioengineering method in managing landslides in Gelangsar Village, Gunungsari, West Lombok. This Community Service activity uses the counseling method and the creation of a pilot project. Counseling was conducted to introduce vetiver grass plants and their cultivation techniques. Making a pilot project (pilot project) have been done in the homes of residents who are partners. The Pilot Project was created as a media for socialization and as a strategy for efficiency and effectiveness for target activities. The community of partners are willing to implement and develop vetiver grass as a biotechnology agent to prevent erosion and also as an economic opportunity. This programme has been able to provide knowledge in mitigation landslide disaster management activities independently. Planting vetiver grass is expected to be able to reduce the risk of erosion at landslide-prone points in the village of Gelangsar, improve the physical condition of the land and increase land productivity.

Application of genipin-crosslinked small intestine submucosa and urine-derived stem cells for the prevention of intrauterine adhesion in a rat model

As a major cause for recurrent miscarriage and secondary infertility, intrauterine adhesion (IUA) is characterized by partial or total obliteration of the uterine cavity and/or cervical canal due to injuries to the basal layer of endometrium caused by improper intrauterine operation and/or infection. While various treatments have been used to prevent the occurrence of IUA, these approaches have shown limited therapeutic efficiency. As a promising bio-engineering method, tissue engineering technology may be used for enhancing endometrial regeneration. Extracellular matrix such as small intestine submucosa (SIS) may provide a favorable microenvironment, whilst stem cells can promote the endometrial regenerative capacity. In this study, genipin-crosslinked SIS (GP-SIS) loaded with urine-derived stem cells (USCs) has been applied for the regeneration of endometrium. The GP-SIS scaffolds have demonstrated sound mechanical properties and excellent biocompatibility and promoted epithelial migration and angiogenesis in vitro. In a rat model for IUA caused by endometrial damage, transplantation of the GP-SIS/USCs has maintained normal luminal structure, promoted endometrial and glandular regeneration, vascularization, inhibited fibrogenesis and improved endometrial receptivity. Our research has therefore provided a new strategy for promoting the recovery of endometrial structure and function, which may be beneficial for the prevention and treatment of IUA in the clinics.

Plant cell culture technologies: A promising alternatives to produce high-value secondary metabolites

Plants have been used for its medicinal values since ancient time. The medicinal properties of plants are based on their phytochemical constituent particularly secondary metabolites which are produced in low amounts by plants. Secondary metabolites have been used as medicines, flavors, colors, and fragrances. In recent time, these natural compounds are gaining enormous attention in pharmaceutical, cosmetics, and nutraceutical industries and are regarded economically valuable products. The production of plant secondary metabolites in plant is largely dependent on the plant species, environmental factors and geographical regions. In addition, the main challenges in their mass production is reported to be the quality and quantity issues during their synthesis. Therefore, enthusiasm has grown for increasing the production of secondary metabolites by employing in vitro plant cell culture technology and bioengineering methods. Such technological advancement, has led to production of a huge number of medicinal herbs and high-value secondary metabolites that are mostly used in pharmaceuticals, cosmetics and nutraceuticals industries. The current mini-review article focuses on applications of plant cell culture system for the production secondary metabolites and recent techniques used to improve metabolite contents. Furthermore, our review emphasizes safety issues of plant cell culture derived products.

Bioengineering to Accelerate Biodiesel Production for a Sustainable Biorefinery.

Biodiesel is an alternative, carbon-neutral fuel compared to fossil-based diesel, which can reduce greenhouse gas (GHGs) emissions. Biodiesel is a product of microorganisms, crop plants, and animal-based oil and has the potential to prosper as a sustainable and renewable energy source and tackle growing energy problems. Biodiesel has a similar composition and combustion properties to fossil diesel and thus can be directly used in internal combustion engines as an energy source at the commercial level. Since biodiesel produced using edible/non-edible crops raises concerns about food vs. fuel, high production cost, monocropping crisis, and unintended environmental effects, such as land utilization patterns, it is essential to explore new approaches, feedstock and technologies to advance the production of biodiesel and maintain its sustainability. Adopting bioengineering methods to produce biodiesel from various sources such as crop plants, yeast, algae, and plant-based waste is one of the recent technologies, which could act as a promising alternative for creating genuinely sustainable, technically feasible, and cost-competitive biodiesel. Advancements in genetic engineering have enhanced lipid production in cellulosic crops and it can be used for biodiesel generation. Bioengineering intervention to produce lipids/fat/oil (TGA) and further their chemical or enzymatic transesterification to accelerate biodiesel production has a great future. Additionally, the valorization of waste and adoption of the biorefinery concept for biodiesel production would make it eco-friendly, cost-effective, energy positive, sustainable and fit for commercialization. A life cycle assessment will not only provide a better understanding of the various approaches for biodiesel production and waste valorization in the biorefinery model to identify the best technique for the production of sustainable biodiesel, but also show a path to draw a new policy for the adoption and commercialization of biodiesel.

Bioengineering Supports Preantral Follicle In Vitro Growth

ecent advances using bioengineering methods (e.g. biomaterials, 3D-printing, microfluidics tec.) has opened the possibility to study female reproductive system and reproductive diseases in a totally new way and with unpredictable perspectives

Diagnostic and Therapeutic Roles of the "Omics" in Hypoxic-Ischemic Encephalopathy in Neonates.

Perinatal asphyxia and neonatal encephalopathy remain major causes of neonatal mortality, despite the improved availability of diagnostic and therapeutic tools, contributing to neurological and intellectual disabilities worldwide. An approach using a combination of clinical data, neuroimaging, and biochemical parameters is the current strategy towards the improved diagnosis and prognosis of the outcome in neonatal hypoxic–ischemic encephalopathy (HIE) using bioengineering methods. Traditional biomarkers are of little use in this multifactorial and variable phenotype-presenting clinical condition. Novel systems of biology-based “omics” approaches (genomics, transcriptome proteomics, and metabolomics) may help to identify biomarkers associated with brain and other tissue injuries, predicting the disease severity in HIE. Biomarker studies using omics technologies will likely be a key feature of future neuroprotective treatment methods and will help to assess the successful treatment and long-term efficacy of the intervention. This article reviews the roles of different omics as biomarkers of HIE and outlines the existing knowledge of our current understanding of the clinical use of different omics molecules as novel neonatal brain injury biomarkers, which may lead to improved interventions related to the diagnostic and therapeutic aspects of HIE.