Tissue Engineering Research Research Articles

Platelet-Rich Fibrin Scaffolds for Cartilage and Tendon Regenerative Medicine: From Bench to Bedside.

Nowadays, research in Tissue Engineering and Regenerative Medicine is focusing on the identification of instructive scaffolds to address the requirements of both clinicians and patients to achieve prompt and adequate healing in case of injury. Among biomaterials, hemocomponents, and in particular Platelet-rich Fibrin matrices, have aroused widespread interest, acting as delivery platforms for growth factors, cytokines and immune/stem-like cells for immunomodulation; their autologous origin and ready availability are also noteworthy aspects, as safety- and cost-related factors and practical aspects make it possible to shorten surgical interventions. In fact, several authors have focused on the use of Platelet-rich Fibrin in cartilage and tendon tissue engineering, reporting an increasing number of in vitro, pre-clinical and clinical studies. This narrative review attempts to compare the relevant advances in the field, with particular reference being made to the regenerative role of platelet-derived growth factors, as well as the main pre-clinical and clinical research on Platelet-rich Fibrin in chondrogenesis and tenogenesis, thereby providing a basis for critical revision of the topic.

Biological properties of a bionic scaffold for esophageal tissue engineering research

Polyurethane is a good matrix material with wide application prospects in tissue engineering because of its adjustable and mechanical properties. A novel biodegradable crosslinked poly(ester urethane) (CPU) with flexible poly(caprolactone) (PCL) and hydrophilic poly(ethylene glycol) (PEG) components has been synthesized using a ferric iron catalyst in our laboratory. In the present study, to promote the interaction between the CPU material and cells, the material was superficially modified by silk fibroin (SF) grafting using an aminolysis and glutaraldehyde crosslinking method to achieve a biocompatible material, CPU-SF. Considering the esophageal-specific architecture, three types of scaffolds were fabricated. S1 was a CPU-SF channel (200 μm in diameter and 30 μm in depth with 30 μm of wall thickness) to support muscle regeneration; S2 was the decellularized matrix of the esophageal mucosa/submucosa obtained by enzyme treatment; and S3 was a combination of S1 and S2, aiming to promote esophageal regeneration with histological structure and function. The biological properties and functions of the materials and scaffolds were investigated by qualitative and quantitative analyses using scanning electron microscopy, immunofluorescence staining, cell adhesion and proliferation measurements, and western blotting technology. The results showed that esophageal smooth muscle cells (SMCs) and epithelial cells (ECs) were very well supported by the scaffolds. In particular, SMCs exhibited guided directional growth and ECs infiltrated the acellular mucosa with retained biological functions when co-cultured on the composite scaffold S3. These findings suggest that the composite bionic scaffold will be a good alternative for esophageal replacement.

Cost‐effective synthesis of polyethylene glycol diacrylate‐acrylic acid (PEGDA‐AA), a tissue engineering hydrogel ideal for C2C12 mouse muscle cells

Current research in tissue engineering focuses on growing and functionalizing cells in vitro to assemble viable constructs that restore, maintain, or improve damaged tissues or whole organs. Specific to the field of tissue engineering, hydrogels have proven successful in the development of multiple cell types with the goal of one day serving as grafts that will replace large areas of lost or damaged tissue. This assembly depends on the presence of a scaffold, or three‐dimensional structure that guides the growth of new tissue, and hydrogel polymers in particular have been known to be an excellent building block for scaffold formation. This research focused on the synthesis and characterization of the hydrogel polyethylene glycol diacrylate‐acrylic acid (PEGDA‐AA), a hydrophilic material that provides a viable environment for cell maturation because of its favorable characteristics for the development and maturation of mouse C2C12 muscle cells. Using modified procedures adapted from Browe et al. (2016) to create a cost‐effective way of synthesizing this precursor, PEGDA was successfully synthesized and characterized through 13C and 1H‐NMR spectroscopy. Current research focuses on analyzing the effects of the hydrogel on the growth and development of C2C12 cells with the long‐term goal of establishing an efficient model for human tissue engineering.Support or Funding InformationHampden‐Sydney College Office of Undergraduate ResearchThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Translational Research of Tissue Engineering in China.

The presented translational research work represents the important progress and evolution of tissue engineering (TE) researches in China from proof-of-concept to actual application of TE technology for health management and biotechnology development in the past 15 years. It has been made with national governmental support of "863" project grant, which gathers multidisciplinary people in China to work together towards the translation of basic/applied research achievements into clinical reality and the fulfillment of unmet medical needs. It also truly reflects the global trend of fast development in this field from China's perspective.

3D bioprinted complex constructs reinforced by hybrid multilayers of electrospun nanofiber sheets

Despite the usefulness of hydrogels for cell-based bioprinting, the fragility of their resulting constructs has hindered their practical applications in tissue engineering research. Here, we suggest a hybrid integration method based on cell-hydrogel bioprinting that includes alternate layering of flexible nanofiber (NF) sheets. Because the bioprinting was implemented on a nanofibrous surface, the hydrogel-based materials could be printed with enhanced shape resolution compared to printing on a bare hydrogel. Furthermore, the insertion of NF sheets was effective for alleviating the shrinkage distortion of the hydrogel construct, which is inherently generated during the crosslinking process, thereby enhancing shape fidelity throughout the three-dimensional (3D) architecture. In addition to the structural precision, the NF-embedded constructs improved the mechanical properties in terms of compressive strength, modulus, and resilience limit (up to four-fold enhancement). With structural and mechanical supports, we could 3D fabricate complex constructs, including fully opened internal channels, which provided a favorable perfusion condition for cell growth. We confirmed the enhanced bioactivity of the NF-embedded bioprinted construct via cell culture experiments with 80% enhanced proliferation compared to the monolithic one. The synergistic combination of the two flexible materials, NFs and hydrogels, is expected to have extensive applicability in soft tissue engineering.

Computational Design and Optimization of Nerve Guidance Conduits for Improved Mechanical Properties and Permeability

Nerve guidance conduits (NGCs) are tubular tissue engineering scaffolds used for nerve regeneration. The poor mechanical properties and porosity have always compromised their performances for guiding and supporting axonal growth. Therefore, in order to improve the properties of NGCs, the computational design approach was adopted to investigate the effects of different NGC structural features on their various properties, and finally, design an ideal NGC with mechanical properties matching human nerves and high porosity and permeability. Three common NGC designs, namely hollow luminal, multichannel, and microgrooved, were chosen in this study. Simulations were conducted to study the mechanical properties and permeability. The results show that pore size is the most influential structural feature for NGC tensile modulus. Multichannel NGCs have higher mechanical strength but lower permeability compared to other designs. Square pores lead to higher permeability but lower mechanical strength than circular pores. The study finally selected an optimized hollow luminal NGC with a porosity of 71% and a tensile modulus of 8 MPa to achieve multiple design requirements. The use of computational design and optimization was shown to be promising in future NGC design and nerve tissue engineering research.

Concise Review: Application of Chemically Modified mRNA in Cell Fate Conversion and Tissue Engineering.

Chemically modified RNA (cmRNA) has potential as a safe and efficient tool for nucleic acid‐based therapies and regenerative medicine. Modifications in the chemistry of mRNA can enhance stability, reduce immunogenicity, and thus facilitate mRNA‐based nucleic acid therapy, which eliminates risk of insertional mutagenesis. In addition to these valuable advantages, the mRNA‐based method showed significantly higher efficacy for reprogramming somatic cells to pluripotency compared with DNA‐ or protein‐based methods. These findings suggest cmRNA can provide a powerful and safe tool for cell programming and reprogramming. Delivery methods, particularly using lipid nanoparticles, provide strategies for cell and organ‐specific targeting. The present study comprehensively compares studies that have used cmRNAs for cell fate conversion and tissue engineering. The information should be useful for investigators looking to choose the most efficient and straightforward cmRNA‐based strategy and protocol for tissue engineering and regenerative medicine research. stem cells translational medicine 2019;8:833&843

Construction of Cell–Extracellular Matrix Microenvironments by Conjugating ECM Proteins on Supported Lipid Bilayers

The cell membrane is an organized and fluid structure that modulates cellular activities in response to specific extracellular signals, and maintains the critical communication, integration and homeostasis between the cytosol and the extracellular matrix (ECM). In recent years, tissue engineering and cell biology research has been rapidly progressed by a remarkable understanding of cell and ECM interfaces. In this review, the design of new biomimetic platforms based on the conjugation of ECM proteins on solid supported lipid bilayers (SLBs) will be summarized. The platforms provide a better system to evaluate cellular responses to specific recognition events, gradient, mechanical property, nanostructures, and inter- and intra-molecular interactions of ECM proteins on a nonfouling and fluid membrane. Moreover, the findings from the molecular interactions and cellular activities will be highlighted to look into the cell-materials mechanisms.

Effect of copper nanoparticles on physico-chemical properties of chitosan and gelatin-based scaffold developed for skin tissue engineering application.

Development of new and effective scaffold continues to be an area of intense research in skin tissue engineering. The objective of this study was to study the effect of copper nanoparticles over physico-chemical properties of the chitosan and gelatin composite scaffolds for skin tissue engineering. The copper-doped scaffolds were prepared using freeze-drying method. Chitosan and gelatin were taken in varied composition with 0.01%, 0.02%, and 0.03% Cu nanoparticles. The physico-chemical properties of the copper nanoparticles and the scaffolds were analyzed using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy. Porosity of the scaffolds was measured by liquid displacement method and hemocompatibility was tested using goat blood. SEM micrographs of the scaffolds displayed the interconnected pores which ranged between 25 and 40µm. This average pore size was later enhanced to 95µm after the addition of copper nanoparticles. Cell viability assay was performed to ensure the growth and proliferation of the skin cells over the scaffolds. FTIR, EDS, and XRD analysis of scaffolds confirmed the presence of copper in the chitosan-based scaffolds. Porosity measurement showed the interconnectivity between pores which ranged between 65 and 88% as required for skin tissue engineering application. The degradation study of the scaffolds was done which depicted that, after the addition of copper nanoparticles with 0.03%, degradation rate was decreased. SEM and cytocompatibility assay on all scaffolds showed the cell adhesion and proliferation on the scaffolds which was not affected after addition of copper nanoparticles. Oxidative stress evaluation was done to study the effect of copper nanoparticles on the cells which showed that there was no such production of ROS in the scaffolds. Hence, scaffolds prepared after doping of copper nanoparticles show suitable physico-chemical and biological properties for skin tissue engineering application.

Disciplinary Learning From an Authentic Engineering Context

This small-scale design study describes disciplinary learning in mathematical modeling and science from an authentic engineeringthemed module. Current research in tissue engineering served as source material for the module, including science content for readings and a mathematical modeling activity in which students work in small teams to design a model in response to a problem from a client. The design of the module was guided by well-established principles of model-eliciting activities (a special class of problem-solving activities deeply studied in mathematics education) and recently published implementation design principles, which emphasize the portability of model-eliciting activities to many classroom settings.

Tissue engineering for the repair of peripheral nerve injury.

Peripheral nerve injury is a common clinical problem and affects the quality of life of patients. Traditional restoration methods are not satisfactory. Researchers increasingly focus on the field of tissue engineering. The three key points in establishing a tissue engineering material are the biological scaffold material, the seed cells and various growth factors. Understanding the type of nerve injury, the construction of scaffold and the process of repair are necessary to solve peripheral nerve injury and promote its regeneration. This review describes the categories of peripheral nerve injury, fundamental research of peripheral nervous tissue engineering and clinical research on peripheral nerve scaffold material, and paves a way for related research and the use of conduits in clinical practice.

Ethics of Using Animal Models as Predictors of Human Response in Tissue Engineering.

Use of outcomes from animal research for prediction of human response in tissue engineering studies has many ethical considerations. This article aims to contribute to the ethical discussion by delineating the framework of animal research and the ethical considerations at play, in particular with respect to cartilage tissue engineering. The history of animal research regulation and the current status of animal research in orthopedic tissue engineering is discussed. Questions addressed include how the proper animal models are chosen, how regulatory bodies ensure animal wellness and safety, and how guidelines are implemented and maintained throughout the life cycle of a project. Finally, we provide examples of both in vitro and in vivo cartilage tissue engineering research where animal models were employed as a predictive model of human response.

Application of injectable hydrogels for cardiac stem cell therapy and tissue engineering.

Cardiovascular diseases are responsible for approximately one-third of deaths around the world. Among cardiovascular diseases, the largest single cause of death is ischemic heart disease. Ischemic heart disease typically manifests as progressive constriction of the coronary arteries, which obstructs blood flow to the heart and can ultimately lead to myocardial infarction. This adversely affects the structure and function of the heart. Conventional treatments lack the ability to treat the myocardium lost during an acute myocardial infarction. Stem cell therapy offers an excellent solution for myocardial regeneration. Stem cell sources such as adult stem cells, embryonic and induced pluripotent stem cells have been the focal point of research in cardiac tissue engineering. However, cell survival and engraftment post-transplantation are major limitations that must be addressed prior to widespread use of this technology. Recently, biomaterials have been introduced as 3D vehicles to facilitate stem cell transplantation into infarct sites. This has shown significant promise with improved cell survival after transplantation. In this review, we discuss the various injectable hydrogels that have been tried in cardiac tissue engineering. Exploring and optimizing these cell-material interactions will guide cardiac tissue engineering towards developing stem cell based functional 3D constructs for cardiac regeneration.

Design and FEM model Analysis of MEMS cantilever structure for detection of Colon Cancer using Mass sensing

Colorectal tumor is the third most regularly analysed malignancy and the fourth driving reason for growth demise in both men and ladies in India. The Indian Cancer Society estimates that 3,79,100 individuals will be determined to have colorectal malignancy and 49,190 individuals will die from the disease in 2018. Traditionally, colon tumor is analysed utilizing infinitesimal investigation of histopathological colon tests. We present a method of detecting Human colorectal carcinoma cells in blood. Cells experience duplication and separation inside multi-cell living beings. The portrayal of physical properties of cells, for example, their mass and solidness has been of awesome intrigue and can have significant ramifications in cell science, tissue engineering, and disease research including cancer. The mechanism of duplication will increase the mass of the infected cells in the blood. In our study, we identify the change in mass of the carcinoma cells in the blood by observing the change in Eigen frequency of the cantilever beam.

Lateral Approach to the Lumbar Spine of Sprague Dawley Rat: Development of a Novel Animal Model for Spine Surgery

Background: Low back pain is a common ailment affecting individuals all around the globe. Animal models are required to study and further explore the treatment modalities. Lumbar spinal surgeries and disc repair is an important tissue engineering research domain. Dorsal and ventral approaches to access rat spine have been traditionally performed but suffer from a number of shortcomings such as higher morbidity, loss of neurology, high postoperative pain, and longer surgery. Methods: We used ten male Sprague Dawly rats, 3 months of age, and weighing an average of 280 gm. The surgeries were performed under dissociative anesthesia (ketamine: 50 mg/kg body weight). The spine was approached by left lateral incision extending from iliac crest and centering the level to be exposed. Skin and subcutaneous tissues were cut, external and internal oblique muscles were split in the direction of fibers, transverse abdominis was split vertically, and psoas was sacrificed. This made the spine and disc levels visible from the left lateral aspect. The muscles were approximated, and skin was closed with nonabsorbable mattress sutures. Postoperative analgesics (meloxicam 5 mg/kg body weight) and antibiotics (ceftriaxone 30 mg/kg body weight) were used. Results: This work has led to the development of a novel in vivo rat model using lateral retroperitoneal approach. This approach provides less pain and faster recovery in the postoperative stage. Moreover, it allows easy exposure and little surgery-related peri- or post-operative complications. Conclusion: Lateral retroperitoneal approach is a novel and safe method of spinal exposure in rats which may pave way for various live rat spine surgery models and experiments in future.

Bone marrow-derived mesenchymal stem cells enhance bone marrow regeneration in dental extraction sockets.

Bone marrow-derived mesenchymal stem cells (BMMSCs) remain the most widely used source of osteogenic cells in bone tissue engineering research. A cell-based treatment for alveolar ridge augmentation has received attention as an alternative to bone grafting. In the present study, BMMSC transplantation into tooth extraction sockets of C57BL/6J mice was evaluated for alveolar ridge regeneration. The first right maxillary molars were extracted, and then BMMSCs (PDGFRα+ Sca-1+ CD45- TER119- cells) isolated from femoral and tibial bone marrow were immediately transplanted into the extraction sockets. A control group underwent the same procedure except for BMMSC transplantation. Bone formation in the sockets was evaluated using micro-computed tomography and histological and immunohistochemical analyses. At 3 weeks, bone formation in the sockets was more advanced in the experimental group than in the control group. Histological analysis at 6 weeks after transplantation showed that the sockets in the experimental group also contained a greater quantity of bone marrow. Interestingly, socket bone mineral density was lower in the experimental group than in the control group at 6 weeks. These findings suggest that BMMSC transplantation accelerates bone healing and augments bone marrow formation in tooth extraction sockets.

The relationship between the umbilical cord length and it's diameter with the total CD34+ and total nucleated cell (TNC) as a parameter of cord blood selection

Background: The stem cell transplantation successful influenced by the quality of the umbilical cord blood which includes the number of CD34 + and Total Nucleated Cells (TNC).
 Objectives: The study aims to determine the correlation between umbilical cord length and it’s diameter, with the number of CD34 + and TNC cells as indicators of the quality of cord blood storage feasibility. Several other factors as maternal age, gestation period, and infant birth weight also examined.
 Methods: Thirty four of umbilical cords from the delivered woman in Dr. Mohammad Hoesin Hospital (RSMH) Palembang were collected from May to June 2018. The length and diameter of the cord were immediately measured after delivery. The evaluated cells were counted with a flow cytometer at Klaster Stem Cell and Tissue Engineering Research Centre (SCTE) IMERI Faculty of Medicine University of Indonesia (FKUI). 
 Results: Spearman correlation test show that there was no correlation between the length and diameter of the umbilical cord, maternal age, gestational period and infant weight, with the number of CD34 + and TNC cells in the cord (p> 0.05). However, the profile analysis indicated that the longer and larger the diameter of the umbilical cord, the higher concentration of the CD34+ and TNC cells.
 Conclusion: This study suggested that the younger maternal age, older gestational age, and higher infant birth weight, also normal hemoglobin level, tend to increase the number of CD34+ and TNC cells in the cord blood.

Microfibrous scaffolds from poly(l-lactide-co-ε-caprolactone) blended with xeno-free collagen/hyaluronic acid for improvement of vascularization in tissue engineering applications

Success of 3D tissue substitutes in clinical applications depends on the presence of vascular networks in their structure. Accordingly, research in tissue engineering is focused on the stimulation of angiogenesis or generation of a vascular network in the scaffolds prior to implantation. A novel, xeno-free, collagen/hyaluronic acid-based poly(l-lactide-co-ε-caprolactone) (PLC/COL/HA) (20/9.5/0.5 w/w/w) microfibrous scaffold was produced by electrospinning. Collagen types I and III, and hyaluronic acid were isolated from human umbilical cords and blended with the GMP grade PLC. When compared with PLC scaffolds the PLC/COL/HA had higher water uptake capacity (103% vs 66%) which may have contributed to the decrease in its Young's Modulus (from 1.31 to 0.89 MPa). The PLC/COL/HA better supported adipose tissue-derived mesenchymal stem cell (AT MSC) adhesion; within 24 h the cell number on the PLC/COL/HA scaffolds was 3 fold higher. Co-culture of human umbilical vein endothelial cells and AT MSCs induced capillary formation on both scaffold types, but the PLC/COL/HA led to formation of interconnected vessels whose total length was 1.6 fold of the total vessel length on PLC. Clinical use of this scaffold would eliminate the immune response triggered by xenogeneic collagen and transmission of animal-borne diseases while promoting a better vascular network formation.

Method for isolation of pancreatic blood vessels, their culture and coculture with islets of langerhans.

The only cure available for Type 1 diabetes involves the transplantation of islets of Langerhans isolated from donor organs. However, success rates are relatively low. Disconnection from vasculature upon isolation and insufficient rate of revascularization upon transplantation are thought to be a major cause, as islet survival and function depend on extensive vascularization. Research has thus turned toward the development of pretransplantation culture techniques to enhance revascularization of islets, so far with limited success. With the aim to develop a technique to enhance islet revascularization, this work proposes a method to isolate and culture pancreas-derived blood vessels. Using a mild multistep digestion method, pancreatic blood vessels were retrieved from whole murine pancreata and cultured in collagen Type 1. After 8 days, 50% of tissue explants had formed anastomosed microvessels which extended up to 300 μm from the explant tissue and expressed endothelial cell marker CD31 but not ductal marker CK19. Cocultures with islets of Langerhans revealed survival of both tissues and insulin expression by islets up to 8 days post-embedding. Microvessels were frequently found to encapsulate islets, however no islet penetration could be detected. This study reports for the first time the isolation and culture of pancreatic blood vessels. The methods and results presented in this work provide a novel explant culture model for angiogenesis and tissue engineering research with relevance to islet biology. It opens the door for in vivo validation of the potential of these pancreatic blood vessel explants to improve islet transplantation therapies. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2745, 2019.

Cartilage Tissue Engineering Using Stem Cells and Bioprinting Technology-Barriers to Clinical Translation.

There is no long-term treatment strategy for young and active patients with cartilage defects. Early and effective joint preserving treatments in these patients are crucial in preventing the development of osteoarthritis. Tissue engineering over the past few decades has presented hope in overcoming the issues involved with current treatment strategies. Novel advances in 3D bioprinting technology have promoted more focus on efficient delivery of engineered tissue constructs. There have been promising in-vitro studies and several animal studies looking at 3D bioprinting of engineered cartilage tissue. However, to date there are still no human clinical trials using 3D printed engineered cartilage tissue. This review begins with discussion surrounding the difficulties with articular cartilage repair and the limitations of current clinical management options which have led to research in cartilage tissue engineering. Next, the major barriers in each of the 4 components of cartilage tissue engineering; cells, scaffolds, chemical, and physical stimulation will be reviewed. Strategies that may overcome these barriers will be discussed. Finally, we will discuss the barriers surrounding intraoperative delivery of engineered tissue constructs and possible solutions.