The in vivo bioreactor is a tissue engineering paradigm that uses bioreactor methodology to grow neotissue in vivo that augments or replaces malfunctioning native tissue. Tissue engineering principles are used to construct a confined, artificial bioreactor space in vivo that hosts a tissue scaffold and key biomolecules necessary for neotissue growth. Said space often requires inoculation with pluripotent or specific stem cells to encourage initial growth, and access to a blood source. A blood source allows for recruitment of stem cells from the body alongside nutrient delivery for continual growth. This delivery of cells and nutrients to the bioreactor eventually results in the formation of a neotissue product.
Conceptually, the in vivo bioreactor was borne from complications in a repair method of bone fracture, bone loss, necrosis, and tumor reconstruction known as bone grafting. Traditional bone grafting strategies require fresh, autologous bone harvested from the iliac crest; this harvest site is limited by the amount of bone that can safely be removed, as well as associated pain and morbidity. [1] Other methods include cadaverous allografts and synthetic options (often made of hydroxyapatite) that have become available in recent years. In response to the question of limited bone sourcing, it has been posited that bone can be grown to fit a damaged region within the body through the application of tissue engineering principles. [2]
Tissue engineering is a biomedical engineering discipline that combines biology, chemistry, and engineering to design neotissue (newly formed tissue) on a scaffold. [3] Tissues scaffolds are functionally identical to the extracellular matrix found, acting as a site upon which regenerative cellular components adsorb to encourage cellular growth. [4] This cellular growth is then artificially stimulated by additive growth factors in the environment that encourage tissue formation. The scaffold is often seeded with stem cells and growth additives to encourage a smooth transition from cells to tissues, and more recently, organs. Traditionally, this method of tissue engineering is performed in vitro , where scaffold components and environmental manipulation recreate in vivo stimuli that direct growth. Environmental manipulation includes changes in physical stimulation, pH, potential gradients, cytokine gradients, and oxygen concentration. [5] The overarching goal of in vitro tissue engineering is to create a functional tissue that is equivalent to native tissue in terms of composition, biomechanical properties, and physiological performance. [6] However, in vitro tissue engineering suffers from a limited ability to mimic in vitro conditions, often leading to inadequate tissue substitutes. Therefore, in vivo tissue engineering has been suggested as a method to circumvent the tedium of environmental manipulation and use native in vivo stimuli to direct cell growth. To achieve in vivo tissue growth, an artificial bioreactor space must be established in which cells may grow. The in vivo bioreactor depends on harnessing the reparative qualities of the body to recruit stem cells into an implanted scaffold, and utilize vasculature to supply all necessary growth components.
Tissue engineering done in vivo is capable of recruiting local cellular populations into a bioreactor space. [2] [7] Indeed a range of neotissue growth has been shown: bone, cartilage, fat, and muscle. [7] [8] [9] [10] In theory, any tissue type could be grown in this manner if all necessary components (growth factors, environmental and physical ques) are met. Recruitment of stem cells require a complex process of mobilization from their niche, [11] though research suggests that mature cells transplanted upon the bioreactor scaffold can improve stem cell recruitment. [12] [13] [14] These cells secrete growth factors that promote repair and can be co-cultured with stem cells to improve tissue formation.
Scaffold materials are designed to enhance tissue formation through control of the local and surrounding environments. [15] [16] [17] Scaffolds are critical in regulating cellular growth and provide a volume in which vascularization and stem cell differentiation can occur. [18] Scaffold geometry significantly affects tissue differentiation through physical growth ques. Predicting tissue formation computationally requires theories that link physical growth ques to cell differentiation. Current models rely on mechano-regulation theory, widely shaped by Prendergast et al. for predicting cell growth. [19] Thus a quantitative analysis of geometry and materials commonly used in tissue scaffolds is capable.
Such materials include:
Initially, focusing on bone growth, subcutaneous pockets were used for bone prefabrication as a simple in vivo bioreactor model. The pocket is an artificially created space between varying levels of subcutaneous fascia. The location provides regenerative ques to the bioreactor implant but does not rely on pre-existing bone tissue as a substrate. Furthermore, these bioreactors may be wrapped with muscle tissue to encourage vascularization and bone growth. Another strategy is through the use of a periosteal flap wrapped around the bioreactor, or the scaffold itself to create an in vivo bioreactor. This strategy utilizes the guided bone regeneration treatment scheme, and is a safe method for bone prefabrication. These 'flap' methods of packing the bioreactor within fascia, or wrapping it in tissue is effective, though somewhat random due to the non-directed vascularization these methods incur. The axial vascular bundle (AVB) strategy requires that an artery and vein are inserted in an in vitro bioreactor to transport growth factors, cells, and remove waste. This ultimately results in extensive vascularization of the bioreactor space and a vast improvement in growth capability. This vascularization, though effective, is limited by the surface contact that it can achieve between the scaffold and the capillaries filling the bioreactor space. Thus, a combination of the flap and AVB techniques can maximize the growth rate and vascular contact of the bioreactor as suggested by Han and Dai, by inserting a vascular bundle into a scaffold wrapped in either musculature or periosteum. [28] If inadequate pre-existing vasculature is present in the growth site due to damage or disease, an arteriovenous loop (AVL) can be used. The AVL strategy requires a surgical connection be made between an artery of vein to form an arteriovenous fistula which is then placed within an in vitro bioreactor space containing a scaffold. A capillary network will form from this loop and accelerate the vascularization of new tissue. [29]
Materials used in the construction of an in vivo bioreactor space vary widely depending on the type of substrate, type of tissue, and mechanical demands of said tissue being grown. At its simplest, a bioreactor space will be created between tissue layers through the use of hydrogel injections to create a bioreactor space. Early models used an impermeable silicone shroud to encase a scaffold, [6] though more recent studies have begun 3D printing custom bioreactor molds to further enhance the mechanical growth properties of the bioreactors. The choice of bioreactor chamber material generally requires that it is nontoxic and medical grade, examples include: "silicon, polycarbonate, and acrylic polymer". [27] Recently both Teflon and titanium have been used in the growth of bone. [27] One study utilized Polymethyl methacrylate as a chamber material and 3D printed hollow rectangular blocks. [30] Yet another study pushed the limits of the in vivo bioreactor by proving that the omentum is suitable as a bioreactor space and chamber. Specifically, highly vascularized and functional bladder tissue was grown within the omentum space. [31]
An example of the implementation of the IVB approach was in the engineering of autologous bone by injecting calcium alginate in a sub-periosteal location. [32] [33] The periosteum is a membrane that covers the long bones, jawbone, ribs and the skull. This membrane contains an endogenous population of pluripotent cells called the periosteal cells, which are a type of mesenchymal stem cells (MSC), which reside in the cambium layer, i.e., the side facing the bone. A key step in the procedure is the elevation of the periosteum without damaging the cambium surface and to ensure this a new technique called hydraulic elevation was developed. [34]
The choice of the sub-periosteum site is used because stimulation of the cambium layer using transforming growth factor–beta resulted in enhanced chondrogenesis, i.e., formation of cartilage. In development the formation of bone can either occur via a Cartilage template initially formed by the MSCs that then gets ossified through a process called endochondral ossification or directly from MSC differentiation to bone via a process termed intra-membranous ossification . Upon exposure of the periosteal cells to calcium from the alginate gel, these cells become bone cells and start producing bone matrix through the intra-membranous ossification process, recapitulating all steps of bone matrix deposition. The extension of the IVB paradigm to engineering autologous hyaline cartilage was also recently demonstrated. [35] In this case, agarose is injected and this triggers local hypoxia, which then results in the differentiation of the periosteal MSCs into articular chondrocytes, i.e. cells similar to those found in the joint cartilage. Since this processes occurs in a relative short period of less than two weeks and cartilage can remodel into bone, this approach might provide some advantages in treatment of both cartilage and bone loss. The IVB concept needs to be however realized in humans and this is currently being undertaken.
Tissue engineering is a biomedical engineering discipline that uses a combination of cells, engineering, materials methods, and suitable biochemical and physicochemical factors to restore, maintain, improve, or replace different types of biological tissues. Tissue engineering often involves the use of cells placed on tissue scaffolds in the formation of new viable tissue for a medical purpose, but is not limited to applications involving cells and tissue scaffolds. While it was once categorized as a sub-field of biomaterials, having grown in scope and importance, it can is considered as a field of its own.
Pullulan is a polysaccharide polymer consisting of maltotriose units, also known as α-1,4- ;α-1,6-glucan'. Three glucose units in maltotriose are connected by an α-1,4 glycosidic bond, whereas consecutive maltotriose units are connected to each other by an α-1,6 glycosidic bond. Pullulan is produced from starch by the fungus Aureobasidium pullulans. Pullulan is mainly used by the cell to resist desiccation and predation. The presence of this polysaccharide also facilitates diffusion of molecules both into and out of the cell.
Endochondral ossification is one of the two essential pathways by which bone tissue is produced during fetal development of the mammalian skeletal system, the other pathway being intramembranous ossification. Both endochondral and intramembranous processes initiate from a precursor mesenchymal tissue, but their transformations into bone are different. In intramembranous ossification, mesenchymal tissue is directly converted into bone. On the other hand, endochondral ossification starts with mesenchymal tissue turning into an intermediate cartilage stage, which is eventually substituted by bone.
Articular cartilage, most notably that which is found in the knee joint, is generally characterized by very low friction, high wear resistance, and poor regenerative qualities. It is responsible for much of the compressive resistance and load bearing qualities of the knee joint and, without it, walking is painful to impossible. Osteoarthritis is a common condition of cartilage failure that can lead to limited range of motion, bone damage and invariably, pain. Due to a combination of acute stress and chronic fatigue, osteoarthritis directly manifests itself in a wearing away of the articular surface and, in extreme cases, bone can be exposed in the joint. Some additional examples of cartilage failure mechanisms include cellular matrix linkage rupture, chondrocyte protein synthesis inhibition, and chondrocyte apoptosis. There are several different repair options available for cartilage damage or failure.
Articular cartilage repair treatment involves the repair of the surface of the articular joint's hyaline cartilage, though these solutions do not perfectly restore the articular cartilage. These treatments have been shown to have positive results for patients who have articular cartilage damage. They can provide some measure of pain relief, while slowing down the accumulation of damage, or delaying the need for joint replacement surgery.
Autologous chondrocyte implantation is a biomedical treatment that repairs damages in articular cartilage. ACI provides pain relief while at the same time slowing down the progression or considerably delaying partial or total joint replacement surgery.
Clemens A. van Blitterswijk is a Dutch tissue engineer who contributed to the use of biomaterials to heal bone injuries, especially using osteoinductive ceramics. In collaboration with Jan de Boer and others, he has contributed to screening microtextures to study cell-biomaterial interactions, an approach termed materiomics.
Nano-scaffolding or nanoscaffolding is a medical process used to regrow tissue and bone, including limbs and organs. The nano-scaffold is a three-dimensional structure composed of polymer fibers very small that are scaled from a Nanometer scale. Developed by the American military, the medical technology uses a microscopic apparatus made of fine polymer fibers called a scaffold. Damaged cells grip to the scaffold and begin to rebuild missing bone and tissue through tiny holes in the scaffold. As tissue grows, the scaffold is absorbed into the body and disappears completely.
A fibrin scaffold is a network of protein that holds together and supports a variety of living tissues. It is produced naturally by the body after injury, but also can be engineered as a tissue substitute to speed healing. The scaffold consists of naturally occurring biomaterials composed of a cross-linked fibrin network and has a broad use in biomedical applications.
Arginylglycylaspartic acid (RGD) is the most common peptide motif responsible for cell adhesion to the extracellular matrix (ECM), found in species ranging from Drosophila to humans. Cell adhesion proteins called integrins recognize and bind to this sequence, which is found within many matrix proteins, including fibronectin, fibrinogen, vitronectin, osteopontin, and several other adhesive extracellular matrix proteins. The discovery of RGD and elucidation of how RGD binds to integrins has led to the development of a number of drugs and diagnostics, while the peptide itself is used ubiquitously in bioengineering. Depending on the application and the integrin targeted, RGD can be chemically modified or replaced by a similar peptide which promotes cell adhesion.
Gordana Vunjak-NovakovicFRSC is a Serbian American biomedical engineer and university professor. She is a University Professor at Columbia University, as well as the Mikati Foundation Professor of Biomedical Engineering and Medical Sciences. She also heads the laboratory for Stem Cells and Tissue Engineering at Columbia University. She is part of the faculty at the Irving Comprehensive Cancer Center and the Center for Human Development, both found at Columbia University. She is also an honorary professor at the Faculty of Technology and Metallurgy at the University of Belgrade, an honorary professor at the University of Novi Sad, and an adjunct professor at the Department of Biomedical Engineering at Tufts University.
Decellularization is the process used in biomedical engineering to isolate the extracellular matrix (ECM) of a tissue from its inhabiting cells, leaving an ECM scaffold of the original tissue, which can be used in artificial organ and tissue regeneration. Organ and tissue transplantation treat a variety of medical problems, ranging from end organ failure to cosmetic surgery. One of the greatest limitations to organ transplantation derives from organ rejection caused by antibodies of the transplant recipient reacting to donor antigens on cell surfaces within the donor organ. Because of unfavorable immune responses, transplant patients suffer a lifetime taking immunosuppressing medication. Stephen F. Badylak pioneered the process of decellularization at the McGowan Institute for Regenerative Medicine at the University of Pittsburgh. This process creates a natural biomaterial to act as a scaffold for cell growth, differentiation and tissue development. By recellularizing an ECM scaffold with a patient’s own cells, the adverse immune response is eliminated. Nowadays, commercially available ECM scaffolds are available for a wide variety of tissue engineering. Using peracetic acid to decellularize ECM scaffolds have been found to be false and only disinfects the tissue.
A 3D cell culture is an artificially created environment in which biological cells are permitted to grow or interact with their surroundings in all three dimensions. Unlike 2D environments, a 3D cell culture allows cells in vitro to grow in all directions, similar to how they would in vivo. These three-dimensional cultures are usually grown in bioreactors, small capsules in which the cells can grow into spheroids, or 3D cell colonies. Approximately 300 spheroids are usually cultured per bioreactor.
Muscle tissue engineering is a subset of the general field of tissue engineering, which studies the combined use of cells and scaffolds to design therapeutic tissue implants. Within the clinical setting, muscle tissue engineering involves the culturing of cells from the patient's own body or from a donor, development of muscle tissue with or without the use of scaffolds, then the insertion of functional muscle tissue into the patient's body. Ideally, this implantation results in full regeneration of function and aesthetic within the patient's body. Outside the clinical setting, muscle tissue engineering is involved in drug screening, hybrid mechanical muscle actuators, robotic devices, and the development of engineered meat as a new food source.
Treena Livingston Arinzeh is an American biomedical engineer and academic.
Antonios Georgios Mikos is a Greek-American biomedical engineer who is the Louis Calder Professor of Bioengineering and Chemical and Biomolecular Engineering at Rice University. He specialises in biomaterials, drug delivery, and tissue engineering.
Nasal chondrocytes (NC) are present in the hyaline cartilage of the nasal septum and in fact are the only cell type within the tissue. Similar to chondrocytes present in articular cartilage, NC express extracellular matrix proteins such as glycosaminoglycans and collagen.
Tissue engineered heart valves (TEHV) offer a new and advancing proposed treatment of creating a living heart valve for people who are in need of either a full or partial heart valve replacement. Currently, there are over a quarter of a million prosthetic heart valves implanted annually, and the number of patients requiring replacement surgeries is only suspected to rise and even triple over the next fifty years. While current treatments offered such as mechanical valves or biological valves are not deleterious to one's health, they both have their own limitations in that mechanical valves necessitate the lifelong use of anticoagulants while biological valves are susceptible to structural degradation and reoperation. Thus, in situ (in its original position or place) tissue engineering of heart valves serves as a novel approach that explores the use creating a living heart valve composed of the host's own cells that is capable of growing, adapting, and interacting within the human body's biological system.
Sarah Harriet Cartmell is a British biomaterials scientist and Professor of Bioengineering at the University of Manchester. She specializes on the potential use of electrical regimes to influence cellular activity for orthopaedic tissue engineering applications.
Craniofacial regeneration refers to the biological process by which the skull and face regrow to heal an injury. This page covers birth defects and injuries related to the craniofacial region, the mechanisms behind the regeneration, the medical application of these processes, and the scientific research conducted on this specific regeneration. This regeneration is not to be confused with tooth regeneration. Craniofacial regrowth is broadly related to the mechanisms of general bone healing.