Tissue Engineering Paradigm Research Articles

RNAi functionalized collagen-chitosan/silicone membrane bilayer dermal equivalent for full-thickness skin regeneration with inhibited scarring

Scar inhibition of dermal equivalent is one of the key issues for treatment of full thickness skin defects. To yield a bioactive RNAi functionalized matrix for skin regeneration with inhibited scarring, collagen-chitosan/silicone membrane bilayer dermal equivalent (BDE) was combined with trimetylchitosan (TMC)/siRNA complexes which could induce suppression of transforming growth factor-β1 (TGF-β1) pathway. The RNAi-BDE functioned as a reservoir for the incorporated TMC/siRNA complexes, enabling a prolonged siRNA release. The seeded fibroblasts in the RNAi-BDE showed good viability, internalized the TMC/siRNA complexes effectively and suppressed TGF-β1 expression constantly until 14 d. Application of the RNAi-BDE on the full-thickness skin defects of pig backs confirmed the in vivo inhibition of TGF-β1 expression by immunohistochemistry, real-time quantitative PCR and western blotting during 30 d post surgery. The levels of other scar-related factors such as collagen type I, collagen type III and α-smooth muscle actin (α-SMA) were also down-regulated. In combination with the ultra-thin skin graft transplantation for 73 d, the regenerated skin by RNAi-BDE had an extremely similar structure to that of the normal one. Our study reflects the latest paradigm of tissue engineering by incorporating the emerging biomolecule siRNA. The 3-D scaffolding materials for siRNA delivery may have general implications in generation of bioactive matrix as well.

Tissue-Engineered Vascular Grafts for Use in the Treatment of Congenital Heart Disease: From the Bench to the Clinic and Back Again

Since the first tissue-engineered vascular graft (TEVG) was implanted in a child over a decade ago, growth in the field of vascular tissue engineering has been driven by clinical demand for improved vascular prostheses with performance and durability similar to an autologous blood vessel. Great strides were made in pediatric congenital heart surgery using the classical tissue engineering paradigm, and cell seeding of scaffolds in vitro remained the cornerstone of neotissue formation. Our second-generation bone marrow cell-seeded TEVG diverged from tissue engineering dogma with a design that induces the recipient to regenerate vascular tissue in situ. New insights suggest that neovessel development is guided by cell signals derived from both seeded cells and host inflammatory cells that infiltrate the graft. The identification of these signals and the regulatory interactions that influence cell migration, phenotype and extracellular matrix deposition during TEVG remodeling are yielding a next-generation TEVG engineered to guide neotissue regeneration without the use of seeded cells. These developments represent steady progress towards our goal of an off-the-shelf tissue-engineered vascular conduit for pediatric congenital heart surgery.

Engineering of dental tissues; reality or distant prospect

Tissue engineering envelops a fast developing multidisciplinary field. Both from a medical and dental social point of view, there is great need to safeguard tissues and organs to overcome the shortage of donor organs and to reduce the health care cost. The ability to replace malfunctioning body parts has long been an issue of fiction and vision throughout the development of medical practice. One of the most striking examples is the legend of St. Cosmas and St. Damien, which describes the replacement of a man’s gangrenous leg with the limb of a dead black man. Apart from this legendary example, the more realistic replacement of damaged body tissue started in dentistry. The French surgeon Ambroise Pare (1510–1590) described the replacement of missing teeth by donor teeth, which were taken form “serfs”. It goes without saying that this kind of early transplantation was prone to failure due to existing lack of knowledge about immunology and sterility as well as the underlying phenomena of wound healing. The discovery of the sterility principles meant an enormous step forward. However, especially, all recent progress, as made in the field of molecular sciences as well as biomaterials and chemical engineering, seem to make the original dream of complete tissue and organ replacement reality. The question is whether this suggestion is indeed true which advantages and disadvantages are associated with this technique and what the final use of tissue engineering will be for the general practitioner. Tissue engineering can best be described as the creation of tissue. Tissue engineering is based on the tissue engineering paradigm, i.e. scaffold materials (a biomaterial in the shape of the newly to be regenerated tissue), stem cells (primordial cells which possess the potency to differentiate and proliferate in all kinds of different tissues), and signaling molecules (growth factors responsible for the differentiation of stem cells). In principle, tissue engineering can be applied for each existing tissue defect. At the moment, bone, periodontal ligament, gingiva and a complete tooth are important applications for dental tissue engineering.

Computed Tomography Detects Tissue Formation in a Stented Engineered Heart Valve

Tissue-engineered heart valves (TEHV) are being explored as an alternative to conventional heart valve prostheses. Using the classic tissue engineering paradigm, a stented tri-leaflet valve is fabricated. Subsequently, the construct is implanted into the pulmonary position in a sheep. Follow-up by means of computed tomography, magnetic resonance imaging, and echocardiography was used to assess tissue formation. After 4 weeks, the scaffold of the TEHV has degraded and new tissue is formed. However, small areas without tissue formation were present at macroscopic inspection. This phenomenon was only visible on computed tomographic images. Therefore, computed tomography appears a promising technique for in vivo follow-up of tissue formation in tissue-engineered heart valves.

Mesenchymal stem cells and bone regeneration: Current status

The enhancement of bone regeneration with biological agents including osteogenic growth factors and mesenchymal stem cells (MSCs) is becoming a clinical reality. Many exciting findings have been obtained following MSC implantation in animal models, and the data demonstrating their clinical efficacy in humans are promising. The overwhelming majority of experimental work has been performed with MSCs “amplified” in vitro. The nature of native MSCs in skeletal tissues however, remains poorly understood. This review summarizes recent findings pertaining to the definition and characterisation of MSCs in skeletal tissues and discusses the mechanisms of their actions in regenerating of bone in vivo. In respect to traditional tissue engineering paradigm, we bring together literature showing that the ways MSCs are extracted, expanded and implanted can considerably affect bone formation outcomes. Additionally, we discuss current animal models used in MSC research and highlight recent experiments showing important contribution of the host, and not only donor MSCs, in bone tissue formation. This knowledge provides a platform for novel therapy development for bone regeneration based on pharmacologically manipulated endogenous MSCs.

Influence of electrospun fiber mesh size on hMSC oxygen metabolism in 3D collagen matrices: Experimental and theoretical evidences

The traditional paradigm of tissue engineering of regenerating in vitro tissue or organs, through the combination of an artificial matrix and a cellular population has progressively changed direction. The most recent concept is the realization of a fully functional biohybrid, where both, the artificial and the biotic phase, concur in the formation of the novel organic matter. In this direction, interest is growing in approaches taking advantage of the control at micro- and nano-scale of cell material interaction based on the realization of elementary tassels of cells and materials which constitute the beginning point for the expansion of 3D more complex structures. Since a spontaneous assembly of all these components is expected, however, it becomes more fundamental than ever to define the features influencing cellular behavior, either they were material functional properties, or material architecture. In this work, it has been investigated the direct effect of electrospun fiber sizes on oxygen metabolism of h-MSC cells, when any other culture parameter was kept constant. To this aim, thin PCL electrospun membranes, with micro- and nano-scale texturing, were layered between two collagen slices up to create a sandwich structure (µC-PCL-C and nC-PCL-C). Cells were seeded on membranes, and the oxygen consumption was determined by a phosphorescence quenching technique. Results indicate a strong effect of the architecture of scaffolds on cell metabolism, also revealed by the increasing of HIF1-α gene expression in nC-PCL-C. These findings offer new insights into the role of materials in specific cell activities, also implying the existence of very interesting criteria for the control of tissue growth through the tuning of scaffold architecture.

An Emerging Paradigm in Tissue Engineering: From Chemical Engineering to Developmental Engineering for Bioartificial Tissue Formation through a Series of Unit Operations that Simulate the In Vivo Successive Developmental Stages†

The field of Tissue Engineering is in a critical stage, with the need to replace the trial-and-error methods (which, so far, are the most-often used) with rational methodologies. The opinions of se...

Noninvasive Transplantation of Bone Marrow Stromal Cells for Ischemic Stroke

Recent studies have indicated that bone marrow stromal cells (BMSCs) have the potential to improve neurological function when transplanted into animal models of cerebral infarct. However, it is still undetermined how the BMSCs should be transplanted to obtain the most efficient therapeutic benefits safely. The aim of this study was to assess whether a thermoreversible gelation polymer (TGP) hydrogel acts as a noninvasive, valuable scaffold in BMSC transplantation for infarct brain. The mice were subjected to permanent middle cerebral artery occlusion. Vehicle, BMSC suspension, or the BMSC-TGP construct was transplanted onto the ipsilateral intact neocortex at 7 days after the insult. Neurological symptoms were assessed throughout the experiments. The fate of the transplanted BMSC was examined 8 weeks after transplantation with immunohistochemistry. TGP hydrogel completely disappeared and provoked no inflammation in the host brain. Many transplanted cells were widely engrafted in the ipsilateral cerebrum, including the dorsal neocortex adjacent to the cerebral infarct in the BMSC-TGP construct-treated mice. Their number was significantly larger than in the BMSC-treated mice. The majority were positive for both NeuN and MAP2 and morphologically simulated the neurons. The findings suggest that surgical transplantation of tissue-engineered BMSCs onto the intact neocortex enhances the engraftment of donor cells around the cerebral infarct. These data may be useful in developing a noninvasive but efficient paradigm in neural tissue engineering. TGP hydrogel can be a promising candidate for valuable scaffolds in BMSC transplantation for central nervous system disorders because of its unique biochemical properties.

Modified polyelectrolyte complex fibrous scaffold as a matrix for 3D cell culture

The paradigm of scaffold tissue engineering relies on the provision of an appropriate environment for cell growth, which includes both structural support and the presentation of cellular signals. In terms of biosignal presentation, fibrous scaffolds by interfacial polyelectrolyte complexation (IPC) offer a clear advantage over other scaffold types as IPC scaffolds are formed using an aqueous-based, room-temperature process compatible with the incorporation of biological molecules. This paper establishes two primary methods for the chemical and biochemical modification of these scaffolds: (i) physical entrapment of the bioactive component, and (ii) covalent binding of the bioactive component. For the first method, extracellular matrix (ECM) proteins, collagen, fibronectin and laminin were drawn into the IPC fiber. For the second method, the cell adhesion peptide, RGD, was chemically conjugated to a thiol-active maleimidylated form of the scaffold. Immobilization of the bioactive components was characterized by confocal fluorescence microscopy, scanning electron microscopy (SEM) and BCA protein assay. The ECM proteins were distributed throughout the bulk and surface of the fiber. The ratio of covalently bound to physisorbed RGD was ∼ 2:3. The performance of the various scaffolds as a matrix to maintain the differentiated function of primary hepatocytes showed that albumin levels in the supernatant were in the order of RGD-modified scaffold > collagen Type I-modified scaffold > fibronectin- or laminin-modified scaffold > unmodified scaffold > plate, while no clear trend in urea production could be discerned. Thus, IPC scaffolds offered a promising platform for the presentation of signals to cells, in this case, to influence their differentiated function.

Soluble and insoluble signals sculpt osteogenesis in angiogenesis.

The basic tissue engineering paradigm is tissue induction and morphogenesis by combinatorial molecular protocols whereby soluble molecular signals are combined with insoluble signals or substrata. The insoluble signal acts as a three-dimensional scaffold for the initiation of de novo tissue induction and morphogenesis. The osteogenic soluble molecular signals of the transforming growth factor-β (TGF-β) supergene family, the bone morphogenetic/osteogenic proteins (BMPs/OPs) and, uniquely in the non-human primate Papio ursinus (P. ursinus), the three mammalian TGF-β isoforms induce bone formation as a recapitulation of embryonic development. In this paper, I discuss the pleiotropic activity of the BMPs/OPs in the non-human primate P. ursinus, the induction of bone by transitional uroepithelium, and the apparent redundancy of molecular signals initiating bone formation by induction including the three mammalian TGF-β isoforms. Amongst all mammals tested so far, the three mammalian TGF-β isoforms induce endochondral bone formation in the non-human primate P. ursinus only. Bone tissue engineering starts by erecting scaffolds of biomimetic biomaterial matrices that mimic the supramolecular assembly of the extracellular matrix of bone. The molecular scaffolding lies at the hearth of all tissue engineering strategies including the induction of bone formation. The novel concept of tissue engineering is the generation of newly formed bone by the implantation of "smart" intelligent biomimetic matrices that per se initiate the ripple-like cascade of bone differentiation by induction without exogenously applied BMPs/OPs of the TGF-β supergene family. A comprehensive digital iconographic material presents the modified tissue engineering paradigm whereby the induction of bone formation is initiated by intelligent smart biomimetic matrices that per se initiate the induction of bone formation without the exogenous application of the soluble osteogenic molecular signals. The driving force of the intrinsic induction of bone formation by bioactive biomimetic matrices is the shape of the implanted substratum. The language of shape is the language of geometry; the language of geometry is the language of a sequence of repetitive concavities, which biomimetizes the remodelling cycle of the primate osteonic bone.

High-Throughput and Combinatorial Technologies for Tissue Engineering Applications

As the field of tissue engineering progresses, new technology is essential to accelerate the identification of potentially translatable approaches for the repair of tissues damaged due to disease or trauma. The development of high-throughput and combinatorial technologies is helping to speed up research that is applicable to all aspects of the tissue engineering paradigm. This diverse technology can be used for both the rapid synthesis of polymers and their characterization with respect to local and bulk properties in a high-throughput fashion. The interactions of cells with many diverse materials in both two- and three-dimensions can be assessed rapidly through the use of microarrays and rapid outcome measures and with microfluidic devices for investigation of soluble factor and material combinations. Finally, small molecule screening of large libraries is being used to identify molecules that exhibit control over cell behavior, including stem cell differentiation. All of these approaches are aimed to move beyond traditional iterative methods to identify unique materials and molecules that would not have been identified otherwise. Although much of this work is only applicable for in vitro studies, future methods may translate into rapid screening of these approaches in vivo.

Human microvasculature fabrication using thermal inkjet printing technology

The current tissue engineering paradigm is that successfully engineered thick tissues must include vasculature. As biological approaches alone, such as VEGF, have fallen short of their promises, one may look for an engineering approach to build microvasculature. Layer-by-layer approaches for customized fabrication of cell/scaffold constructs have shown some potential in building complex 3D structures. With the advent of cell printing, one may be able to build precise human microvasculature with suitable bio-ink. Human microvascular endothelial cells (HMVEC) and fibrin were studied as bio-ink for microvasculature construction. Endothelial cells are the only cells to compose the human capillaries and also form the entire inner lining of cardiovascular system. Fibrin has been already widely recognized as tissue engineering scaffold for vasculature and other cells, including skeleton/smooth muscle cells and chondrocytes. In our study, we precisely fabricated micron-sized fibrin channels using a drop-on-demand polymerization. This printing technique uses aqueous processes that have been shown to induce little, if any, damage to cells. When printing HMVEC cells in conjunction with the fibrin, we found the cells aligned themselves inside the channels and proliferated to form confluent linings. The 3D tubular structure was also found in the printed patterns. We conclude that a combined simultaneous cell and scaffold printing can promote HMVEC proliferation and microvasculature formation.

Monitoring angiogenesis in soft-tissue engineered constructs for calvarium bone regeneration: anin vivolongitudinal DCE-MRI study

Tissue engineering is a promising technique for bone repair and can overcome the major drawbacks of conventional autogenous bone grafting. In this in vivo longitudinal study, we proposed a new tissue-engineering paradigm: inserting a biological soft-tissue construct within the bone defect to enhance angiogenesis for improved bone regeneration. The construct acts as a resorbable scaffold to support desired angiogenesis and cellular activity and as a vector of vascular endothelial growth factor, known to promote both vessel and bone growth. Dynamic contrast- enhanced magnetic resonance imaging was performed to investigate and characterize angiogenesis necessary for bone formation following the proposed paradigm of inserting a VEGF-impregnated tissue-engineered construct within the critical-sized calvarial defect in the membranous parietal bone of the rabbit. Results show that a model-free quantitative approach, the normalized initial area under the curve metric, provides sensitive and reproducible measures of vascularity that is consistent with known temporal evolution of angiogenesis during bone regeneration.

Blood vessel replacement: 50 years of development and tissue engineering paradigms in vascular surgery

The gold standard material in bypass surgery of blood vessels remains the patient's own artery or vein. However, this material may be unavailable, or may suffer vein graft disease. Currently available vascular prostheses, namely polyethylene terephthalate (PET, Dacron) and expanded polytetrafluoroethylene (ePTFE), perform well as large-caliber replacements, but their long-term patency is discouraging in small-caliber applications (<6 mm), such as in coronary, crural or microvessel surgery. This failure is mainly a result of an unfavorable healing process with surface thrombogenicity, due to lack of endothelial cells and anastomotic intimal hyperplasia caused by hemodynamic disturbances. An ideal small-diameter vascular graft has become a major focus of research. Novel biomaterials have been manufactured, and tissue-biomaterial interactions have been optimized. Tissue engineering technology has proven that the concept of partially or totally living blood vessels is feasible. The purpose of this review is to outline the vascular graft materials that are currently being implanted, taking into account cell-biomaterial physiology, tissue engineering approaches and the collective achievements of the authors.

Tissue engineering using autologous microcirculatory beds as vascularized bioscaffolds

Classic tissue engineering paradigms are limited by the incorporation of a functional vasculature and a reliable means for reimplantation into the host circulation. We have developed a novel approach to overcome these obstacles using autologous explanted microcirculatory beds (EMBs) as bioscaffolds for engineering complex three-dimensional constructs. In this study, EMBs consisting of an afferent artery, capillary beds, efferent vein, and surrounding parenchymal tissue are explanted and maintained for 24 h ex vivo in a bioreactor that preserves EMB viability and function. Given the rapidly advancing field of stem cell biology, EMBs were subsequently seeded with three distinct stem cell populations, multipotent adult progenitor cells (MAPCs), and bone marrow and adipose tissue-derived mesenchymal stem cells (MSCs). We demonstrate MAPCs, as well as MSCs, are able to egress from the microcirculation into the parenchymal space, forming proliferative clusters. Likewise, human adipose tissue-derived MSCs were also found to egress from the vasculature and seed into the EMBs, suggesting feasibility of this technology for clinical applications. We further demonstrate that MSCs can be transfected to express a luciferase protein and continue to remain viable and maintain luciferase expression in vivo. By using the vascular network of EMBs, EMBs can be perfused ex vivo and seeded with stem cells, which can potentially be directed to differentiate into neo-organs or transfected to replace failing organs and deficient proteins.

In vivo biocompatibility of ultra-short single-walled carbon nanotube/biodegradable polymer nanocomposites for bone tissue engineering

Scaffolds play a pivotal role in the tissue engineering paradigm by providing temporary structural support, guiding cells to grow, assisting the transport of essential nutrients and waste products, and facilitating the formation of functional tissues and organs. Single-walled carbon nanotubes (SWNTs), especially ultra-short SWNTs (US-tubes), have proven useful for reinforcing synthetic polymeric scaffold materials. In this article, we report on the in vivo biocompatibility of US-tube reinforced porous biodegradable scaffolds in a rabbit model. US-tube nanocomposite scaffolds and control polymer scaffolds were implanted in rabbit femoral condyles and in subcutaneous pockets. The hard and soft tissue response was analyzed with micro-computed tomography (micro CT), histology, and histomorphometry at 4 and 12 weeks after implantation. The porous US-tube nanocomposite scaffolds exhibited favorable hard and soft tissue responses at both time points. At 12 weeks, a three-fold greater bone tissue ingrowth was seen in defects containing US-tube nanocomposite scaffolds compared to control polymer scaffolds. Additionally, the 12 week samples showed reduced inflammatory cell density and increased connective tissue organization. No significant quantitative difference in polymer degradation was observed among the various groups; qualitative differences between the two time points were consistent with expected degradation due to the progression of time. Although no conclusions can be drawn from the present study concerning the osteoinductivity of US-tube nanocomposite scaffolds, the results suggest that the presence of US-tubes may render nanocomposite scaffolds bioactive assisting osteogenesis.

Freeze-dried Tendon Allografts as Tissue-engineering Scaffolds for Gdf5 Gene Delivery

Tendon reconstruction using grafts often results in adhesions that limit joint flexion. These adhesions are precipitated by inflammation, fibrosis, and the paucity of tendon differentiation signals during healing. In order to study this problem, we developed a mouse model in which the flexor digitorum longus (FDL) tendon is reconstructed using a live autograft or a freeze-dried allograft, and identified growth and differentiation factor 5 (Gdf5) as a therapeutic target. In this study we have investigated the potential of rAAV-Gdf5 -loaded freeze-dried tendon allografts as "therapeutically endowed" tissue-engineering scaffolds to reduce adhesions. In reporter gene studies we have demonstrated that recombinant adeno-associated virus (rAAV)-loaded tendon allografts mediate efficient transduction of adjacent soft tissues, with expression peaking at 7 days. We have also demonstrated that the rAAV-Gdf5 vector significantly accelerates wound healing in an in vitro fibroblast scratch model and, when loaded onto freeze-dried FDL tendon allografts, improves the metatarsophalangeal (MTP) joint flexion to a significantly greater extent than the rAAV-lacZ controls do. Collectively, our data demonstrate the feasibility and efficacy of therapeutic tendon allograft processing as a novel paradigm in tissue engineering in order to address difficult clinical problems such as tendon adhesions.

Simultaneous Deposition of Human Microvascular Endothelial Cells and Biomaterials for Human Microvasculature Fabrication Using Inkjet Printing

The current tissue engineering paradigm is that successfully engineered thick tissues must include vasculature. As biological approaches alone such as VGEF have fallen short of their promises, one may look for an engineering approach to build microvasculature. Layer-by-layer approach for customized fabrication of cell/scaffold constructs have shown some potential in building complex 3D structures. With the advent of cell printing, one may be able to build precise human microvasculature with suitable bioink. Human Microvascular Endothelial Cells (HMEC) and fibrin were studied as bioink for microvasculature construction. Endothelial cells are the only cells to compose the human capillaries and also the major cells of blood vessel intima layer. Fibrin has been already widely recognized as tissue engineering scaffold for vasculature and other cells, including skeleton/smooth muscle cells and chondrocytes. In our study, we precisely fabricated micron-sized fibrin channels using a drop-on-demand polymerization. This printing technique uses aqueous processes that have been shown to induce little, if any, damage to cells. When printing HMEC cells in conjunction with the fibrin, we found the cells aligned themselves inside the channels and proliferated to form confluent linings. Current studies to characterize the biology and functionality of these engineered microvascular structures will be presented. The preliminary data suggests that a combined simultaneous cell and scaffold printing can promote HMEC proliferation and microvasculature formation.

Integrating Principles of Developmental Biology in Tissue Engineering of Heart Valves

Approximately eight out of every thousand infants are born with congenital heart disease. The majority of these defects involve malformations or absence of the pulmonary valve and main pulmonary arteries. Although repair early in life is possible, the procedure often requires the use of a replacement valve. Current replacement options include prosthetic and bioprosthetic valves; however, these replacements have limited durability and are subject to calcification, thrombosis and a lack of growth potential [1]. Children, therefore, face an ongoing morbidity due to the limitations of these replacement valves and must endure multiple follow-up operations in order to place progressively larger valves to accommodate their growth. Tissue engineering offers the potential to create a living valve with the capacity for growth and self-repair, and which is resistant to infection. Tissue engineering is defined as an interdisciplinary field that applies the principles of engineering and life sciences toward the development of biological substitutes that restore, maintain or improve function [2]. This approach is based on seeding autologous cells onto biodegradable scaffolds or decelullarized biological matrices in order to provide temporary structural support and organization until the cells synthesize their own extracellular matrix. Previous studies by our group and others have demonstrated in vivo results culminating in the creation of single-valve leaflets [3], vascular grafts [4,5] and trileaflet-valved conduits [6–11]. These tissue-engineered heart valve (TEHV) structures have been created using vascular cells [3,4,6–9], umbilical bloodderived endothelial progenitor cells [11] and noncirculating bone marrow-derived mesenchymal stem cells [12,13] as cell sources. Our group and others have also demonstrated results in preliminary in vivo experiments. However, many questions remain unanswered: • What is the optimal cell source? • How will the scaffold material influence tissue growth and allow favorable scaffold cell and extracellular matrix interactions? • What in vitro conditions provide the most cell growth? • What is the timeframe for in vivo maturation? The ideal heart valve replacement would not only be biocompatible, readily available and incredibly durable, but also have the potential for growth [14]. Although the ultimate goal of TEHV is to recapitulate the matrix and cells found in the native tissue, variabilities exist within the potential strategies and sources of cells. A widely accepted paradigm of tissue engineering comprises of a scaffold that is preseeded with cells, followed by an in vitro stage of tissue formation typically conducted in a bioreactor (that recapitulates a physiological metabolic and mechanical environment) and, following subsequent implantation of the construct, an in vivo stage of tissue growth and remodeling [15].

Functional tissue engineering for tendon repair: A multidisciplinary strategy using mesenchymal stem cells, bioscaffolds, and mechanical stimulation

Over the past 8 years, our group has been continuously improving tendon repair using a functional tissue engineering (FTE) paradigm. This paradigm was motivated by inconsistent clinical results after tendon repair and reconstruction, and the modest biomechanical improvements we observed after repair of rabbit central patellar tendon defects using mesenchymal stem cell-gel-suture constructs. Although possessing a significantly higher stiffness and failure force than for natural healing, these first generation constructs were quite weak compared to normal tendon. Fundamental to the new FTE paradigm was the need to determine in vivo forces to which the repair tissue might be exposed. We first recorded these force patterns in two normal tendon models and then compared these peak forces to those for repairs of central defects in the rabbit patellar tendon model (PT). Replacing the suture with end-posts in culture and lowering the mesenchymal stem cell (MSC) concentration of these constructs resulted in failure forces greater than peak in vivo forces that were measured for all the studied activities. Augmenting the gel with a type I collagen sponge further increased repair stiffness and maximum force, and resulted in the repair tangent stiffness matching normal stiffness up to peak in vivo forces. Mechanically stimulating these constructs in bioreactors further enhanced repair biomechanics compared to normal. We are now optimizing components of the mechanical signal that is delivered in culture to further improve construct and repair outcome. Our contributions in the area of tendon functional tissue engineering have the potential to create functional load-bearing repairs that will revolutionize surgical reconstruction after tendon and ligament injury.