Layered Scaffolds for Osteochondral Tissue Engineering.

Despite huge efforts, tissue engineers and orthopedic surgeons still face a great challenge to functionally repair osteochondral (OC) defects. Nevertheless, over the past decade great progress has been made to find suitables strategies towards OC regeneration. In the clinics, some osteochondral tissue engineering (OCTE) strategies have already been applied although with some incongruous outcomes as OC tissue is complex in its architecture and function. In this chapter, we have summarized current OCTE strategies that are focused on hierarchical scaffold design, mainly layered scaffolds. Most suitable candidates towards functional regeneration of OC tissues are envisaged from monophasic to layered scaffolds. Herein is documented a variety of strategies with their intrinsic properties for further application as bare scaffolds or in combination with biologics. Both in vitro and in vivo approaches have been thoroughly studied aiming at functional OC regeneration. The most noteworthy studies in OC regeneration developed within the past 5 years are herein documented as well as some current clinical trials.

Gellan Gum-Based Hydrogels for Osteochondral Repair.

Gellan gum (GG) is a widely explored natural polysaccharide that has been gaining attention in tissue engineering (TE) and regenerative medicine field, and more recently in osteochondral TE approaches. Taking advantage of its inherent features such as biocompatibility, biodegradability, similarity with the extracellular matrix and easy functionalization, GG-based hydrogels have been studied for their potential for cartilage and bone tissue regeneration. Several preclinical studies describe the successful outcome of GG in cartilage tissue engineering. By its turn, GG composites have also been proposed in several strategies to guide bone formation. The big challenge in osteochondral TE approaches is still to achieve cartilage and bone regeneration simultaneously through a unique integrated bifunctional construct. The potential of GG to be used as polymeric support to reach both bone and cartilage regeneration has been demonstrated. This chapter provides an overview of GG properties and the functionalization strategies employed to tailor its behaviour to a particular application. The use of GG in soft and hard tissues regeneration approaches, as well in osteochondral integrated TE strategies is also revised.

Silk Fibroin-Based Hydrogels and Scaffolds for Osteochondral Repair and Regeneration.

Osteochondral lesions treatment and regeneration demands biomimetic strategies aiming physicochemical and biological properties of both bone and cartilage tissues, with long-term clinical outcomes. Hydrogels and scaffolds appeared as assertive approaches to guide the development and structure of the new osteochondral engineered tissue. Moreover, these structures alone or in combination with cells and bioactive molecules bring the mechanical support after in vitro and in vivo implantation. Moreover, multilayered structures designed with continuous interfaces furnish appropriate features of the cartilage and subchondral regions, namely microstructure, composition, and mechanical properties. Owing the potential as scaffolding materials, natural and synthetic polymers, bioceramics, and composites have been employed. Particularly, significance is attributed to the natural-based biopolymer silk fibroin from the Bombyx mori silkworm, considering its unique mechanical and biological properties. The significant studies on silk fibroin-based structures, namely hydrogels and scaffolds, towards bone, cartilage, and osteochondral tissue repair and regeneration are overviewed herein. The developed biomimetic strategies, processing methodologies, and final properties of the structures are summarized and discussed in depth.

Commercial Products for Osteochondral Tissue Repair and Regeneration.

The osteochondral tissue represents a complex structure composed of four interconnected structures, namely hyaline cartilage, a thin layer of calcified cartilage, subchondral bone, and cancellous bone. Due to the several difficulties associated with its repair and regeneration, researchers have developed several studies aiming to restore the native tissue, some of which had led to tissue-engineered commercial products. In this sense, this chapter discusses the good manufacturing practices, regulatory medical conditions and challenges on clinical translations that should be fulfilled regarding the safety and efficacy of the new commercialized products. Furthermore, we review the current osteochondral products that are currently being marketed and applied in the clinical setting, emphasizing the advantages and difficulties of each one.

Advances for Treatment of Knee OC Defects.

Osteochondral (OC) defects are prevalent among young adults and are notorious for being unable to heal. Although they are traumatic in nature, they often develop silently. Detection of many OC defects is challenging, despite the criticality of early care. Current repair approaches face limitations and cannot provide regenerative or long-standing solution. Clinicians and researchers are working together in order to develop approaches that can regenerate the damaged tissues and protect the joint from developing osteoarthritis. The current concepts of tissue engineering and regenerative medicine, which have brought many promising applications to OC management, are overviewed herein. We will also review the types of stem cells that aim to provide sustainable cell sources overcoming the limitation of autologous chondrocyte-based applications. The various scaffolding materials that can be used as extracellular matrix mimetic and having functional properties similar to the OC unit are also discussed.

Nanoparticles-Based Systems for Osteochondral Tissue Engineering.

Osteochondral lesions represent one of the major causes of disabilities in the world. These defects are due to degenerative or inflammatory arthritis, but both affect the articular cartilage and the underlying subchondral bone. Defects from trauma or degenerative pathology frequently cause severe pain, joint deformity, and loss of joint motion. Osteochondral defects are a significant challenge in orthopedic surgery, due to the cartilage complexity and unique structure, as well as its exposure to high pressure and motion. Although there are treatments routinely performed in the clinical practice, they present several limitations. Tissue engineering can be a suitable alternative for osteochondral defects since bone and cartilage engineering had experienced a notable advance over the years. Allied with nanotechnology, osteochondral tissue engineering (OCTE) can be leveled up, being possible to create advanced structures similar to the OC tissue. In this chapter, the current strategies using nanoparticles-based systems are overviewed. The results of the studies herein considered confirm that advanced nanomaterials will undoubtedly play a crucial role in the design of strategies for treatment of osteochondral defects in the near future.

PRP Therapy.

Osteochondral lesions remain as a clinical challenge despite the advances in orthopedic regenerative strategies. Biologics, in particular, platelet-rich plasma, has been applied for the reparative and regenerative effect in many tissues, and osteochondral tissue is not an exception. Platelet-rich plasma is an autologous concentrate prepared from the collected blood; thus, this safe application is free of immune response or risk of transmission of disease. It has a high potential to promote regeneration, thanks to its content, and can be applied alone or can reinforce a tissue engineering strategy. The relevant works making use of platelet-rich plasma in osteochondral lesions are overviewed herein. The practical success of platelet-rich plasma is uncertain since there are many factors involved including but not limited to its preparation and administration method. Nevertheless, today, the issues and challenges of platelet-rich plasma have been well acknowledged by researchers and clinicians. Thus, it is believed that a consensus will be built it, and then with high-quality randomized controlled trials and standardized protocols, the efficacy of platelet-rich plasma therapy can be better evaluated. HIGHLIGHTS: The need of treating the osteochondral lesions has not been yet met in the clinics. Thanks to being an autologous source of growth factors, interleukins, and other cytokines and relative ease of clinical application, i.e., during a single-step surgical procedure, the use of platelet-rich plasma is of great interest. The high theoretical potential of the role of platelet-rich plasma in the regeneration process of osteochondral lesions is known, and the efficiency needs to be confirmed by high-quality randomized controlled trials for a robust position in the treatments of osteochondral lesions in the clinics.

In Vitro Mimetic Models for the Bone-Cartilage Interface Regeneration.

In embryonic development, pure cartilage structures are in the basis of bone-cartilage interfaces. Despite this fact, the mature bone and cartilage structures can vary greatly in composition and function. Nevertheless, they collaborate in the osteochondral region to create a smooth transition zone that supports the movements and forces resulting from the daily activities. In this sense, all the hierarchical organization is involved in the maintenance and reestablishment of the equilibrium in case of damage. Therefore, this interface has attracted a great deal of interest in order to understand the mechanisms of regeneration or disease progression in osteoarthritis. With that purpose, in vitro tissue models (either static or dynamic) have been studied. Static in vitro tissue models include monocultures, co-cultures, 3D cultures, and ex vivo cultures, mostly cultivated in flat surfaces, while dynamic models involve the use of bioreactors and microfluidic systems. The latter have emerged as alternatives to study the cellular interactions in a more authentic manner over some disadvantages of the static models. The current alternatives of in vitro mimetic models for bone-cartilage interface regeneration are overviewed and discussed herein.

Small Animal Models.

Animal assays represent an important stage between in vitro studies and human clinical applications. These models are crucial for biomedical research and regenerative medicine studies, as these offer precious information for systematically assessing the efficacy and risks of recently created biomaterials, medical devices, drugs, and therapeutic modalities prior to initiation of human clinical trials. Therefore, selecting a suitable experimental model for tissue engineering purposes is essential to establish valid conclusions. However, it remains important to be conscious of the advantages and limitations of the various small and large animal models frequently used for biomedical research as well as the different challenges encountered in extrapolating data obtained from animal studies and the risks of misinterpretation. This chapter discusses the various small animal model strategies used for osteochondral defect repair. Particular emphasis will be placed on analyzing the materials and strategies used in each model.

Bioceramics for Osteochondral Tissue Engineering and Regeneration.

Considerable advances in tissue engineering and regeneration have been accomplished over the last decade. Bioceramics have been developed to repair, reconstruct, and substitute diseased parts of the body and to promote tissue healing as an alternative to metallic implants. Applications embrace hip, knee, and ligament repair and replacement, maxillofacial reconstruction and augmentation, spinal fusion, bone filler, and repair of periodontal diseases. Bioceramics are well-known for their superior wear resistance, high stiffness, resistance to oxidation, and low coefficient of friction. These specially designed biomaterials are grouped in natural bioceramics (e.g., coral-derived apatites), and synthetic bioceramics, namely bioinert ceramics (e.g., alumina and zirconia), bioactive glasses and glass ceramics, and bioresorbable calcium phosphates-based materials. Physicochemical, mechanical, and biological properties, as well as bioceramics applications in diverse fields of tissue engineering are presented herein. Ongoing clinical trials using bioceramics in osteochondral tissue are also considered. Based on the stringent requirements for clinical applications, prospects for the development of advanced functional bioceramics for tissue engineering are highlighted for the future.

Tissue Engineering Strategies for Osteochondral Repair.

Tissue engineering strategies have been pushing forward several fields in the range of biomedical research. The musculoskeletal field is not an exception. In fact, tissue engineering has been a great asset in the development of new treatments for osteochondral lesions. Herein, we overview the recent developments in osteochondral tissue engineering. Currently, the treatments applied in a clinical scenario have shown some drawbacks given the difficulty in regenerating a fully functional hyaline cartilage. Among the different strategies designed for osteochondral regeneration, it is possible to identify cell-free strategies, scaffold-free strategies, and advanced strategies, where different materials are combined with cells. Cell-free strategies consist in the development of scaffolds in the attempt to better fulfill the requirements of the cartilage regeneration process. For that, different structures have been designed, from monolayers to multilayered structures, with the intent to mimic the osteochondral architecture. In the case of scaffold-free strategies, they took advantage on the extracellular matrix produced by cells. The last strategy relies in the development of new biomaterials capable of mimicking the extracellular matrix. This way, the cell growth, proliferation, and differentiation at the lesion site are expedited, exploiting the self-regenerative potential of cells and its interaction with biomolecules. Overall, despite the difficulties associated with each approach, tissue engineering has been proven a valuable tool in the regeneration of osteochondral lesions and together with the latest advances in the field, promises to revolutionize personalized therapies.

Bioreactors and Microfluidics for Osteochondral Interface Maturation.

The cell culture techniques are in the base of any biology-based science. The standard techniques are commonly static platforms as Petri dishes, tissue culture well plates, T-flasks, or well plates designed for spheroids formation. These systems faced a paradigm change from 2D to 3D over the current decade driven by the tissue engineering (TE) field. However, 3D static culture approaches usually suffer from several issues as poor homogenization of the formed tissues and development of a necrotic center which limits the size of in vitro tissues to hundreds of micrometers. Furthermore, for complex tissues as osteochondral (OC), more than recovering a 3D environment, an interface needs to be replicated. Although 3D cell culture is already the reality adopted by a newborn market, a technological revolution on cell culture devices needs a further step from static to dynamic already considering 3D interfaces with dramatic importance for broad fields such as biomedical, TE, and drug development. In this book chapter, we revised the existing approaches for dynamic 3D cell culture, focusing on bioreactors and microfluidic systems, and the future directions and challenges to be faced were discussed. Basic principles, advantages, and challenges of each technology were described. The reported systems for OC 3D TE were focused herein.

Hyaluronic Acid.

In recent times, the field of tissue engineering and regenerative medicine (TERM) has considerably increased the extent of therapeutic strategies for clinical application in orthopedics. However, TERM approaches have its rules and requirements, in the respect of the biologic response of each tissue and bioactive agents which need to be considered, respected, and subject of ongoing studies. Different medical devices/products have been prematurely available on the market and used in clinics with limited success. However, other therapeutics, when used in a serious and evidence-based approach, have achieved considerable success, considering the respect for solid expectations from doctors and patients (when properly informed).Orthobiologics has appeared as a recent technological trend in orthopedics. This includes the improvement or regeneration of different musculoskeletal tissues by means of using biomaterials (e.g., hyaluronic acid), stem cells, and growth factors (e.g., platelet-rich plasma). The potential symbiotic relationship between biologic therapies and surgery makes these strategies suitable to be used in one single intervention.However, herein, the recent clinical studies using hyaluronic acid (HA) in the treatment of orthopedic conditions will mainly be overviewed (e.g., osteochondral lesions, tendinopathies). The possibilities to combine different orthobiologic agents as TERM clinical strategies for treatment of orthopedic problems will also be briefly discussed.

Promising Biomolecules.

The osteochondral defect (OD) comprises the articular cartilage and its subchondral bone. The treatment of these lesions remains as one of the most problematic clinical issues, since these defects include different tissues, requiring distinct healing approaches. Among the growing applications of regenerative medicine, clinical articular cartilage repair has been used for two decades, and it is an effective example of translational medicine; one of the most used cell-based repair strategies includes implantation of autologous cells in degradable scaffolds such as alginate, agarose, collagen, chitosan, chondroitin sulfate, cellulose, silk fibroin, hyaluronic acid, and gelatin, among others. Concerning the repair of osteochondral defects, tissue engineering and regenerative medicine started to design single- or bi-phased scaffold constructs, often containing hydroxyapatite-collagen composites, usually used as a bone substitute. Biomolecules such as natural and synthetic have been explored to recreate the cartilage-bone interface through multilayered biomimetic scaffolds. In this chapter, a succinct description about the most relevant natural and synthetic biomolecules used on cartilage and bone repair, describing the procedures to obtain these biomolecules, their chemical structure, common modifications to improve its characteristics, and also their application in the biomedical fields, is given.

Stem Cells for Osteochondral Regeneration.

Stem cell research plays a central role in the future of medicine, which is mainly dependent on the advances on regenerative medicine (RM), specifically in the disciplines of tissue engineering (TE) and cellular therapeutics. All RM strategies depend upon the harnessing, stimulation, or guidance of endogenous developmental or repair processes in which cells have an important role. Among the most clinically challenging disorders, cartilage degeneration, which also affects subchondral bone becoming an osteochondral (OC) defect, is one of the most demanding. Although primary cells have been clinically applied, stem cells are currently seen as the promising tool of RM-related research because of its availability, in vitro proliferation ability, pluri- or multipotency, and immunosuppressive features. Being the OC unit, a transition from the bone to cartilage, mesenchymal stem cells (MSCs) are the main focus for OC regeneration. Promising alternatives, which can also be obtained from the patient or at banks and have great differentiation potential toward a wide range of specific cell types, have been reported. Still, ethical concerns and tumorigenic risk are currently under discussion and assessment. In this book chapter, we revise the existing stem cell-based approaches for engineering bone and cartilage, focusing on cell therapy and TE. Furthermore, 3D OC composites based on cell co-cultures are described. Finally, future directions and challenges still to be faced are critically discussed.

Clinical Trials and Management of Osteochondral Lesions.

Osteochondral lesions are frequent and important causes of pain and disability. These lesions are induced by traumatic injuries or by diseases that affect both the cartilage surface and the subchondral bone. Due to the limited cartilage ability to regenerate and self-repair, these lesions tend to gradually worsen and progress towards osteoarthritis. The clinical, social, and economic impact of the osteochondral lesions is impressive and although therapeutic alternatives are under discussion, a consensus is not yet been achieved. Over the previous decade, new strategies based on innovative tissue engineering approaches have been developed with promising results. However, in order those products reach the market and help the actual patient in an effective manner, there is still a lot of work to be done. The current state of the implications, clinical aspects, and available treatments for this pathology, as well as the ongoing preclinical and clinical trials are presented in this chapter.

Emerging Concepts in Treating Cartilage, Osteochondral Defects, and Osteoarthritis of the Knee and Ankle.

The management and treatment of cartilage lesions, osteochondral defects, and osteoarthritis remain a challenge in orthopedics. Moreover, these entities have different behaviors in different joints, such as the knee and the ankle, which have inherent differences in function, biology, and biomechanics. There has been a huge development on the conservative treatment (new technologies including orthobiologics) as well as on the surgical approach. Some surgical development upraises from technical improvements including advanced arthroscopic techniques but also from increased knowledge arriving from basic science research and tissue engineering and regenerative medicine approaches. This work addresses the state of the art concerning basic science comparing the knee and ankle as well as current options for treatment. Furthermore, the most promising research developments promising new options for the future are discussed.

Global rotation has high sensitivity in ACL lesions within stress MRI.

PURPOSE: This study aims to objectively compare side-to-side differences of P-A laxity alone and coupled with rotatory laxity within magnetic resonance imaging, in patients with total anterior cruciate ligament (ACL) rupture. METHODS: This prospective study enrolled sixty-one patients with signs and symptoms of unilateral total anterior cruciate ligament rupture, which were referred to magnetic resonance evaluation with simultaneous instrumented laxity measurements. Sixteen of those patients were randomly selected to also have the contralateral healthy knee laxity profile tested. Images were acquired for the medial and lateral tibial plateaus without pressure, with postero-anterior translation, and postero-anterior translation coupled with maximum internal and external rotation, respectively. RESULTS: All parameters measured were significantly different between healthy and injured knees (P < 0.05), with exception of lateral plateau without stress. The difference between injured and healthy knees for medial and lateral tibial plateaus anterior displacement (P < 0.05) and rotation (P < 0.001) was statistically significant. It was found a significant correlation between the global rotation of the lateral tibial plateau (lateral plateau with internal + external rotation) with pivot-shift, and between the anterior global translation of both tibial plateaus (medial + lateral tibial plateau) with Lachman. The anterior global translation of both tibial plateaus was the most specific test with a cut-off point of 11.1 mm (93.8 %), and the global rotation of the lateral tibial plateau was the most sensitive test with a correspondent cut-off point of 15.1 mm (92.9 %). CONCLUSION: Objective laxity quantification of ACL-injured knees showed increased sagittal laxity, and simultaneously in sagittal and transversal planes, when compared to their healthy contralateral knee. Moreover, when measuring instability from anterior cruciate ligament ruptures, the anterior global translation of both tibial plateaus and global rotation of the lateral tibial plateau add diagnostic specificity and sensitivity. This work strengthens the evidence that the anterior cruciate ligament plays an important biomechanical role in controlling the anterior translation, but also both internal and external rotation. The high sensitivity and specificity of this device in objectively identifying and measuring the multiplanar instability clearly guides stability restoration clinical procedures. Level of evidence Cross-sectional study, Level III.

Prevalence of Articular Cartilage Lesions and Surgical Clinical Outcomes in Football (Soccer) Players' Knees: A Systematic Review.

PURPOSE: To systematize the available scientific literature on the prevalence of articular cartilage and/or osteochondral lesions in football (soccer) players' knees, and overview the surgical procedures and functional outcomes and return to sports. METHODS: A comprehensive search using Pubmed, Cochrane Library, SPORTDiscus, and CINAHL databases was carried out until September 30, 2015. All English language studies that assessed the outcomes of a surgical technique for the treatment of articular cartilage lesions in football players' knees, with a minimum follow-up of 12 months, were included. The reference list of the most relevant papers was screened. The main outcomes of interest were the clinical, arthroscopy or imaging primary outcomes and the return to sports rate. The methodological and reporting qualities were assessed according to Coleman methodology score. RESULTS: The search provided 485 titles and abstracts. Five studies were eligible for inclusion (mean Coleman score of 37.2 points), comprising a total of 183 football players with a mean age of 25.7 years. A total of 217 articular cartilage and/or osteochondral lesions were reported, where the medial and lateral femoral condyles were the most common sites of lesion. The surgical procedures investigated were mosaicplasty, microfracture, autologous chondrocyte implantation, and chondral debridement. CONCLUSIONS: No definitive conclusion could be made in respect to the best current surgical technique for articular cartilage and osteochondral lesions. Microfracture and mosaicplasty can provide a faster return to competition and faster clinical and functional results, whereas autologous chondrocyte implantation and/or matrix-induced autologous chondrocytes implantation procedures can enhance longstanding clinical and functional results. LEVEL OF EVIDENCE: Level IV, systematic review of Level III and IV studies.

Hydroxyapatite/alginate/gellan gum inks with osteoconduction and osteogenic potential for bioprinting bone tissue analogues.

There is a growing demand for engineered bone tissues custom-designed to match the patient-specific defect size and in vitro models for studying bone diseases and/or drug screening. Herein, we propose a bioprinted bone tissue construct using SaOs-2 cells within alginate/gellan gum/hydroxyapatite inks. Different ink formulations were developed with varying hydroxyapatite content and then evaluated for viscoelasticity, printability, biomineralization properties, post-printing viability, proliferation, metabolic activity, and osteogenic phenotype of SaOs-2-encapsulated cells. Results indicate that ink formulations exhibit non-Newtonian shear-thinning behaviour, maintaining shape integrity and structural stability post-printing. Ink mineralization rates increase with the hydroxyapatite content, rendering them suitable for bone defect strategies. Post-printed cells in the developed constructs remain live, spreading, and metabolically active but do not proliferate. Osteogenic gene and protein expression, both early and late, show upregulation at day 7 relative to day 1, followed by downregulation at day 14. Lower hydroxyapatite content inks demonstrate up to fourfold upregulation in genes and proteins at most time points. Additionally, these constructs release calcium and phosphate at levels conducive to mineralization. Overall, the tissue-engineered miniaturized constructs not only meet the criteria for early-stage bone defect/fracture regeneration but also serve as a promising platform for drug screening and evaluating potential therapeutic treatments.