Vancomycin-Loaded, Nanohydroxyapatite-Based Scaffold for Osteomyelitis Treatment: In Vivo Rabbit Toxicological Tests and In Vivo Efficacy Tests in a Sheep Model

The treatment for osteomyelitis consists of surgical debridement, filling of the dead space, soft tissue coverage, and intravenous administration of antimicrobial (AM) agents for long periods. Biomaterials for local delivery of AM agents, while providing controllable antibiotic release rates and simultaneously acting as a bone scaffold, may be a valuable alternative; thus, avoiding systemic AM side effects. V-HEPHAPC is a heparinized nanohydroxyapatite (nHA)/collagen biocomposite loaded with vancomycin that has been previously studied and tested in vitro. It enables a vancomycin-releasing profile with an intense initial burst, followed by a sustained release with concentrations above the Minimum Inhibitory Concentration (MIC) for MRSA. In vitro results have also shown that cellular viability is not compromised, suggesting that V-HEPHAPC granules may be a promising alternative device for the treatment of osteomyelitis. In the present study, V-HEPHAPC (HEPHAPC with vancomycin) granules were used as a vancomycin carrier to treat MRSA osteomyelitis. First, in vivo Good Laboratory Practice (GLP) toxicological tests were performed in a rabbit model, assuring that HEPHAPC and V-HEPHAPC have no relevant side effects. Second, V-HEPHAPC proved to be an efficient drug carrier and bone substitute to control MRSA infection and simultaneously reconstruct the bone cavity in a sheep model.This work was financed by FEDER-Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020-Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020; by Portuguese funds through FCT/MCTES in the framework of the project institute for Research and Innovation in Health Sciences (POCI-01-0145-FEDER-007274); by the Project Biotherapies (NORTE-01-0145-FEDER-000012); and by the project HEPHAPC Program RESOLVE, Norte 2020 (NORTE-01-0246-FEDER-000018). The authors would also like to acknowledge the technical support for histology and histochemical studies of Rui Fernandes and Rossana Correia and all the staff from HEMS/i3S, as well as the support of all the staff and students at the Hospital Veterinario-Universidade de Evora

Regular Strength and Sprint Training Counteracts Bone Aging: A 10-Year Follow-Up in Male Masters Athletes

Cross-sectional and interventional studies suggest that high-intensity strength and impact-type training provide a powerful osteogenic stimulus even in old age. However, longitudinal evidence on the ability of high-intensity training to attenuate age-related bone deterioration is currently lacking. This follow-up study assessed the role of continued strength and sprint training on bone aging in 40- to 85-year-old male sprinters (n = 69) with a long-term training background. Peripheral quantitative computed tomography (pQCT)-derived bone structural, strength, and densitometric parameters of the distal tibia and tibia midshaft were assessed at baseline and 10 years later. The groups of well-trained (actively competing, sprint training including strength training ≥2 times/week; n = 36) and less-trained (<2 times/week, no strength training, switched to endurance training; n = 33) athletes were formed according to self-reports at follow-up. Longitudinal changes in bone traits in the two groups were examined using linear mixed models. Over the 10-year period, group-by-time interactions were found for distal tibia total bone mineral content (BMC), trabecular volumetric bone mineral density (vBMD), and compressive strength index, and for mid-tibia cortical cross-sectional area, medullary area, total BMC, and BMC at the anterior and posterior sites (polar mass distribution analysis) (p < 0.05). These interactions reflected maintained (distal tibia) or improved (mid-tibia) bone properties in the well-trained and decreased bone properties in the less-trained athletes over the 10-year period. Depending on the bone variable, the difference in change in favor of the well-trained group ranged from 2% to 5%. The greatest differences were found in distal tibia trabecular vBMD and mid-tibia posterior BMC, which remained significant (p < 0.05) after adjustment for multiple testing. In conclusion, our longitudinal findings indicate that continued strength and sprint training is associated with maintained or even improved tibial properties in middle-aged and older male sprint athletes, suggesting that regular, intensive exercise counteracts bone aging. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research

Supercritical fluid technology as a tool to prepare gradient multifunctional architectures towards regeneration of osteochondral injuries

Platelet lysates (PLs) are a natural source of growth factors (GFs) known for its stimulatory role on stem cells which can be obtained after activation of platelets from blood plasma. The possibility to use PLs as growth factor source for tissue healing and regeneration has been pursued following different strategies. Platelet lysates are an enriched pool of growth factors which can be used as either a GFs source or as a three-dimensional (3D) hydrogel. However, most of current PLs-based hydrogels lack stability, exhibiting significant shrinking behavior. This chapter focuses on the application of supercritical fluid technology to develop three-dimensional architectures of PL constructs, crosslinked with genipin. The proposed technology allows in a single step operation the development of mechanically stable porous structures, through chemical crosslinking of the growth factors present in the PL pool, followed by supercritical drying of the samples. Furthermore gradient structures of PL-based structures with bioactive glass are also presented and are described as an interesting approach to the treatment of osteochondral defects.info:eu-repo/semantics/publishedVersio

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.FROnTHERA (NORTE-01-0145- FEDER-000023), supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). The authors would also like to acknowledge H2020-MSCA-RISE program, as this work is part of developments carried out in BAMOS project, funded by the European Union’s Horizon 2020 research and innovation program under grant agreement N° 734156. The financial support from the Portuguese Foundation for Science and Technology under the program Investigador FCT 2012 and 2015 (IF/00423/2012 and IF/01285/2015)info:eu-repo/semantics/publishedVersio

Functionally graded additive manufacturing to achieve functionality specifications of osteochondral scaffolds

Osteoarthritis (OA) is a degenerative joint disease, characterized by cartilage loss and changes in bone at the interface of a joint resulting in pain, stiffness and reduced mobility. OA is one of the most prevalent chronic conditions as identified in Bone and Joint Decade. According to the World Health Organization, 40% of people over the age of 70 have OA. This joint disease affects around 0.4 billion people with patients in Europe accounting for up to 30%. The figure is set to increase with the ageing problem. Current non-surgical treatments for OA involve non-steroidal anti-inflammatory drug administration. Surgical treatments include osteotomy, abrasion arthroplasty, micro-fracture and autologous chondrocyte implantation (ACI). This is a two-stage surgical procedure with the associate costs and infection being the main concern. For small osteochondral defects, micro-fracture (MF) marrow stimulation and for large cartilaginous defects the autologous chondrocyte implantation are considered as necessary treatments. However, MF produces fibrocartilage not native hyaline cartilage. For defects that have progressed to a stage that affects the subchondral bone, other treatments are no longer effective and joint replacement operation is the only alternative. The demand for innovative therapeutic alternatives for complete healing of OA is significant. The treatment of cartilage and osteochondral (OC) defects remains a challenge since treatments so far have failed to achieve complete restoration of the properties of joint cartilage. Many new technologies, such as osteochondral tissue engineering, have been studied and applied to repair osteochondral defects. Commercially available osteochondral scaffolds have been used in patients with OC defects. However, no products have so far demonstrated to provide biomechanical properties suitable to promote the durable regeneration of large OC defects [1]. The main issue with these commercially available OC scaffolds is poor cartilage fill associated with fibrocartilage formation. The aim of this paper is to define the functionality and performance which would be required for intended clinical applications in the treatment of osteoarthritis and also to show that the capabilities of 3D bioprinting and functionally graded additive manufacturing scaffolds are suitable to meet most of these requirements.The authors would like to thank H2020-MSCARISE programme, as this work is part of developments carried out in BAMOS project, funded from the European Union’s Horizon 2020 research and innovation programme under Grant Agreement No. 734156.info:eu-repo/semantics/publishedVersio

The stiffness of living tissues and its implications for tissue engineering

The past 20 years have witnessed ever- growing evidence that the mechanical properties of biological tissues, from nanoscale to macroscale dimensions, are fundamental for cellular behaviour and consequent tissue functionality. This knowledge, combined with previously known biochemical cues, has greatly advanced the field of biomaterial development, tissue engineering and regenerative medicine. It is now established that approaches to engineer biological tissues must integrate and approximate the mechanics, both static and dynamic, of native tissues. Nevertheless, the literature on the mechanical properties of biological tissues differs greatly in methodology, and the available data are widely dispersed. This Review gathers together the most important data on the stiffness of living tissues and discusses the intricacies of tissue stiffness from a materials perspective, highlighting the main challenges associated with engineering lifelike tissues and proposing a unified view of this as yet unreported topic. Emerging advances that might pave the way for the next decadeâ s take on bioengineered tissue stiffness are also presented, and differences and similarities between tissues in health and disease are discussed, along with various techniques for characterizing tissue stiffness at various dimensions from individual cells to organs.The authors would like to acknowledge financial support from the European Research Council, grant agreement ERC-2012-ADG 20120216-321266 (project ComplexiTE). C.F.G. acknowledges scholarship grant no. PD/BD/135253/2017 from Fundação para a Ciência e Tecnologia (FCT). The authors also thank the peer-reviewers for the constructive comments and suggestions that helped to shape this manuscript

Proceedings of the Frontiers of Retrovirology Conference 2016

The emergence of pandemic retroviral infection in small ruminant