Investigating ice nucleation temperature effects on mesenchymal stem cell recovery from cryostorage [Abstract]

Investigating ice nucleation temperature effects on mesenchymal stem cell recovery from cryostorage [Abstract

The use of PRP injections in the management of knee osteoarthritis

Osteoarthritis (OA) is a degenerative disease involving joint damage, an inadequate healing response and progressive deterioration of the joint architecture that commonly affects the knee and/or hip joints. It is a major world public health problem and is predicted to increase rapidly with an ageing population and escalating rate of obesity. Autologous blood-derived products possess much promise in the repair and regeneration of tissue and have important roles in inflammation, angiogenesis, cell migration and metabolism in pathological conditions, including OA. Utilising platelet-rich plasma (PRP) to treat tendon, ligament and skeletal muscle has shown variable results across many studies with the current evidence base for the efficacy of PRP in treating sports injuries remaining inconclusive. More uniformly positive results have been observed by various studies for PRP in OA knee in comparison to hyaluronic acid, other intra-articular injections and placebo than in other musculoskeletal tissue. However, methodological concerns as well as satisfactory PRP product classification prevent the true characterisation of this treatment. Thus, further research is required to investigate how leukocyte inclusion, activation and platelet concentration affect therapeutic efficacy. Furthermore, the optimisation of timing, dosage, volume, frequency and rehabilitation strategies need to be ascertained. For knee OA management, these concerns must be addressed before this promising treatment can be widely implemented

A critical review of current progress in 3D kidney biomanufacturing: advances, challenges, and recommendations

The widening gap between organ availability and need is resulting in a worldwide crisis, particularly concerning kidney transplantation. Regenerative medicine options are becoming increasingly advanced and are taking advantage of progress in novel manufacturing techniques, including 3D bioprinting, to deliver potentially viable alternatives. Cell-integrated and wearable artificial kidneys aim to create convenient and efficient systems of filtration and restore elements of immunoregulatory function. Whilst preliminary clinical trials demonstrated promise, manufacturing and trial design issues and identification of suitable and sustainable cell sources have shown that more development is required for market progression. Tissue engineering and advances in biomanufacturing techniques offer potential solutions for organ shortages; however, due to the complex kidney structure, previous attempts have fallen short. With the recent development and progression of 3D bioprinting, cell positioning and resolution of material deposition in organ manufacture have never seen greater control. Cell sources for constructing kidney building blocks and populating both biologic and artificial scaffolds and matrices have been identified, but in vitro culturing and/or differentiation, in addition to maintaining phenotype and viability during and after lengthy and immature manufacturing processes, presents additional problems. For all techniques, significant process barriers, clinical pathway identification for translation of models to humans, scaffold material availability, and long-term biocompatibility need to be addressed prior to clinical realisation

Small-scale skeletal muscle constructs for in-vitro musculoskeletal junction preclinical testbed [Abstract]

Small-scale skeletal muscle constructs for in-vitro musculoskeletal junction preclinical testbed [Abstract

Preclinical musculoskeletal junction testbed: Optimisation of a reproducible skeletal muscle construct [Abstract]

Preclinical musculoskeletal junction testbed: Optimisation of a reproducible skeletal muscle construct [Abstract

Preclinical musculoskeletal junction testbed: co-culture of 3D tissue engineered skeletal muscle and bone constructs [Abstract]

Preclinical musculoskeletal junction testbed: co-culture of 3D tissue engineered skeletal muscle and bone constructs [Abstract

Fabrication and characterisation of 3D complex hydroxyapatite scaffolds with hierarchical porosity of different features for optimal bioactive performance

To improve the biological performance of hydroxyapatite scaffolds in bone tissue engineering, graphite was used as porogen to create additional micro/nanoporosity to macroporosity, resulting in hierarchical porosity. For maximum imitation of natural bone structures, scaffolds with different porosity features were fabricated using micron/nano-sized graphite. The sintering profile of graphite treated scaffolds was optimised to reduce the influence of shrinkage. To confirm the porosity features, the micro/nanostructures of scaffolds were characterised by scanning electron microscopy and Brunauer-Emmett-Teller method. Considering that hydroxyapatite is resistant to biodegradation in vivo, the degradation rate of scaffolds in modified simulated body fluid was examined. Furthermore, biological evaluations based on myoblasts were carried out to investigate the influence of porosity features on the essential performance such as adhesion, proliferation and differentiation. The results indicate that the scaffolds with dominant microporosity and little nanoporosity formed inside had high potential for clinical applications due to improved performance in bioactivity

Development of a 3D tissue-engineered skeletal muscle and bone co‐culture system

In vitro three‐dimensional (3D) tissue engineered (TE) structures have been shown to better represent in vivo tissue morphology and biochemical pathways than monolayer culture, and are less ethically questionable than animal models. However, to create systems with even greater relevance, multiple integrated tissue systems should be recreated in vitro. In the present study, the effects and conditions most suitable for the co‐culture of TE skeletal muscle and bone were investigated. High‐glucose Dulbecco's Modified Eagle Medium (HG‐DMEM) supplemented with 20% foetal bovine serum (FBS) followed by HG‐DMEM with 2% horse serum was found to enable proliferation of both C2C12 muscle precursor cells and TE85 human osteosarcoma cells, fusion of C2C12s into myotubes, as well as an up‐regulation of RUNX2/CBFa1 in TE85s. Myotube formation was also evident within indirect contact monolayer cultures. Finally, in 3D co‐cultures, TE85 collagen/hydroxyapatite constructs had significantly greater expression of RUNX2/CBFa1 and osteocalcin/BGLAP in the presence of collagen‐based C2C12 skeletal muscle constructs; however, fusion within these constructs appeared reduced. This work demonstrates the first report of the simultaneous co‐culture and differentiation of 3D TE skeletal muscle and bone, and represents a significant step towards a full in vitro 3D musculoskeletal junction model

Development of tissue-engineered skeletal muscle manufacturing variables

Three-dimensional tissue-engineered structures enable more representative determination of novel drug or material effects on tissue than traditional monolayer cell cultures. This study sought to better understand how key manufacturing variables affect the myotube characteristics of a skeletal muscle model toward reducing resource use and to develop an understanding of scaling on model consistency. C2C12 murine myoblasts were seeded in a tethered collagen scaffold from which directional myotubes form in response to lines of tension and a change in medium. Collagen polymerizing area length-to-width ratios greater than one were found to reduced cell–matrix attachment and remodeling forces significantly (p <.05) correlating to a reduction in cell fusion potential. Following this, utilizing a factorial design of experiment, 4 million C2C12s/ml, with a polymerizing area width 150% of the anchor point, produced the most favorable myotube characteristics and dramatically reduced the incidence of rupture. Scaled constructs showed no significant differences when compared to larger models. Approximately 20 myotubes with a variation in the alignment of <25° in the central region were consistently observed in the final models. This demonstrates the influence of initial manufacturing variables on tissue formation and has produced a benchmark model for consistent production across scaled constructs for future optimization and as a potential cost-effective preclinical testbed