Systematic investigation of oxygen and growth factors in clinically valid ex vivo expansion of cord blood CD34+ hematopoietic progenitor cells

Background aims. Cord blood is considered to be a superior source of hematopoietic stem and progenitor cells for transplantation, but clinical use is limited primarily because of the low numbers of cells harvested. Ex vivo expansion has the potential to provide a safe, effective means of increasing cell numbers. However, an absence of consensus regarding optimum expansion conditions prevents standard implementation. Many studies lack clinical applicability, or have failed to investigate the combinational effects of different parameters.

Methods. This is the first study to characterize systematically the effect of growth factor combinations across multiple oxygen levels on the ex vivo expansion of cord blood CD34 hematopoietic cells utilizing clinically approvable reagents and methodologies throughout.

Results. Optimal fold expansion, as assessed both phenotypically and functionally, was greatest with thrombopoietin, stem cell factor, Flt-3 ligand and interleukin-6 at an oxygen level of 10%. With these conditions, serial expansion showed continual target population expansion and consistently higher expression levels of self-renewal associated genes.

Conclusions. This study has identified optimized fold expansion conditions, with the potential for direct clinical translation to increase transplantable cell dose and as a baseline methodology against which future factors can be tested.

X-ray velocimetry within the ex vivo carotid artery

X-ray velocimetry offers a non-invasive method by which blood flow, blood velocity and wall shear stress can be measured in arteries prone to atherosclerosis. Analytical tools for measuring haemodynamics in artificial arteries have previously been developed and here the first quantification of haemodynamics using X-ray velocimetry in a living mammalian artery under physiologically relevant conditions is demonstrated. Whole blood seeded with a clinically used ultrasound contrast agent was pumped with a steady flow through live carotid arterial tissue from a rat, which was kept alive in a physiological salt solution. Pharmacological agents were then used to produce vascular relaxation. Velocity measurements were acquired with a spatial resolution of 14 µm × 14 µm and at a rate of 5000 acquisitions per second. Subtle velocity changes that occur are readily measurable, demonstrating the ability of X-ray velocimetry to sensitively and accurately measure haemodynamics ex vivo. Future applications and possible limitations of the technique are discussed, which allows for detailed living tissue investigations to be carried out for various disease models, including atherosclerosis and diabetic vasculopathy.

Understanding in-vivo abrasion fatigue or common suture materials used in arthroscopic and open shoulder surgery

In orthopaedic surgery the reattachment of tendon to bone requires suture materials that have stable and durable properties to allow healing at the tendon-bone interface. Failure rates of this type of surgery can be as high as 25%. While the tissue suture interface is a weak link, proportions of these failures are caused by in-vivo abrasion of the suture with bone and suture anchor materials. Abrasion of the suture material results from the movement of the suture through the eyelet by the surgeon during surgery, or with limb movement after surgery as the suture is not rigidly restrained within the eyelet. During movement the suture is subjected to bending and frictional forces that can lead to fatigue induced failure. This paper investigates the mechanism of bending abrasion fatigue induced failure of number two grade braided sheath only and braided sheath/multifilament core sutures. Sutures were oscillated over a stainless steel wire at low frequency under load in a dry state to simulate the bending and frictional forces between suture and eyelet. Failure mechanism was determined by video microscopy of the suture during abrasion combined with optical microscopy analysis of partially and fully abraded sutures. Braided only structures had high friction loading on the small number of fibres at the abrasion interface. This caused rapid single fibre breakages that accumulate to cause suture failure. The addition of ultra-high molecular weight polyethylene core fibres to a braided suture distributed the applied load across multiple fibres at the abrasion interface. This improved abrasion resistance by 15-20 times that of braided sheath alone.