Evaluation of the analytical performance of the PC100 platelet counter.

INTRODUCTION: Platelet count can be altered in various diseases and treatments and measuring it may provide better insight into the expected outcome. So far, quantification of platelet count is done within laboratory conditions by using established hematology analyzers, whereas a point-of-care device could be used for this purpose outside of the clinical laboratories. AIM: Our aim was to assess the closeness of agreement between a newly developed point-of-care PC100 platelet counter and two reference methods (Sysmex® XP-300, Sysmex® XN-9000) in measuring platelet counts in whole blood and platelet-rich-plasma (PRP). METHOD: Whole blood was obtained from 119 individuals, of which 74 were used to prepare PRP samples. Whole blood platelet count was measured by the two reference methods and the PC100 platelet counter. PRP was prepared from the whole blood and platelet count was adjusted to the range of 250-3600 × 103/µl and measured with the PC100 platelet counter and Sysmex® XP-300. RESULTS: A median difference of - 1.35% and - 2.98% occurred in whole blood platelet count between the PC100 platelet counter and the Sysmex® XP-300 and Sysmex® XN-9000, respectively. A strong linear correlation (r ≥ 0.98) was seen in both cases and regression equations indicated neither a constant nor a proportional bias between the methods. Direct comparison of the two reference methods revealed a median difference of - 1.15% and a strongly linear relationship (r = 0.99). Platelet count in PRP resulted in a median difference of 1.42% between the PC100 platelet counter and the reference method, Sysmex® XP-300. While the difference between two methods increased with concentration of platelets in PRP, a strong linear relationship remained throughout the whole measuring interval indicated by the high correlation coefficient (r = 0.99). Assessment of the predicted bias at predefined platelet counts showed that the bias in platelet counts falls within the acceptance criterion for both whole blood and PRP measurements. CONCLUSIONS: Our results show that the PC100 platelet counter can be used interchangeably with the reference methods for determining platelet counts.

Alteration of structural and mechanical properties of the temporomandibular joint disc following elastase digestion.

The temporomandibular joint disc is a fibrocartilaginous structure, composed of collagen fibers, elastin fibers, and proteoglycans. Despite the crucial role of elastin fibers in load-bearing properties of connective tissues, its contribution in temporomandibular joint disc biomechanics has been disregarded. This study attempts to characterize the structural-functional contribution of elastin in the temporomandibular joint disc. Using elastase, we selectively perturbed the elastin fiber network in porcine temporomandibular joint discs and investigated the structural, compositional, and mechanical regional changes through: (a) analysis of collagen and elastin fibers by immunolabeling and transmission electron microscopy; (b) quantitative analysis of collagen tortuosity, cell shape, and disc volume; (c) biochemical quantification of collagen, glycosaminoglycan and elastin content; and (d) cyclic compression test. Following elastase treatment, microscopic examination revealed fragmentation of elastin fibers across the temporomandibular joint disc, with a more pronounced effect in the intermediate regions. Also, biochemical analyses of the intermediate regions showed significant depletion of elastin (50%), and substantial decrease in collagen (20%) and glycosaminoglycan (49%) content, likely due to non-specific activity of elastase. Degradation of elastin fibers affected the homeostatic configuration of the disc, reflected in its significant volume enlargement accompanied by remarkable reduction of collagen tortuosity and cell elongation. Mechanically, elastase treatment nearly doubled the maximal energy dissipation across the intermediate regions while the instantaneous modulus was not significantly affected. We conclude that elastin fibers contribute to the restoration and maintenance of the disc resting shape and actively interact with collagen fibers to provide mechanical resilience to the temporomandibular joint disc.

The dynamic mechanical viscoelastic properties of the temporomandibular joint disc: The role of collagen and elastin fibers from a perspective of polymer dynamics.

The temporomandibular joint disc is a structure, characterized as heterogeneous fibrocartilage, and is composed of macromolecular biopolymers. Despite a large body of characterization studies, the contribution of matrix biopolymers on the dynamic viscoelastic behavior of the disc is poorly understood. Given the high permeability and low concentration of glycosaminoglycans in the disc, it has been suggested that poro-elastic behavior can be neglected and that the intrinsic viscoelastic nature of solid matrix plays a dominant role in governing its time-dependent behavior. This study attempts to quantify the contribution of collagen and elastin fibers to the viscoelastic properties of the disc. Using collagenase and elastase, we perturbed the collagen and elastin fibrillar network in porcine temporomandibular joint discs and investigated the changes of dynamic viscoelastic properties in five different regions of the disc. Following both treatments, the storage and loss moduli of these regions were reduced dramatically up to the point that the tissue was no longer mechanically heterogeneous. However, the proportion of changes in storage and loss moduli were different for each treatment, reflected in the decrease and increase of the loss tangent for collagenase and elastase treated discs, respectively. The reduction of storage and loss moduli of the disc correlated with a decrease of biopolymer length. The present study indicates that the compositional and structural changes of collagen and elastin fibers alter the viscoelastic properties of the disc consistent with polymer dynamics.

Diffusion of charged and uncharged contrast agents in equine mandibular condylar cartilage is not affected by an increased level of sugar-induced collagen crosslinking.

Nutrition of articular cartilage relies mainly on diffusion and convection of solutes through the interstitial fluid due to the lack of blood vessels. The diffusion is controlled by two factors: steric hindrance and electrostatic interactions between the solutes and the matrix components. Aging comes with changes in the cartilage structure and composition, which can influence the diffusion. In this study, we treated fibrocartilage of mandibular condyle with ribose to induce an aging-like effect by accumulating collagen crosslinks. The effect of steric hindrance or electrostatic forces on the diffusion was analyzed using either charged (Hexabrix) or uncharged (Visipaque) contrast agents. Osteochondral plugs from young equine mandibular condyles were treated with 500 mM ribose for 7 days. The effect of crosslinking on mechanical properties was then evaluated via dynamic indentation. Thereafter, the samples were exposed to contrast agents and imaged using contrast-enhanced computed tomography (CECT) at 18 different time points up to 48 h to measure their diffusion. Normalized concentration of contrast agents in the cartilage and contrast agent diffusion flux, as well as the content of crosslink level (pentosidine), water, collagen, and glycosaminoglycan (GAG) were determined. Ribose treatment significantly increased the pentosidine level (from 0.01 to 7.6 mmol/mol collagen), which resulted in an increase in tissue stiffness (~1.5 fold). Interestingly, the normalized concentration and diffusion flux did not change after the induction of an increased level of pentosidine either for Hexabrix or Visipaque. The results of this study strongly suggest that sugar-induced collagen crosslinking in TMJ condylar cartilage does not affect the diffusion properties.