Increased chemotaxis and activity of circulatory myeloid progenitor cells may contribute to enhanced osteoclastogenesis and bone loss in the C57BL/6 mouse model of collagen-induced arthritis.

Our study aimed to determine the functional activity of different osteoclast progenitor (OCP) subpopulations and signals important for their migration to bone lesions, causing local and systemic bone resorption during the course of collagen-induced arthritis in C57BL/6 mice. Arthritis was induced with chicken type II collagen (CII), and assessed by clinical scoring and detection of anti-CII antibodies. We observed decreased trabecular bone volume of axial and appendicular skeleton by histomorphometry and micro-computed tomography as well as decreased bone formation and increased bone resorption rate in arthritic mice in vivo. In the affected joints, bone loss was accompanied with severe osteitis and bone marrow hypercellularity, coinciding with the areas of active osteoclasts and bone erosions. Flow cytometry analysis showed increased frequency of putative OCP cells (CD3- B220- NK1.1- CD11b-/lo CD117+ CD115+ for bone marrow and CD3- B220- NK1.1- CD11b+ CD115+ Gr-1+ for peripheral haematopoietic tissues), which exhibited enhanced differentiation potential in vitro. Moreover, the total CD11b+ population was expanded in arthritic mice as well as CD11b+ F4/80+ macrophage, CD11b+ NK1.1+ natural killer cell and CD11b+ CD11c+ myeloid dendritic cell populations in both bone marrow and peripheral blood. In addition, arthritic mice had increased expression of tumour necrosis factor-α, interleukin-6, CC chemokine ligand-2 (Ccl2) and Ccl5, with increased migration and differentiation of circulatory OCPs in response to CCL2 and, particularly, CCL5 signals. Our study characterized the frequency and functional properties of OCPs under inflammatory conditions associated with arthritis, which may help to clarify crucial molecular signals provided by immune cells to mediate systemically enhanced osteoresorption.

Packing of collagen molecules modified with 2-propanol.

X-ray diffraction data of collagen molecules modified with 2-propanol favour a quasi-hexagonal lateral packing over a quasi-tetragonal one.

Structural dynamic of native tendon collagen.

The dynamic behaviour of collagen fibrils is revealed by time-resolved X-ray investigations of native rat tail tendon fibres in tensile tests.

Transformation of the structure of collagen. A time-resolved analysis of mechanochemical processes using synchrotron radiation.

Mechanochemically induced molecular transformations of collagen fibres were analysed using time-resolved small-angle diffraction spectra and histomechanical measurements. In particular, the influence of aqueous and methanolic perchlorate solutions was examined. According to a transformation continuing from the periphery towards the centre, the macroscopic contraction that is completed less than five minutes after incubation with perchlorate is caused by peripherally transformed fibrils only, whereas the centrally situated fibrils first undergo an accordion-like folding, but after more than 20 minutes are transformed similarly. The triple-helical transformation is preceded by a structure-breaking effect on structural water that can be monitored in time-resolved diffraction spectra. The combined loss of meridional low-angle reflections and cross-striated fibrils in micrographs is irreversible. By dialysis of colloidally dissolved collagen against a solution of ATP, however, segment-long spacing aggregates are obtained. Under isometric conditions, an instantaneous transformation of intermittent regions leads to an increase in the long period of adjacent, still structured regions of the same fibril that is correlated with a delayed increase in tension in the fibre. Increase of tension under isometric conditions as well as the flow-properties of a fibre relaxed in perchlorate are interpreted in terms of the parallel sliding of subunits of varying lengths, which has been demonstrated by diffraction analysis.

Stress-induced molecular rearrangement in tendon collagen.

Tension-induced molecular rearrangements in wet native fibres of rat-tail tendons and human finger flexor tendons are registered with the help of time-resolved diffraction spectra using synchrotron radiation. The tension-induced increase of the 67 nm D period is combined with changes in the intensities of some orders of the meridional small angle reflection. Both effects are reversible when unloading the fibre, but are preserved when the load is held constant until the fibre tears. The increase in the D period is partly due to a sliding of the triple helices relative to each other and partly due to a stretching of the triple helices themselves. The sliding of the triple helices results in an alteration of the D stagger, leading to a change in the length of the gap and overlap regions, and to a stretching of the cross-linked telopeptides. This interpretation is supported by comparison with the relative intensities derived from a model with varying length of gap and overlap regions, as well as by comparison with model calculations that include the telopeptides.

Twisted fibrils are a structural principle in the assembly of interstitial collagens, chordae tendineae included.

X-ray diffraction analysis of connective tissue samples, which contain type I and type III collagen shows that twisted collagen fibrils are a general principle of assembly. The occurrence of twisted fibrils in native wet Chordae tendineae, skin and Aorta is combined with a shorter axial periodicity of about 65 nm. This shorter D period is shown to be directly related to the tilt of the molecules, which have to be curved to build-up twisted fibrils.

The macromolecular structure of collagen in tendon fibres of dermatosparactic animals.

Small-angle x-ray diffraction spectra of dermatosparactic tendon collagen show a decreased intensity of the first order reflection. We interprete this finding to be due to the N-terminal propeptide which fills the intermolecular gap region partially.

Ehlers-Danlos syndrome type IV (EDS IV) as model of a defective biopolymer composite material.

The connective tissue of a lethal EDS IV case was investigated for the reasons of the manifested disturbances of the arterial wall. This functional disorder was attributed to the mechanical decoupling of elastin and collagen, with the premise of a composite material consisting of cellular, fibrillar, lamellar and other matrix components. A conceivable relation between the manifested deficiency of type III collagen and a disturbed anchoring of elastin is shown. These findings are supported by biochemical, morphological, x-ray and mechanical data.

New aspects of the etiology of tendon rupture. An analysis of time-resolved dynamic-mechanical measurements using synchrotron radiation.

Native collagen fibers were exposed to different dynamic loads to simulate damage to tendons and ligaments relevant clinically and for sports medicine. The results suggest that the rupture of a tendon is caused at the submicroscopic fibrillar level. Not only slow or very fast elongation, but also very fast unloading of stretched fibers seems to be responsible for disseminated damage, which reduces the stability of a fiber. This damage is induced by intrafibrillar sliding processes, which occur only a few seconds before macroscopic slippage takes place. The significance of these events for the beginning and progress of repair in vivo is discussed. The conclusions are supported by simultaneous mechanical and radiological measurements, as well as by light- and electron-microscopic results.