Elevated solute transport at sites of diffuse matrix damage in cortical bone: Implications on bone repair.

Diffuse matrix damage in rat cortical bone has been observed to self-repair efficiently in 2 weeks without activating bone remodeling, and unlike the case with linear cracks, the local osteocytes at the sites of diffuse damage remain healthy. However, the reason(s) for such high efficiency of matrix repair remains unclear. We hypothesized that transport of minerals and other compounds essential for damage repair is enhanced at the damaged sites and further increased by the application of tensile loading. To test our hypothesis, diffuse damage was introduced in notched bovine wafers under cyclic tensile loading and unloading. Using the Fluorescence Recovery After Photobleaching (FRAP) approach, we measured the transport of a small fluorescent tracer (sodium fluorescein, 376 Da) in damaged versus undamaged regions and under varying tensile load magnitudes (0.2 N, 10 N, 20 N, and 30 N), which corresponded to nominal strains of 12.5, 625, 1,250, and 1,875 microstrains, respectively. We found a 37% increase in transport of fluorescein in damaged regions relative to undamaged regions and a further ∼18% increase in transport under 20 N and 30 N tension compared to the non-loaded condition, possibly due to the opening of the cracking surfaces. The elevated transport of minerals and other adhesive proteins may, at least partially, account for the highly effective repair of diffuse damage observed in vivo. CLINICAL SIGNIFICANCE: Diffuse damage adversely affects bone's fracture resistance and this study provided quantitative data on elevated transport, which may be involved in repairing diffuse damage in vivo. 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:692-698, 2018.

Role of matrix behavior in compressive fracture of bovine cortical bone.

In compressive fracture of dry plexiform bone, we examine the individual roles of overall mean porosity, the connectivity of the porosity network, and the elastic as well as the failure properties of the nonporous matrix, using a random spring network model (RSNM). Porosity network structure is shown to reduce the compressive strength by up to 30%. However, the load-bearing capacity increases with an increase in either of the matrix properties-the elastic modulus or the failure strain threshold. To validate the porosity-based RSNM model with available experimental data, bone-specific failure strain thresholds for the ideal matrix of similar elastic properties were estimated to be within 60% of each other. Further, we observe the avalanche size exponents to be independent of the bone-dependent parameters as well as the structure of the porosity network.

Avaliação morfológica e por imagem radiográfica da matriz óssea mineralizada heteróloga fragmentada e metilmetacrilato, preservados em glicerina para reparação de falhas ósseas em tíbias de coelhos / Morphological evaluation and radiographic image of fragmented heterolog mineralized bone matrix and methylmethacrylate preserved in glycerin to repair osseous defects in tibias of rabbits

Os traumas que resultam em fraturas ósseas, principalmente as cominutivas, têm uma importância muito grande na rotina clínico-cirúrgica veterinária e humana. Foi realizada falha segmentar de 6mm na região metafisária medial da tíbia esquerda de 12 coelhos, a qual foi preenchida com implante constituído de matriz óssea mineralizada heteróloga fragmentada e metilmetacrilato, preservados em glicerina (98%) para a sua reconstrução. Foi realizada avaliação morfológica e radiológica aos 30, 60, 90 e 120 dias e observou-se a incorporação do implante ao leito receptor, em 100% dos casos, mostrando ser biologicamente compatível, pois promoveu a reparação das falhas ósseas, sem sinais de infecção, migração e/ou rejeição, sendo uma opção de substituto para preencher defeitos ósseos.

The traumas that result in bone fractures, especially comminuted, have high importance in veterinary and human surgical routine. A 6mm of segmental defect was performed at the medial metaphyseal region of the left tibia of 12 rabbits and an association of fragmented heterologue mineralized bone matrix and methylmethacrylate conserved in glycerin (98%) was used as a graft to fill the bone defect. To evaluate the procedure morphological and radiological exams were performed after 30, 60, 90 and 120 days. There was gradual integration of the bone graft in the receptor bed in 100% of the cases showing that the material is biologically compatible as it promotes bone defect reparation without signs of infection, migration and/or rejection and can be considered one more option to be used as a substitute to fill bone defects.

[A comparative study on effect of different defect diameters on healing in middle 1/3 tibia monolayer cortical bone defect mouse model].

OBJECTIVE: To compare the effect of different defect diameters on healing in the middle 1/3 tibia monolayer cortical bone defect mouse model so as to establish an animal model for bone tissue engineering study, mechanism study on bone defect repair, and gene therapy research. METHODS: Ten 8-week-old C57BL/6J mice, weighing (20 +/- 2) g, were randomly divided into 2 groups, 5 mice in each group. The middle 1/3 tibiae monolayer cortical bone defect model of 0.8 mm (group A) or 1.0 mm (group B) in diameter was established with burr drill. At 7, 21, and 28 days after modeling, the molybdenum target X-ray radiography was used to observe the defect repair; at 28 days, Micro CT and three-dimensional imaging were used to evaluate bone defect repair, and tibia specimens were harvested for HE staining. RESULTS: At 7 days after modeling, tibia fracture occurred in 5 mice in group B, no fracture in group A. X-ray films, Micro CT scan, and HE staining showed bony union in group A at 28 days. The quantitative analysis of trabecular bone by Micro CT showed that trabecular number, connectivity density, and bone volume in group A were significantly greater than those in group B (P < 0.05), mean of segmented region--mean 2 was significantly less than that in group B (P < 0.05), but no significant difference was found in trabecular separation and trabecular thickness between 2 groups (P > 0.05). CONCLUSION: The middle 1/3 tibia monolayer cortical bone defect mouse model of 0.8 mm in diameter is the ideal animal model for study repair mechanism of tibia defect or bone tissue engineering.

The low level laser therapy effect on the remodeling of bone extracellular matrix.

The low level laser therapy (LLLT) has been used as an option to accelerate the regeneration of bone tissue. In this study, both femurs of male Wistar rats (30 animals) were injured with a drill and the effect of LLLT using a laser diode (100 mW at 660 nm) in the bone matrix on the left paw measured. LLLT effect on the healing bone tissue matrix was evaluated by a combination of immunohistochemical histomorphometry, confocal immunofluorescence microscopy and isolation and characterization of glycosaminoglycans. Histomorphometric analysis showed that LLLT increased bone matrix and showing more organized. Alcian Blue and PAS staining seems to suggest differential glycosaminoglycans and glycoproteins. The data showed increased expression of chondroitin sulfate and hyaluronic acid, after reduction as the LLLT and mature bone, resembling the expression of osteonectin and biglycan. The difference in expression of siblings (DMP-1, OPN and BSP) is in accordance with the repair accelerated bone formation after the application of LLLT as compared with control. The expression of osteonectin and osteocalcin supports their role in bone mineralization protein, indicating that LLLT accelerates this process. The overall data show that LLLT bone changes dynamic array, shortening the time period involved in the bone repair.

Effect of fatigue loading and associated matrix microdamage on bone blood flow and interstitial fluid flow.

Functional adaptation of bone to cyclic fatigue involves a complex physiological response that is targeted to sites of microdamage. The mechanisms that regulate this process are not understood, although lacunocanalicular interstitial fluid flow is likely important. We investigated the effect of a single period of cyclic fatigue on bone blood flow and interstitial fluid flow. The ulnae of 69 rats were subjected to cyclic fatigue unilaterally using an initial peak strain of -6000 muepsilon until 40% loss of stiffness developed. Groups of rats (n=23 per group) were euthanized immediately after loading, at 5 days, and at 14 days. The contralateral ulna served as a treatment control, and a baseline control group (n=23) that was not loaded was also included. After euthanasia, localization of intravascular gold microspheres within the ulna (n=7 rats/group) and tissue distribution of procion red tracer were quantified (n=8 rats/group). Microcracking, modeling, and remodeling (Cr.S.Dn, microm/mm(2), Ne.Wo.B.T.Ar, mm(2), and Rs.N/T.Ar, #/mm(2) respectively) were also quantified histologically (n=8 rats/group). Cyclic fatigue loading induced hyperemia of the loaded ulna, which peaked at 5 days after loading. There was an associated overall decrease in procion tracer uptake in both the loaded and contralateral control ulnae. Tracer uptake was also decreased in the periosteal region, when compared with the endosteal region of the cortex. Pooling of tracer was seen in microdamaged bone typically adjacent to an intracortical stress fracture at all time points after fatigue loading; in adjacent bone tracer uptake was decreased. New bone formation was similar at 5 days and at 14 days, whereas formation of resorption spaces was increased at 14 days. These data suggest that a short period of cyclic fatigue induces bone hyperemia and associated decreased lacunocanalicular interstitial fluid flow, which persists over the time period in which osteoclasts are recruited to sites of microdamage for targeted remodeling. Matrix damage and development of stress fracture also interfere with normal centrifugal fluid flow through the cortex. Changes in interstitial fluid flow in the contralateral ulna suggest that functional adaptation to unilateral fatigue loading may include a more generalized neurovascular response.

Emdogain: últimos avances en regeneración periodontal / Emdogain: an update in periodontal regeneration

Emdogain es un compuesto de proteínas derivadas de la matriz del esmalte, capaz de inducir la regeneración verdadera del aparato de inserción. Como principal indicación destaca el tratamiento de defectos infraóseos, ganancia de hueso y reducción de la profundidad de sondaje con mínima recesión gingival. Es un procedimiento técnicamente simple, con poco riesgo y menos invasivo que las técnicas de regeneración convencionales. La cuidada selección del paciente, el empleo de una técnica adecuada así como el riguroso control postoperatorio son factores importantes para el éxito del tratamiento (AU)

Emdogain is a compound of proteins derived from the enamel matrix which are a crucial factor in initiating the formation of a cellular root cementum and stimulate the development of the periodontal ligament and alveolar bone. The main indication for the application of EMD is the intrabony defects treatment with significant clinical attachment level gains, probing depht reductions and minimal gingival recession. The application of EMD is a simple procedure with less risk than other techniques and less invasive than conventional guided tissue regeneration. A careful selection of the patient, the use of an adecuate technique and the strict postoperatory control are all very important factors for the treatment success (AU)

Cell death after cartilage impact occurs around matrix cracks.

The damage from rapid high energy impacts to cartilage may contribute to the development of osteoarthritis (OA). Understanding how and when cells are damaged during and after the impact may provide insight into how these lesions progress. Mature bovine articular cartilage on the intact patella was impacted with a flat impacter to 53 MPa in 250 ms. Cell viability was determined by culturing the cartilage with nitroblue tetrazolium for 18 h or for 4 days in medium containing 5% serum before labeling (5-day sample) and compared to adjacent, non-impacted tissue as viable cells per area. There was a decrease in viable cell density only in specimens with macroscopic cracks and the loss was localized primarily near matrix cracks, which were in the upper 25% of the tissue. This was confirmed using confocal microscopy with a fluorescent live/dead assay, using 5'-chloromethylfluorescein diacetate and propidium iodide. Cell viability in the impacted regions distant from visible cracks was no different than the non-impacted control. At 5 days, viable cell density decreased in the surface layer in both the control and impacted tissue, but there was no additional impact-related change. In summary, cell death after the impaction of cartilage on bone occurred around impact induced cracks, but not in impacted areas without cracks. If true in vivo, early stabilization of the damaged area may prevent late sequelae that lead to OA.

Matrix and cell injury due to sub-impact loading of adult bovine articular cartilage explants: effects of strain rate and peak stress.

Mechanical overloading of cartilage has been implicated in the initiation and progression of osteoarthrosis. Our objectives were to identify threshold levels of strain rate and peak stress at which sub-impact loads could induce cartilage matrix damage and chondrocyte injury in bovine osteochondral explants and to explore relationships between matrix damage, spatial patterns of cell injury, and applied loads. Single sub-impact loads characterized by a constant strain rate between 3 x 10(-5) and 0.7 s(-1) to a peak stress between 3.5 and 14 MPa were applied, after which explants were maintained in culture for four days. At the higher strain rates, matrix mechanical failure (tissue cracks) and cell deactivation were most severe near the cartilage superficial zone and were associated with sustained increased release of proteoglycan from explants. In contrast, low strain rate loading was associated with cell deactivation in the absence of visible matrix damage. Furthermore, cell activity and proteoglycan synthesis were suppressed throughout the cartilage depth, but in a radially dependent manner with the most severe effects at the center of cylindrical explants. Results highlight spatial patterns of matrix damage and cell injury which depend upon the nature of injurious loading applied. These patterns of injury may also differ in terms of their long-term implications for progression of degradative disease and possibilities for cartilage repair.

Aging and matrix microdamage accumulation in human compact bone.

Bone matrix microdamage in bone matrix, evidenced as microcracks, occurs consequent to cyclic loading. Microdamage caused by in vivo loading has been described in human rib cortex; however, the existence and extent of microcracks in human long bone cortices are largely unknown. Using histomorphometric methods to examine the incidence and localization of microcracks in human femoral compact bone specimens, we found that the amount of microdamage present in femoral compact bone increases dramatically with increasing age. Least squares regression analysis showed that in males, microcrack density (Cr.De., #/mm2) increases exponentially with age (r2 = 0.70). In females, Cr.De. also increases as an exponential function of increasing age (r2 = 0.79), at a significantly higher rate than in male specimens (p < 0.001). The current studies indicate that with increasing age, bone microdamage accumulates more rapidly than intrinsic processes can effect its repair. A combination of cumulative loading history, focal changes in material properties and alteration in the ability of the tissue to perceive and/or react to microcracks may all play role in this accumulation of bone microdamage with aging. This accumulation of microdamage in bone will contribute to decreased strength and stiffness. In addition, and perhaps most significantly for understanding aging and increased bone fragility, matrix microdamage in composite materials like bone will result in a profoundly reduced resistance to fracture. The importance of this accumulation of matrix microdamage in human bone with increasing age in contributing to the increased fragility of the aging skeleton is discussed.