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Chicken eggshell (ES) waste is a rich source of calcium carbonate (CaCO 3 ); however, the potential of ES as dietary calcium (Ca) in old laying hens has not been explored. This study compared the effects of feeding limestone, cockle shell, oyster shell, fine ES, and coarse ES as the sole Ca source on production performance, egg quality, blood biochemical constituents, and tibia characteristics in old laying hens. A total of 450 ISA-Brown laying hens at 73 wk of age with similar egg production rate (EPR) were randomly assigned to 5 treatment groups (90 hens/group, 9 hens/replicate) for 7 wk. Dietary treatment groups comprised a corn-soybean meal based diet containing different Ca sources: (i) limestone (LS; < 2 mm and 2–4 mm mixed in the ratio of 3:7) as control, (ii) cockle shell (CS; 1–4 mm), (iii) oyster shell (OS; 3–16 mm), (iv) ES fine particles (ESF; < 1 mm), and (v) ES coarse particles (ESC; 3–5 mm). Results indicated that dietary inclusion of coarse ES particles significantly increased average egg weight ( P  < 0.001) and daily egg mass ( P  < 0.05), and decreased feed conversion ratio ( P  < 0.001) as compared with the other treatments. However, no significant differences in EPR, feed intake, cracked egg proportion, and mortality were observed among the dietary treatments ( P  > 0.05). Notably, the use of ESF led to a lower proportion of cracked eggs than ESC ( P  < 0.05). ESC fed hens produced the heaviest eggs whereas CS fed hens produced the lightest ( P  < 0.001); the particle size of ES also affected the egg weight ( P  < 0.05). The eggs from OS and ESC fed hens showed a greater albumen height in comparison to eggs from CS group ( P  < 0.05); but no significant difference was observed among the LS, OS, ESF, and ESC groups (P > 0.05). The yolk color was darker in the eggs of group ESF as compared with other dietary groups ( P  < 0.01). However, no significant effects on Haugh units and shell properties were observed among the treatments ( P  > 0.05). The blood biochemistry results were not affected by the dietary Ca ( P  > 0.05) except for lower levels of high-density lipoprotein percentage (HDL %) in OS and ESC fed hens ( P  < 0.05). The tibia characteristics including weight, length, width, and breaking strength did not differ among the dietary groups ( P  > 0.05). However, the ESC and OS fed hens showed higher tibia bone mineral density (BMD) than the other groups ( P  < 0.001). In conclusion, coarse ES as a sole Ca source had beneficial effects on the production performance, egg quality, and tibia BMD in old laying hens.

The peripheral nervous system may play an important role in normal bone maintenance and remodeling. Substance P (SP) is a neuropeptide associated with bone loss and formation that may mediate the effects of the nervous system. The purpose of this study is to determine if treadmill running combined with electro-acupuncture at Jiaji acupoints (Jiaji-EA) affects tibial bone mass and SP expression in rabbits with sciatic nerve injury. Twenty-four juvenile male New Zealand white rabbits were randomly assigned to one of 4 groups: sham injury control (sham), sciatic never crush control (SNCr), treadmill running (treadmill), and Jiaji-EA combined with treadmill running (ET group). The SNCr, treadmill, and ET groups all had an induced sciatic never crush injury of approximately 2mm. Control groups received no intervention; the treadmill and ET groups were trained by treadmill; the ET group also received Jiaji-EA. After the 4 weeks of treatment, toe-spreading index (TSI), BMD, bone strength, and SP expression in the tibia were significantly lower in the nerve injury groups (SNCr, treadmill, and ET) compared to the sham groups (p<0.05). Treatment (treadmill and ET groups) increased all measures compared to the SNCr group (p<0.05). Further, TSI, BMD, bone strength, and SP expression in the ET group were higher than the treadmill group (p<0.05). Our results indicate that treadmill therapy combined with electro-acupuncture at Jiaji acupoints prevents bone loss in rabbit tibias after sciatic nerve injury. This may occur in two ways: indirectly in association with axon regeneration and directly via loading on the bone mediated through increased SP expression. This study provides important evidence for the clinical treatment of bone loss after peripheral nerve injury.

Purpose Platelet rich plasma (PRP) is derived from the patient’s own blood. The activated blood platelets release a cocktail of growth factors, some of which are thought to initiate and stimulate repair. We compared two groups to investigate whether the use of PRP mixed with bone chips improves bone healing in patients with a skeletal defect. Methods In total, 41 patients were observed. One group underwent a high tibial osteotomy with the addition of PRP and bone chips in the open wedge. The other group underwent the same procedure without the addition of PRP. Six patients had to be excluded because of insufficient data or they were lost to follow-up. Bone healing was studied using computed tomography scanning. The blood was sequestered and PRP was produced using a blood cell separator with a PRP software program (Electa, Sorin Group, Mirandola, Italy). Results Analysis focused on the remaining 35 patients. At baseline, there were no differences between the two groups for age, sex and side of operation. At one week postoperatively, the bone density under (p = 0.02) and above the wedge was significantly lower in the PRP group than the control group (p = 0.24). At six weeks postoperatively, no significant differences between the treatment groups were found. At 12 weeks, the PRP group had significantly lower bone density under the wedge compared to the control group (p = 0.01). Conclusions We found that patients with a skeletal defect did not benefit from the application of PRP mixed with an allograft regarding bone healing.

The bone marrow adjusts blood cell production to meet physiological demands in response to insults. The spatial organization of normal and stress responses are unknown owing to the lack of methods to visualize most steps of blood production. Here we develop strategies to image multipotent haematopoiesis, erythropoiesis and lymphopoiesis in mice. We combine these with imaging of myelopoiesis 1 to define the anatomy of normal and stress haematopoiesis. In the steady state, across the skeleton, single stem cells and multipotent progenitors distribute through the marrow enriched near megakaryocytes. Lineage-committed progenitors are recruited to blood vessels, where they contribute to lineage-specific microanatomical structures composed of progenitors and immature cells, which function as the production sites for each major blood lineage. This overall anatomy is resilient to insults, as it was maintained after haemorrhage, systemic bacterial infection and granulocyte colony-stimulating factor (G-CSF) treatment, and during ageing. Production sites enable haematopoietic plasticity as they differentially and selectively modulate their numbers and output in response to insults. We found that stress responses are variable across the skeleton: the tibia and the sternum respond in opposite ways to G-CSF, and the skull does not increase erythropoiesis after haemorrhage. Our studies enable in situ analyses of haematopoiesis, define the anatomy of normal and stress responses, identify discrete microanatomical production sites that confer plasticity to haematopoiesis, and uncover unprecedented heterogeneity of stress responses across the skeleton. This study develops a method for spatially resolving multipotent haematopoiesis, erythropoiesis and lymphopoiesis in mice and uncovers heterogeneous haematopoietic stress responses in different bones.

Little is known about tibial bone remodeling with TKA and its clinical relevance. We performed a randomized clinical trial to compare tibial bone density changes in cemented components with different bearing designs. Bone density changes were assessed using quantitative computed tomography (qCT)-assisted osteodensitometry. Twenty-eight rotating-platform and 26 fixed-platform cemented TKAs were included. The nonoperated contralateral side was used as a control. CT scans were performed postoperatively and 1 year and 2 years after the index operation. Cancellous bone density loss (up to 12.6% at 2 years) was observed in all proximal tibial regions in both cohorts. In contrast, we found lower cortical bone density loss (up to 3.6% at 2 years). We found no differences in bone loss between fixed- and rotating-platform implants. The decrease of cancellous bone density after TKA suggests stress transfer to the cortical bone.

Purpose Forty patients were enrolled in a prospective randomised study using conventional method or “Trabecular Metal Cone” (TM Cone) (Zimmer inc., Warsaw, USA) for reconstruction of bone loss of the proximal tibia during revision total knee arthroplasty (rTKA). The aim was to evaluate changes in bone mineral density (BMD) at the proximal tibia. Material and methods Thirty-six patients [median 67 years (range: 40–85 years)] received rTKA with NexGen® (Zimmer Warsaw, USA) revision system. Knee Society´s Knee Scoring System and the Anderson Orthopaedic Research Institute (AORI) bone classification was used. Changes in BMD were measured by dual energy X-ray absorptiometry (DEXA). Results Knee and function score improved in both groups. No significant changes between the groups were found. Changes in BMD within the two groups were quite similar. Overall decreases in BMD of 0.1 - 5.4 % were found in both groups (ROI 1–6) postoperative to 12 months of follow-up, except that ROI 7 showed an increase in BMD (0.8 - 1.3 %). After 24 months of follow-up, an increase in BMD was found along the stem (ROI 2–5) of 1.9 - 6.3 % , with significant changes in the TM Cone Group (ROI 3, 4, 5) . No significant changes in BMD between the groups were found. Conclusions The bone remodelling pattern was almost the same in the two groups after two years.

Vascularization is fundamental for bone formation and bone tissue homeostasis. However, in human subjects, a direct molecular relationship has not been identified between angiogenesis and agents that promote bone disease or factors related to age. Osteopenia is a condition in which bone mineral density is lower than normal, and it represents a sign of normal aging. Here we tested whether the type H vessel, which was recently identified as strongly positive for CD31 and Endomucin (CD31 hi Emcn hi ) in mice, is an important indicator of aging and osteopenia in human subjects. We found that age-dependent losses of type H vessels in human bone sections conform to the observations in aged mice. The abundance of human type H vessels and osteoprogenitors may be relevant to changes in the skeletal microarchitecture and advanced osteopenia. Furthermore, ovariectomized mice, a widely used model for postmenopausal osteoporosis, exhibited significantly reduced type H vessels accompanied by reduced osteoprogenitors, which is consistent with impaired bone microarchitecture and osteoporosis, suggesting that this feature is an indicator of bone mass independent of aging. More importantly, administration of desferrioxamine led to significantly increased bone mass via enhanced angiogenesis and increased type H vessels in ovariectomized mice. Altogether, these data represent a novel finding that type H vessels are regulated in aged and osteopenia subjects. The abundance of human type H vessels is an early marker of bone loss and represents a potential target for improving bone quality via the induction of type H vessels.

Spaceflight is known to induce severe systemic bone loss and muscle atrophy of astronauts due to the circumstances of microgravity. We examined the influence of artificially produced 2G hypergravity on mice for bone and muscle mass with newly developed centrifuge device. We also analyzed the effects of microgravity (mostly 0G) and artificial produced 1G in ISS (international space station) on mouse bone mass. Experiment on the ground, the bone mass of humerus, femur and tibia was measured using micro-computed tomography (μCT), and the all bone mass was significantly increased in 2G compared with 1G control. In tibial bone, the mRNA expression of bone formation related genes such as Osx and Bmp2 was elevated. The volume of triceps surae muscle was also increased in 2G compared with 1G control, and the mRNA expression of myogenic factors such as Myod and Myh1 was elevated by 2G. On the other hand, microgravity in ISS significantly induced the loss of bone mass on humerus and tibia, compared with artificial 1G induced by centrifugation. Here, we firstly report that bone and muscle mass are regulated by the gravity with loaded force in both of positive and negative on the ground and in the space.

The anterior cruciate ligament is anchored to the femur and tibia via specialized interfaces known as entheses. These play a critical role in ligament homeostasis and joint stability by transferring forces, varying in magnitude and direction between structurally and functionally dissimilar tissues. However, the precise structural and mechanical characteristics underlying the femoral and tibial entheses and their intricate interplay remain elusive. In this study, two thin-graduated mineralization regions in the femoral enthesis (~21 μm) and tibial enthesis (~14 μm) are identified, both exhibiting distinct biomolecular compositions and mineral assembly patterns. Notably, the femoral enthesis interface exhibits progressively maturing hydroxyapatites, whereas the mineral at the tibial enthesis interface region transitions from amorphous calcium phosphate to hydroxyapatites with increasing crystallinity. Proteomics results reveal that Matrix Gla protein uniquely enriched at the tibial enthesis interface, may stabilize amorphous calcium phosphate, while C-type lectin domain containing 11 A, enriched at the femoral enthesis interface, could facilitate the interface mineralization. Moreover, the finite element analysis indicates that the femoral enthesis model exhibited higher resistance to shearing, whereas the tibial enthesis model contributes to tensile resistance, suggesting that the discrepancy in biomolecular expression and the corresponding mineral assembly heterogeneities collectively contribute to the superior mechanical properties of both the femoral enthesis and tibial enthesis models. These findings provide novel perspectives on the structure-function relationships of anterior cruciate ligament entheses, paving the way for improved management of anterior cruciate ligament injury and regeneration. Here, the authors examine the human anterior cruciate femoral and tibial entheses and identify two thin-graduated mineralization regions with distinct biomolecular compositions and mineral assembly patterns to adapt to their respective mechanical functions.

BACKGROUND:Opening-wedge high tibial osteotomy by hemicallotasis for osteoarthritis in the medial compartment of the knee requires external fixation for a long time, until callus maturation is complete. The aim of this study was to determine if low-intensity pulsed ultrasound would accelerate callus maturation when applied after distraction to limbs treated with opening-wedge high tibial osteotomy by hemicallotasis. METHODS:Twenty-one patients with symmetric grades of osteoarthritis and similar degrees of varus deformity in the two knees underwent bilateral one-stage opening-wedge high tibial osteotomy by hemicallotasis. After completion of distraction, the bone mineral density of the distraction callus was measured. Then, one randomly selected limb was subjected to ultrasound treatment for twenty minutes daily until removal of the external fixator. The contralateral limb was left untreated to serve as the control. After four weeks of treatment, bone mineral density was measured again. RESULTS:During the four-week treatment period, the mean increase in callus bone mineral density was significantly greater in the ultrasound-treated tibiae (0.20 ± 0.12 g/cm) than in the control tibiae (0.13 ± 0.10 g/cm) (p = 0.02, unpaired t test). In eighteen patients the increase in the bone mineral density was greater in the ultrasound-treated limb than in the control limb, whereas in three patients the increase was greater in the control limb. CONCLUSIONS:We found that low-intensity pulsed ultrasound applied during the consolidation phase of distraction osteogenesis accelerates callus maturation after opening-wedge high tibial osteotomy by hemicallotasis in elderly patients. LEVEL OF EVIDENCE:Therapeutic study, Level I-1a (randomized controlled trial [significant difference]). See Instructions to Authors for a complete description of levels of evidence.