Mechanical induction of osteoanabolic Wnt1 promotes osteoblast differentiation via Plat.

Physical activity-induced mechanical stimuli play a crucial role in preserving bone mass and structure by promoting bone formation. While the Wnt pathway is pivotal for mediating the osteoblast response to loading, the exact mechanisms are not fully understood. Here, we found that mechanical stimulation induces osteoblastic Wnt1 expression, resulting in an upregulation of key osteogenic marker genes, including Runx2 and Sp7, while Wnt1 knockdown using siRNA prevented these effects. RNAseq analysis identified Plat as a major target through which Wnt1 exerts its osteogenic influence. This was corroborated by Plat depletion using siRNA, confirming its positive role in osteogenic differentiation. Moreover, we demonstrated that mechanical stimulation enhances Plat expression, which, in turn leads to increased expression of osteogenic markers like Runx2 and Sp7. Notably, Plat depletion by siRNA prevented this effect. We have established that Wnt1 regulates Plat expression by activating ß-Catenin. Silencing Wnt1 impairs mechanically induced ß-Catenin activation, subsequently reducing Plat expression. Furthermore, our findings showed that Wnt1 is essential for osteoblasts to respond to mechanical stimulation and induce Runx2 and Sp7 expression, in part through the Wnt1/ß-Catenin/Plat signaling pathway. Additionally, we observed significantly reduced Wnt1 and Plat expression in bones from ovariectomy (OVX)-induced and age-related osteoporotic mouse models compared with non-OVX and young mice, respectively. Overall, our data suggested that Wnt1 and Plat play significant roles in mechanically induced osteogenesis. Their decreased expression in bones from OVX and aged mice highlights their potential involvement in post-menopausal and age-related osteoporosis, respectively.

Impact of hyaluronic acid injection on the knee joint friction.

PURPOSE: The purpose of this in vitro study was to investigate whether or not hyaluronic acid supplementation improves knee joint friction during osteoarthritis progression under gait-like loading conditions. METHODS: Twelve human cadaveric knee joints were equally divided into mild and moderate osteoarthritic groups. After initial conservative preparation, a passive pendulum setup was used to test the whole joints under gait-like conditions before and after hyaluronic acid supplementation. The friction-related damping properties given by the coefficient of friction µ and the damping coefficient c (in kg m2/s) were calculated from the decaying flexion-extension motion of the knee. Subsequently, tibial and femoral cartilage and meniscus samples were extracted from the joints and tested in an established dynamic pin-on-plate tribometer using synthetic synovial fluid followed by synthetic synovial fluid supplemented with hyaluronic acid as lubricant. Friction was quantified by calculating the coefficient of friction. RESULTS: In the pendulum tests, the moderate OA group indicated significantly lower c0 values (p < 0.05) under stance phase conditions and significantly lower µ0 (p = 0.01) values under swing phase conditions. No degeneration-related statistical differences were found for µend or cend. Friction was not significantly different (p > 0.05) with regard to mild and moderate osteoarthritis in the pin-on-plate tests. Additionally, hyaluronic acid did not affect friction in both, the pendulum (p > 0.05) and pin-on-plate friction tests (p > 0.05). CONCLUSION: The results of this in vitro study suggested that the friction of cadaveric knee joint tissues does not increase with progressing degeneration. Moreover, hyaluronic acid viscosupplementation does not lead to an initial decrease in knee joint friction.

Role of Mast-Cell-Derived RANKL in Ovariectomy-Induced Bone Loss in Mice.

Mast cells may contribute to osteoporosis development, because patients with age-related or post-menopausal osteoporosis exhibit more mast cells in the bone marrow, and mastocytosis patients frequently suffer from osteopenia. We previously showed that mast cells crucially regulated osteoclastogenesis and bone loss in ovariectomized, estrogen-depleted mice in a preclinical model for post-menopausal osteoporosis and found that granular mast cell mediators were responsible for these estrogen-dependent effects. However, the role of the key regulator of osteoclastogenesis, namely, receptor activator of NFκB ligand (RANKL), which is secreted by mast cells, in osteoporosis development has, to date, not been defined. Here, we investigated whether mast-cell-derived RANKL participates in ovariectomy (OVX)-induced bone loss by using female mice with a conditional Rankl deletion. We found that this deletion in mast cells did not influence physiological bone turnover and failed to protect against OVX-induced bone resorption in vivo, although we demonstrated that RANKL secretion was significantly reduced in estrogen-treated mast cell cultures. Furthermore, Rankl deletion in mast cells did not influence the immune phenotype in non-ovariectomized or ovariectomized mice. Therefore, other osteoclastogenic factors released by mast cells might be responsible for the onset of OVX-induced bone loss.

Ready-To-Use and Rapidly Biodegradable Magnesium Phosphate Bone Cement: In Vivo Evaluation in Sheep.

In clinical practice, hydroxyapatite (HA) cements for bone defect treatment are frequently prepared by mixing a powder component and a liquid component shortly before implantation in the operation theater, which is time-consuming and error-prone. In addition, HA cements are only slightly resorbed, that is, cement residues can still be found in the bone years after implantation. Here, these challenges are addressed by a prefabricated magnesium phosphate cement paste based on glycerol, which is ready-to-use and can be directly applied during surgery. By using a trimodal particle size distribution (PSD), the paste is readily injectable and exhibits a compressive strength of 9-14 MPa after setting. Struvite (MgNH4 PO4 ·6H2 O), dittmarite (MgNH4 PO4 ·H2 O), farringtonite (Mg3 (PO4 )2 ), and newberyite (MgHPO4 ·3H2 O) are the mineral phases present in the set cement. The paste developed here features a promising degradation of 37% after four months in an ovine implantation model, with 25% of the implant area being newly formed bone. It is concluded that the novel prefabricated paste improves application during surgery, has a suitable degradation rate, and supports bone regeneration.

Oestrogen and Vibration Improve Intervertebral Disc Cell Viability and Decrease Catabolism in Bovine Organ Cultures.

Postmenopausal women are at an increased risk for intervertebral disc degeneration, possibly due to the decrease in oestrogen levels. Low-magnitude, high-frequency vibration (LMHFV) is applied as a therapeutic approach for postmenopausal osteoporosis; however, less is known regarding possible effects on the intervertebral disc (IVD) and whether these may be oestrogen-dependent. The present study investigated the effect of 17ß-oestradiol (E2) and LMHFV in an IVD organ culture model. Bovine IVDs (n = 6 IVDs/group) were treated with either (i) E2, (ii) LMHFV or (iii) the combination of E2 + LMHFV for 2 or 14 days. Minor changes in gene expression, cellularity and matrix metabolism were observed after E2 treatment, except for a significant increase in matrix metalloproteinase (MMP)-3 and interleukin (IL)-6 production. Interestingly, LMHFV alone induced cell loss and increased IL-6 production compared to the control. The combination of E2 + LMHFV induced a protective effect against cell loss and decreased IL-6 production compared to the LMHFV group. This indicates possible benefits of oestrogen therapy for the IVDs of postmenopausal women undergoing LMHFV exercises.

Activation function 2 (AF2) domain of estrogen receptor-α regulates mechanotransduction during bone fracture healing in estrogen-competent mice.

External mechanostimulation applied by whole-body low-magnitude high-frequency vibration (LMHFV) was demonstrated to cause no or negative effects on fracture healing in estrogen-competent rodents, while in ovariectomized (OVX), estrogen-deficient rodents bone formation after fracture was improved. Using mice with an osteoblast-specific deletion of the estrogen receptor α (ERα), we demonstrated that ERα signaling in osteoblasts is required for both the anabolic and catabolic effects of LMHFV during bone fracture healing in OVX and non-OVX mice, respectively. Because the vibration effects mediated by ERα were strictly dependent on the estrogen status, we hypothesized different roles of ligand-dependent and -independent ERα signaling. To investigate this assumption in the present study, we used mice with a deletion of the C-terminal activation function (AF) domain-2 of the ERα receptor, which mediated ligand-dependent ERα signaling (ERαAF-20). OVX and non-OVX ERαAF-20 animals were subjected to femur osteotomy followed by vibration treatment. We revealed that estrogen-competent mice lacking the AF-2 domain were protected from LMHFV-induced impaired bone regeneration, while the anabolic effects of vibration in OVX mice were not affected by the AF-2 knockout. RNA sequencing further showed that genes involved in Hippo/Yap1-Taz and Wnt signaling were significantly downregulated upon LMHFV in the presence of estrogen in vitro. In conclusion, we demonstrated that the AF-2 domain is crucial for the negative effects of vibration during bone fracture healing in estrogen-competent mice suggesting that the osteoanabolic effects of vibration are rather mediated by ligand-independent ERα signaling.

Functional Loss of Terminal Complement Complex Protects Rabbits from Injury-Induced Osteoarthritis on Structural and Cellular Level.

The terminal complement complex (TCC) has been described as a potential driver in the pathogenesis of posttraumatic osteoarthritis (PTOA). However, sublytic TCC deposition might also play a crucial role in bone development and regeneration. Therefore, we elucidated the effects of TCC on joint-related tissues using a rabbit PTOA model. In brief, a C6-deficient rabbit breed was characterized on genetic, protein, and functional levels. Anterior cruciate ligament transection (ACLT) was performed in C6-deficient (C6-/-) and C6-sufficient (C6+/-) rabbits. After eight weeks, the progression of PTOA was determined histologically. Moreover, the structure of the subchondral bone was evaluated by µCT analysis. C6 deficiency could be attributed to a homozygous 3.6 kb deletion within the C6 gene and subsequent loss of the C5b binding site. Serum from C6-/- animals revealed no hemolytic activity. After ACLT surgery, joints of C6-/- rabbits exhibited significantly lower OA scores, including reduced cartilage damage, hypocellularity, cluster formation, and osteophyte number, as well as lower chondrocyte apoptosis rates and synovial prostaglandin E2 levels. Moreover, ACLT surgery significantly decreased the trabecular number in the subchondral bone of C6-/- rabbits. Overall, the absence of TCC protected from injury-induced OA progression but had minor effects on the micro-structure of the subchondral bone.

Autologous semitendinosus meniscus graft significantly improves knee joint kinematics and the tibiofemoral contact after complete lateral meniscectomy.

PURPOSE: The purpose of this study was to investigate the potential of a doubled semitendinosus (ST) and a single gracilis tendon (GT) lateral meniscus autograft to restore the knee joint kinematics and tibiofemoral contact after total lateral meniscectomy (LMM). METHODS: Fourteen human knee joints were tested intact, after LMM and after ST and GT meniscus autograft treatment under an axial load of 200 N during full range of motion (0°-120°) and four randomised loading situations: without external moments, external rotation, valgus stress and a combination of external rotation and valgus stress using a knee joint simulator. Non-parametric statistical analyses were performed on joint kinematics and on the tibiofemoral contact mechanics. RESULTS: LMM led to significant rotational instability of the knee joints (p < 0.02), which was significantly improved after ST autograft application (p < 0.04), except for knee joint flexions > 60°. The GT autograft failed to restore the joint kinematics. LMM significantly increased the tibiofemoral contact pressure (p < 0.03), while decreasing the contact area (p < 0.05). The ST autograft was able to restore the contact mechanics after LMM (p < 0.02), while the GT replacement displayed only an improvement trend. CONCLUSION: The doubled ST lateral meniscus autograft improved the knee joint kinematics significantly and restored the tibiofemoral contact mechanics almost comparable to the native situation. Thus, from a biomechanical point of view, ST meniscus autografts might be a potential treatment alternative for patients who are indicated for meniscus allograft transplantation.

Complement receptor C5aR1 on osteoblasts regulates osteoclastogenesis in experimental postmenopausal osteoporosis.

In recent years, evidence has accumulated that the complement system, an integral part of innate immunity, may be involved in the regulation of bone homeostasis as well as inflammatory bone loss, for example, in rheumatoid arthritis and periodontitis. Complement may also contribute to osteoporosis development, but investigation of the mechanism is limited. Using mice with a conditional deletion of the complement anaphylatoxin receptor C5aR1, we here demonstrated that C5aR1 in osteoblasts (C5aR1 Runx2-Cre mice) or osteoclasts (C5aR1 LysM-Cre mice) did not affect physiological bone turnover or age-related bone loss in either sex, as confirmed by micro-computed tomography, histomorphometry, and biomechanical analyses of the bone and by the measurement of bone turnover markers in the blood serum. When female mice were subjected to ovariectomy (OVX), a common model for postmenopausal osteoporosis, significant bone loss was induced in C5aR1 fl/fl and C5aR1 LysM-Cre mice, as demonstrated by a significantly reduced bone volume fraction, trabecular number and thickness as well as an increased trabecular separation in the trabecular bone compartment. Confirming this, the osteoclast number and the receptor activator of nuclear factor k-B (RANK) ligand (RANKL) serum level were significantly elevated in these mouse lines. By contrast, C5aR1 Runx2-Cre mice were protected from bone loss after OVX and the serum RANKL concentration was not increased after OVX. These data suggested that bone cell-specific C5aR1 may be redundant in bone homeostasis regulation under physiological conditions. However, C5aR1 on osteoblasts was crucial for the induction of bone resorption under osteoporotic conditions by stimulating RANKL release, whereas C5aR1 on osteoclasts did not regulate OVX-induced bone loss. Therefore, our results implicate C5aR1 on osteoblasts as a potential target for treating postmenopausal osteoporosis.

Myeloid cell-derived catecholamines influence bone turnover and regeneration in mice.

Catecholamine signaling is known to influence bone tissue as reuptake of norepinephrine released from sympathetic nerves into bone cells declines with age leading to osteoporosis. Further, ß-adrenoceptor-blockers like propranolol provoke osteoprotective effects in osteoporotic patients. However, besides systemic adrenal and sympathetic catecholamine production, it is also known that myeloid cells can synthesize catecholamines, especially under inflammatory conditions. To investigate the effects of catecholamines produced by CD11b+ myeloid cells on bone turnover and regeneration, a mouse line with specific knockout of tyrosine hydroxylase, the rate-limiting enzyme of catecholamine synthesis, in CD11b+ myeloid cells (THflox/flox/CD11b-Cre+, referred to as THCD11b-Cre) was generated. For bone phenotyping, male mice were sacrificed at eight and twelve weeks of age and harvested bones were subjected to bone length measurement, micro-computed tomography, fluorescence-activated cell sorting of the bone marrow, gene expression analysis, histology and immunohistochemistry. Support for an age-dependent influence of myeloid cell-derived catecholamines on bone homeostasis is provided by the fact that twelve-week-old, but not eight-week-old THCD11b-Cre mice, developed an osteopenic phenotype and showed increased numbers of neutrophils and T lymphocytes in the bone marrow, while CCL2, IL-6, IL-4 and IL-10 mRNA expression was reduced in sorted myeloid bone marrow cells. To investigate the influence of myeloid cell-derived catecholamines on fracture healing, mice received a diaphyseal femur osteotomy. Three days post-fracture, immunohistochemistry revealed an increased number of macrophages, neutrophils and cytotoxic T lymphocytes in the fracture hematoma of THCD11b-Cre mice. Micro-computed tomography on day 21 showed a decreased tissue mineral density, a reduced bone volume and less trabeculae in the fracture callus indicating delayed fracture healing, probably due to the increased presence of inflammatory cells in THCD11b-Cre mice. This indicates a crucial role of myeloid cell-derived catecholamines in immune cell-bone cell crosstalk and during fracture healing.

Correction: Steppe et al. Bone Mass and Osteoblast Activity Are Sex-Dependent in Mice Lacking the Estrogen Receptor α in Chondrocytes and Osteoblast Progenitor Cells. Int. J. Mol. Sci. 2022, 23, 2902.

The authors would like to make corrections to the reference citations in the original article [...].

Bone Mass and Osteoblast Activity Are Sex-Dependent in Mice Lacking the Estrogen Receptor α in Chondrocytes and Osteoblast Progenitor Cells.

While estrogen receptor alpha (ERα) is known to be important for bone development and homeostasis, its exact function during osteoblast differentiation remains unclear. Conditional deletion of ERα during specific stages of osteoblast differentiation revealed different bone phenotypes, which were also shown to be sex-dependent. Since hypertrophic chondrocytes can transdifferentiate into osteoblasts and substantially contribute to long-bone development, we aimed to investigate the effects of ERα deletion in both osteoblast and chondrocytes on bone development and structure. Therefore, we generated mice in which the ERα gene was inactivated via a Runx2-driven cyclic recombinase (ERαfl/fl; Runx2Cre). We analyzed the bones of 3-month-old ERαfl/fl; Runx2Cre mice by biomechanical testing, micro-computed tomography, and cellular parameters by histology. Male ERαfl/fl; Runx2Cre mice displayed a significantly increased cortical bone mass and flexural rigidity of the femurs compared to age-matched controls with no active Cre-transgene (ERαfl/fl). By contrast, female ERαfl/fl; Runx2Cre mice exhibited significant trabecular bone loss, whereas in cortical bone periosteal and endosteal diameters were reduced. Our results indicate that the ERα in osteoblast progenitors and hypertrophic chondrocytes differentially contributes to bone mass regulation in male and female mice and improves our understanding of ERα signaling in bone cells in vivo.

G6b-B regulates an essential step in megakaryocyte maturation.

G6b-B is a megakaryocyte lineage-specific immunoreceptor tyrosine-based inhibition motif-containing receptor, essential for platelet homeostasis. Mice with a genomic deletion of the entire Mpig6b locus develop severe macrothrombocytopenia and myelofibrosis, which is reflected in humans with null mutations in MPIG6B. The current model proposes that megakaryocytes lacking G6b-B develop normally, whereas proplatelet release is hampered, but the underlying molecular mechanism remains unclear. We report on a spontaneous recessive single nucleotide mutation in C57BL/6 mice, localized within the intronic region of the Mpig6b locus that abolishes G6b-B expression and reproduces macrothrombocytopenia, myelofibrosis, and osteosclerosis. As the mutation is based on a single-nucleotide exchange, Mpig6bmut mice represent an ideal model to study the role of G6b-B. Megakaryocytes from these mice were smaller, displayed a less-developed demarcation membrane system, and had a reduced expression of receptors. RNA sequencing revealed a striking global reduction in the level of megakaryocyte-specific transcripts, in conjunction with decreased protein levels of the transcription factor GATA-1 and impaired thrombopoietin signaling. The reduced number of mature MKs in the bone marrow was corroborated on a newly developed Mpig6b-null mouse strain. Our findings highlight an unexpected essential role of G6b-B in the early differentiation within the megakaryocytic lineage.

Mast Cells Trigger Disturbed Bone Healing in Osteoporotic Mice.

Mast cells are important tissue-resident sensor and effector immune cells but also play a major role in osteoporosis development. Mast cells are increased in numbers in the bone marrow of postmenopausal osteoporotic patients, and mast cell-deficient mice are protected from ovariectomy (OVX)-induced bone loss. In this study, we showed that mast cell-deficient Mcpt5-Cre R-DTA mice were protected from OVX-induced disturbed fracture healing, indicating a critical role for mast cells in the pathomechanisms of impaired bone repair under estrogen-deficient conditions. We revealed that mast cells trigger the fracture-induced inflammatory response by releasing inflammatory mediators, including interleukin-6, midkine (Mdk), and C-X-C motif chemokine ligand 10 (CXCL10), and promote neutrophil infiltration into the fracture site in OVX mice. Furthermore, mast cells were responsible for reduced osteoblast and increased osteoclast activities in OVX mice callus, as well as increased receptor activator of NF-κB ligand serum levels in OVX mice. Additional in vitro studies with human cells showed that mast cells stimulate osteoclastogenesis by releasing the osteoclastogenic mediators Mdk and CXCL10 in an estrogen-dependent manner, which was mediated via the estrogen receptor alpha on mast cells. In conclusion, mast cells negatively affect the healing of bone fractures under estrogen-deficient conditions. Hence, targeting mast cells might provide a therapeutic strategy to improve disturbed bone repair in postmenopausal osteoporosis. © 2021 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

Are Knotted or Knotless Techniques Better for Reconstruction of Full-Thickness Tears of the Superior Portion of the Subscapularis Tendon? A Study in Cadavers.

BACKGROUND: Knotted and knotless single-anchor reconstruction techniques are frequently performed to reconstruct full-thickness tears of the upper portion of subscapularis tendon. However, it is unclear whether one technique is superior to the other. QUESTIONS/PURPOSES: (1) When comparing knotless and knotted single-anchor reconstruction techniques in full-thickness tears of the upper subscapularis tendon, is there a difference in stiffness under cyclic load? (2) Are there differences in cyclic gapping between knotless and knotted reconstructions? (3) Are there differences in the maximal stiffness, yield load, and ultimate load to failure? (4) What are the modes of failure of knotless and knotted reconstruction techniques? METHODS: Eight matched pairs of human cadaveric shoulders were dissected, and a full-thickness tear of the subscapularis tendon (Grade 3 according to the Fox and Romeo classification) was created. The cadavers all were male specimens, with a median (range) age of 69 years (61 to 75). Before biomechanical evaluation, the specimens were randomized into two equal reconstruction groups: knotless single anchor and knotted single anchor. All surgical procedures were performed by a single orthopaedic surgeon who subspecializes in sports orthopedics and shoulder surgery. With a customized set up that was integrated in a dynamic material testing machine, the humeri were consecutively loaded from 10 N to 60 N, from 10 N to 100 N, and from 10 N to 180 N for 50 cycles. Furthermore, the gapping behavior of the tear was analyzed using a video tracking system. Finally, the stiffness, gapping, maximal stiffness, yield loads, and maximum failure loads of both reconstruction groups were statistically analyzed. Failure was defined as retearing of the reconstructed gap threshold due to rupture of the tendon and/or failure of the knots or anchors. After biomechanical testing, bone quality was measured at the footprint of the subscapularis using microCT in all specimens. Bone quality was equal between both groups. To detect a minimum 0.15-mm difference in gap formation between the two repair techniques (with a 5% level of significance; α = 0.05), eight matched pairs (n = 16 in total) were calculated as necessary to achieve a power of at least 90%. RESULTS: The first study question can be answered as follows: for stiffness under cyclic load, there were no differences with the numbers available between the knotted and knotless groups at load stages of 10 N to 60 N (32.7 ± 3.5 N/mm versus 34.2 ± 5.6 N/mm, mean difference 1.5 N/mm [95% CI -6.43 to 3.33]; p = 0.55), 10 N to 100 N (45.0 ± 4.8 N/mm versus 45.2 ± 6.0 N/mm, mean difference 0.2 N/mm [95% CI -5.74 to 6.04]; p = 0.95), and 10 N to 180 N (58.2 ± 10.6 N/mm versus 55.2 ± 4.7 N/mm, mean difference 3 N/mm [95% CI -5.84 to 11.79]; p = 0.48). In relation to the second research question, the following results emerged: For cyclic gapping, there were no differences between the knotted and knotless groups at any load levels. The present study was able to show the following with regard to the third research question: Between knotted and knotless repairs, there were no differences in maximal load stiffness (45.3 ± 8.6 N/mm versus 43.5 ± 10.2 N/mm, mean difference 1.8 [95% CI -11.78 to 8.23]; p = 0.71), yield load (425.1 ± 251.4 N versus 379.0 ± 169.4 N, mean difference 46.1 [95% CI -276.02 to 183.72]; p = 0.67), and failure load (521.1 ± 266.2 N versus 475.8 ± 183.3 N, mean difference 45.3 [95% CI -290.42 to 199.79]; p = 0.69). Regarding the fourth question concerning the failure modes, in the knotted repairs, the anchor tore from the bone in 2 of 8, the suture tore from the tendon in 6 of 8, and no suture slipped from the eyelet; in the knotless repairs, the anchor tore from the bone in 2 of 8, the suture tore from the tendon in 3 of 8, and the threads slipped from the eyelet in 3 of 8. CONCLUSION: With the numbers available, we found no differences between single-anchor knotless and knotted reconstruction techniques used to repair full-thickness tears of the upper portion of subscapularis tendon. CLINICAL RELEVANCE: The reconstruction techniques we analyzed showed no differences in terms of their primary stability and biomechanical properties at the time of initial repair and with the numbers available. In view of these experimental results, it would be useful to conduct a clinical study in the future to verify the translationality of the experimental data of the present study.

Influence of Menisci on Tibiofemoral Contact Mechanics in Human Knees: A Systematic Review.

Purpose: Menisci transfer axial loads, while increasing the load-bearing tibiofemoral contact area and decreasing tibiofemoral contact pressure (CP). Numerous clinical and experimental studies agree that an increased CP is one predominant indicator for post-traumatic osteoarthritis (PTOA) of the knee joint. However, due to the immense variability in experimental test setups and wide range of treatment possibilities in meniscus surgery, it is difficult to objectively assess their impact on the CP determination, which is clearly crucial for knee joint health. Therefore, the aim of this systematic review is to investigate the influence of different meniscal injuries and their associated surgical treatments on the CP. Secondly, the influence of different test setups on CP measurements is assessed. On the basis of these results, we established the basis for recommendations for future investigations with the aim to determine CPs under different meniscal states. Methods: This review was conducted in accordance with the PRISMA guidelines. Studies were identified through a systematic literature search in Cochrane, PubMed and Web of Science databases. Literature was searched through pre-defined keywords and medical subject headings. Results: This review indicates a significant increase of up to 235% in peak CP when comparing healthy joints and intact menisci with impaired knee joints, injured or resected menisci. In addition, different test setups were indicated to have major influences on CP: The variety of test setups ranged from standard material testing machines, including customized setups via horizontal and vertical knee joint simulators, through to robotic systems. Differences in applied axial knee joint loads ranged from 0 N up to 2,700 N and resulted unsurprisingly in significantly different peak CPs of between 0.1 and 12.06 MPa. Conclusion: It was shown that untreated traumatic meniscal tears result in an increased CP. Surgical repair intervention were able to restore the CP comparable to the healthy, native condition. Test setup differences and particularly axial joint loading variability also led to major CP differences. In conclusion, when focusing on CP measurements in the knee joint, transparent and traceable in vitro testing conditions are essential to allow researchers to make a direct comparison between future biomechanical investigations.

Estrogen Receptor α Signaling in Osteoblasts is Required for Mechanotransduction in Bone Fracture Healing.

Biomechanical stimulation by whole-body low-magnitude high-frequency vibration (LMHFV) has demonstrated to provoke anabolic effects on bone metabolism in both non-osteoporotic and osteoporotic animals and humans. However, preclinical studies reported that vibration improved fracture healing and bone formation in osteoporotic, ovariectomized (OVX) mice representing an estrogen-deficient hormonal status, but impaired bone regeneration in skeletally healthy non-OVX mice. These effects were abolished in general estrogen receptor α (ERα)-knockout (KO) mice. However, it remains to be elucidated which cell types in the fracture callus are targeted by LMHFV during bone healing. To answer this question, we generated osteoblast lineage-specific ERα-KO mice that were subjected to ovariectomy, femur osteotomy and subsequent vibration. We found that the ERα specifically on osteoblastic lineage cells facilitated the vibration-induced effects on fracture healing, because in osteoblast lineage-specific ERα-KO (ERαfl/fl; Runx2Cre) mice the negative effects in non-OVX mice were abolished, whereas the positive effects of vibration in OVX mice were reversed. To gain greater mechanistic insights, the influence of vibration on murine and human osteogenic cells was investigated in vitro by whole genome array analysis and qPCR. The results suggested that particularly canonical WNT and Cox2/PGE2 signaling is involved in the mechanotransduction of LMHFV under estrogen-deficient conditions. In conclusion, our study demonstrates a critical role of the osteoblast lineage-specific ERα in LMHFV-induced effects on fracture healing and provides further insights into the molecular mechanism behind these effects.

Persistent JunB activation in fibroblasts disrupts stem cell niche interactions enforcing skin aging.

Fibroblasts residing in the connective tissues constitute the stem cell niche, particularly in organs such as skin. Although the effect of fibroblasts on stem cell niches and organ aging is an emerging concept, the underlying mechanisms are largely unresolved. We report a mechanism of redox-dependent activation of transcription factor JunB, which, through concomitant upregulation of p16INK4A and repression of insulin growth factor-1 (IGF-1), initiates the installment of fibroblast senescence. Fibroblast senescence profoundly disrupts the metabolic and structural niche, and its essential interactions with different stem cells thus enforces depletion of stem cells pools and skin tissue decline. In fact, silencing of JunB in a fibroblast-niche-specific manner-by reinstatement of IGF-1 and p16 levels-restores skin stem cell pools and overall skin tissue integrity. Here, we report a role of JunB in the control of connective tissue niche and identified targets to combat skin aging and associated pathologies.

Differences in Fracture Healing Between Female and Male C57BL/6J Mice.

BACKGROUND: Mice are increasingly used in fracture healing research because of the opportunity to use transgenic animals. While both, male and female mice are employed, there is no consensus in the literature whether fracture healing differs between both sexes. Therefore, the aim of the present study was to analyze diaphyseal fracture healing in female and male C57BL/6J mice, a commonly used mouse strain in bone research. METHODS: For that purpose, 12-week-old Female (17-20 g) and Male mice (22-26 g) received a standardized femur midshaft osteotomy stabilized by an external fixator. Mice were euthanized 10 and 21 days after fracture and bone healing was analyzed by biomechanical testing, µCT, histology, immunohistochemistry and qPCR. RESULTS: Ten days after fracture, Male mice displayed significantly more cartilage but less fibrous tissue in the fracture callus compared to Female mice, whereas the amount of bone did not differ. At day 21, Male mice showed a significantly larger fracture callus compared to Female mice. The relative amount of bone in the fracture callus did not significantly differ between both sexes, whereas its tissue mineral density was significantly higher in Male mice on day 21, indicating more mature bone and slightly more rapid fracture healing. These results were confirmed by a significantly greater absolute bending stiffness of the fractured femurs of Male mice on day 21. On the molecular level, Male mice displayed increased active ß-catenin expression in the fracture callus, whereas estrogen receptor α (ERα) expression was lower. CONCLUSION: These results suggest that Male mice display more rapid fracture healing with more prominent cartilaginous callus formation. This might be due to the higher weight of Male mice, resulting in increased mechanical loading of the fracture. Furthermore, Male mice displayed significantly greater activation of osteoanabolic Wnt/ß-catenin signaling, which might also contribute to more rapid bone regeneration.

Neuromapping of the Capsuloligamentous Knee Joint Structures.

PURPOSE: To investigate neuromuscular electromyographic response of the of the upper and lower leg muscles after the application of an intraoperative, isolated mechanical stimulus of the capsuloligamentous structures, including the anterior (ACL) and posterior cruciate ligaments (PCL), lateral (LM) and medial menisci (MM), plica mediopatellaris (PM), and Hoffa's fat pat (HFP). METHODS: The electromyographic response of the upper and lower leg muscles (M. rectus femoris; M. vastus medialis; M. semitendinosus; M. biceps femoris; M. gastrocnemius lateralis) of 15 male patients were measured after an isolated mechanical stimulus of the capsuloligamentous structures during an arthroscopic intervention using a customized intraoperative setup. Target parameters were the short (SLR; <30 milliseconds) and medium latency responses (MLR; >30 milliseconds) after the mechanically-induced trigger. RESULTS: The ACL, PCL, LM, and MM displayed high interindividual reproducibility of >76%. The MM was the only structure indicating both an SLR and MLR for all muscles. Although signals could be detected, there was no reproducibility in electromyographic signal activation for the HFP. The most rapid MLR was observed for the PM (quadriceps: 37 milliseconds). CONCLUSIONS: Each stimulated structure displayed an individual MLR response, which allowed us to create neuromapping combining the anatomical and quantitative representations of the individual muscular activation patterns after isolated mechanical stimulation of the capsuloligamentous knee joint structures, corroborating our hypothesis. LEVEL OF EVIDENCE: Diagnostic - Level II.