Disruption of the c-terminal serine protease domain of Fam111a does not alter calcium homeostasis in mice.

FAM111A gene mutations cause Kenney-Caffey syndrome (KCS) and Osteocraniostenosis (OCS), conditions characterized by short stature, low serum ionized calcium (Ca2+), low parathyroid hormone (PTH), and bony abnormalities. The molecular mechanism mediating this phenotype is unknown. The c-terminal domain of FAM111A harbors all the known disease-causing variations and encodes a domain with high homology to serine proteases. However, whether this serine protease domain contributes to the maintenance of Ca2+ homeostasis is not known. We hypothesized the disruption of the serine protease domain of FAM111A would disrupt Ca2+ homeostasis. To test this hypothesis, we generated with CRISPR/Cas9, mice with a frameshift insertion (c.1450insA) or large deletion (c.1253-1464del) mutation in the Fam111a serine protease domain. Serum-ionized Ca2+ and PTH levels were not significantly different between wild type, heterozygous, or homozygous Fam111a mutant mice. Additionally, there were no significant differences in fecal or urine Ca2+ excretion, intestinal Ca2+ absorption or overall Ca2+ balance. Only female homozygous (c.1450insA), but not heterozygous mice displayed differences in bone microarchitecture and mineral density compared to wild-type animals. We conclude that frameshift mutations that disrupt the c-terminal serine protease domain do not induce a KCS or OCS phenotype in mice nor alter Ca2+ homeostasis.

Three-dimensional morphometric analysis of cranial sutures - A novel approach to quantitative analysis.

Objective: Differences in complexity of cranial suture forms on the endocranial (i.e., deep) and ectocranial (i.e., superficial) skull surfaces have been noted in the literature, indicating through thickness three-dimensional (3D) suture variability depending on the chosen section and necessity for considering the complete 3D structure in many cases. This study aims to evaluate the variability of suture morphology through the skull thickness using a rat model, and to provide more robust metrics and methodologies to analyze suture morphology. Design: X-ray micro-computed tomographic (µCT) imaging methods were utilized in order to provide internal structure information. Methods were developed to isolate and analyze sutures widths and linear interdigitation index (LII) values on each adjacent offset transverse plane of the µCT datasets. LII was defined as the curved path length of the suture divided by the linear length between the ends of the region of interest. Scans were obtained on 15 female rats at ages of 16, 20, and 24 weeks (n = 5/age). Samples were imaged at 18 µm resolutions with 90 kV source voltage, 278 µA source amperage, and 0.7° increments. Suture widths and LII values were compared using a Kruskal-Wallis test. Results: 3D variability in local suture widths within individuals, as well as through thickness variabilities in planar widths and LII was observed. Kruskal-Wallis tests for bulk through thickness averaged suture widths and LII were found to be statistically insignificant, despite clear geometric differences through suture thicknesses. Conclusion: Although the bulk morphometric variability between age groups was found to be statistically insignificant, the 3D variability within individuals point to the importance of analyzing suture form using 3D metrics when studying suture development, response to functional activity, or morphometry in general.

Therapeutic Potential and Predictive Pharmaceutical Modeling of Stilbenes in Cannabis sativa.

Cannabis sativa is a plant used for recreational and therapeutic purposes; however, many of the secondary metabolites in the plant have not been thoroughly investigated. Stilbenes are a class of compounds with demonstrated anti-inflammatory and antioxidant properties and are present in cannabis. Many stilbenes present in cannabis have been investigated for their therapeutic effects. Fourteen stilbenes have been identified to be present in cannabis, all of which are structurally dihydrostilbenoids, with half possessing a prenylated moiety. The stilbenes summarized in this analysis show varying degrees of therapeutic benefits ranging from anti-inflammatory, antiviral, and anti-cancer to antioxidant effects. Many of the identified stilbenes have been researched to a limited extent for potential health benefits. In addition, predictive in silico modeling was performed on the fourteen identified cannabis-derived stilbenes. This modeling provides prospective activity, pharmacokinetic, metabolism, and permeability data, setting the groundwork for further investigation into these poorly characterized compounds.

The effect of morphometric and geometric indices of the human calvarium on mechanical response.

BACKGROUND: When developing a surrogate model of the human skull, there is a multitude of morphometric and geometric properties to consider when constructing the model. To simplify this approach, it is important to identify only the properties that have a significant influence on the mechanical response of the skull. The objective of this study was to identify which morphometric and geometric properties of the calvarium were significant predictors of mechanical response. METHODS: Calvarium specimens (N = 24) were micro-computed tomography scanned to determine morphometric and geometric properties. The specimens were assumed to be Euler-Bernoulli beams and were subject to 4-point quasi-static bending to determine mechanical response. Univariate linear regressions were performed whereby the morphometric and geometric properties were independent or predictor variables and the mechanical responses were dependent or outcome variables. FINDINGS: Nine significant linear regression models were established (p < 0.05). In the diploë, trabecular bone pattern factor was a significant predictor of force and bending moment at fracture. The inner cortical table had more significant predictors (thickness, tissue mineral density, and porosity) of mechanical response compared to the outer cortical table and diploë. INTERPRETATION: Morphometric and geometric properties had a key influence on the calvarium's biomechanics. Trabecular bone pattern factor and the morphometry and geometry of the cortical tables must be considered when evaluating the mechanical response of the calvarium. These properties can aid the design of surrogate models of the skull that seek to mimic its mechanical response for head impact simulation.

The Mechanical Characterization and Comparions of Male and Female Calvaria Under Four-Point Bending Impacts.

The circumstances in which we mechanically test and critically assess human calvarium tissue would find relevance under conditions encompassing real-world head impacts. These conditions include, among other variables, impact velocities, and strain rates. Compared to quasi-static loading on calvaria, there is less reporting on the impact loading of the calvaria and consequently, there are relatively fewer mechanical properties on calvaria at relevant impact loading rates available in the literature. The purpose of this work was to report on the mechanical response of 23 human calvarium specimens subjected to dynamic four-point bending impacts. Impacts were performed using a custom-built four-point impact apparatus at impact velocities of 0.86-0.89 m/s resulting in surface strain rates of 2-3/s-representative of strain rates observed in vehicle collisions and blunt impacts. The study revealed comparable effective bending moduli (11-15 GPa) to the limited work reported on the impact mechanics of calvaria in the literature, however, fracture bending stress (10-47 MPa) was relatively less. As expected, surface strains at fracture (0.21-0.25%) were less compared to studies that performed quasi-static bending. Moreover, the study revealed no significant differences in mechanical response between male and female calvaria. The findings presented in this work are relevant to many areas including validating surrogate skull fracture models in silico or laboratory during impact and optimizing protective devices used by civilians to reduce the risk of a serious head injury.

Cranial suture morphometry and mechanical response to loading: 2D vs. 3D assumptions and characterization.

Cranial sutures are complex soft tissue structures whose mechanics are often studied due to their link with bone growth in the skull. Researchers will often use a cross-sectional two-dimensional slice to define suture geometry when studying morphometry and/or mechanical response to loading. However, using a single cross section neglects the full suture complexity and may introduce significant errors when defining their form. This study aims to determine trends in suture path variability through skull thickness in a swine model and the implications of using a 'representative' cross section on mechanical modeling. To explore these questions, a mixture of quantitative analysis of computed tomography images and finite element models was used. The linear interdigitation and width of coronal and sagittal sutures were analyzed on offset transverse planes through the skull thickness. It was found that sagittal suture width and interdigitation were largely consistent through the skull thickness, whereas the coronal suture showed significant variation in both. The finite element study found that average values of displacement and strain were similar between the two-dimensionally variable and three-dimensionally variable models. Larger ranges and more complex distributions of strain were found in the three-dimensionally variable model. Outcomes of this study indicate that the appropriateness of using a representative cross section to describe suture morphometry and predict mechanical response should depend on specific research questions and goals. Two-dimensional approximations can be sufficient for less-interdigitated sutures and when bulk site mechanics are of interest, while taking the true three-dimensional geometry into account is necessary when considering spatial variability and local mechanical response.

Claudin-2 and claudin-12 form independent, complementary pores required to maintain calcium homeostasis.

Calcium (Ca2+) homeostasis is maintained through coordination between intestinal absorption, renal reabsorption, and bone remodeling. Intestinal and renal (re)absorption occurs via transcellular and paracellular pathways. The latter contributes the bulk of (re)absorption under conditions of adequate intake. Epithelial paracellular permeability is conferred by tight-junction proteins called claudins. However, the molecular identity of the paracellular Ca2+ pore remains to be delineated. Claudins (Cldn)-2 and -12 confer Ca2+ permeability, but deletion of either claudin does not result in a negative Ca2+ balance or increased calciotropic hormone levels, suggesting the existence of additional transport pathways or parallel roles for the two claudins. To test this, we generated a Cldn2/12 double knockout mouse (DKO). These animals have reduced intestinal Ca2+ absorption. Colonic Ca2+ permeability is also reduced in DKO mice and significantly lower than single-null animals, while small intestine Ca2+ permeability is unaltered. The DKO mice display significantly greater urinary Ca2+ wasting than Cldn2 null animals. These perturbations lead to hypocalcemia and reduced bone mineral density, which was not observed in single-KO animals. Both claudins were localized to colonic epithelial crypts and renal proximal tubule cells, but they do not physically interact in vitro. Overexpression of either claudin increased Ca2+ permeability in cell models with endogenous expression of the other claudin. We find claudin-2 and claudin-12 form partially redundant, independent Ca2+ permeable pores in renal and colonic epithelia that enable paracellular Ca2+ (re)absorption in these segments, with either one sufficient to maintain Ca2+ balance.

Experimental repeatability, sensitivity, and reproducibility of force and strain measurements from within the periodontal ligament space during ex vivo swine tooth loading.

The Periodontal Ligament (PDL) is a complex connective tissue that anchors a tooth to the surrounding alveolar bone. The small size and complex geometry of the PDL space within an intact tooth-PDL-bone complex (TPBC) limits strain measurements. An in-fiber Bragg grating (FBG) sensor offers potential for such measurements due to its small size. This work defines an experimental procedure where strain and force were measured during quasi-static, apically directed, displacement-controlled tests on swine premolar crowns. Specifically, the: inter-TPBC, intra-TPBC, and long-term repeatability after a preconditioned state was objectively identified; sensitivity to preload magnitude, TPBC alignment, and sensor depth; and reproducibility within a TPBC was determined. Data clustering was used to determine the appropriate number of preconditioning trials, ranging from one to seven. Strain and force measurements showed intra-TPBC repeatability with average adjusted root mean square from the median of 28.9% of the peak strain and 4.5% of the peak force measurement. A Mann-Whitney U test generally found statistically significant differences in peak strain and force measurements between the left and right sides, suggesting a lack of inter-TPBC repeatability. Using a Friedman test, it was shown that peak strain measures were sensitive to the TPBC alignment and sensor depth, while peak force measures were sensitive to the preload and TPBC alignment. A Friedman test suggested reproducible strain and force measurements when the FBG was replaced within the same TPBC and the preload, alignment, and sensor depth were controlled.

Cortical and trabecular morphometric properties of the human calvarium.

There is currently a gap in the literature that quantitatively describes the complex bone microarchitecture within the diploë (trabecular bone) and cortical layers of the human calvarium. The purpose of this study was to determine the morphometric properties of the diploë and cortical tables of the human calvarium in which key interacting factors of sex, location on the calvarium, and layers of the sandwich structure were considered. Micro-computed tomography (micro-CT) was utilized to capture images at 18 µm resolution of male (n = 26) and female (n = 24) embalmed calvarium specimens in the frontal and parietal regions (N = 50). All images were post-processed and analyzed using vendor bundled CT-Analyzer software to determine the morphometric properties of the diploë and cortical layers. A two-way mixed (repeated measures) analysis of variance (ANOVA) was used to determine diploë morphometric properties accounting for factors of sex and location. A three-way mixed ANOVA was performed to determine cortical morphometric properties accounting for factors of cortical layer (inner and outer table), sex, and location. The study revealed no two-way interaction effects between sex and location on the diploë morphometry except for fractal dimension. Trabecular thickness and separation in the diploë were significantly greater in the male specimens; however, females showed a greater number of trabeculae and fractal dimension on average. Parietal specimens revealed a greater porosity, trabecular separation, and deviation from an ideal plate structure, but a lesser number of trabeculae and connectivity compared to the frontal location. Additionally, the study observed a lower density and greater porosity in the inner cortical layer than the outer which may be due to clear distinctions between each layer's physiological environment. The study provides valuable insight into the quantitative morphometry of the calvarium in which finite element modelers of the skull can refer to when designing detailed heterogenous or subject-specific skull models to effectively predict injury. Furthermore, this study contributes towards the recent developments on physical surrogate models of the skull which require approximate measures of calvarium bone architecture in order to effectively fabricate a model and then accurately simulate a traumatic head impact event.

Tooth attachment and pleurodont implantation in lizards: Histology, development, and evolution.

Squamates present a unique challenge to the homology and evolution of tooth attachment tissues. Their stereotypically pleurodont teeth are fused in place by a single "bone of attachment", with seemingly dubious homology to the three-part tooth attachment system of mammals and crocodilians. Despite extensive debate over the interpretations of squamate pleurodonty, its phylogenetic significance, and the growing evidence from fossil amniotes for the homology of tooth attachment tissues, few studies have defined pleurodonty on histological grounds. Using a sample of extant squamate teeth that we organize into three broad categories of implantation, we investigate the histological and developmental properties of their dental tissues in multiple planes of section. We use these data to demonstrate the specific soft- and hard-tissue features of squamate teeth that produce their disparate tooth implantation modes. In addition, we describe cementum, periodontal ligaments, and alveolar bone in pleurodont squamates, dental tissues that were historically thought to be restricted to extant mammals and crocodilians. Moreover, we show how the differences between pleurodonty and thecodonty do not relate to the identity of the tooth attachment tissues, but rather the arrangements of homologous tissues around the teeth.

Mechanistic understanding of underperforming enteric coated products: Opportunities to add clinical relevance to the dissolution test.

Over the last 70 years several cases of in vivo failure of enteric coated (EC) formulations have been reported. The observed failures seem to be due to the slower than expected in vivo performance of EC products. Upon reaching the intestinal lumen, the dosage form is exposed to a bicarbonate buffered environment at much lower interfacial buffering capacity compared to those applied in compendial phosphate buffers. Hence, there is an urgent need to understand the behavior of EC products in bicarbonate buffer (BCB) and to revaluate the current dissolution methods used for such products. The current pilot study mechanistically investigated the performance of five EC products available in the Canadian market. The evaluated parameters were the buffer system (bicarbonate buffer vs. phosphate buffer), buffer capacity and medium pH. We hypothesized that the performance of EC products in BCB would be different compared with compendial phosphate buffer, giving more physiological insight, and that API properties would impact the dissolution behavior in BCB. The objective of this study was to examine the effect of the aforementioned parameters on the drug release applying physiologically relevant conditions (bicarbonate buffer at low molarities). A first step towards making the use of bicarbonate-based systems more feasible in a quality control setting is also reported. All formulations displayed a fast release in phosphate buffer and complied with the compendial performance specifications. On the other hand, they all had a much slower drug release in bicarbonate buffer and failed the USP acceptance criteria. Also, the nature of the drug (acid vs base) impacted the dissolution behavior in BCB. This pilot study indicates that compendial dissolution test for enteric coated tablets lacks physiological relevance and it needs to be reevaluated. Thus, an in vivo relevant performance method for EC products is needed.

The iliosacral joint in lizards: an osteological and histological analysis.

The development of the iliosacral joint (ISJ) in tetrapods represented a crucial step in the evolution of terrestrial locomotion. This structure is responsible for transferring forces between the vertebral column and appendicular skeleton, thus supporting the bodyweight on land. However, most research dealing with the water-to-land transition and biomechanical studies in general has focused exclusively on the articulation between the pelvic girdle and femur. Our knowledge about the contact between the pelvic girdle and vertebral column (i.e. the ISJ) at a tissue level is restricted so far to human anatomy, with little to no information available on other tetrapods. This lack of data limits our understanding of the development and evolution of such a key structure, and thus on the pattern and processes of the evolution of terrestrial locomotion. Therefore, we investigated the macro- and microanatomy of the ISJ in limb-bearing squamates that, similar to most non-mammalian, non-avian tetrapods, possess only two sacral ribs articulating with the posterior process of the ilium. Using a combination of osteology, micro-computed tomography and histology, we collected data on the ISJ apparatus of numerous specimens, sampling different taxa and different ontogenetic stages. Osteologically, we recorded consistent variability in all three processes of the ilium (preacetabular, supracetabular and posterior) and sacral ribs that correlate with posture and locomotion. The presence of a cavity between the ilium and sacral ribs, abundant articular cartilage and fibrocartilage, and a surrounding membrane of dense fibrous connective tissue allowed us to define this contact as a synovial joint. By comparison, the two sacral ribs are connected to each other mostly by dense fibrous tissue, with some cartilage found more distally along the margins of the two ribs, defining this joint as a combination of a syndesmosis and synchondrosis. Considering the intermediary position of the ISJ between the axial and appendicular skeletons, the shape of the articular surfaces of the sacral ribs and ilium, and the characteristics of the muscles associated with this structure, we argue that the mobility of the ISJ is primarily driven by the movements of the hindlimb during locomotion. We hypothesize that limited torsion of the ilium at the ISJ happens when the hip is abducted, and the joint is likely able to absorb the compressional and extensional forces related to the protraction and retraction of the femur. The mix of fibres and cartilage between the two sacral ribs instead serves primarily as a shock absorber, with the potential for limited vertical translation during locomotion.

Nanomaterials for bone tissue regeneration: updates and future perspectives.

Joint replacement and bone reconstructive surgeries are on the rise globally. Current strategies for implants and bone regeneration are associated with poor integration and healing resulting in repeated surgeries. A multidisciplinary approach involving basic biological sciences, tissue engineering, regenerative medicine and clinical research is required to overcome this problem. Considering the nanostructured nature of bone, expertise and resources available through recent advancements in nanobiotechnology enable researchers to design and fabricate devices and drug delivery systems at the nanoscale to be more compatible with the bone tissue environment. The focus of this review is to present the recent progress made in the rationale and design of nanomaterials for tissue engineering and drug delivery relevant to bone regeneration.

Cranial ontogeny of Thamnophis radix (Serpentes: Colubroidea) with a re-evaluation of current paradigms of snake skull evolution.

Accurate knowledge of skeletal ontogeny in extant organisms is crucial in understanding important morpho-functional systems and in enabling inferences of the ontogenetic stage of fossil specimens. However, detailed knowledge of skeletal ontogeny is lacking for most squamates, including snakes. Very few studies have discussed postnatal development in snakes, with none incorporating data from all three major ontogenetic stages-embryonic, juvenile and adult. Here, we provide the first analysis encompassing these three ontogenetic stages for any squamate, using the first complete micro-computed tomography (micro-CT)-based segmentations of any non-adult snake, based on fresh specimens of Thamnophis radix. The most significant ontogenetic changes involve the feeding apparatus, with major elongation of the tooth-bearing elements and jaw suspensorium causing a posterior shift in the jaw articulation. This shift enables macrostomy (large-gaped feeding in snakes) and occurs in T. radix via a different developmental trajectory than in most other macrostomatans, indicating that the evolution of macrostomy is more complex than previously thought. The braincase of T. radix is also evolutionarily unique among derived snakes in lacking a crista circumfenestralis, a phenomenon considered herein to represent paedomorphic retention of the embryonic condition. We thus present numerous important challenges to current paradigms regarding snake cranial evolution.

Increased FoxO3a expression prevents osteoblast differentiation and matrix calcification.

Forkhead Box O transcription factors play important roles in bone metabolism by defending against oxidative stress and apoptosis. FoxO3a is of special interest as it is the predominant isoform expressed in bone. In osteoblasts, the administration of 1,25 dihydroxyvitamin D3 (1,25D3) increases FoxO3a expression, and alters calcium handling. We therefore queried whether FoxO3a participates in vitamin D-mediated regulation of calcium transport pathways or matrix calcification, independent of reactive oxygen species (ROS) formation. To examine this possibility, we differentiated MC3T3-E1 cells into mature osteoblast-like cells over 7 days. This coincided with an increased ability to mineralize extracellular matrix. FoxO3a expression increased throughout differentiation. 1,25D3 enhanced both FoxO3a mRNA and protein expression. Immunofluorescence microscopy found increased FoxO3a nuclear localization with differentiation and after treatment with 1,25D3. Live cell ratiometric imaging with Fura-2AM identified significant L-type calcium channel mediated calcium uptake that was enhanced by 1,25D3. We observed expression of both Cav1.2 and Cav1.3, although expression decreased throughout differentiation and was not altered by 1,25D3 treatment. FoxO3a overexpression reduced calcium uptake and calcium deposition. FoxO3a overexpression also prevented alterations in calcium channel expression and the cell differentiation associated decrease in expression of Runx2 and increased expression of osteocalcin, findings consistent with a failure for the cells to differentiate. Based on both our expression and functional data, we suggest that high levels of FoxO3a prevent osteoblast differentiation and matrix calcification.

Triple-threat activity of PEDF in bone tumors: Tumor inhibition, tissue preservation and cardioprotection against doxorubicin.

Pigment epithelium-derived factor (PEDF) is known for its osteogenic properties, but its effects against primary and secondary bone tumors have not comprehensively been demonstrated. We show the ubiquitous expression of PEDF in murine embryonic tissue. Continuous administration of PEDF in pregnant mice for five days did not adversely affect foetal health, despite PEDF's known potent antiangiogenic properties. In the case of the devastating childhood bone cancer osteosarcoma, PEDF has direct anticancer activity per se, and protects against the toxicity of doxorubicin in the heart, small intestine and testes. PEDF demonstrated anti-proliferative and pro-apoptotic effects against human prostate and breast cancer cells, tumors which are known to metastasize to bone as the preferred secondary site. Caspase-2 was activated in both tumor cell types by PEDF. In models of prostate and breast cancer in bone, PEDF significantly reduced tumor volumes. When combined with zoledronic acid, continuously-administered PEDF significantly reduced breast tumor volume at the bone, and was able to preserve the quality of bone better than the combination therapy. These multiple positive findings make PEDF an ideal endogenous and safe biological for possible future clinical testing.

Advanced In Vitro HepaRG Culture Systems for Xenobiotic Metabolism and Toxicity Characterization.

Several HepaRG three-dimensional (3D) in vitro model systems have been developed to improve the predictability of xenobiotic metabolism and toxicity. In this review, we present a detailed summary and critique of the performance of various HepaRG 3D models compared to the conventional 2D monolayer culture. HepaRG 3D models can be broadly categorized into (1) scaffold-free, (2) scaffold-based, and (3) bioartificial liver (BAL) models. With respect to the scaffold-free configurations, the hanging drop model closely mimics the normal physiological function and metabolic profile of the liver. The micromold model is suitable for high-throughput multiplexed assays and exhibits higher accuracy when predicting drug-induced liver toxicity risk in both acute and chronic culture. Scaffold- and BAL-based models also present improved precision and accuracy for hepatotoxic drug screening in addition to allowing improved model control to closely mimic physiological assay conditions. Overall, all 3D HepaRG models exhibit improved cellular function, metabolic activity, and toxicity screening ability compared to the conventional 2D monolayer culture. These improvements reported in 3D models may be due to a higher degree of differentiation and cell polarity. Nevertheless, the expression and functions of various phase II, phase III, and nuclear receptors need to be further characterized in these 3D models.

DOX-Vit D, a Novel Doxorubicin Delivery Approach, Inhibits Human Osteosarcoma Cell Proliferation by Inducing Apoptosis While Inhibiting Akt and mTOR Signaling Pathways.

Doxorubicin (DOX) is a very potent and effective anticancer agent. However, the effectiveness of DOX in osteosarcoma is usually limited by the acquired drug resistance. Recently, Vitamin D (Vit-D) was shown to suppress the growth of many human cancer cells. Taken together, we synthesized DOX-Vit D by conjugating Vit-D to DOX in order to increase the delivery of DOX into cancer cells and mitigate the chemoresistance associated with DOX. For this purpose, MG63 cells were treated with 10 µM DOX or DOX-Vit D for 24 h. Thereafter, MTT, real-time PCR and western blot analysis were used to determine cell proliferation, genes and proteins expression, respectively. Our results showed that DOX-Vit D, but not DOX, significantly elicited an apoptotic signal in MG63 cells as evidenced by induction of death receptor, Caspase-3 and BCLxs genes. Mechanistically, the DOX-Vit D-induced apoptogens were credited to the activation of p-JNK and p-p38 signaling pathway and the inhibition of proliferative proteins, p-Akt and p-mTOR. Our findings propose that DOX-Vit D suppressed the growth of MG63 cells by inducing apoptosis while inhibiting cell survival and proliferative signaling pathways. DOX-Vit D may serve as a novel drug delivery approach to potentiate the delivery of DOX into cancer cells.

Alginate Bead-Encapsulated PEDF Induces Ectopic Bone Formation In Vivo in the Absence of Co-Administered Mesenchymal Stem Cells.

BACKGROUND: Bone defects can be severely debilitating and reduce quality of life. Osteoregeneration can alleviate some of the complications in bony defects. For therapeutic use in future, a single factor that can cause potent bone regeneration is highly preferred as it will be more costeffective, any off-target effects will be more easily monitored and potentially managed, and for ease of administration which would lead to better patient compliance and satisfaction. OBJECTIVE: We demonstrate that pigment epithelium-derived factor (PEDF), one such factor that is known to be potent against angiogenesis, promotes osteoblastogenesis in mesenchymal stem cells in vitro, but does not need co-encapsulation of cells in alginate bead scaffolds for osteogeneration in vivo. RESULTS: Osteogenic differentiation by PEDF in vitro was confirmed with immunoblotting and immunocytochemical staining for bone markers (alkaline phosphatase, osteocalcin, osteopontin, collagen I), calcified mineral deposition, and assay for alkaline phosphatase activity. PEDF-mediated bone formation in a muscle pocket in vivo model was confirmed by microcomputed tomography (microCT), histology (haematoxylin and eosin, Alcian blue staining), immunostaining for bone markers and for collagen I-processing proteins (heat shock protein 47 and membrane type I matrix metalloproteinase). CONCLUSION: PEDF therefore presents itself as a promising biological for osteogeneration.

Urinary Bone Turnover Markers as Target Indicators for Monitoring Bisphosphonate Drug Treatment in the Management of Osteoporosis.

BACKGROUND: Osteoporosis is a debilitating disease characterized by bone micro-architecture degradation contributing to fragility fractures. Currently, determining bone mineral density (BMD) via dual-energy X-ray absorptiometry (DXA) is the most reliable form of diagnosing osteoporosis and managing pharmacological treatment regimens. However, changes in BMD occur slowly (i.e., several months) and DXA does not reflect the metabolic rate of bone turnover. Alternatively, biochemical bone turnover markers are metabolic indicators released into serum and/or urine, and their quantity reflects the metabolic activity of bone. bone turnover markers show a rapid response following antiresorptive drug administration, and enzyme-linked immunosorbent assays (ELISA) have been established and used in clinical trials to help assess and predict fracture risk independent of BMD. OBJECTIVE: This review highlights various established bone turnover markers that have found utility in the clinic as reliable and standardized indicators of bone turnover, with attention to those used to assess efficacy of bisphosphonate drug therapy - particularly in monitoring medication adherence in patients with postmenopausal osteoporosis. RESULTS: We posit that the use of urinary bone turnover markers values determined by immunoassay or ELISA at routine clinic visits might serve as valuable feedback to healthcare professionals and patients to help monitor the efficacy and adherence of bisphosphonate therapy and disease progression. CONCLUSION: Our belief is that when assessed in combination with an algorithm of independent risk factors, measuring urinary bone turnover markers using a point of care kit may find utility in the osteoporosis clinic as an accessible, non-invasive and cost-effective alternative for the routine assessment of efficacy of bisphosphonate therapies.