TiO2 Nanocoatings with Controllable Crystal Type and Nanoscale Topography on Zirconia Implants to Accelerate Bone Formation

AUTHORS

Nan Li, Zhichao Liu, Guanqi Liu, Zhi Wang, Xianwei Guo, Chuanbin Guo, Jianmin Han

ABSTRACT

In dentistry, zirconia implants have emerged as a promising alternative for replacing missing teeth due to their superior aesthetic performance and chemical stability. To improve the osseointegration of zirconia implants, modifying their surface with hierarchical micro/nanotopography and bioactive chemical composition are two effective ways. In this work, a microscale topography was prepared on a zirconia surface using hydrofluoric acid etching, and then a 50 nm TiO2 nanocoating was deposited via atomic layer deposition (ALD). Subsequently, an annealing treatment was used to transform the TiO2 from amorphous to anatase and simultaneously generate nanoscale topography. Various investigations into the coating surface morphology, topography, wettability, and chemical composition were carried out using scanning electron microscopy, white light interferometry, contact-angle measurement, X-ray diffraction, and X-ray photoelectron spectroscopy. In addition, in vitro cytocompatibility and osteogenic potential performance of the coatings were evaluated by human bone marrow mesenchymal stem cells (hBMSCs), and in vivo osseointegration performance was assessed in a rat femoral condyle model. Moreover, the possible mechanism was also investigated. The deposition of TiO2 film with/without annealing treatment did not alter the microscale roughness of the zirconia surface, whereas the nanotopography changed significantly after annealing. The in vitro studies revealed that the anatase TiO2 coating with regular wavelike nanostructure could promote the adhesion and proliferation of osteoblasts and further improve the osteogenic potential in vitro and osseointegration in vivo. These positive effects may be caused by nanoscale topography via the canonical Wnt/β-catenin pathway. The results suggest that using ALD in combination with annealing treatment to fabricate a nanotopographic TiO2 coating is a promising way to improve the osteogenic properties of zirconia implants.

Reduced bone mass in collagen prolyl 4-hydroxylase P4ha1+/-;P4ha2-/- compound mutant mice

AUTHORS

Jussi-Pekka Tolonen, Antti M. Salo, Mikko Finnilä, Ellinoora Aro, Emma Karjalainen, Veli-Pekka Ronkainen, Kati Drushinin, Christophe Merceron, Valerio Izzi, Ernestina Schipani, Johanna Myllyharju

ABSTRACT

Proper deposition of the extracellular matrix and its major components, the collagens, is essential for endochondral ossification and bone mass accrual. Collagen prolyl 4-hydroxylases (C-P4Hs) hydroxylate proline residues in the -X-Pro-Gly- repeats of all known collagen types. Their product, 4-hydroxyproline, is essential for correct folding and thermal stability of the triple-helical collagen molecules in physiological body temperatures. We have previously shown that inactivation of the mouse P4ha1 gene, which codes for the catalytic α subunit of the major C-P4H isoform, is embryonic lethal, while inactivation of the P4ha2 gene produced only a minor phenotype. Instead, mice with a haploinsufficiency of the P4ha1 gene combined with a homozygous deletion of the P4ha2 gene present with a moderate chondrodysplasia due to transient cell death of the growth plate chondrocytes. Here, to further characterize the bone phenotype of the P4ha1+/-;P4ha2-/- mice, we have carried out gene expression analyses at whole tissue and single cell levels, biochemical analyses, microcomputed tomography and histomorphometric analyses and second harmonic generation microscopy to show that C-P4H α subunit expression peaks early and that the C-P4H deficiency leads to reduced collagen amount, a reduced rate of bone formation and a loss of trabecular and cortical bone volume in the long bones. The total osteoblast number in the proximal P4ha1+/-;P4ha2-/- tibia and the C-P4H activity in primary P4ha1+/-;P4ha2-/- osteoblasts were reduced, while the population of osteoprogenitor colony forming-unit fibroblasts was increased in the P4ha1+/-;P4ha2-/- marrow. Thus, the P4ha1+/-;P4ha2-/- mouse model recapitulates key aspects of a recently recognized congenital connective tissue disorder with short stature and bone dysplasia caused by bi-allelic variants of the human P4HA1 gene. Altogether, the data demonstrate the allele-dose dependent importance of the C-P4Hs to the developing organism and a threshold effect of C-P4H activity in the proper production of bone matrix.

Bone volume, mineral density, and fracture risk after kidney transplantation

AUTHORS

Satu Keronen, Leena Martola, Patrik Finne, Inari S. Burton, Xiaoyu F. Tong, Heikki Kröger, Eero Honkanen

ABSTRACT

Background

Disordered mineral metabolism reverses incompletely after kidney transplantation in numerous patients. Post-transplantation bone disease is a combination of pre-existing chronic kidney disease and mineral disorder and often evolving osteoporosis. These two frequently overlapping conditions increase the risk of post-transplantation fractures.

Material and methods

We studied the prevalence of low bone volume in bone biopsies obtained from kidney transplant recipients who were biopsied primarily due to the clinical suspicion of persistent hyperparathyroidism between 2000 and 2015 at the Hospital District of Helsinki and Uusimaa. Parameters of mineral metabolism, results of dual-energy x-ray absorptiometry scans, and the history of fractures were obtained concurrently.

One hundred nine bone biopsies taken at a median of 31 (interquartile range, IQR, 18–70) months after transplantation were included in statistical analysis. Bone turnover was classified as high in 78 (72%) and normal/low in 31 (28%) patients. The prevalence of low bone volume (n = 47, 43%) was higher among patients with low/normal turnover compared to patients with high turnover [18 (58%) vs. 29 (37%), P = 0.05]. Thirty-seven fragility fractures in 23 (21%) transplant recipients corresponding to fracture incidence 15 per 1000 person-years occurred during a median follow-up 9.1 (IQR, 6.3–12.1) years. Trabecular bone volume did not correlate with incident fractures. Accordingly, low bone mineral density at the lumbar spine correlated with low trabecular bone volume, but not with incident fractures. The cumulative corticosteroid dose was an important determinant of low bone volume, but not of incident fractures.

Conclusions

Despite the high prevalence of trabecular bone loss among kidney transplant recipients, the number of fractures was limited. The lack of association between trabecular bone volume and fractures suggests that the bone cortical compartment and quality are important determinants of bone strength and post-transplantation fracture.

Conditional loss of IKKα in Osterix + cells has no effect on bone but leads to age-related loss of peripheral fat

AUTHORS

Jennifer L. Davis, Nitin Kumar Pokhrel, Linda Cox, Nidhi Rohatgi, Roberta Faccio & Deborah J. Veis

ABSTRACT

NF-κB has been reported to both promote and inhibit bone formation. To explore its role in osteolineage cells, we conditionally deleted IKKα, an upstream kinase required for non-canonical NF-κB activation, using Osterix (Osx)-Cre. Surprisingly, we found no effect on either cancellous or cortical bone, even following mechanical loading. However, we noted that IKKα conditional knockout (cKO) mice began to lose body weight after 6 months of age with severe reductions in fat mass and lower adipocyte size in geriatric animals. qPCR analysis of adipogenic markers in fat pads of cKO mice indicated no difference in early differentiation, but instead markedly lower leptin with age. We challenged young mice with a high fat diet finding that cKO mice gained less weight and showed improved glucose metabolism. Low levels of recombination at the IKKα locus were detected in fat pads isolated from old cKO mice. To determine whether recombination occurs in adipocytes, we examined fat pads in Osx-Cre;TdT reporter mice; these showed increasing Osx-Cre-mediated expression in peripheral adipocytes from 6 weeks to 18 months. Since Osx-Cre drives recombination in peripheral adipocytes with age, we conclude that fat loss in cKO mice is most likely caused by progressive deficits of IKKα in adipocytes

Skeletal muscle mitoribosomal defects are linked to low bone mass caused by bone marrow inflammation in male mice

AUTHORS

Jingwen Tian, Hyo Kyun Chung, Ji Sun Moon, Ha Thi Nga, Ho Yeop Lee, Jung Tae Kim, Joon Young Chang, Seul Gi Kang, Dongryeol Ryu, Xiangguo Che, Je-Yong Choi, Masayuki Tsukasaki, Takayoshi Sasako, Sang-Hee Lee, Minho Shong, Hyon-Seung Yi

ABSTRACT

Background

Mitochondrial oxidative phosphorylation (OxPhos) is a critical regulator of skeletal muscle mass and function. Although muscle atrophy due to mitochondrial dysfunction is closely associated with bone loss, the biological characteristics of the relationship between muscle and bone remain obscure. We showed that muscle atrophy caused by skeletal muscle-specific CR6-interacting factor 1 knockout (MKO) modulates the bone marrow (BM) inflammatory response, leading to low bone mass.

Methods

MKO mice with lower muscle OxPhos were fed a normal chow or high-fat diet and then evaluated for muscle mass and function, and bone mineral density. Immunophenotyping of BM immune cells was also performed. BM transcriptomic analysis was used to identify key factors regulating bone mass in MKO mice. To determine the effects of BM-derived CXCL12 (C–X–C motif chemokine ligand 12) on regulation of bone homeostasis, a variety of BM niche-resident cells were treated with recombinant CXCL12. Vastus lateralis muscle and BM immune cell samples from 14 patients with hip fracture were investigated to examine the association between muscle function and BM inflammation.

Results

MKO mice exhibited significant reductions in both muscle mass and expression of OxPhos subunits but increased transcription of mitochondrial stress response-related genes in the extensor digitorum longus (P < 0.01). MKO mice showed a decline in grip strength and a higher drop rate in the wire hanging test (P < 0.01). Micro-computed tomography and von Kossa staining revealed that MKO mice developed a low mass phenotype in cortical and trabecular bone (P < 0.01). Transcriptomic analysis of the BM revealed that mitochondrial stress responses in skeletal muscles induce an inflammatory response and adipogenesis in the BM and that the CXCL12–CXCR4 (C–X–C chemokine receptor 4) axis is important for T-cell homing to the BM. Antagonism of CXCR4 attenuated BM inflammation and increased bone mass in MKO mice. In humans, patients with low body mass index (BMI = 17.2 ± 0.42 kg/m2) harboured a larger population of proinflammatory and cytotoxic senescent T-cells in the BMI (P < 0.05) and showed reduced expression of OxPhos subunits in the vastus lateralis, compared with controls with a normal BMI (23.7 ± 0.88 kg/m2) (P < 0.01).

Conclusions

Defects in muscle mitochondrial OxPhos promote BM inflammation in mice, leading to decreased bone mass. Muscle mitochondrial dysfunction is linked to BM inflammatory cytokine secretion via the CXCL12–CXCR4 signalling axis, which is critical for inducing low bone mass.

Probabilistic model for cattail and canola fibers: effect of environmental conditions, structural parameters, fiber length, and estimators

AUTHORS

Md Shadhin, Danny Mann, Mashiur Rahman

ABSTRACT

Biomass fibers are being widely investigated for industrial applications as an alternative to synthetic fibers using a standard humidity condition. In this study, the mechanical properties of two waste biomass fibers – canola and cattail – have been investigated when subjected to different environmental conditions, fiber length, and type of estimators used during analysis. The effect of different environmental conditions and structural variations were investigated by measuring the tensile properties after exposing them to eight different relative humidity conditions using a fixed fiber length of 25 mm. Further investigation was conducted using fiber lengths of 25, 35 and 45 mm using the most conservative relative humidity condition. The data were analyzed by a Weibull distribution model using four different estimators. The results revealed that Weibull strength (σavg) and modulus (Eavg) closely followed experimental values for cattail and canola fibers. The different relative humidity conditions and fiber lengths resulted in different Weibull parameters with 11% relative humidity and the mean rank estimator predicted the most conservative tensile strength for both waste biomass fibers. The experimental and characteristic Weibull strength decreased when fiber gauge length increased from 25 to 45 mm. The tensile strength and modulus of both waste biomass fibers at 50% reliability lie within the range of average experimental values. However, these values are reduced to 155 MPa (strength) and 20 GPa (modulus) for cattail fiber at 90% reliability. The survival probability of the tensile strength and modulus were found to be the highest at 75% and 100% relative humidity for cattail and canola fibers, respectively.