Cigarette smoke-associated inflammation impairs bone remodeling through NFκB activation

AUTHORS

Yi Lu, Yuanpu Peter Di, Ming Chang, Xin Huang, Qiuyan Chen, Ni Hong, Beth A. Kahkonen, Marissa E. Di, Chunyan Yu, Evan T. Keller & Jian Zhang

ABSTRACT

Background

Cigarette smoking constitutes a major lifestyle risk factor for osteoporosis and hip fracture. It is reported to impair the outcome of many clinical procedures, such as wound infection treatment and fracture healing. Importantly, although several studies have already demonstrated the negative correlation between cigarette consume and impaired bone homeostasis, there is still a poor understanding of how does smoking affect bone health, due to the lack of an adequately designed animal model. Our goal was to determine that cigarette smoke exposure impairs the dynamic bone remodeling process through induction of bone resorption and inhibition of bone formation.

Methods

We developed cigarette smoke exposure protocols exposing mice to environmental smoking for 10 days or 3 months to determine acute and chronic smoke exposure effects. We used these models, to demonstrate the effect of smoking exposure on the cellular and molecular changes of bone remodeling and correlate these early alterations with subsequent bone structure changes measured by microCT and pQCT. We examined the bone phenotype alterations in vivo and ex vivo in the acute and chronic smoke exposure mice by measuring bone mineral density and bone histomorphometry. Further, we measured osteoclast and osteoblast differentiation gene expression levels in each group. The function changes of osteoclast or osteoblast were evaluated.

Results

Smoke exposure caused a significant imbalance between bone resorption and bone formation. A 10-day exposure to cigarette smoke sufficiently and effectively induced osteoclast activity, leading to the inhibition of osteoblast differentiation, although it did not immediately alter bone structure as demonstrated in mice exposed to smoke for 3 months. Cigarette smoke exposure also induced DNA-binding activity of nuclear factor kappaB (NFκB) in osteoclasts, which subsequently gave rise to changes in bone remodeling-related gene expression.

Conclusions

Our findings suggest that smoke exposure induces RANKL activation-mediated by NFκB, which could be a “smoke sensor” for bone remodeling.

Simvastatin loaded chitosan guided bone regeneration membranes stimulate bone healing

AUTHORS

Vishnu Priya Murali, Fernanda D. Guerra, Najib Ghadri, James M. Christian, Sidney H. Stein, Jessica A. Jennings, Richard A. Smith, Joel D. Bumgardner

ABSTRACT

Background and Objective

Electrospun chitosan membranes (ESCM) modified with short-chain fatty acids have the ability to control the release of simvastatin (SMV), an anti-cholesterol drug with osteogenic potential, for guided bone regeneration (GBR) applications. This study evaluated in vivo osteogenic effects of rapid short release of SMV (4 weeks) vs long sustained release (8 weeks) from acetic anhydride (AA)—and hexanoic anhydride (HA)-modified ESCMs, respectively.

Methods

AA ESCMs loaded with 10 or 50 µg SMV and HA ESCMs loaded with 50 µg SMV were evaluated for biocompatibility and bone formation at 4 and 8 weeks, in 5 mm critical size rat calvarial defects, using histological evaluation and micro-CT analysis.

Results

No severe inflammatory response was noticed around the ESCMs. Less hydrophobic AA membranes showed signs of resorption by week 4 and were almost completely resorbed by week 8 whereas the more hydrophobic HA membranes resorbed slowly, remaining intact over 8 weeks. In micro-CT analysis, 10 µg SMV-loaded AA membranes did not show significant bone formation as compared to non-loaded AA membranes at either evaluation time points. 50 µg SMV-loaded AA membranes stimulated significantly more bone formation than non-loaded AA membranes by week 4 (%bone = 31.0 ± 5.9% (AA50) vs 18.5 ± 13.7% (AA0)) but showed no difference at week 8. HA membranes with 50 µg SMV showed significantly more bone formation as compared to corresponding non-loaded membranes by week 8 (%bone = 61.7 ± 8.9% (HA50) vs 33.9 ± 29.7% (HA0)), though such an effect was not significant at week 4.

Conclusion

These results indicate that modified ESCMs may be used to control the release of SMV and promote bone healing in GBR applications.

The Effects of Photobiomodulation on Leukocyte and Platelet-Rich Fibrin as Barrier Membrane on Bone Regeneration: An Experimental Animal Study

AUTHORS

Seren Surmeli Baran, Andy Temmerman, Fariz Salimov, Onur Ucak Turer, Tugce Sapmaz, Mehmet Cenk Haytac, and Mustafa Ozcan

ABSTRACT

Objective: To compare the effects of leukocyte and platelet-rich fibrin (L-PRF) and photobiomodulation therapy (PBMT)-applied L-PRF (PBMT/L-PRF) as barrier membranes on new bone formation (BV/TV) for the treatment of critical-sized bone defects.

Materials and methods: The right iliac crests of five sheep were used in this experimental animal study. Eight critical-sized defects were surgically created in each sheep and a total of 40 defects were obtained. A deproteinized bovine bone graft was placed in all defects, and the defects were divided into four groups to be covered with L-PRF membrane, PBMT/L-PRF membrane, collagen membrane, or left uncovered as controls. Animals were sacrificed at 1 month. The sections obtained were histomorphometrically analyzed.

Results: The results showed that the collagen group presented significantly higher values for main bone healing parameters (BV/TV, bone volume, and bone surface; p < 0.05). The PBMT/L-PRF group presented higher values than the L-PRF group and controls for these parameters though not statistically significant (p > 0.05).

Conclusions: The findings show that PBMT may provide additional regenerative properties to L-PRF when used as barrier membranes. However, these results did not reach the collagen membranes, which warrants further studies for adapting the laser parameters to increase regenerative capacity of L-PRF.

Collagenated Porcine Heterologous Bone Grafts: Histomorphometric Evaluation of Bone Formation Using Different Physical Forms in a Rabbit Cancellous Bone Model

AUTHORS

Rui I. Falacho, Paulo J. Palma, Joana A. Marques, Maria H. Figueiredo, Francisco Caramelo, Isabel Dias, Carlos Viegas, Fernando Guerra

ABSTRACT

Collagenated porcine-derived bone graft materials exhibit osteoconductive properties and the development of different formulations intends to enhance bone regeneration. This study aims to evaluate bone healing in a rabbit cancellous bone defect in response to grafting with different physicochemical forms of heterologous porcine bone. Twenty-six adult male New Zealand White rabbits received two critical size femoral bone defects per animal (n = 52), each randomly assigned to one of the five tested materials (Apatos, Gen-Os, mp3, Putty, and Gel 40). Animals were sacrificed at 15- and 30-days post-surgery. Qualitative and quantitative (new bone, particle and connective tissue percentages) histological analyses were performed. Histomorphometry showed statistically significant differences in all evaluated parameters between mp3 and both Putty and Gel 40 groups, regardless of the timepoint (p < 0.05). Moreover, statistical differences were observed between Apatos and both Putty (p = 0.014) and Gel 40 (p = 0.007) groups, at 30 days, in regard to particle percentage. Within each group, regarding new bone formation, mp3 showed significant differences (p = 0.028) between 15 (40.93 ± 3.49%) and 30 (52.49 ± 11.04%) days. Additionally, intragroup analysis concerning the percentage of particles revealed a significant reduction in particle occupied area from 15 to 30 days in mp3 and Gen-Os groups (p = 0.009). All mp3, Gen-Os and Apatos exhibited promising results in terms of new bone formation, thus presenting suitable alternatives to be used in bone regeneration.

Ablation of Enpp6 Results in Transient Bone Hypomineralization

AUTHORS

Scott Dillon, Karla Suchacki, Shun-Neng Hsu, Louise A Stephen, Rongling Wang, William P Cawthorn, Alan J Stewart, Fabio Nudelman, Nicholas M Morton, Colin Farquharson

ABSTRACT

Biomineralization is a fundamental process key to the development of the skeleton. The phosphatase orphan phosphatase 1 (PHOSPHO1), which likely functions within extracellular matrix vesicles, has emerged as a critical regulator of biomineralization. However, the biochemical pathways that generate intravesicular PHOSPHO1 substrates are currently unknown. We hypothesized that the enzyme ectonucleotide pyrophosphatase/phosphodiesterase 6 (ENPP6) is an upstream source of the PHOSPHO1 substrate. To test this, we characterized skeletal phenotypes of mice homozygous for a targeted deletion of Enpp6 (Enpp6−/−). Micro-computed tomography of the trabecular compartment revealed transient hypomineralization in Enpp6−/− tibias (p < 0.05) that normalized by 12 weeks of age. Whole-bone cortical analysis also revealed significantly hypomineralized proximal bone in 4- but not 12-week-old Enpp6−/− mice (p < 0.05) compared with WT animals. Back-scattered SEM revealed a failure in 4-week-old trabecular bone of mineralization foci to propagate. Static histomorphometry revealed increased osteoid volume (p > 0.01) and osteoid surface (p < 0.05), which recovered by 12 weeks but was not accompanied by changes in osteoblast or osteoclast number. This study is the first to characterize the skeletal phenotype of Enpp6−/− mice, revealing transient hypomineralization in young animals compared with WT controls. These data suggest that ENPP6 is important for bone mineralization and may function upstream of PHOSPHO1 as a novel means of generating its substrates inside matrix vesicles.