Difference In Bone Formation Research Articles

Protein phosphatase SCP4 regulates cartilage development and endochondral osteogenesis via FoxO3a dephosphorylation.

The regulatory mechanisms involved in embryonic development are complex and yet remain unclear. SCP4 represents a novel nucleus-resident phosphatase identified in our previous study. The primary aim of this study was to elucidate the function of SCP4 in the progress of cartilage development and endochondral osteogenesis. SCP4-/- and SCP4Col2ER mice were constructed to assess differences in bone formation using whole skeleton staining. ABH/OG staining was used to compare chondrocyte differentiation and cartilage development. Relevant biological functions were analysed using RNA-sequencing and GO enrichment, further validated by immunohistochemical staining, Co-IP and Western Blot. Global SCP4 knockout led to abnormal embryonic development in SCP4-/- mice, along with delayed endochondral osteogenesis. In parallel, chondrocyte-specific removal of SCP4 yielded more severe embryonic deformities in SCP4Col2ER mice, including limb shortening, reduced chondrocyte number in the growth plate, disorganisation and cell enlargement. Moreover, RNA-sequencing analysis showed an association between SCP4 and chondrocyte apoptosis. Notably, Tunnel-positive cells were indeed increased in the growth plates of SCP4Col2ER mice. The deficiency of SCP4 up-regulated the expression levels of pro-apoptotic proteins both in vivo and in vitro. Additionally, phosphorylation of FoxO3a (pFoxO3a), a substrate of SCP4, was heightened in chondrocytes of SCP4Col2ER mice growth plate, and the direct interaction between SCP4 and pFoxO3a was further validated in chondrocytes. Our findings underscore the critical role of SCP4 in regulating cartilage development and endochondral osteogenesis during embryonic development partially via inhibition of chondrocytes apoptosis regulated by FoxO3a dephosphorylation.

Impact of PEG sensitization on the efficacy of PEG hydrogel-mediated tissue engineering

While poly(ethylene glycol) (PEG) hydrogels are generally regarded as biologically inert blank slates, concerns over PEG immunogenicity are growing, and the implications for tissue engineering are unknown. Here, we investigate these implications by immunizing mice against PEG to stimulate anti-PEG antibody production and evaluating bone defect regeneration after treatment with bone morphogenetic protein-2-loaded PEG hydrogels. Quantitative analysis reveals that PEG sensitization increases bone formation compared to naive controls, whereas histological analysis shows that PEG sensitization induces an abnormally porous bone morphology at the defect site, particularly in males. Furthermore, immune cell recruitment is higher in PEG-sensitized mice administered the PEG-based treatment than their naive counterparts. Interestingly, naive controls that were administered a PEG-based treatment also develop anti-PEG antibodies. Sex differences in bone formation and immune cell recruitment are also apparent. Overall, these findings indicate that anti-PEG immune responses can impact tissue engineering efficacy and highlight the need for further investigation.

Comparative analysis of the in vivo kinetic properties of various bone substitutes filled into a peri-implant canine defect model.

Deproteinized bovine bone or synthetic hydroxyapatite are 2 prevalent bone grafting materials used in the clinical treatment of peri-implant bone defects. However, the differences in bone formation among these materials remain unclear. This study evaluated osteogenesis kinetics in peri-implant defects using 2 types of deproteinized bovine bone (Bio-Oss® and Bio-Oss/Collagen®) and 2 types of synthetic hydroxyapatite (Apaceram-AX® and Refit®). We considered factors including newly generated bone volume; bone, osteoid, and material occupancy; and bone-to-implant contact. A beagle model with a mandibular defect was created by extracting the bilateral mandibular third and fourth premolars. Simultaneously, an implant was inserted into the defect, and the space between the implant and the surrounding bone walls was filled with Bio-Oss, Bio-Oss/Collagen, Apaceram-AX, Refit, or autologous bone. Micro-computed tomography and histological analyses were conducted at 3 and 6 months postoperatively (Refit and autologous bone were not included at the 6-month time point due to their rapid absorption). All materials demonstrated excellent biocompatibility and osteoconductivity. At 3 months, Bio-Oss and Apaceram-AX exhibited significantly greater volumes of formation than the other materials, with Bio-Oss having a marginally higher amount. However, this outcome was reversed at 6 months, with no significant difference between the 2 materials at either time point. Apaceram-AX displayed notably slower bioresorption and the largest quantity of residual material at both time points. In contrast, Refit had significantly greater bioresorption, with complete resorption and rapid maturation involving cortical bone formation at the crest at 3 months, Refit demonstrated the highest mineralized tissue and osteoid occupancy after 3 months, albeit without statistical significance. Overall, the materials demonstrated varying post-implantation behaviors in vivo. Thus, in a clinical setting, both the properties of these materials and the specific conditions of the defects needing reinforcement should be considered to identify the most suitable material.

Dietary restriction inhibits bone formation in alveolar bone modeling and remodelling

Objective: Methods: Balb/C mice weaned at the age of 21 days were assigned to one of the following groups: control group fed a regular hard diet ad libitum (food consumption was measured daily), and dietary restricted group (DR) received 75% of the amount of food consumed by the respective control mice the previous day. Body weight of all animals was recorded throughout. Mice were euthanized in groups of ten at 25 and 60 days of experience. Mandibles were dissected, descalcified in EDTA and embedded in paraffin. Buccolingual sections of the mesial root of the first lower molar were stained with hematoxylin-eosin and submitted to histomorphometric studies. Results: At 25 days, DR mice shows bone formation values lower than control animals for modeling and remodeling sides, coupled to an increase with bone at rest values in the modeling side. At 60 days, DR mice shows bone formation values lower than control animals in the modeling side linked with a similar increase in values corresponding to bone resorption and bone at rest. In the remodeling side, no differences in bone formation were observed between control and DR mice. However, high bone resorption and a decrease in bone at rest areas were observed. Conclusions: Dietery restriction impaires bone formation in physiologic alveolar bone modeling and remodeling.

Bone Structure and Turnover in Postmenopausal Women With Long-Standing Type 1 Diabetes.

Compromised bone structural and mechanical properties are implicated in the increased fracture risk in type 1 diabetes (T1D). We investigated bone structure and turnover by histomorphometry in postmenopausal women with T1D and controls without diabetes using tetracycline double-labeled transiliac bone biopsy. After in vivo tetracycline double labeling, postmenopausal women with T1D of at least 10 years and without diabetes underwent transiliac bone biopsy. An expert blinded to the study group performed histomorphometry. Static and dynamic histomorphometry measurements were performed and compared between the two groups. The analysis included 9 postmenopausal women with T1D (mean age 58.4 ± 7.1 years with 37.9 ± 10.9 years of diabetes and HbA1c 7.1% ± 0.4%) and 7 postmenopausal women without diabetes (mean age 60.9 ± 3.3 years and HbA1c 5.4% ± 0.2%). There were no significant differences in serum PTH (38.6 ± 8.1 versus 51.9 ± 23.9 pg/mL), CTX (0.4 ± 0.2 versus 0.51 ± 0.34 ng/mL), or P1NP (64.5 ± 26.2 versus 87.3 ± 45.3 ng/mL). Serum 25-hydroxyvitamin D levels were higher in T1D than in controls (53.1 ± 20.8 versus 30.9 ± 8.2 ng/mL, p < 0.05). Bone structure metrics (bone volume, trabecular thickness, trabecular number, and cortical thickness) were similar between the groups. Indices of bone formation (osteoid volume, osteoid surface, and bone formation rate) were 40% lower in T1D and associated with lower activation frequency. However, the differences in bone formation were not statistically significant. Long-standing T1D may affect bone turnover, mainly bone formation, without significantly affecting bone structure. Further research is needed to understand bone turnover and factors affecting bone turnover in people with T1D. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC. on behalf of American Society for Bone and Mineral Research.

Melatonin is more effective on bone metabolism when given at early night than during the day in ovariectomized rats

Melatonin has diverse effects, and has been reported to promote bone formation in addition to regulating the sleep–wake cycle. In the present study, we investigated the effects of melatonin on bone metabolism using ovariectomized (OVX) rats; a model of postmenopausal osteoporosis. Here, we focused on the differences in bone formation when melatonin was subcutaneous injected at day or early night. The OVX rats were injected with melatonin once daily (0.8 or 8 mg/head) between 11:00 to 14:00 or 18:00 to 19:30 for the day or early night, respectively, for six weeks. After completion of the injection, the femur and tibia in the OVX rats were dissected under general anesthesia and examined by quantitative computed tomography (pQCT) and histological analysis, respectively. Interestingly, the trabecular bone mineral density in the femur metaphysis of the OVX rats receiving 8 mg/head melatonin at early night was higher than those receiving melatonin during the day and they recovered to a similar level as the rats with sham treatment. In the diaphysis, the pQCT analysis results indicated that there was no significant difference in bone mineral density between the day and early night melatonin-injected OVX rats. Histological analysis of the secondary trabecular bone in the tibia of the OVX rats, revealed that the bone matrix area of the group receiving 8 mg/head melatonin at early night was higher compared with that of the day group and had a significant difference compared with OVX treatment rats. Taken together, the subcutaneous melatonin injection in OVX rats at early night was found to promote trabecular bone formation better than melatonin injection during the day. The timing of melatonin injection is a crucial factor when examining the influence of bone metabolism.

The combined effect of zinc and calcium on the biodegradation of ultrahigh-purity magnesium implants

Magnesium (Mg)-based implants are promising candidates for orthopedic interventions, because of their biocompatibility, good mechanical features, and ability to degrade completely in the body, eliminating the need for an additional removal surgery. In the present study, we synthesized and investigated two Mg-based materials, ultrahigh-purity ZX00 (Mg–Zn–Ca; <0.5 wt% Zn and <0.5 wt% Ca, in wt%; Fe-content <1 ppm) and ultrahigh-purity Mg (XHP-Mg, >99.999 wt% Mg; Fe-content <1 ppm), in vitro and in vivo in juvenile healthy rats to clarify the effect of the alloying elements Zn and Ca on mechanical properties, microstructure, cytocompatibility and degradation rate. Potential differences in bone formation and bone in-growth were also assessed and compared with state-of-the-art non-degradable titanium (Ti)-implanted, sham-operated, and control (non-intervention) groups, using micro-computed tomography, histology and scanning electron microscopy. At 6 and 24 weeks after implantation, serum alkaline phosphatase (ALP), calcium (Ca), and Mg level were measured and bone marrow stromal cells (BMSCs) were isolated for real-time PCR analysis. Results show that ZX00 implants have smaller grain size and superior mechanical properties than XHP-Mg, and that both reveal good biocompatibility in cytocompatibilty tests. ZX00 homogenously degraded with an increased gas accumulation 12 and 24 weeks after implantation, whereas XHP-Mg exhibited higher gas accumulation already at 2 weeks. Serum ALP, Ca, and Mg levels were comparable among all groups and both Mg-based implants led to similar relative expression levels of Alp, Runx2, and Bmp-2 genes at weeks 6 and 24. Histologically, Mg-based implants are superior for new bone tissue formation and bone in-growth compared to Ti implants. Furthermore, by tracking the sequence of multicolor fluorochrome labels, we observed higher mineral apposition rate at week 2 in both Mg-based implants compared to the control groups.Our findings suggest that (i) ZX00 and XHP-Mg support bone formation and remodeling, (ii) both Mg-based implants are superior to Ti implants in terms of new bone tissue formation and osseointegration, and (iii) ZX00 is more favorable due to its lower degradation rate and moderate gas accumulation.

&lt;i&gt;In Vivo&lt;/i&gt; Evaluation of Calcium-Phosphate Ceramics with Highly-Interconnected Pores Using Porcine Tibia Defect Model

The aim of this study was to compare the difference in bone formation between hydroxyapatite (HAp) and β-tricalcium phosphate (β-TCP) ceramics with similar porous structure. Porous HAp and β-TCP ceramics (total porosity: ~70%) with highly-interconnected structure were fabricated by firing calcium-phosphate fiber mixed with carbon beads (~150 μm). The size of highly-interconnected pores was measured by mercury intrusion technique. Each sample (ϕ4 mm × 8 mm) was implanted into the tibial bone defect (ϕ4.2 mm × 8 mm) of a pig (2 years old, male). After 6 weeks of implantation, undecalcified section was prepared and stained with hematoxylin and eosin (HE) stain. New bone area and remaining material area were evaluated.Interconnection sizes of HAp and β-TCP ceramics were 23.0 µm and 15.6 µm, respectively. New bone area of β-TCP ceramics was significantly higher compared to HAp ceramics. Remaining material ratio was 77.1 ± 3.3% for β-TCP ceramics whereas HAp ceramics remained stable. Bone formation in HAp ceramics was inhomogeneous compared to β-TCP ceramics in HE stained section. Although the β-TCP ceramics interconnection size was smaller, higher resorption may have played a positive role in homogeneous bone formation. According to the result of the study, it is suggested that highly-interconnected porous ceramics made by β-TCP could be a suitable material for bone regeneration.

Comparison of Macro-and Micro-porosity of a Titanium Mesh for Guided Bone Regeneration: An In Vivo Experimental Study.

Guided bone regeneration (GBR) is one of the surgical methods used for vertical ridge augmentation prior to dental implant placements. Titanium meshes have been used for osteogenic space maintenance in GBR sites by clinicians. We aimed to compare the influence of micropores and macropores in a titanium mesh on bone regeneration in a rat calvarial vertical GBR model. The calvaria of nine rats were exposed, and plastic cylinders were set bilaterally. Eighteen surgical sites were randomly allocated into three groups according to the materials of titanium lid and bone substitutes: microporous titanium lid+deproteinized bovine bone mineral (DBBM), macroporous titanium lid +DBBM, microporous titanium lid+carbonate apatite. Newly generated bone inside the cylinders was evaluated using micro-computed tomography (micro-CT). Furthermore, bone regeneration and angiogenesis were evaluated histologically at 12 weeks. Quantitative volumetric analyses using micro-CT showed a gradual increase in bone volume inside the cylinders in all three groups. Histological observation confirmed vigorous bone regeneration in the microporous groups compared to that in the macroporous group. In the upper part of the cylinders, soft tissue invaded the GBR site by passing through the pores of the macroporous mesh. The blood vessels in the upper part of the cylinders were smaller in the microporous groups than in the macroporous group. There was no difference in bone formation between cylinders filled with DBBM or carbonate apatite. Microvasculature penetrates 50-μm diameter micropores and accelerates bone formation inside the cylinder, which was set on rat calvaria. The microporous titanium mesh can facilitate angiogenesis from both the dura mater and periosteal in vertical ridge augmentation. Our data showed superiority of microporous titanium vascular permeability and osteoconductivity, supporting bone growth.

In vivo evaluation of bone regeneration behavior of novel β-tricalcium phosphate/layered double hydroxide nanocomposite granule as bone graft substitutes.

This study was based on in vivo assessment of bone regeneration capacity of synthesized porous β-tricalcium phosphate (β-TCP) nanocomposite granules and aimed to explore the effects of fabricated β-TCP granules reinforced with layered double hydroxides (LDH) nanoclay compared to β-TCP granules, in terms of osteoconductivity and biodegradability. Granules with diameters of 2-3 mm were implanted into cavities drilled in rabbit distal femur and were left in situ for up to 3months. The mechanical study demonstrated that the presence of LDH nanoparticles in β-TCP granules resulted in a significant increase in compressive modulus from 174.4 to 231.4MPa, while the porosity was constant at 76%-80%. The results revealed that the obtained granules showed no cytotoxicity. In this study, x-ray radiographic, micro-computed tomography, and histological staining analysis were taken to evaluate the percentage of bone ingrowth and biodegradability of the porous granules. The results exhibited that both granules support bone regeneration and also the amount of new bone formation in the bone defect filled with both granules was almost six times higher than the empty defects. Although no significant difference in bone formation for two different granules was observed, a higher biodegradability was detected in β-TCP granules in comparison to β-TCP/LDH granules. Overall, the addition of LDH nanoclay (10%) enhanced the physicochemical and mechanical properties of β-TCP granules while it is biological and osteoconductity properties have been maintained and its biodegradation rate has been decreased.

The sole and combined effects of gelatin-thrombin matrix and freeze-dried bone allograft on early bone healing.

Gelatin-thrombin matrix (GTM) is a hemostatic agent with applications in maxillofacial surgery consisting of human-derived thrombin and bovine-derived gelatin matrix. The aim of this study was to evaluate the efficacy of GTM alone or with freeze-dried bone allograft (FDBA) in improving early bone healing. Forty-six adult male Sprague-Dawley rats were used. All animals were randomly assigned to a control group (n = 4) and three study groups (n = 14). Each group was divided into two subgroups for histomorphometric and histological analyses at weeks 1 and 4. The new bone formation, inflammation, fibrosis, necrosis, foreign body reaction, and bone healing scores were evaluated based on the histopathological findings. Multiple comparisons were performed using the Kruskal-Wallis test. Variables that were not normally distributed were evaluated using the Mann-Whitney U test. At 1 week, the GTM + FDBA group showed less bone formation (mean ± SD: 0.08 ± 0.03 mm2), compared with the study and control groups (FDBA: 0.15 ± 0.06 mm2; GTM: 0.13 ± 0.06 mm2). At 4 weeks, the GTM group (0.48 ± 0.1 mm2) showed more bone formation than the GTM + FDBA group (0.33 ± 0.17 mm2). Foreign body reactions were observed at weeks 1 and 4 in all GTM-containing groups. Within the limitations of this study, GTM group did not show a significant difference in bone formation compared with the control group. GTM did not inhibit bone healing at 1 and 4 weeks, and no significant difference was observed compared with the control groups. GTM was more effective for bone healing when administered without FDBA. GTM is considered safe when bone hemorrhage is encountered.

Human osteoblasts obtained from distinct periarticular sites demonstrate differences in biological function in vitro.

AimsAccumulated evidence indicates that local cell origins may ingrain differences in the phenotypic activity of human osteoblasts. We hypothesized that these differences may also exist in osteoblasts harvested from the same bone type at periarticular sites, including those adjacent to the fixation sites for total joint implant components.MethodsHuman osteoblasts were obtained from the acetabulum and femoral neck of seven patients undergoing total hip arthroplasty (THA) and from the femoral and tibial cuts of six patients undergoing total knee arthroplasty (TKA). Osteoblasts were extracted from the usually discarded bone via enzyme digestion, characterized by flow cytometry, and cultured to passage three before measurement of metabolic activity, collagen production, alkaline phosphatase (ALP) expression, and mineralization.ResultsOsteoblasts from the acetabulum showed lower proliferation (p = 0.034), cumulative collagen release (p < 0.001), and ALP expression (p = 0.009), and produced less mineral (p = 0.006) than those from the femoral neck. Osteoblasts from the tibia produced significantly less collagen (p = 0.021) and showed lower ALP expression than those from the distal femur.ConclusionWe have demonstrated for the first time an anatomical regional variation in the biological behaviours of osteoblasts on either side of the hip and knee joint. The lower osteoblast proliferation, matrix production, and mineralization from the acetabulum compared to those from the proximal femur may be reflected in differences in bone formation and implant fixation at these sites.Cite this article: Bone Joint Res 2021;10(9):611–618.

Biological Evaluation of a New Sodium-Potassium Silico-Phosphate Glass for Bone Regeneration: In Vitro and In Vivo Studies.

In vitro and in vivo studies are fundamental steps in the characterization of new implantable materials to preliminarily assess their biological response. The present study reports the in vitro and in vivo characterizations of a novel experimental silicate bioactive glass (BG) (47.5B, 47.5SiO2-10Na2O-10K2O-10MgO-20CaO-2.5P2O5 mol.%). Cytocompatibility tests were performed using human mature osteoblasts (U2OS), human mesenchymal stem cells (hMSCs) and human endothelial cells (EA.hy926). The release of the early osteogenic alkaline phosphatase (ALP) marker suggested strong pro-osteogenic properties, as the amount was comparable between hMSCs cultivated onto BG surface and cells cultivated onto polystyrene control. Similarly, real-time PCR revealed that the osteogenic collagen I gene was overexpressed in cells cultivated onto BG surface without biochemical induction. Acute toxicity tests for the determination of the median lethal dose (LD50) allowed classifying the analyzed material as a slightly toxic substance with LD50 = 4522 ± 248 mg/kg. A statistically significant difference in bone formation was observed in vivo through comparing the control (untreated) group and the experimental one, proving a clear osteogenic effect induced by the implantation at the defect site. Complete resorption of 47.5B powder was observed after only 3 months in favor of newly formed tissue, thus confirming the high osteostimulatory potential of 47.5B glass.

Bone Conduction Capacity of Highly Porous 3D-Printed Titanium Scaffolds Based on Different Pore Designs.

In porous titanium scaffolds manufactured via 3D printing, the differences in bone formation according to pore design and implantation period were studied. Titanium scaffolds with three types of different pore structures (Octadense, Gyroid, and Dode) were fabricated via 3D printing using the selective laser melting method. Mechanical properties of scaffolds were investigated. Prepared specimens were inserted into both femurs of nine rabbits and their clinical characteristics were observed. Three animals were sacrificed at the 2nd, 4th, and 6th weeks, and the differences in bone formation were radiologically and histologically analyzed. The percentage of new bone and surface density in the pore structure were observed to be approximately 25% and 8 mm2/mm3, respectively. There was no difference in the amount of newly formed bone according to the pore design at 2, 4, and 6 weeks. In addition, no differences in the amount of newly formed bone were observed with increasing time within the same pore design for all three designs. During the 6-week observation period, the proportion of new bones in the 3D-printed titanium scaffold was approximately 25%. Differences in bone formation according to the pore design or implantation period were not observed.

Physical characterization of biphasic bioceramic materials with different granulation sizes and their influence on bone repair and inflammation in rat calvaria

Biphasic calcium phosphate bioceramics (BCP) consist of a mixture of hydroxyapatite (HA) and beta-tricalcium phosphate (β-TCP) within the same particle. Due to their osteoconductive properties, biocompatibility and resemblance to natural bone, these materials have become a promising and suitable alternative to autologous bone grafting. First, the topography characteristics, specific surface area, and total pore volume of BCP were evaluated using scanning electron microscopy and the BET and BJH methods. Next, this study aimed to evaluate the intensity of the inflammatory process and the bone neoformation capacity of various particle sizes of BCP in the repair of critical defects in the calvaria of rats. A xenogeneic biomaterial was used in the control group. After 30, 60, and 90 days, the animals were euthanized, followed by the processing of the samples to measure the intensity of inflammatory infiltrates and the areas of bone neoformation. Our results indicate that no considerable differences were observed in the inflammatory scores in sites treated with distinct BCP grain sizes. A greater area of bone neoformation was measured in the xenogeneic group at all analysis times, with no substantial differences in bone formation between the BCP particle size in the range of 250–500 µm and 500–1000 µm.

Local administration of allogeneic or autologous bone marrow-derived mesenchymal stromal cells enhances bone formation similarly in distraction osteogenesis

Background aimsDistraction osteogenesis (DO) is a surgical technique to promote bone regeneration that requires a long time for bone healing. Bone marrow-derived mesenchymal stromal cells (MSCs) have been applied to accelerate bone formation in DO. Allogeneic MSCs are attractive, as they could be ready to use in clinics. Whether allogeneic MSCs would have an effect similar to autologous MSCs with regard to promoting bone formation in DO is still unknown. This study compares the effect of autologous MSCs versus allogeneic MSCs on bone formation in a rat DO model. MethodsRat bone marrow-derived MSCs were isolated, characterized and expanded in vitro. Adult rats were subjected to right tibia transverse osteotomy. On the third day of distraction, each rat received one injection of phosphate-buffered saline (PBS), autologous MSCs or allogeneic MSCs at the distraction site. Tibiae were harvested after 28 days of consolidation for micro-computed tomography examination, mechanical test and histological analysis. ResultsResults showed that treatment with both allogeneic and autologous MSCs promoted bone formation, with significantly higher bone mass, mechanical properties and mineral apposition rate as well as expression of angiogenic and bone formation markers at the regeneration sites compared with the PBS-treated group. No statistical difference in bone formation was found between the allogeneic and autologous MSC treatment groups. ConclusionsThis study indicates that allogeneic and autologous MSCs have a similar effect on promoting bone consolidation in DO. MSCs from an allogeneic source could be used off-the-shelf with DO to achieve early bone healing.

Partial Replacement of Animal Proteins with Plant Proteins for 12 Weeks Accelerates Bone Turnover Among Healthy Adults: A Randomized Clinical Trial

BackgroundPlant-based diets may reduce the risk of chronic diseases, but can also lead to low calcium and vitamin D intakes, posing a risk for bone health. ObjectivesWe investigated whether partial replacement of animal proteins with plant-based proteins using a whole-diet approach affects bone and mineral metabolism in healthy adults in 3 groups fed diets differing in protein composition. MethodsThis 12-week clinical trial was comprised of 107 women and 29 men (20–69 years old; BMI mean ± SD, 24.8 ± 3.9) randomly assigned to consume 1 of 3 diets designed to provide 17 energy percent (E%) protein: “animal” (70% animal protein, 30% plant protein of total protein intake), “50/50” (50% animal, 50% plant), and “plant” (30% animal, 70% plant) diets. We examined differences in bone formation [serum intact procollagen type I amino-terminal propeptide (S-iPINP)], bone resorption [serum collagen type 1 cross-linked C-terminal telopeptide (S-CTX)], mineral metabolism markers (primary outcomes), and nutrient intakes (secondary outcomes) by ANOVA/ANCOVA. ResultsS-CTX was significantly higher in the plant group (mean ± SEM, 0.44 ± 0.02 ng/mL) than in the other groups (P values < 0.001 for both), and differed also between the animal (mean ± SEM, 0.29 ± 0.02 ng/mL) and 50/50 groups (mean ± SEM, 0.34 ± 0.02 ng/mL; P = 0.018). S-iPINP was significantly higher in the plant group (mean ± SEM, 63.9 ± 1.91 ng/mL) than in the animal group (mean ± SEM, 55.0 ± 1.82 ng/mL; P = 0.006). In a subgroup without a history of vitamin D supplement use, plasma parathyroid hormone was significantly higher in the plant than in the animal group (P = 0.018). Vitamin D and calcium intakes were below recommended levels in the plant group (mean ± SEM, 6.2 ± 3.7 μg/d and 733 ± 164 mg/d, respectively). ConclusionsPartial replacement of animal proteins with plant-based proteins for 12 weeks increased the markers of bone resorption and formation among healthy adults, indicating a possible risk for bone health. This is probably caused by lower vitamin D and calcium intakes from diets containing more plant-based proteins, but it is unclear whether differences in protein intake or quality play a major role. This trial was registered at http://www.clinicaltrials.gov as NCT03206827.

Leukocyte and Platelet-Rich Fibrin Have Same Effect as Blood Clot in the 3-Dimensional Alveolar Ridge Preservation. A Split-Mouth Randomized Clinical Trial

Leukocyte- and platelet-rich fibrin (L-PRF) has been used for alveolar ridge preservation (ARP) in postextraction tooth sockets. However, current reports have measured its effectiveness in linear measurements of 3-dimensional ridge preservation. The purpose of this study was to determine the effectiveness of the use of L-PRF filling versus natural clot blood healing in ARP according to the clinical, radiographic, and volumetric measurements of postextraction tooth sockets. A split-mouth randomized clinical trial was designed. Healthy patients who needed bilateral extraction of upper third molars were selected. After the tooth extraction, the socket was filled and distributed randomly with L-PRF and the contralateral socket only with the blood clot. The dimensional change of soft tissue healing around the sockets, and the length, depth, and difference of bone formation were examined using standardized periapical radiographs. Volumetric measurement variation of the sockets was evaluated by 3-dimensional scanning of dental casts. Changes of all measures were analyzed at 7 days (initial) and 3 months (final) after the tooth extraction and compared between both groups (t test; P<.05). Sixteen patients (aged 24.75±3.53years; 56.25% women) participated. Measurements of wound healing and the length, depth, and difference of bone formation were similar for both study groups at initial and final times. The calculation of initial-final volumetric socket variation was 15.45±13.12μL using L-PRF and 14.12±11.23μL using blood clot (P=.78). L-PRF filling showed the same dimensional and volumetric behavior as normal blood clot healing in the ARP of postextraction tooth sockets. Future investigations will have to analyze the use of surgical models and digital instruments in ARP techniques.

Evaluation of the bone morphology around four types of porous metal implants placed in distal femur of ovariectomized rats

BackgroundTo compare structural features of the femoral bone of ovariectomized and non-ovariectomized rats after implantation of porous materials (TANTALUM, CONCELOC, TTM, ATLANT).MethodsExperiments were carried out on 56 white laboratory female rats aged 6 months. Rats were randomly assigned into groups: sham-operated control group (SH) or ovariectomy group (OVX). Four different commercial implant materials (TTM, CONCELOC, TANTALUM, ATLANT) were placed into the defects (diameter 2.5 mm, depth 3.0 mm) in the distal metaphysis of femurs. Rats were sacrificed 45 days after surgery. Histological study was performed and the percentage of the bone area (BA%) around the implant at a distance of 500 μm in the cancellous area was measured.ResultsFormation of mature bone tissue of varying degrees around all of the implants was detected. In OVX rats cancellous bone defect zone was characterized by a high density of osteocytes on the surface. In the SH group, no differences in BA% among implant materials were found. In OVX rats, the BA% around ATLANT implants was 1.5-time less (p = 0.002) than around TANTALUM. The BA% around the rest of the materials was not statistically different.ConclusionsBone formation around the studied porous titanium and tantalum materials in the osteoporosis model was lower than in normal bone. There were differences in bone formation around the different materials in the osteoporosis model, while in the normal bone model, these differences were absent.

3D-printed Biomimetic Bioactive Glass Scaffolds for Bone Regeneration in Rat Calvarial Defects.

The pore geometry of scaffold intended for the use in the bone repair or replacement is one of the most important parameters in bone tissue engineering. It affects not only the mechanical properties of the scaffold but also the amount of bone regeneration after implantation. Scaffolds with five different architectures (cubic, spherical, x, gyroid, and diamond) at different porosities were fabricated with bioactive borate glass using the selective laser sintering (SLS) process. The compressive strength of scaffolds with porosities ranging from 60% to 30% varied from 1.7 to 15.5 MPa. The scaffold’s compressive strength decreased significantly (up to 90%) after 1-week immersion in simulated body fluids. Degradation of scaffolds is dependent on porosity, in which the scaffold with the largest surface area has the largest reduction in strength. Scaffolds with traditional cubic architecture and biomimetic diamond architecture were implanted in 4.6 mm diameter full-thickness rat calvarial defects for 6 weeks to evaluate the bone regeneration with or without bone morphogenetic protein 2 (BMP-2). Histological analysis indicated no significant difference in bone formation in the defects treated with the two different architectures. However, the defects treated with the diamond architecture scaffolds had more fibrous tissue formation and thus have the potential for faster bone formation. Overall, the results indicated that borate glass scaffolds fabricated using the SLS process have the potential for bone repair and the addition of BMP-2 significantly improves bone regeneration.