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Platelet-rich fibrin, the coagulated plasma fraction of blood, is commonly used to support natural healing in clinical applications. The rat calvaria defect is a standardized model to study bone regeneration. It remains, however, unclear if the rat calvaria defect is appropriate to investigate the impact of human PRF (Platelet-Rich Fibrin) on bone regeneration. To this end, we soaked Bio-Gide® collagen membranes in human or rat liquid concentrated PRF before placing them onto 5 mm calvarial defects in Sprague Dawley rats. Three weeks later, histology and micro-computed tomography (μCT) were performed. We observed that the collagen membranes soaked with rat PRF show the characteristic features of new bone and areas of mineralized collagen matrix, indicated by a median mineralized volume of 1.5 mm3 (range: 0.9; 5.3 mm3). Histology revealed new bone growing underneath the membrane and hybrid bone where collagen fibers are embedded in the new bone. Moreover, areas of passive mineralization were observed. The collagen membranes soaked with human PRF, however, were devoid of histological features of new bone formation in the center of the defect; only occasionally, new bone formed at the defect margins. Human PRF (h-PRF) caused a median bone volume of 0.9 mm3 (range: 0.3–3.3 mm3), which was significantly lower than what was observed with rat PRF (r-PRF), with a BV median of 1.2 mm3 (range: 0.3–5.9 mm3). Our findings indicate that the rat calvaria defect model is suitable for assessing the effects of rat PRF on bone formation, but caution is warranted when extrapolating conclusions regarding the efficacy of human PRF. [ABSTRACT FROM AUTHOR]

Objective: Pigs are emerging as a preferred experimental in vivo model for bone regeneration. The study objective was to answer the focused PEO question: in the pig model (P), what is the capacity of experimental alveolar bone defects (E) for spontaneous regeneration in terms of new bone formation (O)? Methods: Following PRISMA guidelines, electronic databases were searched for studies reporting experimental bone defects or extraction socket healing in the maxillae or mandibles of pigs. The main inclusion criteria were the presence of a control group of untreated defects/sockets and the assessment of regeneration via 3D tomography [radiographic defect fill (RDF)] or 2D histomorphometry [new bone formation (NBF)]. Random effects meta‐analyses were performed for the outcomes RDF and NBF. Results: Overall, 45 studies were included reporting on alveolar bone defects or extraction sockets, most frequently in the mandibles of minipigs. Based on morphology, defects were broadly classified as 'box‐defects' (BD) or 'cylinder‐defects' (CD) with a wide range of healing times (10 days to 52 weeks). Meta‐analyses revealed pooled estimates (with 95% confidence intervals) of 50% RDF (36.87%–63.15%) and 43.74% NBF (30.47%–57%) in BD, and 44% RDF (16.48%–71.61%) and 39.67% NBF (31.53%–47.81%) in CD, which were similar to estimates of socket‐healing [48.74% RDF (40.35%–57.13%) and 38.73% NBF (28.57%–48.89%)]. Heterogeneity in the meta‐analysis was high (I2 > 90%). Conclusion: A substantial body of literature revealed a high capacity for spontaneous regeneration in experimental alveolar bone defects of (mini)pigs, which should be considered in future studies of bone regeneration in this animal model. [ABSTRACT FROM AUTHOR]

Bio-Gide® is a collagen membrane routinely used in guided bone regeneration. Recent studies have shown that this collagen membrane has osteoconductive properties, meaning that it can support the growth of new bone. However, it has also been observed that the collagen membrane has areas of mineralized fibers which can occur spontaneously and independently of osteoblasts. To better understand how this works, we established a model using minced collagen membranes to reduce the active mineralization of intact collagen membranes in favor of passive mineralization. We thus compared the original intact membrane with a minced collagen membrane in a 5 mm calvarial defect model in Sprague Dawley rats. After three weeks of healing, histology and microcomputed tomography (μCT) were performed. Histological analysis confirmed the osteoconductive properties, with new bone growing inside the intact collagen membrane. However, in minced collagen membranes, the osteoconductive properties were restricted to the defect margins. Interestingly, histology revealed large mineralized areas indicating passive mineralization with no signs of bone formation. In the μCT analysis, the intact collagen membranes caused a higher median mineralized volume (1.5 mm3) compared with the minced group (0.4 mm3), but this lacked significance (p = 0.09). The μCT analysis needs to be interpreted carefully, particularly in defects filled with minced membranes, considering that the mineralized tissue may not necessarily be bone but also the result of passive mineralization. Taken together, the findings suggest that Bio-Gide® collagen membranes support bone formation while also exhibiting potential for passive mineralization. [ABSTRACT FROM AUTHOR]

Background: There is growing evidence that extracellular vesicles (EVs) play a crucial role in the paracrine mechanisms of transplanted human mesenchymal stem cells (hMSCs). Little is known, however, about the influence of microenvironmental stimuli on the osteogenic effects of EVs. This study aimed to investigate the properties and functions of EVs derived from undifferentiated hMSC (Naïve-EVs) and hMSC during the early stage of osteogenesis (Osteo-EVs). A further aim was to assess the osteoinductive potential of Osteo-EVs for bone regeneration in rat calvarial defects. Methods: EVs from both groups were isolated using size-exclusion chromatography and characterized by size distribution, morphology, flow cytometry analysis and proteome profiling. The effects of EVs (10 µg/ml) on the proliferation, migration, and osteogenic differentiation of cultured hMSC were evaluated. Osteo-EVs (50 µg) or serum-free medium (SFM, control) were combined with collagen membrane scaffold (MEM) to repair critical-sized calvarial bone defects in male Lewis rats and the efficacy was assessed using µCT, histology and histomorphometry. Results: Although Osteo- and Naïve-EVs have similar characteristics, proteomic analysis revealed an enrichment of bone-related proteins in Osteo-EVs. Both groups enhance cultured hMSC proliferation and migration, but Osteo-EVs demonstrate greater efficacy in promoting in vitro osteogenic differentiation, as evidenced by increased expression of osteogenesis-related genes, and higher calcium deposition. In rat calvarial defects, MEM with Osteo-EVs led to greater and more consistent bone regeneration than MEM loaded with SFM. Conclusions: This study discloses differences in the protein profile and functional effects of EVs obtained from naïve hMSC and hMSC during the early stage of osteogenesis, using different methods. The significant protein profile and cellular function of EVs derived from hMSC during the early stage of osteogenesis were further verified by a calvarial bone defect model, emphasizing the importance of using differentiated MSC to produce EVs for bone therapeutics. [ABSTRACT FROM AUTHOR]

Based on the evidence of a significant communication and connection pathway between the bone and immune systems, a new science has emerged: osteoimmunology. Indeed, the immune system has a considerable impact on bone health and diseases, as well as on bone formation during grafts and its stability over time. Chronic inflammation induces the excessive production of oxidants. An imbalance between the levels of oxidants and antioxidants is called oxidative stress. This physio-pathological state causes both molecular and cellular damage, which leads to DNA alterations, genetic mutations and cell apoptosis, and thus, impaired immunity followed by delayed or compromised wound healing. Oxidative stress levels experienced by the body affect bone regeneration and maintenance around teeth and dental implants. As the immune system and bone remodeling are interconnected, bone loss is a consequence of immune dysregulation. Therefore, oral tissue deficiencies such as periodontitis and peri-implantitis should be regarded as immune diseases. Bone management strategies should include both biological and surgical solutions. These protocols tend to improve immunity through antioxidant production to enhance bone formation and prevent bone loss. This narrative review aims to highlight the relationship between inflammation, oxidation, immunity and bone health in the oral cavity. It intends to help clinicians to detect high-risk situations in oral surgery and to propose biological and clinical solutions that will enhance patients' immune responses and surgical treatment outcomes. [ABSTRACT FROM AUTHOR]

Objectives: Autologous bone is considered the gold standard for grafting, yet it suffers from a tendency to undergo resorption over time. While the exact mechanisms of this resorption remain elusive, osteocytes have been shown to play an important role in stimulating osteoclastic activity through their expression of receptor activator of NF‐κB (RANK) ligand (RANKL). The aim of this study was to assess the function of osteocyte‐derived RANKL in bone graft remodeling. Materials and Methods: In Tnfsf11fl/fl;Dmp1‐Cre mice without osteocyte‐specific RANKL as well as in Dmp1‐Cre control mice, 2.6 mm calvarial bone disks were harvested and transplanted into mice with matching genetic backgrounds either subcutaneously or subperiosteally, creating 4 groups in total. Histology and micro‐computed tomography of the grafts and the donor regions were performed 28 days after grafting. Results: Histology revealed marked resorption of subcutaneous control Dmp1‐Cre grafts and new bone formation around subperiosteal Dmp1‐Cre grafts. In contrast, Tnfsf11fl/fl;Dmp1‐Cre grafts showed effectively neither signs of bone resorption nor formation. Quantitative micro‐computed tomography revealed a significant difference in residual graft area between subcutaneous and subperiosteal Dmp1‐Cre grafts (p <.01). This difference was not observed between subcutaneous and subperiosteal Tnfsf11fl/fl;Dmp1‐Cre grafts (p =.17). Residual graft volume (p =.08) and thickness (p =.13) did not differ significantly among the groups. Donor area regeneration was comparable between Tnfsf11fl/fl;Dmp1‐Cre and Dmp1‐Cre mice and restricted to the defect margins. Conclusions: The results suggest an active function of osteocyte‐derived RANKL in bone graft remodeling. [ABSTRACT FROM AUTHOR]

Bone allografts are widely used as osteoconductive support to guide bone regrowth. Bone allografts are more than a scaffold for the immigrating cells as they maintain some bioactivity of the original bone matrix. Yet, it remains unclear how immigrating cells respond to bone allografts. To this end, we have evaluated the response of mesenchymal cells exposed to acid lysates of bone allografts (ALBA). RNAseq revealed that ALBA has a strong impact on the genetic signature of gingival fibroblasts, indicated by the increased expression of IL11, AREG, C11orf96, STC1, and GK—as confirmed by RT-PCR, and for IL11 and STC1 by immunoassays. Considering that transforming growth factor-β (TGF-β) is stored in the bone matrix and may have caused the expression changes, we performed a proteomics analysis, TGF-β immunoassay, and smad2/3 nuclear translocation. ALBA neither showed detectable TGF-β nor was the lysate able to induce smad2/3 translocation. Nevertheless, the TGF-β receptor type I kinase inhibitor SB431542 significantly decreased the expression of IL11, AREG, and C11orf96, suggesting that other agonists than TGF-β are responsible for the robust cell response. The findings suggest that IL11, AREG, and C11orf96 expression in mesenchymal cells can serve as a bioassay reflecting the bioactivity of the bone allografts. [ABSTRACT FROM AUTHOR]

Purpose: Compaction of particulated grafts is done manually; thus, the effect of compression force on bone regeneration remains unclear. The aim of this study was to evaluate the impact of two different compression forces on the consolidation of particulated bovine hydroxyapatite. Materials and Methods: Two titanium cylinders were fixed on the calvarium of eight New Zealand rabbits. Both defects were filled with particulated bovine hydroxyapatite subjected to a compression force of 0.7 kg/cm2 or 1.6 kg/cm2 before being covered with a resorbable collagen membrane. A handheld device that uses a spring to control the compression force applied by the plugger was used. At 6 weeks, histomorphometry of the area immediately adjacent to the calvaria bone and to the collagen membrane was performed. Results: It was shown that next to the calvaria, the bone volume per tissue volume (BV/TV) was 29.0% ± 8.8% and 27.6% ± 8.2% at low and high compression force, respectively; the bone-to-biomaterial contact (BBC) was 58.2% ± 25.0% and 69.3% ± 22.9%, respectively (P > .05). In the corresponding area next to the collagen membrane, BV/TV was 4.9% ± 5.1% and 5.7% ± 4.7%, and the BBC was 18.3% ± 20.8% and 20.1% ± 15.9%, respectively (P > .05). In addition, the number and area of blood vessels were not significantly affected by compression force. Conclusion: Both compression forces applied resulted in similar consolidation of bovine hydroxyapatite expressed by new bone formation and vascularization based on a rabbit calvaria augmentation model. [ABSTRACT FROM AUTHOR]

A static magnetic field generated by neodymium–iron–boron (NdFeB) magnets placed in the inner cavity of dental implants can enhance bone regeneration in rabbits. It is, however, unknown whether static magnetic fields support osseointegration in a canine model. We therefore determined the potential osteogenic effect of implants carrying NdFeB magnets inserted in the tibia of six adult canines in the early stages of osseointegration. Here, we report that after 15 days of healing, magnetic and regular implants showed a high variation with a median new bone-to-implant contact (nBIC) in the cortical (41.3% and 7.3%) and the medullary (28.6% and 44.8%) region, respectively. Consistently, the median new bone volume/tissue volume (nBV/TV) in the cortical (14.9% and 5.4%) and the medullary (22.2% and 22.4%) region were not significantly different. One week of healing only resulted in negligible bone formation. These findings suggest that considering the large variation and the pilot nature of this study, magnetic implants failed to support peri-implant bone formation in a canine model. [ABSTRACT FROM AUTHOR]

Aim: It has been proposed that platelet‐rich fibrin (PRF) can be used to support bone regeneration during alveolar ridge augmentation. The aim of this study was to determine whether an approach utilizing PRF provides similar performance to the established guided bone regeneration (GBR) procedure. Materials and Methods: Two‐wall defects were surgically created in beagle dogs and treated in three experimental groups: (i) a sticky bone (SB) substitute prepared using liquid PRF and deproteinized porcine bone mineral (DPBM); (ii) SB covered with solid PRF compressed into a membrane; and (iii) GBR performed using DPBM covered by a collagen membrane. Quantitative reverse‐transcription polymerase chain reaction was applied to the specimen after 1 week of healing, and microcomputed tomography (micro‐CT) and histological outcomes were analysed after 8 weeks of healing. Results: Compared with GBR, PRF resulted in a moderate increase in the expression levels of osteoblast and osteoclast markers, osteocalcin, and calcitonin receptor. Moreover, PRF modestly increased angiogenesis and the inflammation markers vascular endothelial growth factor (VEGF) and IL‐6. Micro‐CT and histological analyses confirmed the expected increased alveolar ridge area, with no significant differences between the three groups. Consistently, graft consolidation, as indicated by new bone formation at the defect site, did not differ significantly between groups. Conclusions: The present results demonstrate that PRF‐based approaches perform comparably to the established GBR procedure in terms of the consolidation of DPBM in two‐wall alveolar defects. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the sinus membrane perforation and implant survival rate after crestal minimally invasive sinus floor augmentation using hydraulic pressure and vibrations. Materials and Methods: In this retrospective single cohort study, all patients who underwent minimally invasive sinus floor augmentation between 2007 and 2015 using hydraulic pressure and vibrations were included. The sinus membrane is elevated by physiologic saline at 1.5 bar. The fluid is then set into vibration to further separate the sinus membrane from the bony floor. The endpoints were sinus membrane perforation and the survival rate of implants. Results: The hydraulic pressure and vibration technique was applied in 156 patients. Seven patients with perforations of the sinus membrane were treated with the lateral window approach and excluded from the follow-up analysis. In the remaining 149 patients, 184 crestal sinus floor augmentations were performed and 184 implants were placed. In 10 of these 184 cases, a perforation was suspected in the postoperative computed tomography (CT) scan. In total, the perforation rate was 8.9% (17/191). Nineteen implants were lost during the follow-up period ranging from 0.2 to 8.4 years with a median of 2.3 years. The cumulative implant survival rates after 1, 3, and 5 years were 94.4%, 87.7%, and 87.7%, respectively. No severe perioperative complications were noted. Conclusion: The hydraulic pressure and vibration technique allows a minimally invasive crestal sinus augmentation with a perforation rate less than 10% and implant survival rates of approximately 90%. [ABSTRACT FROM AUTHOR]

Simvastatin embedded into poly(lactic-co-glycolic acid) (PLGA)-based scaffolds can stimulate bone regeneration in preclinical models. However, the ideal pharmacological dose has not been evaluated. This systematic review reports on the simvastatin doses used in preclinical studies and evaluates the regeneration of critical-sized bone defects. References were selected in a two-phase process. Electronic databases (Embase, LILACS, LIVIVO, PubMed, SCOPUS, and Web of Science) and grey literature databases (Google Scholar, Open Grey, and ProQuest) were searched until September 2022. The risk of bias was considered to be low based on the SYRCLE tool. We identified four studies in rat, two in parietal and two in calvaria bone, one in mouse parietal bone, and one in rabbit femur bone. Simvastatin, ranging from 8 to 100 µg, significantly increased bone formation in five studies, as compared to the scaffold alone based on µ-computed tomography, histomorphometric, and radiography analysis. The median increase in bone formation caused by simvastatin was 2.1-fold compared to the PLGA-based scaffold alone. There was, however, no significant correlation between the relative bone gain and the doses of simvastatin (p = 0.37). The data suggest that relatively lower doses of simvastatin can consistently promote preclinical bone regeneration. However, the interpretation of these data must consider the heterogenicity of the PLGA-scaffolds, the defect anatomy, the observation period, and the evaluation method. [ABSTRACT FROM AUTHOR]

Demineralized bone matrix (DBM) is commonly used for the reconstruction of bone defects. Early graft consolidation involves a transient inflammatory process. It is, however, unclear whether DBM can modulate this process. To test this possibility, we prepared acid lysates of demineralized ground cortical (DGC) and moldable demineralized fibers (MDF). Murine RAW 264.7 and primary bone marrow macrophages were exposed to acid lysates of DGC and MFD prior to provoking an inflammatory response with lipopolysaccharide (LPS). Similarly, murine ST2 mesenchymal cells were exposed to DGC and MFD with and without interleukin 1β (IL1) and TNFα. We show here that acid lysates of DGC and MFD reduced the expression of IL1 and IL6 in RAW 264.7 macrophages, as determined by RT-PCR and, for IL6, by immunoassay. This response was confirmed with primary macrophages. Likewise, desalted acid lysates exert anti-inflammatory properties on RAW 264.7 cells and in ST2 cells, the forced expression of IL6, inducible nitric oxide synthase (iNOS) and chemokine ligand 5 (CCL5) was reduced. These in vitro findings suggest that DGC and MFD lower the inflammation-induced expression of inflammatory mediators in murine cell-based bioassays. [ABSTRACT FROM AUTHOR]

Ginseng, a herbal plant, is rich in pharmacologically active ginsenosides capable of promoting bone regeneration and of reducing inflammatory osteolysis. Ginseng was therefore proposed to reduce the catabolic changes during periodontitis. Here, we tested the capability of ginseng to modulate orthodontic tooth movement (OTM). To this aim, 55 male Sprague Dawley rats were randomly distributed into five groups: (I) a normal group without any interventions; (II) an untreated OTM serving as a control; and (III, IV, and V) treated OTMs receiving daily oral administrations of 75, 150, and 300 mg/kg of a standardized extract from the roots of Korean Panax ginseng G115 for three weeks, respectively. The molar tooth was moved towards the incisor during three weeks followed by measurements of the distance between the first and the second molars. Moreover, the impact of OTM and ginseng extracts on body weight was determined. Our data showed that, compared with the OTM control, 150 and 300 mg/kg of G115 ginseng extract significantly decreased the OTM from 0.87 mm (min 0.69; max 0.96) to 0.53 (min 0.42, max 0.62; p = 0.002) and 0.36 (min 0.27, max 0.43; p < 0.0001), respectively. Moreover, 150 and 300 mg/kg of G115 significantly lowered the body weights of the rats when compared with the respective controls (p = 0.002 and p < 0.0001, respectively). These findings suggest that extracts from Panax ginseng are capable of reducing orthodontic tooth movement in rats and is associated with a decrease in body weight. [ABSTRACT FROM AUTHOR]

Objectives: Chronic liver disease increases the risk for periodontal disease and osteoporotic fractures, but its impacts on bone regeneration remain unknown. Herein, we studied the impact of liver cirrhosis on peri‐implant bone formation. Material and Methods: A total of 20 male Wistar rats were randomly divided into two groups: one with the common bile duct ligated (BDL) and the respective sham‐treated control group (SHAM). After four weeks of disease induction, titanium mini‐screws were inserted into the tibia. Successful induction of liver cirrhosis was confirmed by the presence of clinical symptoms. Another four weeks later, peri‐implant bone volume per tissue volume (BV/TV) and bone‐to‐implant contact (BIC) were determined by histomorphometric analysis. Results: Peri‐implant bone formation was not significantly different between the SHAM and BDL groups. In the cortical compartment, the median percentage of peri‐implant new bone was 10.1% (95% CI of mean 4.0–35.7) and 22.5% (13.8–30.6) in the SHAM and BDL groups, respectively (p =.26). Consistently, the new bone in direct contact with the implant was 18.1% (0.4–37.8) and 23.3% (9.2–32.8) in SHAM and BDL groups, respectively (p =.38). When measuring the medullary compartment, the new bone area was 7.1% (4.8–10.4) and 10.4% (7.2–13.5) in the SHAM and BDL groups, respectively (p =.17). Medullary new bone in direct contact with the implant was 10.0% (1.2–50.4) and 20.6% (16.8–35.3) in SHAM and BDL groups, respectively, and thus comparable between the two groups (p =.46). Conclusions: Bile duct ligation has no significant impact on the early stages of peri‐implant bone formation. [ABSTRACT FROM AUTHOR]

Aim: To determine the healing outcome following grafting with deproteinized porcine bone mineral (DPBM) with or without collagen membrane coverage in two‐wall (both buccal and lingual)‐damaged extraction sockets. Materials and methods: Distal roots of three mandibular premolars in six beagle dogs were extracted, and the whole buccal and lingual bony walls were surgically removed. Three treatment protocols were then applied according to the following group allocation: no graft (None), grafting DPBM (BG), and grafting DPBM with coverage by a collagen membrane (BG + M). Two observational periods (2 and 8 weeks) were used with the split‐mouth design, and quantitative and qualitative analyses were performed by microcomputed tomography and histology. Results: The dimensions of the alveolar ridge at both grafted sites (BG and BG + M) remained similar to those of the pristine ridge in the histologic and radiographic analyses, whereas the ungrafted sites (None) collapsed both vertically and horizontally. Both grafting protocols produced substantial bony regeneration, but the addition of a covering membrane enhanced the proportion of mineralized tissue within the augmented area, and the BG + M group also showed a significantly larger area of regenerated ridge than the None group (p <.05). Conclusions: Bone grafting with collagen membrane can maintain the alveolar ridge dimensions with substantial bone regeneration in a two‐wall‐damaged extraction socket. [ABSTRACT FROM AUTHOR]

Background: Bilayer collagen membranes are routinely used in guided bone/tissue regeneration to serve as osteoconductive scaffolds and prevent the invasion of soft tissues. It is recommended to place the membranes with their dense layer towards the soft tissue and their porous layer towards the bony defect area. However, evidence supporting this recommendation is lacking. This study aimed to determine whether the alignment of bilayer collagen membranes has an effect on bone regeneration. Methods: In two groups of ten male Sprague-Dawley rats each, a 5-mm calvarial defect was created. Thereafter, the defect was randomly covered with a bilayer, resorbable, pure type I and III collagen membrane placed either regularly or upside-down (i.e., dense layer towards bone defect). After 4 weeks of healing, micro-computed tomography (μCT), histology, and histomorphometry of the inner cylindrical region of interest (4.5 mm in diameter) were performed to assess new bone formation and the consolidation of the collagen membrane in the defect area. Results: Quantitative μCT showed similar bone volume (median 8.0 mm3, interquartile range 7.0–10.0 vs. 6.2 mm3, 4.3–9.4, p = 0.06) and trabecular thickness (0.21 mm, 0.19–0.23 vs. 0.18 mm, 0.17–0.20, p = 0.03) between upside-down and regular placement, both leading to an almost complete bony coverage. Histomorphometry showed comparable new bone areas between the upside-down and regularly placed membranes, 3.9 mm2 (2.7–5.4) vs. 3.8 mm2 (2.2–4.0, p = 0.31), respectively. Both treatment groups revealed the same regeneration patterns and spatial distribution of bone with and without collagen fibers, as well as residual collagen fibers. Conclusions: Our data support the osteoconductive properties of collagen membranes and suggest that bone regeneration is facilitated regardless of membrane layer alignment. [ABSTRACT FROM AUTHOR]

Fas ligand (FasL) is a member of the tumor necrosis factor (TNF) superfamily involved in the activation of apoptosis. Assuming that apoptosis is initiated after tooth extraction it is reasonable to suggest that FasL may play a pivotal role in the healing of extraction sockets. Herein, we tested the hypothesis of whether the lack of FasL impairs the healing of extraction sockets. To this end, we extracted upper right incisors of FasL knockout (KO) mice and their wildtype (WT) littermates. After a healing period of two weeks, bone volume over total volume (BV/TV) via µCT and descriptive histological analyses were performed. µCT revealed that BV/TV in the coronal region of the socket amounted to 39.4% in WT and 21.8% in KO, with a significant difference between the groups (p=0.002). Likewise, in the middle region of the socket, BV/TV amounted to 50.3% in WT and 40.8% in KO (p<0.001). In the apical part, however, no difference was noticed. Consistently, WT mice displayed a significantly higher median trabecular thickness and a lower trabecular separation when compared to the KO group at the coronal and central region of the socket. There was the overall tendency that in both, female and male mice, FasL affects bone regeneration. Taken together, these findings suggest that FasL deficiency may reduce bone regeneration during the healing process of extraction sockets. [ABSTRACT FROM AUTHOR]

Acid bone lysates (ABLs) represent the growth factors and other molecules released during autologous graft resorption. However, the impact of these bone‐derived growth factors on the healing of bone defects has not yet been investigated. The aim of the present study was, therefore, to examine the impact of ABLs adsorbed to collagen membranes on bone regeneration. To this end, in 16 female Sprague Dawley rats, a standardized 5‐mm‐diameter critical size defect on the calvarial bone was created. The defects were covered with collagen membranes that had been soaked either in serum‐free media or ABLs followed by lyophilization. After a healing period of 4 weeks, micro‐computed tomography (μCT) and histological analyses by means of undecalcified thin ground sections were performed. μCT analysis of the inner 4 mm of the calvaria defect showed a greater bone defect coverage in the control group when compared to ABL group, 29.8% (confidence interval [CI]: 17.7–50.3) versus 5.6% (CI: 1.0–29.8, p =.03), respectively. Moreover, we found significantly more absolute bone volume (BV) in the control group when compared to ABL group, 0.59 mm3 (CI: 0.27–1.25) versus 0.07 mm3 (CI: 0.06–0.59, p =.04), respectively. Histomorphometry confirmed these findings with a relative BV in the central compartment of 14.1% (CI: 8.4–20.6) versus 5.6% (CI: 3.4–7.9, p =.004), respectively. These findings indicate that bone‐derived growth factors contained in ABLs are able to attenuate bone regeneration within collagen membranes. [ABSTRACT FROM AUTHOR]

Purpose: Autografts are considered to support bone regeneration. Paracrine factors released from cortical bone might contribute to the overall process of graft consolidation. The aim of this study was to characterize the paracrine factors by means of proteomic analysis. Materials and Methods: Bone-conditioned medium (BCM) was prepared from fresh bone chips of porcine mandibles and subjected to proteomic analysis. Proteins were categorized and clustered using the bioinformatic tools UNIPROT and PANTHER, respectively. Results: Proteomic analysis showed that BCM contains more than 150 proteins, of which 43 were categorized into “secreted” and “extracellular matrix.” Growth factors that are not only detectable in BCM, but potentially also target cellular processes involved in bone regeneration, eg, pleiotrophin, galectin-1, transforming growth factor beta (TGF-β)-induced gene (TGFBI), lactotransferrin, insulin-like growth factor (IGF)–binding protein 5, latency-associated peptide forming a complex with TGF-β1, and TGF-β2, were discovered. Conclusion: The present results demonstrate that cortical bone chips release a large spectrum of proteins with the possibility of modulating cellular aspects of bone regeneration. The data provide the basis for future studies to understand how these paracrine factors may contribute to the complex process of graft consolidation. [ABSTRACT FROM AUTHOR]

Background: MicroRNAs (miRNAs) are small noncoding RNAs demonstrated as critical post-transcriptional modulators in dental tissues and bone regeneration, particularly miR-21-5p. However, the role of miR-21-5p in the healing of alveolar sockets following tooth extraction remains unknown. In this study we evaluated the influence of miR-21-5p in the healing of alveolar socket after tooth extraction.Methods: Eight miR-21-5p knockout mice and eight littermate controls underwent tooth extraction of the upper right incisor. After a healing period of 14 days microCT and histological analyses were performed.Results: MicroCT analysis showed that the percentage of bone in the extraction socket was significantly higher in the control group than in the miR-21 knockout mice; either in the coronal (39.0%, CI 31.8 to 48.0 versus 23.0%, CI 17.8 to 35.2, P = 0.03) or in the middle part of the alveolar socket (56.0%, CI 50.9 to 62.5 versus 43.5% CI 28.6 to 54.6, P = 0.03). These differences were not noted in the apical part of the extraction socket. Histological analysis supported the microCT findings. Newly bone volume per tissue volume (BV/TV) was significantly higher in the control group when compared to miR-21 knockout mice, 27.4% (CI 20.6 to 32.9) versus 19.0% (CI 14.7 to 21.5, P < 0.05), respectively. Surprisingly, no evident signs of buccal bone resorption were observed in both groups.Conclusion: Despite the limitation of one observation period, these findings suggest that miR-21-5p delays the early healing of alveolar socket following tooth extraction. Whether miR-21-5p is essential for healing of alveolar sockets remains to be elucidated. [ABSTRACT FROM AUTHOR]

Purpose: Osteoporosis and diabetes are physiologic determinants that affect the process of osseointegration. The demand for dental implants is high in these patients, who have benefited from progress in preclinical research. Osseointegration has been extensively studied in rat models of osteoporosis and diabetes. The aim of this study is to present an assessment of the published data and provide consideration for future studies. Materials and Methods: The present study summarizes information through a systematic review of the literature from 1997 to 2008 available through PUBMED. The authors found 20 and 13 articles that report on osseointegration of implants in osteoporotic and diabetic rats, respectively. The survey was extended to recent publications on relevant aspects related to species, strains, and quality of endpoint data. Results: Differences in experimental variables such as timing of implant insertion, location of implants, or duration of disease prior to implant placement were reported and can be justified based on the experimental questions. Moreover, heterogeneity was further caused by variations in the age of rats and in reporting of the main parameters of osseointegration in the cortical and medullary compartments. Regardless of these differences, the evidence indicates consistently that ovariectomy results in reduced osseointegration, while there is only moderate evidence for impaired osseointegration in diabetic rats. Conclusions: Selection of the appropriate protocol is critical when studying implant osseointegration in rats. This overview highlights the demand for quality endpoint data related to animal numbers, data collection, and the way to establish osseointegration. Because of the differences in research questions, general recommendations for uniform models for osteoporosis and diabetes cannot be made. [ABSTRACT FROM AUTHOR]

Purpose: Osteogenic cells contribute to the process of osseointegration and graft consolidation. However, whether the cells survive low oxygen tension and maintain their responsiveness to natural and therapeutic growth and differentiation factors remains unknown. Materials and Methods: To determine the effects of low oxygen tension on osteogenic cell viability and responsiveness in vitro, human bone cells were placed into plastic pouches intended to create anaerobic conditions and were either simultaneously or subsequently exposed to supernatants from activated platelets or recombinant bone morphogenetic protein (BMP)-6. Results: Bone cells cultured for up to 72 hours under hypoxia moderately decreased their metabolic activity, which was paralleled by morphologic changes but not by cleavage of the apoptosis markers caspase-3 and poly(ADP)ribose polymerase. Hypoxia suppressed the mitogenic response of bone cells to platelet-released supernatant and the expression of osteogenic differentiation markers alkaline phosphatase and osteocalcin upon incubation with BMP-6. Stimulation of bone cells with platelet-released supernatant and BMP-6 immediately after re-establishment of normoxia caused a moderate cellular response. However, when bone cells were allowed to recover for 7 days under normoxia, their responsiveness was equal to that of cells not previously exposed to low oxygen tension. Conclusions: These findings suggest that osteogenic cells can survive transient hypoxia and retain their potential to respond to growth and differentiation factors once normoxia is re-established. The data also implicate that reoxygenation, and thus blood vessel formation, may be an important determinant for the process of osseointegration and graft consolidation. [ABSTRACT FROM AUTHOR]

Aim: Osteoimmunology covers the cellular and molecular mechanisms responsible for inflammatory osteolysis that culminates in the degradation of alveolar bone. Osteoimmunology also focuses on the interplay of immune cells with bone cells during bone remodelling and regeneration. The aim of this review was to provide insights into how osteoimmunology affects alveolar bone health and disease. Method: This review is based on a narrative approach to assemble mouse models that provide insights into the cellular and molecular mechanisms causing inflammatory osteolysis and on the impact of immune cells on alveolar bone regeneration. Results: Mouse models have revealed the molecular pathways by which microbial and other factors activate immune cells that initiate an inflammatory response. The inflammation‐induced alveolar bone loss occurs with the concomitant suppression of bone formation. Mouse models also showed that immune cells contribute to the resolution of inflammation and bone regeneration, even though studies with a focus on alveolar socket healing are rare. Conclusions: Considering that osteoimmunology is evolutionarily conserved, osteolysis removes the cause of inflammation by provoking tooth loss. The impact of immune cells on bone regeneration is presumably a way to reinitiate the developmental mechanisms of intramembranous and endochondral bone formation. [ABSTRACT FROM AUTHOR]

Background and Aims: To describe the biology of alveolar bone regeneration. Material and Methods: Four comprehensive reviews were performed on (a) mesenchymal cells and differentiation factors leading to bone formation; (b) the critical interplay between bone resorbing and formative cells; (c) the role of osteoimmunology in the formation and maintenance of alveolar bone; and (d) the self‐regenerative capacity following bone injury or tooth extraction were prepared prior to the workshop. Results and Conclusions: This summary information adds to the fuller understanding of the alveolar bone regenerative response with implications to reconstructive procedures for patient oral rehabilitation. The group collectively formulated and addressed critical questions based on each of the reviews in this consensus report to advance the field. The report concludes with identified areas of future research. [ABSTRACT FROM AUTHOR]

Objectives: Osteoblasts lay down new bone on implant surfaces. The underlying cellular mechanism and the spatio‐temporal mode of action, however, remain unclear. It can be proposed that growth factors released upon acidification by osteoclasts adsorb to the implant surface and control the early stages of osseointegration. Methods: To simulate bone lysis by osteoclasts, titanium discs were exposed to acid bone lysate (ABL) followed by vigorous washing and seeding of oral fibroblasts. The expression of TGF‐β target genes interleukin 11 (IL11) and NADPH oxidase 4 (NOX4) was evaluated by reverse transcriptase polymerase chain reaction and IL11 ELISA. TGF‐β signaling activation was assessed via Smad2/3 immunofluorescence. The impact of ABL on osteogenic differentiation was determined with murine ST2 mesenchymal stromal cells. Results: We report here that ABL‐conditioned titanium discs, independent of turned or rough surface, increased the expression of IL11 and NOX4. This increase was blocked by the TGF‐β receptor 1 antagonist SB431542. Further support for the TGF‐β signaling activation came from the translocation of Smad2/3 into the nucleus of oral fibroblasts. Moreover, titanium discs exposed to ABL decreased alkaline phosphatase and osteopontin in ST2 cells. Conclusions: These in vitro findings suggest that titanium can adsorb TGF‐β from ABLs. The data provide a strong impetus for studies on the protein adsorption on implant surfaces in vitro and in vivo, specifically for growth factors including bone‐derived TGF‐β during successful and failed osseointegration. [ABSTRACT FROM AUTHOR]

Objective: To assess the effect of platelet‐rich plasma (PRP) on implant dentistry. The primary focused question was as follows: What are the clinical, histological, and radiographic outcomes of PRP administration for bone regeneration and implant therapy? Methods: A literature search was conducted involving three databases: MEDLINE, EMBASE and Cochrane database followed by a hand search of relevant scientific journals. Human studies using PRP for bone regeneration and implant therapy were considered and articles published up to December 31, 2017 were included. Eligible studies were selected based on the inclusion criteria, and quality assessments were conducted. Results: In total, out from the 9,497 titles meeting the original search criteria, 22 fulfilled the inclusion criteria and were chosen for data extraction. Among them were 15 randomized controlled trials (RCT) and seven controlled clinical trials (CCT). Overall, the risk of bias was moderate to high. A total of seven studies showed superior outcomes when PRP was added during sinus floor elevation and five showed no superior outcome. Three studies found a significant advantage of PRP for alveolar bone regeneration and another three studies for soft tissue healing. Three studies reported on beneficial effects of PRP directly during implant placement while another study failed to find significant differences. Due to the heterogeneity of study designs, no meta‐analysis could be performed. Summary and Conclusions: Despite the lack of consistent evidence supporting the clinical benefit of PRP in healthy patients, PRP might have a positive effect on wound healing and bone regeneration in compromised patients. [ABSTRACT FROM AUTHOR]

Objective: To assess the impact of platelet‐rich fibrin (PRF) on implant dentistry. The primary focused question was as follows: What are the clinical, histological, and radiographic outcomes of PRF administration for bone regeneration and implant therapy? Method: A systematic literature search comprised three databases: MEDLINE, EMBASE, and Cochrane followed by a hand search of relevant scientific journals. Human studies using PRF for bone regeneration and implant therapy were considered and articles published up to December 31, 2017 were included. Eligible studies were selected based on the inclusion criteria. Randomized controlled trials (RCT) and controlled clinical trials (CCT) were included. Results: In total, 5,963 titles were identified with the search terms and by hand search. A total of 12 randomized controlled trials (RCT) met the inclusion criteria and were chosen for data extraction. Included studies focused on alveolar ridge preservation after tooth extraction, osseointegration process, soft tissue management, bone augmentation, bone regeneration after sinus floor elevation and surgical peri‐implantitis treatment. Overall, the risk of bias was moderate or unclear. Nine studies showed superior outcomes for PRF for any of the evaluated variables, such as ridge dimension, bone regeneration, osseointegration process, soft tissue healing. Three studies failed to show any beneficial effects of PRF. No meta‐analysis could be performed due to the heterogeneity of study designs. Conclusions: There is moderate evidence supporting the clinical benefit of PRF on ridge preservation and in the early phase of osseointegration. It remains unclear whether PRF can reduce pain and improve soft tissue healing. More research support is necessary to comment on the role of PRF to improve other implant therapy outcomes. [ABSTRACT FROM AUTHOR]

Abstract: Objectives: Collagen membranes are not limited to be occlusive barriers as they actively support bone regeneration. However, the impact of bone‐derived growth factors on their osteoconductive competence has not been examined. Methods: Twenty adult Sprague Dawley rats were included in the study. Calvaria defects with a diameter of five millimeter were created. The defect was covered with one layer of a collagen membrane previously soaked in conditioned medium of porcine bone chips or in culture medium alone. After 4 weeks, microcomputed tomography was performed. Undecalcified thin‐ground sections were subjected to light and scanning electron microscopy. Primary outcome parameter was the bone volume in the defect. Unit of analysis was the bone‐conditioned medium (BCM). Results: In the central defect area of the control and the BCM group, median new bone connected to the host bone was 0.54 and 0.32 mm³, respectively (p = .10). In the ectocranial defect area, the control group showed significantly more bone than the BCM group (0.90 and 0.26 mm³; p = .02). Based on an exploratory interpretation, the control group had smaller bony islands than the BCM group. Scanning electron microscopy and histology indicate the formation of bone but also the collagen membrane to be mineralized in the defect site. Conclusions: These results demonstrate that the commercial collagen membrane holds an osteoconductive competence in a rat calvaria defect model. Soaking collagen membranes with BCM shifts bone formation toward the formation of bony islands rather than new bone connected to the host bone. [ABSTRACT FROM AUTHOR]

This overview provides insights into the molecular and cellular mechanisms involved in guided bone regeneration, in particular focusing on aspects presented in the 3D movie, Cell-To-Cell Communication in Guided Bone Regeneration. The information presented here is based almost exclusively on genetic mouse models in which single genes can be deleted or overexpressed, even in a specific cell type. This information needs to be extrapolated to humans and related to aspects relevant to graft consolidation under the clinical parameters of guided bone regeneration. The overview follows the ground tenor of the Cell-To-Cell Communication series and focuses on aspects of cell-to-cell communication in bone regeneration and guided bone regeneration. Here, we discuss (1) the role of inflammation during bone regeneration, including (2) the importance of the fibrin matrix, and (3) the pleiotropic functions of macrophages. We highlight (4) the origin of bone-forming osteoblasts and bone-resorbing osteoclasts as well as (5) what causes a progenitor cell to mature into an effector cell. (6) We touch on the complex bone adaptation and maintenance after graft consolidation and (7) how osteocytes control this process. Finally, we speculate on (8) how barrier membranes and the augmentation material can modulate graft consolidation. [ABSTRACT FROM AUTHOR]

Background: Supplementation of bone substitutes with recombinant platelet-derived growth factor-BB (PDGF-BB) can enhance bone regeneration. The aim of the study was to evaluate the effect of PDGF-BB on bone formation in the presence of β-tricalcium phosphate and bovine bone mineral matrix in a rat calvaria defect model. Methods: The authors examined 5 mm rat calvarial defects treated with β-tricalcium phosphate (TCP) or demineralized bovine bone mineral (DBBM) with and without 0.3 mg/ml recombinant PDGF-BB. Calvaria defects were randomly divided into the following treatment groups (n = 5); TCP; TCP plus PDGF-BB; DBBM; DBBM plus PDGF-BB; and untreated empty control. After 45 days, bone formation was evaluated by histomorphometry and fluorescence microscopy. Results: The authors report that the area of newly formed bone was similar between the empty controls and the two bone substitutes, TCP and DBBM. Supplementation of TCP and DBBM with PDGF-BB had no significant impact on bone formation. Fluorochrome staining revealed no visible changes in the pattern of bone formation in defects filled with TCP and DBBM, irrespective of PDGF-BB. Furthermore, supplementation with PDGF-BB did not influence biomaterial degradation. Conclusions: The authors concluded that PDGF-BB had no impact on bone formation and degradation of bone substitutes in the respective rodent models. Thus, possible beneficial effects of PDGF-BB may require other model situations. Keywords: Bone substitutes, Bone formation, Bone regeneration, Bone augmentation, Platelet-derived growth factor-BB, Rat calvaria defects, β-tricalcium phosphate, Demineralized bovine bone mineral, Histomorphometry, Graft consolidation [ABSTRACT FROM AUTHOR]

Background: Preclinical studies support the assumption that connective tissue grafts preserve the alveolar bone from resorption; the underlying cellular mechanisms, however, remain unknown. The cellular mechanisms may be attributed to the paracrine activity of the palatal fibroblasts. It was thus reasonable to suggest that palatal connective tissue grafts reduce the formation of osteoclasts. Methods: To test this hypothesis, human palatal fibroblasts were examined for their capacity to modulate the formation of osteoclasts in murine bone marrow cultures exposed to RANKL, M-CSF and TGF-β1. Osteoclastogenesis was determined by tartrate-resistant acid phosphatase (TRAP) staining and gene expression analysis. The formation of antigen presenting cells was based on the expression of CD14 and costimmulatory molecules of antigen presenting cells. The paracrine interaction of fibroblasts and the bone marrow was modeled in vitro with inserts of cell-occlusive membranes. Results: In cocultures without cell-to-cell contact, palatal fibroblasts caused a decrease in the expression of the osteoclast marker genes in bone marrow cells; calcitonin receptors, cathepsin K, TRAP, and osteoclast-associated receptor. Also the number of TRAP positive multinucleated cells was decreased in the presence of fibroblasts. Notably, palatal fibroblasts increased the expression of CD14 and the co-stimulatory proteins CD40, CD80, and CD86 in bone marrow cells. Bone marrow cells had no considerable impact on fibroblast viability and proliferation marker genes. With regard to cell distribution, osteoclasts were most prominent in the center of the membranes, while fibroblasts accumulated immediately adjacent to the border of the insert forming a ring-like structure on the surface of the culture plate. Conclusion: The data suggest that palatal fibroblasts provide a paracrine environment that reduces osteoclastogenesis and increases markers of antigen presenting cells. Morover, the paracrine model revealed a joint activity between palatal fibroblasts and bone marrow cells visualized by the characteristic cell distribution in the two separated compartments. [ABSTRACT FROM AUTHOR]

Objectives Osteoclasts rapidly form on the surface of bone chips at augmentation sites. The underlying molecular mechanism, however, is unclear. Soluble factors released from bone chips in vitro have a robust impact on mesenchymal cell differentiation. Whether these soluble factors change the differentiation of hematopoietic cells into osteoclasts remains unknown. Methods Osteoclastogenesis, the formation of tartrate-resistant acid phosphatase-positive multinucleated cells, was studied with murine bone marrow cultures exposed to RANKL and M- CSF, and conditioned medium from fresh ( BCM) and demineralized bone matrix ( DCM). Histochemical staining, gene and protein expression, as well as viability assays were performed. Results This study shows that BCM had no impact on osteoclastogenesis. However, when BCM was heated to 85°C ( BCMh), the number of tartrate-resistant acid phosphatase-positive multinucleated cells that developed in the presence of RANKL and M- CSF approximately doubled. In line with the histochemical observations, there was a trend that BCMh increased expression of osteoclast marker genes, in particular the transcription factor c-fos. The expression of c-fos was significantly reduced by the TGF-β receptor I antagonist SB431542. DCM significantly stimulated osteoclastogenesis, independent of thermal processing. Conclusions These data demonstrate that activated BCM by heat and DBM are able to stimulate osteoclastogenesis in vitro. These in vitro results support the notion that the resorption of autografts may be supported by as yet less defined paracrine mechanisms. [ABSTRACT FROM AUTHOR]

Objective Signaling molecules derived from osteocytes have been proposed as a mechanism by which autografts contribute to bone regeneration. However, there have been no studies that determined the role of osteocytes in bone grafts. Material and method Herein, it was examined whether bone chips and demineralized bone matrix release sclerostin and FGF-23, both of which are highly expressed by osteocytes. Results Bone grafts from seven donors were placed in culture medium. Immunoassay showed that bone chips released sclerostin (median 1.0 ng/ml) and FGF-23 (median 9.8 relative units/ml) within the first day, with declining levels overtime. Demineralized bone matrix also released detectable amounts of sclerostin into culture medium, while FGF-23 remained close to the detection limit. In vitro expanded isolated bone cells failed to release detectable amounts of sclerostin and FGF-23. Conclusion These results suggest that autografts but also demineralized bone matrix can release signaling molecules that are characteristically produced by osteocytes. [ABSTRACT FROM AUTHOR]

Background and Purpose Autografts are used for bone reconstruction in regenerative medicine including oral and maxillofacial surgery. Bone grafts release paracrine signals that can reach mesenchymal cells at defect sites. The impact of the paracrine signals on osteogenic, adipogenic, and chondrogenic differentiation of mesenchymal cells has remained unclear. Material and Methods Osteogenesis, adipogenesis, and chondrogenesis were studied with murine ST2 osteoblast progenitors, 3 T3- L1 preadipocytes, and ATDC5 prechondrogenic cells, respectively. Primary periodontal fibroblasts from the gingiva, from the periodontal ligament, and from bone were also included in the analysis. Cells were exposed to bone-conditioned medium ( BCM) that was prepared from porcine cortical bone chips. Results BCM inhibited osteogenic and adipogenic differentiation of ST2 and 3 T3- L1 cells, respectively, as shown by histological staining and gene expression. No substantial changes in the expression of chondrogenic genes were observed in ATDC5 cells. Primary periodontal fibroblasts also showed a robust decrease in alkaline phosphatase and peroxisome proliferator-activated receptor gamma ( PPARγ) expression when exposed to BCM. BCM also increased collagen type 10 expression. Pharmacologic blocking of transforming growth factor ( TGF)-β receptor type I kinase with SB431542 and the smad-3 inhibitor SIS3 at least partially reversed the effect of BCM on PPARγ and collagen type 10 expression. In support of BCM having TGF-β activity, the respective target genes were increasingly expressed in periodontal fibroblasts. Conclusions The present work is a pioneer study on the paracrine activity of bone grafts. The findings suggest that cortical bone chips release soluble signals that can modulate differentiation of mesenchymal cells in vitro at least partially involving TGF-β signaling. [ABSTRACT FROM AUTHOR]

Objectives The occurrence of multinucleated giant cells ( MNGCs) on bone substitute materials has been recognized for a long time. However, there have been no studies linking material characteristics with morphology of the MNGCs. The aim was to analyze the qualitative differences of MNGCs on two commercially available calcium phosphate bone substitute materials retrieved from bone defects. Material and methods Six defects were prepared bilaterally in the mandibular body of three mini pigs. The defects were randomly grafted with either deproteinized bovine bone mineral ( DBBM) or biphasic calcium phosphate ( BCP). After a healing period of four weeks, bone blocks were embedded in LR White resin. Three consecutive sections per defect were analyzed as follows: two with light microscopy using toluidine blue and tartrate-resistant acid phosphatase ( TRAP) staining and one with transmission electron microscopy. Results Multinucleated giant cells appeared on both biomaterials. On BCP, MNGCs had a flat morphology and were not observed in resorption lacunae. On DBBM, the MNGCs appeared more round and were often found in shallow concavities. MNGCs on both biomaterials demonstrated a varying degree of TRAP staining, with a tendency toward higher staining intensity of MNGCs on BCP. At the ultrastructural level, signs of superficial dissolution of BCP together with phagocytosis of minor fragments were observed. MNGCs on the surface of DBBM demonstrated sealing zones and ruffled borders, both features of mature osteoclasts. Conclusion MNGCs demonstrated distinctly different histological features depending on the bone substitute material used. Further research is warranted to understand the clinical implications of these morphological observations. [ABSTRACT FROM AUTHOR]

Aim Pharmacological inhibitors of prolyl hydroxylases, also termed hypoxia-mimetic agents ( HMAs), when repeatedly injected can support angiogenesis and bone regeneration. However, the possible role of HMA loaded onto bone substitutes to support angiogenesis and bone regeneration under diabetic condition is unknown. The capacity of HMA loaded onto deproteinized bovine bone mineral ( DBBM) to support angiogenesis and bone formation was examined in diabetic Wistar rats. Methods Diabetes was induced by intraperitoneal injection of streptozotocin. The HMA dimethyloxalylglycine ( DMOG) and desferrioxamine ( DFO) were lyophilized onto DBBM. Calvarial defects were created with a trephine drill and filled with the respective bone substitutes. After 4 weeks of healing, the animals were subjected to histological and histomorphometric analysis. Results In this report, we provide evidence that DMOG loaded onto DBBM can support angiogenesis in vivo. Specifically, we show that DMOG increased the vessel area in the defect site to 2.4% ± 1.3% compared with controls 1.1% ± 0.48% ( P = 0.012). There was a trend toward an increased vessel number in the defect site with 38.6 ± 17.4 and 31.0 ± 10.3 in the DMOG and the control group ( P = 0.231). The increase in angiogenesis, however, did not translate into enhanced bone formation in the defect area with 9.2% ± 7.1% and 8.4% ± 5.6% in DMOG and control group, respectively. No significant changes were caused by DFO. Conclusions The results suggest that DMOG loaded onto DBBM can support angiogenesis, but bone formation does not increase accordingly in a type 1 diabetic rat calvarial defect model at the indicated time point. [ABSTRACT FROM AUTHOR]

Aim Vitamin D deficiency is considered to diminish bone regeneration. Yet, raising the serum levels takes months. A topic application of the active vitamin D metabolite, calcitriol, may be an effective approach. Thus, it becomes important to know the effect of vitamin D deficiency and local application on alveolar bone regeneration. Material and Methods Sixty rats were divided into three groups; two vitamin depletion groups and a control group. Identical single defects (2 mm diameter) were created in the maxilla and mandible treated with calcitriol soaked collagen in one deficiency group while in the other two groups not. Histomorphometric analysis and micro CTs were performed after 1 and 3 weeks. Serum levels of 25( OH)D3 and PTH were determined. Results Bone formation rate significantly increased within the observation period in all groups. Bone regeneration was higher in the maxilla than in the mandible. However, bone regeneration was lower in the control group compared to vitamin depletion groups, with no significant effects by local administration of calcitriol (micro CT mandible p = 0.003, maxilla p < 0.001; histomorphometry maxilla p = 0.035, mandible p = 0.18). Conclusion Vitamin D deficiency not necessarily impairs bone regeneration in the rat jaw and a single local calcitriol application does not enhance healing. [ABSTRACT FROM AUTHOR]

Background: Contour augmentation around early-placed implants (Type 2 placement) using autogenous bone chips combined with deproteinized bovine bone mineral (DBBM) and a collagen barrier membrane has been documented to predictably provide esthetically satisfactory clinical outcomes. In addition, recent data from cone beam computed tomography studies have shown the augmented volume to be stable long-term. However, no human histologic data are available to document the tissue reactions to this bone augmentation procedure. Methods: Over an 8-year period, 12 biopsies were harvested 14 to 80 months after implant placement with simultaneous contour augmentation in 10 patients. The biopsies were subjected to histologic and histomorphometric analysis. Results: The biopsies consisted of 32.0% ± 9.6% DBBM particles and 40.6%± 14.6%mature bone. 70.3%± 14.5%ofthe DBBM particle surfaces were covered with bone. On the remaining surface, multinucleated giant cells with varying intensity of tartrate-resistant acid phosphatase staining were regularly present. No signs of inflammation were visible, and no tendency toward a decreasing volume fraction of DBBM over time was observed. Conclusions: The present study confirms previous findings that osseointegrated DBBM particles do not tend to undergo substitution over time. This low substitution rate may be the reason behind the clinically and radiographically documented long-term stability of contour augmentation using a combination of autogenous bone chips, DBBM particles, and a collagen membrane. [ABSTRACT FROM AUTHOR]

Objectives Osteocytes, the most common cells of the bone, are buried in lacunae. Density and area of the osteocyte lacunae change with increasing maturation of the newly formed bone. Evaluation of osteocyte lacunae can therefore provide insights into the process of graft consolidation. Materials and Methods Here, we determined the osteocyte lacunar density (number of osteocyte lacunae per bone area; N.Ot/ BAr) and the osteocyte lacunar area in μm2 (Lac.Ar) in histological specimens 6 and 12 weeks after the sinuses of 10 minipigs were augmented with Bio-Oss®, a deproteinized bovine bone mineral, and Ostim®, an aqueous paste of synthetic nanoparticular hydroxyapatite. The region of interest was defined by the following criteria: (i) >1 mm from the host bone, (ii) >0.5 mm from the sinus mucosa, (iii) minimum area of 0.2 mm2, and (iv) bone tissue spanning at least two bone substitute particles. Results The overall osteocyte lacunar density was significantly higher in the Bio-Oss® group than in the Ostim® group and decreased during the observation period at a similar range in both groups. The osteocyte lacunar area was smaller in the Bio-Oss® group than the Ostim® group but there was no significant change within the groups over time. Conclusions These results suggest that bone substitutes affect the osteocyte lacunar density and the osteocyte lacunar area in the newly formed bone within the augmented sinus in this particular model situation. These measures can provide insights into the maturation of newly formed bone in the augmented sinus. [ABSTRACT FROM AUTHOR]

Objective Crohn's disease is a chronic inflammatory process that has recently been associated with a higher risk of early implant failure. Herein we provide information on the impact of colitis on peri-implant bone formation using preclinical models of chemically induced colitis. Methods Colitis was induced by intrarectal instillation of 2,4,6-trinitro-benzene-sulfonic-acid ( TNBS). Colitis was also induced by feeding rats dextran-sodium-sulfate ( DSS) in drinking water. One week after disease induction, titanium miniscrews were inserted into the tibia. Four weeks after implantation, peri-implant bone volume per tissue volume ( BV/ TV) and bone-to-implant contacts ( BIC) were determined by histomorphometric analysis. Results Cortical histomorphometric parameters were similar in the control ( n = 10), DSS ( n = 10) and TNBS ( n = 8) groups. Cortical BV/ TV was 92.2 ± 3.7%, 92.0 ± 3.0% and 92.6 ± 2.7%. Cortical BIC was 81.3 ± 8.8%, 83.2 ± 8.4% and 84.0 ± 7.0%, respectively. No significant differences were observed when comparing the medullary BV/ TV and BIC (19.5 ± 6.4%, 16.2 ± 5.6% and 15.4 ± 9.0%) and (48.8 ± 12.9%, 49.2 ± 6.2 and 41.9 ± 11.7%), respectively. Successful induction of colitis was confirmed by loss of body weight and colon morphology. Conclusions The results suggest bone regeneration around implants is not impaired in chemically induced colitis models. Considering that Crohn's disease can affect any part of the gastrointestinal tract including the mouth, our model only partially reflects the clinical situation. [ABSTRACT FROM AUTHOR]

Aim Vitamin D deficiency is highly prevalent in the population and associated with impaired peri-implant bone regeneration. Yet, there is a gap in understanding the impact of vitamin D supplementation on the process of osseointegration. In this study, the effect of vitamin D supplementation on peri-implant bone regeneration was investigated. Methods Fifty ovariectomized Sprague-Dawley rats were divided into three groups. The depletion group was fed a vitamin D-free diet for 8 weeks. The repletion group received vitamin D-free diet for 6 weeks, before animals were switched to standard diet containing 2400 IU/kg vitamin D. The control group was fed the standard diet. Two titanium mini-implants were placed in the tibia. All groups remained on their previous diet until sacrifice. Blood sample testing and histomorphometric analysis were performed. Results Vitamin D depletion caused a significant reduction in 25-hydroxvitamin D in rat serum that returned to control levels in the repletion group. This vitamin deficiency was associated with a decrease in bone-to-implant contact in the cortical area, which was leveled to controls in the repletion group. No significant changes by vitamin D depletion were noticed in the medullar compartment. Moreover, also the peri-implant bone area and the mineral apposition rate remained unchanged upon vitamin D depletion. Conclusion These results indicate that vitamin D deficiency has a negative impact on cortical peri-implant bone formation in ovariectomized rats, which can be compensated by vitamin D supplementation. This study provides first insight into the potential beneficial effect of vitamin D supplementation in implant dentistry. [ABSTRACT FROM AUTHOR]

Objectives: The original aim of the study was to determine the osteoconductive capacity of deproteinized bovine bone mineral (DBBM) of different particle sizes underneath acrylic hemispheres in vivo. However, the model failed and allowed us to report on the resorption of DBBM. Material and methods: Acrylic hemispheres were filled with and without a DBBM at a small particle size of 125-250 μm and at the regular particle size of 250-1000 μm. The hemispheres were positioned on the calvaria of eight minipigs. Histological and histomorphometric analysis was performed after 12 weeks. Results: We found that the acrylic hemispheres were displaced and a dense fibrous capsule sequestered the augmented area. Histology showed severe resorption activity and the presence of multinucleated cells on the surface of DBBM particles in areas adjacent to the fibrous capsule. Histomorphometric analysis revealed that only less than half of the originally augmented area, which was approximately 30 mm2, remained after 12 weeks. The amount of residual DBBM (median 0.9 and 3.49 mm2) and bone (median 7.22 and 7.51 mm2) in the augmented area was similar in the small and the regular particle size group. Conclusion: The model represents a pathologic situation of excess resorption of DBBM and bone in an augmented area. The underlying cellular mechanisms remain to be uncovered. To cite this article: Busenlechner D, Tangl S, Arnhart C, Redl H, Schuh C, Watzek G, Gruber R. Resorption of deproteinized bovine bone mineral in a porcine calvaria augmentation model. Clin. Oral Impl. Res. 23, 2012; 95-99. doi: 10.1111/j.1600-0501.2011.02198.x [ABSTRACT FROM AUTHOR]

Implant placement frequently depends on bone augmentation. However, the impact of systemic metabolic diseases on the consolidation of bone substitutes remains poorly understood. Our goal is to study the impact of ovariectomy and hyperglycemia on graft consolidation in rat calvaria. We determined a rat model in which methacrylate hemispheres filled with deproteinized bovine bone mineral were fixed on the calvaria. The first group received streptozotocin (STZ) to induce diabetes. The second group of animals underwent ovariectomy (OVX), causing osteoporosis. Control animals remained untreated, only receiving vehicle injections (STZ-control) but not sham operation, respectively. Specimens were assessed by histomorphometry and μCT. Graft consolidation was similar between the two groups. The rate of new bone formation after 4 weeks was 0.61±0.53% in the STZ group ( n=10) and 0.69±0.91% in the control group ( n=8). After 8 weeks, the rates of new bone formation were 4.98±3.16% in the OVX group ( n=7) and 2.35±1.30% in the control group ( n=10). The volume occupied by the bone substitute was not affected by STZ or OVX treatment. The low amount of newly formed bone could not be quantified by μCT. We conclude that neither STZ nor OVX altered the early phase of graft consolidation. Our findings are limited by the weak osteogenic potential of the rat calvaria in this augmentation model. Fuegl A, Tangl S, Keibl C, Watzek G, Redl H, Gruber R. The impact of ovariectomy and hyperglycemia on graft consolidation in rat calvaria. Clin. Oral Impl. Res. , 2011; 524-529. doi: 10.1111/j.1600-0501.2010.02048.x [ABSTRACT FROM AUTHOR]

Diabetes is considered a risk factor in the osseointegration of dental implants, which suggests that these patients might benefit from anabolic therapies. Preclinical studies, including investigations by this research group, revealed that intermittent administration of parathyroid hormone (PTH) stimulates bone formation on the surface of titanium implants under physiological conditions. However, the anabolic effect of PTH on osseointegration under the hyperglycemic condition of diabetes is unknown. The ability of PTH to stimulate osseointegration was investigated in 40 female Wistar rats that were randomly divided into the following treatment groups: diabetes, diabetes plus PTH, control, and control plus PTH. Diabetes was induced by intraperitoneal injection of streptozotocin (45 mg/kg) at 1 week before implantation. Rats received PTH at a dose of 60 μg/kg or a vehicle by subcutaneous injection starting at the day of implant insertion into the tibia. Histomorphometric analysis was performed after 4 weeks. The medullary peri-implant bone area significantly increased in rats receiving PTH in comparison with the control group (41±12% to 20±12%; P<0.01). Moreover, there was an increased bone-to-implant contact (BIC) area in animals treated with PTH (47±18% to 27±16%; P<0.05). In contrast, diabetic rats failed to benefit from the anabolic treatment. A similar peri-implant bone area occurred in the diabetes group, independent of treatment with PTH (13±9% to 15±6%; P>0.05). Moreover, PTH did not affect the BIC area under hyperglycemic conditions (16±12% to 16±8%; P>0.05). No significant changes were observed in the cortical compartment of all groups. These results demonstrate that the metabolic characteristics of the diabetic rats produced a condition that was unable to respond to PTH treatment. These findings led us to hypothesize that metabolic control of diabetes might be a critical determinant when diabetic patients are undergoing anabolic therapy to enhance osseointegration. Kuchler U, Spilka T, Baron K, Tangl S, Watzek G, Gruber R. Intermittent parathyroid hormone fails to stimulate osseointegration in diabetic rats. Clin. Oral Impl. Res. , 2011; 518-523 doi: 10.1111/j.1600-0501.2010.02047.x [ABSTRACT FROM AUTHOR]

Objective: Collagen barrier membranes (CBM) are used for guided bone regeneration to support the process of graft consolidation. It remains, unknown however, whether CBM can affect the consolidation of bone grafts by controlling the differentiation of progenitor cells into bone-resorbing osteoclasts and bone-forming osteoblasts. Material and Methods: To gain an insight into the underlying mechanisms, we performed in vitro bone marrow cultures on CBM (Bio-Gide®) under conditions that favor osteoclastogenesis and osteoblastogenesis, respectively. Measures of osteoclastogenesis were based on the number of tartrate-resistant acid-phosphatase-positive (TRAP+) multinucleated cells. Resorption assays revealed the activity of mature osteoclasts. Osteoblastogenesis was determined by alkaline-phosphatase activity. Viability was investigated utilizing the MTT assay. Results: Cultivation of murine bone marrow on CBM reduced the number of TRAP+ multinucleated cells compared with cultures on tissue culture plates. Inhibition of osteoclastogenesis was observed on the porous and the dense CBM surfaces. The majority of TRAP+ cells were mononucleated and the decreased osteoclastogenesis was not due to changes in cell viability. Furthermore, CBM are inert regarding the resorptive activity of mature osteoclasts. Moreover, osteoblastogenesis was not reduced when bone marrow cells were grown on the surface of CBM. Conclusions: These in vitro findings demonstrate that CBM can reduce the formation but not the activity of multinucleated osteoclasts. Our data further reveal that the formation of osteogenic cells from their progenitors is not reduced by the CBM. Overall, our results suggest that the beneficial effects of CBM during graft consolidation may involve their direct impact on osteoclastogenesis. To cite this article: Agis H, Magdalenko M, Stögerer K, Watzek G, Gruber R. Collagen barrier membranes decrease osteoclastogenesis in murine bone marrow cultures. Clin. Oral Impl. Res. 21, 2010; 656–661. doi: 10.1111/j.1600-0501.2009.01888.x [ABSTRACT FROM AUTHOR]