Enhance Bone Healing Research Articles

Novel Fracture Site Targeting Drug Improves Fracture Healing with Pain Relief: Preclinical Evaluation of MAK123

Non-union bone fracture occurs in 5-10% of fracture injuries. Interventions include surgery with local implantation of autograft, allograft, demineralized bone matrix, and/or bone morphogenetic proteins. These types of fracture injuries are also accompanied by acute and chronic pain states. In most instances, opioids are provided to injured patients during and after surgery. With the opioid crisis, identifying new analgesic therapies that could reduce or eliminate opioid use, while also improving bone healing is important. Here we show the ability of a novel compound, MAK123, to both enhance bone healing and reduce pain behavior in a surgically induced femoral fracture mouse model. Briefly, 20 male C57BL/6 mice underwent a surgically induced femoral fracture and then were treated with 0, 2, 6, or 20 mg/kg, 3X/week for the 3 week study duration. Weekly X-rays were used to examine healing progression. Prior to euthanasia, mice underwent behavioral testing to measure evoked pain behaviors. Upon euthanasia, ex vivo µCT imaging and analysis was completed to assess fracture callus size and composition. While all doses of MAK123 tested resulted in improved healing, the 6mg/kg dose resulted in accelerated bone healing and a significant increase in mineralized callus volume (p<0.05). Similarly, while all doses of MAK123 reduced evoked responses to tactile stimulus as demonstrated by increased paw withdrawal thresholds, 6 mg/kg of MAK123 resulted in a more robust and significant improvement (p<0.05). We postulate that optimization of the dosing schedule/concentration could further improve both bone healing and behavioral measures thought to represent pain in rodents. That said, these promising pre-clinical data warrant further evaluation as MAK123 may prove to be a unique tool for orthopaedic surgery usage whereby it could both improve bone healing and reduce clinical pain, improving overall patient outcomes.

Bone Regeneration with Platelet-Rich Fibrin Following Surgical Management of Periapical Lesion: Achieving Success in a Resource-limited Area. A Case Report

Background: Intra-oral bony defects following periradicular surgery have been documented to have high selfregenerating potential (especially when small). However, early permanent rehabilitation of tooth to function is important post-surgery. Thus, there is a need to enhance bone healing through regeneration due to the prolonged natural bone healing process. Many materials have been proposed and used but the Platelet-rich fibrin (PRF); a second-generation platelet with fibrin membranes enriched with platelets growth factors is an autologous material, easy to produce and a cost-effective way of regenerating bony defects. Aim: To assess and report the regenerative potential of PRF following periradicular surgery. Methodology: A case report of a 60-year old female who presented with a chronic periapical abscess in relation to the upper right incisors. A periapical pathology of 30mm in its widest dimension was present on the radiograph. Following root canal treatment, the patient had periradicular surgery to remove the apical cystic lesion. The bony defect was filled with autologous platelet-rich fibrin (produced through centrifuging of peripheral blood) to stimulate bone regeneration, and the tooth was reviewed for one year. Result: The one-year review post periradicular surgery showed good healing and bone regeneration. Conclusion: The report showed the use of PRF in regeneration of intra bony defect post degranulation of periapical lesion (after removal of cystic lesion) as an effective and cheap way of achieving regeneration.

Sex Differences in Mesenchymal Stem Cell Therapy With Gelatin-Based Microribbon Hydrogels in a Murine Long Bone Critical-Size Defect Model.

Mesenchymal stem cell (MSC)-based therapy and novel biomaterials are promising strategies for healing of long bone critical size defects. Interleukin-4 (IL-4) over-expressing MSCs within a gelatin microribbon (µRB) scaffold was previously shown to enhance the bridging of bone within a critical size femoral bone defect in male Balb/c mice. Whether sex differences affect the healing of this bone defect in conjunction with different treatments is unknown. In this study, we generated 2-mm critical-sized femoral diaphyseal bone defects in 10–12-week-old female and male Balb/c mice. Scaffolds without cells and with unmodified MSCs were implanted immediately after the primary surgery that created the bone defect; scaffolds with IL-4 over-expressing MSCs were implanted 3 days after the primary surgery, to avoid the adverse effects of IL-4 on the initial inflammatory phase of fracture healing. Mice were euthanized 6 weeks after the primary surgery and femurs were collected. MicroCT (µCT), histochemical and immunohistochemical analyses were subsequently performed of the defect site. µRB scaffolds with IL-4 over-expressing MSCs enhanced bone healing in both female and male mice. Male mice showed higher measures of bone bridging and increased alkaline phosphatase (ALP) positive areas, total macrophages and M2 macrophages compared with female mice after receiving scaffolds with IL-4 over-expressing MSCs. Female mice showed higher Tartrate-Resistant Acid Phosphatase (TRAP) positive osteoclast numbers compared with male mice. These results demonstrated that sex differences should be considered during the application of MSC-based studies of bone healing.

The Effects of SRT1720 Treatment on Endothelial Cells Derived from the Lung and Bone Marrow of Young and Aged, Male and Female Mice.

Angiogenesis is critical for successful fracture healing. Age-related alterations in endothelial cells (ECs) may cause impaired bone healing. Therefore, examining therapeutic treatments to improve angiogenesis in aging may enhance bone healing. Sirtuin 1 (SIRT1) is highly expressed in ECs and its activation is known to counteract aging. Here, we examined the effects of SRT1720 treatment (SIRT1 activator) on the growth and function of bone marrow and lung ECs (BMECs and LECs, respectively), derived from young (3–4 month) and old (20–24 month) mice. While aging did not alter EC proliferation, treatment with SRT1720 significantly increased proliferation of all LECs. However, SRT1720 only increased proliferation of old female BMECs. Vessel-like tube assays showed similar vessel-like structures between young and old LECs and BMECs from both male and female mice. SRT1720 significantly improved vessel-like structures in all LECs. No age, sex, or treatment differences were found in migration related parameters of LECs. In males, old BMECs had greater migration rates than young BMECs, whereas in females, old BMECs had lower migration rates than young BMECs. Collectively, our data suggest that treatment with SRT1720 appears to enhance the angiogenic potential of LECs irrespective of age or sex. However, its role in BMECs is sex- and age-dependent.

Osteoprotective Effect of Enamel Matrix Derivatives on the Regeneration of Mandibular Defects in Experimentally Glucocorticoid-Induced Osteoporosis.

Purpose Osteoporosis is a progressive systematic skeletal illness characterized by low bone mineral density and susceptibility to fracture caused by bone resorption. Aim of the Study. This study intended to evaluate the possible role of emdogain in combination with calcitonin on the healing of surgically induced mandibular defects performed on osteoporotic rats. Materials and Methods Forty healthy female white albino rats were included in this study and divided into four groups. In group I (negative control), 10 rats received a vehicle injection after which a unilateral mandibular defect was created in each rat of all groups. Three groups were subjected to induction of osteoporosis by subcutaneous injection of 0.1 mg/kg/day dexamethasone for 60 days. In group II, rats were kept without treatment. In group III, rats were treated with daily intramuscular injection of 2.5 IU/kg of synthetic salmon calcitonin. In group IV, rats were handled as group III, and the created cavity was filled with emdogain. Rats were euthanized at 2nd and 4th week postsurgically. Hematoxylin and eosin, Masson's trichrome, NF-κB (nuclear factor of activated B cells), and immunohistochemical stains were used, followed by statistical analysis. Results Group I showed normal stages of bone defects healing. Group II revealed the formation of granulation tissue with dilated blood vessels, while groups III and IV showed enhanced bone healing and proper collagen fibers. The percentage area of newly formed collagen fibers was significantly higher in group IV at 2nd week (13.96 ± 0.020%) and 4th week (16.95 ± 0.024%) than in group II (8.75 ± 0.015% and 10.29 ± 0.015%, respectively) and group III (12.93 ± 0.015% and 14.61 ± 0.021%, respectively), but was lower than that in group I (15.75 ± 0.015% and 17.49 ± 0.015%, respectively). Conclusion The local application of emdogain combined with systemically injected calcitonin improves bone healing in surgically induced bone defects in osteoporotic rats.

Flunixin Meglumine Enhanced Bone Fracture Healing in Rabbits Associated with Activation of Early Collagen Deposition and Enhancement of Vascular Endothelial Growth Factor Expression.

Simple SummaryPostfracture treatment with nonsteroidal anti-inflammatory drugs might result in delayed healing. This study was aimed to evaluate and compare the effects of flunixin meglumine (FM) and ketoprofen on bone fracture healing in rabbits. A simple unilateral diaphyseal fracture was made and followed by fixation by K-wire. Healing was evaluated with radiography, histopathology, and immunohistochemistry. Interestingly, the results revealed that FM enhanced bone fracture healing combined with the activation of early collagen deposition, marked angiogenesis, and enhanced vascular endothelial growth factor. However, ketoprofen delayed bone fracture healing. These findings provide novel baseline information about the potential beneficial effects of FM on bone fracture healing cases.Nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most commonly used postoperative analgesics, antipyretics, and anti-inflammatories, and they help prevent blood clotting. However, most NSAIDs delay bone healing. This study was aimed to investigate bone healing in a rabbit animal model by assessing the ability of flunixin meglumine (FM) and ketoprofen to induce fracture healing by examining histology, radiological changes, and vascular endothelial growth factor (VEGF) immunostaining during bone healing. For this purpose, 24 New Zealand rabbits were assigned to three groups: the control group, the FM group, and the ketoprofen group. Our results revealed that there were no intraoperative complications, and all surviving rabbits achieved full-weight bearing. Significant periosteal reaction and callus formation were confirmed at 2 postoperative weeks. Interestingly, FM enhanced callus formation, bone union, and remodeling in the FM group compared to the control and ketoprofen groups. FM enhanced bone healing through early collagen deposition and marked angiogenesis process activation by increasing the expression of VEGF. Our findings demonstrated, for the first time, the potential imperative action of FM in the bone healing process rather than other NSAIDs in animals.

Bi-directional gene activation and repression promote ASC differentiation and enhance bone healing in osteoporotic rats

Calvarial bone healing is challenging, especially for individuals with osteoporosis because stem cells from osteoporotic patients are highly prone to adipogenic differentiation. Based on previous findings that chondrogenic induction of adipose-derived stem cells (ASCs) can augment calvarial bone healing, we hypothesized that activating chondroinductive Sox Trio genes (Sox5, Sox6, Sox9) and repressing adipoinductive genes (C/ebp-α, Ppar-γ) in osteoporotic ASCs can reprogram cell differentiation and improve calvarial bone healing after implantation. However, simultaneous gene activation and repression in ASCs is difficult. To tackle this problem, we built a CRISPR-BiD system for bi-directional gene regulation. Specifically, we built a CRISPR-AceTran system that exploited both histone acetylation and transcription activation for synergistic Sox Trio activation. We also developed a CRISPR interference (CRISPRi) system that exploited DNA methylation for repression of adipoinductive genes. We combined CRISPR-AceTran and CRISPRi to form the CRISPR-BiD system, which harnessed three mechanisms (transcription activation, histone acetylation, and DNA methylation). After delivery into osteoporotic rat ASCs, CRISPR-BiD significantly enhanced chondrogenesis and invitro cartilage formation. Implantation of the engineered osteoporotic ASCs into critical-sized calvarial bone defects significantly improved bone healing in osteoporotic rats. These results implicated the potential of the CRISPR-BiD system for bi-directional regulation of cell fate and regenerative medicine.

Can low-intensity pulsed ultrasound (LIPUS) accelerate bone healing after intramedullary screw fixation for proximal fifth metatarsal stress fractures? A retrospective study

BackgroundIntramedullary screw fixation is considered the standard treatment for proximal fifth metatarsal stress fractures. Low-intensity pulsed ultrasound (LIPUS) is a well-known bone-healing enhancement device. However, to the best of our knowledge, no clinical study has focused on the effect of LIPUS for postoperative bone union in proximal fifth metatarsal stress fractures. This study aimed to investigate the effect of LIPUS treatment after intramedullary screw fixation for proximal fifth metatarsal stress fractures.MethodsBetween January 2015 and March 2020, patients who underwent intramedullary screw fixation for proximal fifth metatarsal stress fractures were investigated retrospectively. All patients underwent intramedullary screw fixation using a headless compression screw with autologous bone grafts from the base of the fifth metatarsal. The time to restart running and return to sports, as well as that for radiographic bone union, were compared between groups with or without LIPUS treatment. LIPUS treatment was initiated within 3 weeks of surgery in all cases.ResultsOf the 101 ft analyzed, 57 ft were assigned to the LIPUS treatment group, and 44 ft were assigned to the non-LIPUS treatment group. The mean time to restart running and return to sports was 6.8 and 13.7 weeks in the LIPUS treatment group and was 6.2 and 13.2 weeks in the non-LIPUS treatment group, respectively. There were no significant differences in these parameters between groups. In addition, the mean time to radiographic bone union was not significantly different between the LIPUS treatment group (11.9 weeks) and the non-LIPUS treatment group (12.0 weeks). The rate of postoperative nonunion in the LIPUS treatment group was 0% (0/57), while that in the non-LIPUS treatment group was 4.5% (2/44). However, this difference was not statistically significant.ConclusionsThere were no statistically significant differences regarding the time to start running, return to sports, and radiographic bone union in patients with or without LIPUS treatment after intramedullary screw fixation for proximal fifth metatarsal stress fractures. Therefore, we cannot recommend the routine use of LIPUS to shorten the time to bone union after intramedullary screw fixation for proximal fifth metatarsal stress fractures.

Optimal intermittent administration interval of parathyroid hormone 1-34 for bone morphogenetic protein-induced bone formation in a rat spinal fusion model.

IntroductionBoth bone morphogenetic protein 2 (BMP‐2) and teriparatide (parathyroid hormone [PTH] 1‐34) are used to enhance bone healing. There is still no established opinion regarding the optimum dose and administration method. We investigated the optimal administration method for the combination of BMP‐2 and PTH 1‐34 in a rat spinal fusion model.MethodsGroup I was implanted with a control carrier. Groups II, III, and IV were implanted with a carrier containing 3 μg of recombinant human BMP‐2 (rhBMP‐2). In addition, following implantation, PTH 1‐34 injections were administered to Group III thrice a week (total, 180 μg/kg/week) and Group IV six times a week (total, 180 μg/kg/week). The rats were euthanized after 8 weeks, and their spines were explanted; assessed by manual palpation, radiographs, and high‐resolution micro‐computed tomography (micro‐CT); and subjected to histological analysis. Serum markers of bone metabolism were also analyzed.ResultsManual palpation tests showed that the fusion rates in Groups III and IV were considerably higher than those in Group I. They also had higher radiographic scores than Group I and II. Micro‐CT analysis revealed Tb.Th in the Group IV had higher values than that in the Group I, II, III with significant differences and Tb.Sp in the Group IV had lower values than that in the Group I, II, III with significant differences. Serum marker analysis revealed that Group IV had higher osteocalcin and lower tartrate‐resistant acid phosphatase‐5b than Group III. Histological analysis indicated that Group IV had enhanced trabecular bone structure.ConclusionsFrequent administration of PTH may be better in making thicker and strengthening the trabecular bone structure in newly formed bone in the rat spinal fusion model using insufficient BMP‐2.

Adipose-Derived Stromal Cells and Mineralized Extracellular Matrix Delivery by a Human Decellularized Amniotic Membrane in Periodontal Tissue Engineering.

Periodontitis is a prevalent disease characterized by the loss of periodontal supporting tissues, bone, periodontal ligament, and cementum. The application of a bone tissue engineering strategy with Decellularized Human Amniotic Membrane (DAM) with adipose-derived stromal cells (ASCs) has shown to be convenient and valuable. This study aims to investigate the treatments of a rat periodontal furcation defect model with DAM, ASCs, and a mineralized extracellular matrix (ECM). Rat ASCs were expanded, cultivated on DAM, and with a bone differentiation medium for four weeks, deposited ECM on DAM. Periodontal healing for four weeks was evaluated by micro-computed tomography and histological analysis after treatments with DAM, ASCs, and ECM and compared to untreated defects on five consecutive horizontal levels, from gingival to apical. The results demonstrate that DAM preserves its structure during cultivation and healing periods, supporting cell attachment, permeation, bone deposition on DAM, and periodontal regeneration. DAM and DAM+ASCs enhance bone healing compared to the control on the gingival level. In conclusion, DAM with ASC or without cells and the ECM ensures bone tissue healing. The membrane supported neovascularization and promoted osteoconduction.

Simultaneous Treatment of Photobiomodulation and Demineralized Bone Matrix With Adipose-Derived Stem Cells Improve Bone Healing in an osteoporotic bone defect.

Introduction: The ability of simultaneous treatment of critical-sized femoral defects (CSFDs) with photobiomodulation (PBM) and demineralized bone matrix (DBM) with or without seeded adipose-derived stem cells (ASCs) to induce bone reconstruction in ovariectomized induced osteoporotic (OVX) rats was investigated. Methods: The OVX rats with CSFD were arbitrarily separated into 6 groups: control, scaffold (S, DBM), S + PBM, S + alendronate (ALN), S + ASCs, and S + PBM + ASCs. Each group was assessed by cone beam computed tomography (CBCT) and histological examinations. Results: In the fourth week, CBCT and histological analyses revealed that the largest volume of new bone formed in the S + PBM and S + PBM + ASC groups. The S + PBM treatment relative to the S and S + ALN treatments remarkably reduced the CSFD (Mann-Whitney test, P = 0.009 and P = 0.01). Furthermore, S + PBM + ASCs treatment compared to the S and S + ALN treatments significantly decreased CSFD (Mann Whitney test, P = 0.01). In the eighth week, CBCT analysis showed that extremely enhanced bone regeneration occurred in the CSFD of the S + PBM group. Moreover, the CSFD in the S + PBM group was substantially smaller than S, S + ALN and S + ASCs groups (Mann Whitney test, P = 0.01, P = 0.02 and P = 0.009). Histological observations showed more new bone formation in the treated CSFD of S + PBM + ASCs and S + PBM groups. Conclusion: The PBM plus DBM with or without ASCs significantly enhanced bone healing in the CSFD in OVX rats compared to control, DBM alone, and ALN plus DBM groups. The PBM plus DBM with or without ASCs significantly decreased the CSFD area compared to either the solo DBM or ALN plus DBM treatments.

Men who stare at bone: multimodal monitoring of bone healing.

Knowledge of the physiological and pathological processes, taking place in bone during fracture healing or defect regeneration, is essential in order to develop strategies to enhance bone healing under normal and critical conditions. Preclinical testing allows a wide range of imaging modalities that may be applied both simultaneously and longitudinally, which will in turn lower the number of animals needed to allow a comprehensive assessment of the healing process. This work provides an up-to-date review on morphological, functional, optical, biochemical, and biophysical imaging techniques including their advantages, disadvantages and potential for combining them in a multimodal and multiscale manner. The focus lies on preclinical testing of biomaterials modified with artificial extracellular matrices in various animal models to enhance bone remodeling and regeneration.

Does Needle Design Affect the Regenerative Potential of Bone Marrow Aspirate? An In Vitro Study.

While autologous bone is still the gold standard for treatment of bone defects, its availability is limited. Sufficient numbers of mesenchymal stroma cells (MSC) may be an alternative. Small volumes of bone marrow aspirate (BMA) were harvested with two different needle systems comparing the yield and regenerative potency of the MSCs. BMA (10 mL) was aspirated from the posterior iliac crest of 12 patients with degenerative spinal disc disease using both needle systems in each patient: the Jamshidi needle (JAM) and on the contralateral side the Marrow Cellution® Needle (AMC). Number of mononuclear cells (MNCs) and regeneration capacity (colony-forming unit/CFU) were determined. MSCs were characterized for surface markers and their differentiation into trilineages. There was no significant difference between the two harvesting needles regarding the quantity of MNCs in BMA: 5.2 ± 1.8 × 109 MNC/mL for AMC vs. 4.8 ± 2.5 × 109 MNC/mL for JAM, p = 0.182. The quantity of CFUs per ml BMA was similar for both groups: 3717 ± 5556 for AMC and 4305 ± 5507 for JAM (p = 0.695). The potency of MSCs expressed as colony-forming potential per 106 MNC resulted in 0.98 ± 1.51 for AMC and 1.00 ± 0.96 for JAM (p = 0.666). Regardless of the needle design, 10 mL bone marrow aspirate contains a sufficient number of about 40,000 MSCs that can be used to enhance bone healing.

Biological and biomechanical basis of long-bone diaphyseal fractures: from fracture to non-union

Bone healing of diaphyseal fractures is a complex biological process that can often be adversely affected by patient-related and fracture-related factors and eventually end in delayed union and non-union. Surgical and non-surgical approaches have been widely applied, according to the fracture pattern and patient characteristics. For humeral diaphyseal fractures, plate fixation provided excellent results in terms of healing rate and time to union. For femoral and tibial shaft fractures, locked intramedullary nailing is considered the technique of choice. If impaired, the reparative process after intramedullary nailing can be enhanced through different surgical techniques, such as dynamization or exchange nailing. Moreover, the mechanical stability of the fracture site can be improved through augmentation plating, bone grafting or external fixation techniques, with satisfactory results. This article aims to review the biomechanical principles of reparative osteogenesis in long bone fractures after conservative and surgical treatment. Moreover, the evidence on the current options for bone healing enhancement, and treatment and prevention of delayed union and non-union will be discussed.

Brd4 Inactivation Increases Adenoviral Delivery of BMP2 for Paracrine Stimulation of Osteogenic Differentiation as a Gene Therapeutic Concept to Enhance Bone Healing.

ABSTRACTBromodomain (BRD) proteins are histone code interpreters that recognize acetylated lysines and link the dynamic state of chromatin with the transcriptional machinery. Here, we demonstrate that ablation of the Brd4 gene in primary mouse bone marrow–derived mesenchymal stem cells via a conditional Brd4fl/fl allele suppresses osteogenic lineage commitment. Remarkably, loss of Brd4 function also enhances expression of genes in engineered adenoviral vectors, including Cre recombinase and green fluorescent protein (GFP). Similarly, vector‐based expression of BMP2 mRNA and protein levels are enhanced upon Brd4 depletion in cells transduced with an adenoviral vector that expresses BMP2 (Ad‐BMP2). Importantly, Brd4 depletion in MC3T3‐E1 and human adipose‐derived mesenchymal stem cells (AMSCs) transduced with Ad‐BMP2 enhances osteogenic differentiation of naïve MC3T3‐E1 cells via paracrine mechanisms based on transwell and conditioned medium studies. Our studies indicate that Brd4 depletion enhances adenoviral transgene expression in mammalian cells, which can be leveraged as a therapeutic strategy to improve viral vector‐based gene therapies. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Low-level laser therapy (LLLT) improves alveolar bone healing in rats.

The main objective of the present study was to evaluate the effect of low-level laser therapy (LLLT) in enhancing bone healing in irradiated alveolus post-tooth extraction. Sixty male Wistar rats (180 ± 10g) were used in the present study. The left maxillary first molars were extracted, and the alveolar region was irradiated by diode laser device (GaAlAs) immediately after extraction and for more 3-day daily applications. The animals were randomly assigned into two groups: control group (n = 30, with left maxillary molar extraction-CG) and experimental group (n = 30, with tooth extraction and low-level laser therapy applied to the dental alveolus for 42s-EG). These groups were divided into subgroups (five rats per subgroup) according to the observation time point-1, 2, 3, 5, 7, and 10days-post-tooth extraction. The maxillary bone was separated, and the specimens were stained with hematoxylin and eosin, Masson's trichrome, and picrosirius red and immunohistochemistry for RUNX-2. Parametric and nonparametric tests were used with a significance level of 5%. LLLT accelerated bone healing with mature collagen fiber bundles and early new bone formation. Histomorphometric analysis revealed an increase of osteoblast (RUNX-2) and osteoclast (TRAP) activity and in the area percentage of cancellous bone in the lased alveolus compared to the control group. This increase was statistically significant (p < 0.05). Application of LLLT with a GaAlAs diode laser device enhanced bone healing and mineralization on alveolar region.

Local Delivery of Therapeutic Boron for Bone Healing Enhancement.

To evaluate if local delivery of boron can accelerate bone healing and examine if the bioactive salt impacts the osteogenic response of bone-derived osteoclasts and osteoblasts by the regulation of the Wnt/β-catenin pathway. Bilateral femoral cortical defects were created in 32 skeletally mature C57 mice. On the experimental side, boric acid (8 mg/kg concentration) was injected locally, whereas on the control side, saline was used. Mice were euthanized at 7, 14, and 28 days. MicroCT was used to quantify bone regeneration at the defect. Histological staining for alkaline phosphatase and tartrate-resistant acid phosphatase was used to quantify osteoblast and osteoclast activity, respectively. Immunohistochemical antibodies, β-catenin, and CD34 were used to quantify active β-catenin levels and angiogenesis, respectively. The boron group exhibited higher bone volume and trabecular thickness at 28 days on microCT. Both alkaline phosphatase activity and β-catenin activity was significantly higher in the boron group at 7 days. In addition, CD34 staining revealed increased angiogenesis at 14 days in boron-treated groups. We found boron to have no association with osteoclast activity. This study shows that local delivery of boron is associated with an increase in osteoblast activity at early phases of healing. The corresponding increase in β-catenin likely supports that boron increases osteoblast activity by the Wnt/β-catenin pathway. Increased angiogenesis at 14 days could be a separate mechanism of increasing bone formation that is independent of Wnt/β-catenin activation.

English

BACKGROUND In recent days a high success rate has been documented for dental implants as a treatment option for missing teeth replacement. Low level laser therapy (LLLT) is a novel therapeutic modality reported to enhance bone healing. Few studies reported that LLLT enhances proliferation and differentiation of osteoblasts and also reduces postoperative pain. The purpose of this study was to assess if post-operative lowlevel laser irradiation of the implant site has any influence on crestal bone loss and pain reduction around dental implants. METHODS In this split mouth randomised study, 9 subjects with bilateral edentulous sites in the mandibular posterior region willing for replacement were taken into consideration. A total of 30 implants were placed on either side of the mandible followed by Low Level Laser irradiation on the test side immediately after implant placement, on 4th and 7th day. Crestal bone level assessment was done radiographically at 3 months and post-operative pain was assessed using visual analogue scale (VAS) scoring by patients from baseline to 6 days. Statistical analysis was done using unpaired t test for inter group crestal bone loss and Mann-Whitney U test for intergroup pain reduction scores. RESULTS The mean crestal bone loss of control group was 1.40 with a standard deviation of 0.39 and for test group it was 0.76 with a standard deviation of 0.26. A significant difference was observed between control and test groups with respect to crestal bone loss scores at 3 months. On comparing VAS pain scores between the two groups from day 1 to 6, significant difference between the groups was observed on day 2, 3 and 4 with P-values 0.042, 0.0011, 0.0036 respectively. CONCLUSIONS Low level irradiation around the dental implants showed a significant reduction in crestal bone loss and post-operative pain. KEY WORDS Dental Implant, Low Level Laser Therapy, Crestal Bone Loss, Post-Operative Pain

Functionalization of Synthetic Bone Substitutes.

Bone substitutes have been applied to treat osseous defects for a long time. To prevent implant related infection (IRI) and enhance bone healing functionalized biomaterials, antibiotics and osteoinductive substances have been introduced. This study gives an overview of the current available surface-coated bone substitutes and provides an outlook for future perspectives.

Improved activity of MC3T3-E1 cells by the exciting piezoelectric BaTiO3/TC4 using low-intensity pulsed ultrasound

Developing bioactive materials for bone implants to enhance bone healing and bone growth has for years been the focus of clinical research. Barium titanate (BT) is an electroactive material that can generate electrical signals in response to applied mechanical forces. In this study, a BT piezoelectric ceramic coating was synthesized on the surface of a TC4 titanium alloy, forming a BT/TC4 material, and low-intensity pulsed ultrasound (LIPUS) was then applied as a mechanical stimulus. The combined effects on the biological responses of MC3T3-E1 cells were investigated. Results of scanning electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray diffraction showed that an uniform nanospheres -shaped BT coating was formed on TC4 substrate. Piezoelectric behaviors were observed using piezoelectric force microscopy with the piezoelectric coefficient d33 of 0.42 pC/N. Electrochemical measures indicated that LIPUS-stimulated BT/TC4 materials could produce a microcurrent of approximately 10 μA/cm2. In vitro, the greatest osteogenesis (cell adhesion, proliferation, and osteogenic differentiation) was found in MC3T3-E1 cells when BT/TC4 was stimulated using LIPUS. Furthermore, the intracellular calcium ion concentration increased in these cells, possibly because opening of the L-type calcium ion channels was promoted and expression of the CaV1.2 protein was increased. Therefore, the piezoelectric BT/TC4 material with LIPUS loading synergistically promoted osteogenesis, rending it a potential treatment for early stage formation of reliable bone-implant contact.