BMP-2

Background: Extracorporeal shock wave therapy (ESWT) exerts its principal therapeutic effects through mechanotransduction rather than direct mechanical disruption. The absence of a unified conceptual framework linking its biophysical parameters to a coherent regenerative mechanism has impeded protocol standardization and formal classification within orthobiology. Hypothesis: We propose OLTHEShockw — Orthobiology by Local Transient Hyperbaric Effect with Shock Waves (Spanish: OHTLOc) — which posits that ESWT generates focal transient high-pressure microenvironments at tissue impedance interfaces, reaching peak pressures of 10–100 MPa in microseconds. These events exceed classical hyperbaric chamber pressures (0.1–0.2 MPa) by 50- to 1,000-fold, constituting a form of mechanical, pulsed, focal hyperbaria that activates mechanotransduction cascades releasing VEGF, BMP-2, TGF-β, eNOS/NO, and IGF-1. Significance: OLTHEShockw provides a unifying metrological criterion — peak tissue pressure (MPa) — reconciling disparate ESWT measurement systems (bar for radial devices, mJ/mm² for focused devices) and formally positioning focused ESWT as a biophysical orthobiologic. This framework supports evidence-based protocol standardization and regulatory classification. Keywords: OLTHEShockw; OHTLOc; HTL; LTHE; extracorporeal shock wave therapy; orthobiology; mechanotransduction; local transient hyperbaric effect; protocol standardization; VEGF; BMP-2; regenerative medicine.
Keywords: OLTHEShockw, OHTLOc, HTL, LTHE,
extracorporeal shock wave therapy, orthobiology,
mechanotransduction, local transient hyperbaric effect,
protocol standardization, VEGF, BMP-2,
regenerative medicine, Zenodo Preprint 2026

The preclinical study aimed to compare the healing of segmental bone defects treated with biodegradable hyaluronic acid and tricalcium phosphate-based hydrogel with the established autologous spongioplasty. Another aim was to evaluate the hydrogel as a scaffold for osteoinductive growth factor of bone morphogenetic protein-2 (BMP-2) and stem cells.

Bone morphogenetic proteins (BMPs) are responsible for bone formation during embryogenesis and bone regeneration and remodeling. The osteoinductive action of BMPs, especially BMP-2 and BMP-7, has led to their use in a range of insurmountable treatments where intervention is required for effective bone regeneration. Introduction of BMP products to the market, however, was not without reports of multiple complications and side effects. Aiming for optimization of the therapeutic efficacy and safety, efforts have been focused on improving the delivery of BMPs to lower the administered dose, localize the protein, and prolong its retention time at the site of action. A major challenge with these efforts is that the protein stability should be maintained. With this review we attempt to shed light on how the stability of BMPs can be affected in the formulation and delivery processes. We first provide a short overview of the current standing of the complications experienced with BMP products. We then discuss the different delivery parameters studied in association with BMPs, and their influence on the efficacy and safety of BMP treatments. In particular, the literature addressing the stability of BMPs and their possible interactions with components of the delivery system as well as their sensitivity to conditions of the formulation process is reviewed. In summary, recent developments in the fields of bioengineering and biopharmaceuticals suggest that a good understanding of the relationship between the formulation/delivery conditions and the stability of growth factors such as BMPs is a prerequisite for a safe and effective treatment.

Background:  Mandibular fracture is one of the most common fractures. The most common treatment for mandibular fractures is fixation. Therefore, xenogeneic agents such as platelet-rich fibrin (PRF) and Biodentine accelerate the reparative process. Biodentine is an interesting active ingredient that can induce bone regeneration. PRF and Biodentine can promote bone healing, but no literature discusses the differences between PRF and Biodentine osteoinduction mechanisms in treating mandibular fractures. Objective:  This article aimed to compare the effect of osteoinductive PRF with Biodentine for mandibular fractures. Material and Method: The research was conducted as a scoping review by performing a thorough search of the PubMed, Scopus, Science Direct, Elsevier, and Google Scholar databases. The study was obtained based on literature studies in the form of journals and textbooks in the last ten years (2013-2023). Result: The osteoinductive effect and mechanism of Biodentine in enhancing bone repair are likely correlated with releasing biologically active ions from calcium silicate cement and stimulating gene expression Runx-2. PRF has an osteoinductive role, causing the mechanism of releasing growth factors such as PDGF, VEGF, TGF-β, and IGF that promote the osteogenic process. Conclusion: There was no significant difference in the osteoinduction effect of PRF and Biodentine because these materials have different mechanisms of action for bone repair.

The bone morphogenic proteins(BMPs) are multifunctional growth factors that belong to the TGF-β superfamily. Among BMPs, BMP-2 is known as the most effective cytokine. The present study evaluated effects of anorganic bovine bone graft material coated with synthetic peptide which corresponds to residues 73-92 of the knuckle epitope of hBMP-2 on bone formation in rabbit calvarial defects. The residues 73-92 of hBMP-2 was absorbed on anorganic bone mineral(ABM) by incubating ABM for 24 hrs. The rabbit calvarial defects were not filled at all(control group), and were filled ABM only(ABM group), ABM with residues 73-92 derived from hBMP-2(ABM/hBMP-2P group) and ABM combined with hBMP-2(ABM/rhBMP-2 group). The groups were evaluated using histologic observation and histomorphometrc analysis at 4 and 8 weeks after healing. The higher osteoconductivity of the ABM/hBMP-2P and ABM/rhBMP-2 groups than the control and ABM only groups. Compared to the control group at 4 weeks after healing, the defects grafted with ABM/hBMP-2P and ABM/rhBMP-2 showed more active bone formation with a statistical significance(p<0.05). And ABM/hBMP-2P and ABM/rhBMP-2 showed a higher percentage bone length from the defect margins than control and ABM only groups at 4 weeks of healing with a statistical significance(p<0.05). ABM/hBMP-2P and ABM/rhBMP-2 may involve mainly initial bone healing and residues 73-92 of hBMP-2 could attribute to new bone formation. Based on the results of this study, anorganic bovine bone maybe a good carrier for peptide and anorganic bovine combined with residues 73-92 of hBMP-2 can increase the new bone formation in early stage in rabbit calvaria.

Calcifications are not only one of the most important early diagnostic markers of breast cancer, but are also increasingly believed to aggravate the proliferation of cancer cells and invasion of surrounding tissue. Moreover, this influence appears to vary with calcification composition. Despite this, remarkably little is known about the composition and crystal structure of the most common type of breast calcifications, and how this differs between benign and malignant lesions. We sought to determine how the phase composition and crystallographic parameters within calcifications varies with pathology, using synchrotron X-ray diffraction. This is the first time crystallite size and lattice parameters have been measured in breast calcifications, and we found that these both parallel closely the changes in these parameters with age observed in fetal bone. We also discovered that these calcifications contain a small proportion of magnesium whitlockite, and that this proportion increases ...

Ovarian cancer is one of the most common causes of death in the female population. Among malignant tumors, ovarian cancer ranks fourth after neoplasms of the body, cervix, and mammary gland. High mortality rates in malignant ovarian neoplasms are primarily due to the asymptomatic course, the ineffectiveness of screening diagnostic methods, and the progression of the tumor process. One of the clinical and morphological features of ovarian cancer is pathological biomineralization. The purpose of the work is to establish the morphological features and phase composition of pathological biominerals. Also to investigate the potential diagnostic value of mineral formations of ovarian cancer. We have analyzed 60 samples of ovarian cancer by histopathology (hematoxylineosin and von Kossa staining), immunohistochemistry, scanning electron microscopy with EDX and transmission electron microscopy. Results. We detected hyperechoic formations of a rounded shape with smooth edges and precise contours during the ultrasound diagnosis of ovarian cancer. The size ranged from 2 to 5 mm. Histological analysis demonstrated the presence of psammoma bodies (Pbs) in the tumor tissue of the ovary with a variation in the amount from 1 to 200 units. PBs were mainly localized at the base of papillary growths of ovarian serous carcinoma and tumor tissue detritus. The sizes of these formations varied from 12.6 to 493.7 μm. Psammoma bodies and their fragments in the tumor tissue were stained black with von Kossa staining. This indicates the presence of calcium phosphate salts in the composition of these bodies. Immunohistochemical Chapter «Medical sciences» examination of ovarian cancer tissue demonstrated the accumulation of OPN and OC on the surface of mineral formations. This indicates the presence of hydroxyapatite and proteins in the structural composition of PBs. Scanning electron microscopy established that the calcifications have different sizes, rounded shapes, and fragile structures. The main lines of Ca and P were present in the EDX spectra. Their intensity ratio reflects the characteristic features of hydroxyapatite Ca 10 (PO 4) 6 (OH) 2. Transmission electron microscopy demonstrated the poly-and monodispersity of apatite crystals. The electron diffraction (ED) pattern indicates the polycrystalline nature of these mineral inclusions. Crystalline particles ranged in size from 5 to 15 nm to 40 to 50 nm, which illustrates the polydisperse morphology of nanosized crystals of the pathological deposit. Therefore, studying the structure, physicochemical, and phase composition of ovarian cancer calcifications, and their imaging features is essential because of the possible practical application of this pathological phenomenon for the early diagnosis of ovarian cancer and other malignant neoplasms with biomineralization. The structural properties of biomineral deposits of tumor tissue make it possible to increase the accuracy of early diagnosis, and the informativeness of the final result at the pre-and postoperative stages, which can improve the prognosis and preserve the quality of life of patients.

Identification of predictive biomarkers for ovarian cancer (OC) treatment, particularly in the platinum-resistant/refractory setting, is highly relevant for clinical management. E-cadherin, vimentin, and osteopontin (OPN) are proteins associated with tumor microenvironment (TME) remodelling that play key roles in cancer. This study aimed to evaluate the association between the staining patterns of these proteins with survival outcomes in a series of OC patients, namely in patients with platinum-resistant/refractory disease. Low E-cadherin expression and high vimentin expression in all patient groups (as well as for E-cadherin in the platinum-resistant arm) were significantly associated with longer overall survival (OS). Low cytoplasmic OPN expression (and cytoplasmic and membrane OPN in the platinum-resistant arm) were significantly associated with longer OS. In patients that responded to treatment (pegylated liposomal doxorubicin (PLD) or other), low cytoplasmic OPN expression was ...

This study aims to investigate the possible different roles of the BMP-2 variants, cytoplasmic and nuclear variant, in both epithelial to mesenchymal transition and in microcalcifications origin in human breast cancers. To this end, the in situ expression of cytoplasmic and nuclear BMP-2 was associated with the expression of the main epithelial to mesenchymal transition biomarkers (e-cadherin and vimentin) and molecules involved in bone metabolisms (RUNX2, RANKL, SDF-1) by immunohistochemistry. In addition, the expression of cytoplasmic and nuclear BMP-2 was associated with the presence of microcalcifications. Our data showed a significant association among the number of cytoplasmic BMP-2-positive cells and the number of both vimentin (positive association) and e-cadherin (negative association) positive breast cells. Conversely, no associations were found concerning the nuclear BMP-2-positive breast cells. Surprisingly, the opposite result was obtained by analyzing the variants of B...

The Long Pentraxin 3 (PTX3) is a multifunctional glycoprotein released by peripheral blood leukocytes and myeloid dendritic cells in response to primary pro-inflammatory stimuli, that acts as a non-redundant component of the humoral arm of innate immunity. In addition to the primary role in the acute inflammatory response, PTX3 seems to be involved in other physiological and pathological processes. Indeed, PTX3 seems to play a pivotal role in the deposition and remodeling of bone matrix during the mineralization process, promoting osteoblasts differentiation and activity. Recently, PTX3 was seen to be involved in the ectopic calcifications’ formation in breast cancer disease. In this regard, it has been observed that breast cancer tumors characterized by high expression of PTX3 and high amount of Breast Osteoblast Like Cells (BOLCs) showed several Hydroxyapatite (HA) microcalcifications, suggesting a likely role for PTX3 in differentiation and osteoblastic activity in both bone and ...

The detection of microcalcifications in the breast by mammography is of great importance for the early diagnostics of breast cancer. This study aimed to establish the basic morphological and crystal-chemical properties of microscopic calcifications and their impact on breast cancer tissue. During the retrospective study, 55 out of 469 breast cancer samples had microcalcifications. The expression of the estrogen and progesterone receptors and Her2-neu showed no significant difference from the non-calcified samples. An in-depth study of 60 tumor samples revealed a higher expression of osteopontin in the calcified breast cancer samples (p ˂ 0.01). The mineral deposits had a hydroxyapatite composition. Within the group of calcified breast cancer samples, we detected six cases of colocalization of oxalate microcalcifications together with biominerals of the usual “hydroxyapatite” phase composition. The simultaneous presence of calcium oxalate and hydroxyapatite was accompanied by a diffe...

Despite the high prevalence of ischemic heart diseases worldwide, no antibody-based treatment currently exists. Starting from the evidence that a specific isoform of the Bone Morphogenetic Protein 1 (BMP1.3) is particularly elevated in both patients and animal models of myocardial infarction, here we assess whether its inhibition by a specific monoclonal antibody reduces cardiac fibrosis. We find that this treatment reduces collagen deposition and cross-linking, paralleled by enhanced cardiomyocyte survival, both in vivo and in primary cultures of cardiac cells. Mechanistically, we show that the anti-BMP1.3 monoclonal antibody inhibits Transforming Growth Factor β pathway, thus reducing myofibroblast activation and inducing cardioprotection through BMP5. Collectively, these data support the therapeutic use of anti-BMP1.3 antibodies to prevent cardiomyocyte apoptosis, reduce collagen deposition and preserve cardiac function after ischemia.

Background: Complex bone defects could be challenging and implant placement difficult after a complicated extraction. Treatment planning should include implant placement, bone augmentation and soft tissue grafting in horizontal vertical and transversal relationship. The aim of this study is to evaluate clinically the result of a staged-treatment planning for bone augmentation and soft tissue management before placing the implant and after implantation. Methods: A case is report during a period of 5 years. Showing staged treatment planning. Results: Clinical success was achieved with this protocol of bone augmentation and connective tissue grafting for soft tissue augmentation. Conclusions: To restore the anatomy of the smile of a patient, and achieved an esthetically acceptable result is important to evaluate and planned for not only the implant placement but also for restoring the tissues supporting and surrounding the implant. Multiple procedures are necessary to obtain this result.

Identification of predictive biomarkers for ovarian cancer (OC) treatment, particularly in the platinum-resistant/refractory setting, is highly relevant for clinical management. E-cadherin, vimentin, and osteopontin (OPN) are proteins associated with tumor microenvironment (TME) remodelling that play key roles in cancer. This study aimed to evaluate the association between the staining patterns of these proteins with survival outcomes in a series of OC patients, namely in patients with platinum-resistant/refractory disease. Low E-cadherin expression and high vimentin expression in all patient groups (as well as for E-cadherin in the platinum-resistant arm) were significantly associated with longer overall survival (OS). Low cytoplasmic OPN expression (and cytoplasmic and membrane OPN in the platinum-resistant arm) were significantly associated with longer OS. In patients that responded to treatment (pegylated liposomal doxorubicin (PLD) or other), low cytoplasmic OPN expression was ...

Microcalcifications serve as diagnostic markers for breast cancer, yet their formation pathway(s) and role in cancer progression are debated due in part to a lack of relevant 3D culture models that allow studying the extent of cellular regulation over mineralization. Previous studies have suggested processes ranging from dystrophic mineralization associated with cell death to bone-like mineral deposition. Here, we evaluated microcalcification formation in 3D multicellular spheroids, generated from non-malignant, pre-cancer, and invasive cell lines from the MCF10A human breast tumor progression series. The spheroids with greater malignancy potential developed necrotic cores, thus recapitulating spatially distinct viable and non-viable areas known to regulate cellular behavior in tumors in vivo. The spatial distribution of the microcalcifications, as well as their compositions, were characterized using nanoCT, electron-microscopy, and X-ray spectroscopy. Apatite microcalcifications we...

Background: The aim of this study was to determine the osteoconductivity of biphasic calcium phosphate collagen composite (BCPC) in rabbit calvarial defect model by comparing with biphasic calcium phosphate (BCP). Four 8 mm diameter bicortical calvarial defects were made in ten rabbits. Each of the defects was randomly assigned and filled with 1) collagen sponge, 2) BCP, 3) BCPC, and 4) nothing as control. The animals were sacrificed at either 2 weeks (n = 5) or 8 weeks (n = 5) healing period. Results: All groups showed wedge shaped new bone formation limited to the area of the defect margin at both healing periods. The amounts of new bone and defect closure were similar among all groups. In the control and collagen sponge group, the center of the defect was depressed by surrounding tissues. In contrast, in BCP and BCPC group, the center of the defect did not depressed and the grafted materials maintained the space. And the augmented area was significantly higher in BCP and BCPC group compared to the control and collagen sponge group at both healing periods (p < 0.05). Conclusions: The BCPC and BCP demonstrated proper space maintaining capacity and osteoconductive property, suggesting BCPC can be efficiently utilized in various clinical situations.

Background The bone morphogenetic proteins (BMPs) belong to a unique group of proteins that includes the growth factor TGF-β. BMPs play important roles in cell differentiation, cell proliferation, and inhibition of cell growth. They also participate in the maturation of several cell types, depending on the microenvironment and interactions with other regulatory factors. Depending on their concentration gradient, the BMPs can attract various types of cells and act as chemotactic, mitogenic, or differentiation agents. BMPs can interfere with cell proliferation and the formation of cartilage and bone. In addition, BMPs can induce the differentiation of mesenchymal progenitor cells into various cell types, including chondroblasts and osteoblasts. The aim of this study was to analyze the effects of treatment with rhBMP-2 on the proliferation of canine mesenchymal stem cells (cMSCs) and the tumor suppression properties of rhBMP-2 in canine osteocarcoma (OST) cells. Osteosarcoma cell lines...

Introduction: A variety of materials have been used to construct scaffolds for tissue-engineering applications, but their pressfitting properties in complex defects are seldom reported. A novel elastomeric nanosized hydroxyapatite-incorporated poly(ester-urethane) (nHA/PU) scaffold was developed and shown to adsorb proteins [1] and promote angiogenesis [2-4] within the scaffold. However, limited in vivo studies have been performed to investigate whether nHA/PU scaffolds can effectively enhance bone regeneration. The objective of this study is to determine the effectiveness of nHA/PU scaffold as a carrier of rhBMP-2 (R&D Systems) for guided bone regeneration. We hypothesize that the presence of rhBMP-2 on the nHA/PU scaffold surface will enhance bone repair. Methods: Scaffold preparation. The scaffold was prepared using a saltleaching/phase-inverse process as previously described [5]. The scaffold contains 10% by weight nHA, and 90% w:w PU synthesized by polycondensation of 1,6-hexamethylene diisocyanate, poly(ε-caprolactone) diol and 1,4,3,6-dianhydro-D-sorbitol. For this study, the nHA/PU scaffolds were prepared with a circular dimension of 5.5 mm and 1 mm thickness, with pore sizes of 90 to 300 μm. Surgical preparation. All animal procedures were carried out in accordance with the guidelines of the Canadian Council on Animal Care (CCAC). A 5-mm circular defect was made on each parietal bone of anaesthetized Sprague-Dawley rats (aged 11-12 weeks) as described in other studies [6, 7]. No-protein control scaffolds (nHA/PU) or nHA/PU scaffolds coated with 1 µg of rhBMP-2 (nHA/PU/rhBMP-2) were inserted into the calvarial defects. Analysis. At intervals of 2 weeks, the rats were imaged with a live-animal microCT scanner (GE eXplore Locus) to quantify mineral deposition within the defect sites. In addition, at 6 and 12 weeks, implants and surrounding bone were also recovered and fixed for histological analysis. Mechanical testing. At 6 and 12 weeks, specimens were prepared for micro-indentation and push-out tests to determine the mechanical properties and osteointegration of the newly formed bone. Statistics. Quantitative data were expressed as means ± standard error. Statistical analysis among groups was performed using two-way analysis of variance (ANOVA) and p<0.05 was considered to be statistically significant. Results: At 12 weeks, the nHA/PU/rhBMP-2 implants were filled with apparent new bone, whereas for the nHA/PU group apparent bone formation was minimal (Fig 1). Using quantitative microCT analysis, addition of rhBMP-2 to the scaffolds enhanced bone formation (approximately 4-fold) compared to control (Fig 2). Histological sections stained with H&E and Masson trichrome also agree with microCT data showing higher bone formation in the nHA/PU/rhBMP-2 group compared to control (data not shown). Furthermore, the newly formed bone in the nHA/PU/rhBMP-2 group has a higher stiffness and required more load to fracture the implant-bone interface compared to the control group. However, these values are still lower compared to native bone (Fig 3). Discussion: The addition of rhBMP-2 to nHA/PU scaffolds significantly enhanced in vivo bone repair. Based on microCT and histological analysis, the new in-growth bone appears to form from the host bone adjacent to the dura membrane. No major evidence of inflammation was seen in any of the groups, which indicate that the nHA/PU scaffold is biocompatible and can be used as a bioactive protein carrier. Mechanically, however, the properties of the newly formed bone do not reach that of undamaged bone in the 12 weeks post-surgery period. Of note, nHA/PU scaffolds bind rhBMP-2 well and slowly release the growth factor over time (85% of the rhBMP-2 was still bound to the scaffold at 72 h; data not shown). Significance: This study demonstrates that nHA/PU scaffolds are an effective carrier for rhBMP-2 for use in bone-regeneration therapy. Our ongoing study will look at the effectiveness of the scaffold as a carrier for various bioactive proteins and whether addition of mesenchymal stromal cells further enhances bone repair.

This thesis focuses on the fabrication of complex microcapsules that can be deliver therapeutic drugs on-demand using ultrasound waves. These microcapsules are composed of a water-based core and a biologically inert shell into which particles are embedded. Upon the application of ultrasound, these embedded particles (like corks on a bottle) are popped out to release the "corks" from the shell, creating pores from which the drug in the microcapsule core can be released. In the absence of ultrasound signals, the microcapsules do not release any of their contents, making these effective for "on-demand" release. These microcapsules are made using a modified process based on electrospraying which allows very precise control over the microcapsules' physical properties, incorporating a key modification that overcomes an inherent issue with the general technique. These microcapsules also improve on currently used ultrasound triggered drug delivery systems by requiring shorter periods of ultrasound and/or enabling better control over the dynamics of drug release following the ultrasound pulse.

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Despite the high regenerative capacity of bone tissue, there are some cases where bone repair is insufficient for a complete functional and structural recovery after damage. Current surgical techniques utilize natural and synthetic bone grafts for bone healing, as well as collagen sponges loaded with drugs. However, there are certain disadvantages associated with these techniques in clinical usage. To improve the therapeutic efficacy of bone tissue regeneration, a number of drug delivery systems based on biodegradable natural and synthetic polymers were developed and examined in in vitro and in vivo studies. Recent studies have demonstrated that biodegradable polymers play a key role in the development of innovative drug delivery systems and tissue engineered constructs, which improve the treatment and regeneration of damaged bone tissue. In this review, we discuss the most recent advances in the field of polymer-based drug delivery systems for the promotion of bone tissue regenerat...

Several methods are used to enhance bone repair and new bone formation. Bone matrix gelatin is recently introduced and calcium phosphate has gained importance in orthopaedics as repair materials for bone defects. The objective of this study was to evaluate of effects of a mixture of calcium phosphate granules and allogeneic bone matrix gelatin on bone repair in rabbit. 30 male rabbits were divided into three groups. After induction of general anesthesia, a segmental bone defect of 5 mm in length was created in the middle of the left radius shaft. In control group defect was left untreated, in group 2, calcium phosphate granules were used and in group 3, a mixture of calcium phosphate granules and bone matrix gelatin were used to fill the bone defect. Rabbits of each group were euthanized at 45 days intervals for evaluation of histopathological and biomechanical changes. Histopathology finding in control group, defect seemed to be filled with connective tissue and bone marrow spaces ...

Antioxidant agents stimulate new bone formation during the process of repair of bone defects. This study aimed to examine the effects of systemic administration of resveratrol, an antioxidant agent, on new bone formation in a rat model of experimentally induced bone defects. The study consisted of four groups. Each group was divided into 2 subgroups on the basis of the date on which the animals were sacrificed. We formed a 5-mm bone defect in the right mandible of the animals in each group. Control group (C) received no further treatment, first experimental group (G) received a bovine bone graft, second experimental group (R) received systemic administration of resveratrol (10 mg/kg), third experimental group (GR) received systemic administration of resveratrol (10 mg/kg) with a bovine bone graft. The results of histopathological examination on the 14 th and 28 th day showed significant differences in the degree of ossification, fibrosis, angiogenesis, and the density of inflammator...

The dentist faces a difficult challenge in preserving the quantity and quality of gum and bone tissues. Furthermore, the processes adopted should meet these objectives while posing no substantial future issues. In current dental and maxillofacial surgery, a variety of materials as Morphogenic Protein of Bone (BMP) and Platelet Rich Fibrins (PRF) are employed to repair and reconstruct bone tissue. The purpose of this review was to compare the efficiency of both BMP’s and PRF’s in Socket preservation. Reviewing the literature was conducted by using the following search strategies: Bone Morphogenic Proteins, Platelet Rich Fibrins, socket preservation approach, comparison of BMPs and PRFs, Bone Morphogenic Proteins and effectiveness in socket preservation and Platelet Rich Fibrins’ effectiveness in socket preservation. In conclusion, BMPs have been studied more completely than PRFs, and they have higher impacts on hard tissue regeneration and repair, according to the research. The fact ...

A porous calcium phosphate ceramic, which induced bone formation in soft tissues of dogs, was termed as osteoinductive biomaterial and studied as a carrier of bone morphogenetic protein (rhBMP-2). Cylinder implants (laser spot 4 x 5 mm) impregnated with 0, 1, 10 and 40 microg rhBMP-2 were implanted in dorsal muscles of rabbits for five weeks. Histological observation and histomorphometric analysis were performed on thin un-decalcified sections. No bone formation was detected in the implants without rhBMP-2, while mature lamellar bone was found inside the implants with 1 microg rhBMP-2, both on the outer surface and inside the implants with 10 microg and 40 microg rhBMP-2. Little difference in formed bone was found between 1 microg and 10 microg rhBMP-2, but no difference was found between 10 microg and 40 microg rhBMP-2. A significant difference in bone marrow formation was found among 1, 10 and 40 microg rhBMP-2. The more rhBMP-2, the more bone marrow formed. The present results in...

INTRODUCTION: Comminuted fractures of the mandible are an important traumatism, in which the mandibular bone is splintered or crushed. Treatment of these types of fractures had always been a challenge to surgeons, considering both the severity of the trauma and the presence of discontinuity defects which require replacement and augmentation in order to treat the bone loss. Titanium mesh was proved to be particularly useful for reconstruction of mandibular discontinuity defects also it can be used as a scaffold for bone grafts. Electrophoretic deposition (EPD) is a coating technique used to increase the osseointegration and increases bone regeneration. OBJECTIVES: To histologically evaluate the effect of titanium mesh coated with hydroxyapatite nanocrystals using electrophoretic deposition on bone osseointegration and healing following comminuted mandibular fractures. MATERIALS AND METHODS: A controlled induced comminuted fracture on the inferior border of the mandible extending from the 2 nd premolar to the 2 nd molar was performed on 12 mongrel dogs. Six dogs were treated using titanium mesh coated with hydroxyapatite nanocrystals (study group) and the other six using uncoated titanium mesh (control group). The dogs were sacrificed at 2, 4 and 6 weeks and the specimens were dissected to be histologically and radiographically evaluated for new bone formation. RESULTS: The histological outcome showed superior bone healing in the bony defects in the resembling comminuted fracture in the study group than the control group. CONCLUSIONS: The nano-hydroxyapatite coated titanium mesh is a suitable method for reconstruction of comminuted fractures with bony defects.

Autologous bone grafts are considered the gold standard grafting material for the treatment of nonunion, but in very large bone defects, traditional autograft alone is insufficient to induce repair. Recombinant human bone morphogenetic protein 2 (rhBMP-2) can stimulate bone regeneration and enhance the healing efficacy of bone grafts. The delivery of rhBMP-2 may even enable engineered synthetic scaffolds to be used in place of autologous bone grafts for the treatment of critical size defects, eliminating risks associated with autologous tissue harvest. We here demonstrate that an osteoinductive scaffold, fabricated by combining a 3D printed rigid polymer/ceramic composite scaffold with an rhBMP-2-eluting collagen sponge can treat extremely large-scale segmental defects in a pilot feasibility study using a new sheep metatarsus fracture model stabilized with an intramedullary nail. Bone regeneration after 24 weeks was evaluated by micro-computed tomography, mechanical testing, and his...

This study trialled the controlled delivery of growth factors within a biodegradable scaffold in a large segmental bone defect model. We hypothesised that co-delivery of vascular endothelial growth factor (VEGF) and platelet derived growth factor (PDGF) followed by bone morphogenetic protein-2 (BMP-2) could be more effective in stimulating bone repair than the delivery of BMP-2 alone. Poly(lactic-co-glycolic acid) (PLGA) based microparticles were used as a delivery system to achieve a controlled release of growth factors within a medical-grade Polycaprolactone (PCL) scaffold. The scaffolds were assessed in a well-established preclinical ovine tibial segmental defect measuring 3 cm. After six months, mechanical properties and bone tissue regeneration were assessed. Mineralised bone bridging of the defect was enhanced in growth factor treated groups. The inclusion of VEGF and PDGF (with BMP-2) had no significant effect on the amount of bone regeneration at the six-month time point in comparison to BMP-2 alone. However, regions treated with VEGF and PDGF showed increased vascularity. This study demonstrates an effective method for the controlled delivery of therapeutic growth factors in vivo, using microparticles.

Background: Mammographic density (MD) and malignant-appearing microcalcifications (MAMCs) represent the earliest mammographic findings of non-palpable breast carcinomas. Matrix proteoglycans versican and decorin are frequently over-expressed in various malignancies and are differently involved in the progression of cancer. In the present study, we have evaluated the expression of versican and decorin in non-palpable breast carcinomas and their association with high risk mammographic findings and tumor characteristics. Methods: Three hundred and ten patients with non-palpable suspicious breast lesions, detected during screening mammography, were studied. Histological examination was carried out and the expression of decorin, versican, estrogen receptor α (ERα), progesterone receptor (PR) and c-erbB2 (HER-2/neu) was assessed by immunohistochemistry. Results: Histological examination showed 83 out of 310 (26.8%) carcinomas of various subtypes. Immunohistochemistry was carried out in 62/83 carcinomas. Decorin was accumulated in breast tissues with MD and MAMCs independently of the presence of malignancy. In contrast, versican was significantly increased only in carcinomas with MAMCs (median ± SE: 42.0 ± 9.1) and MD (22.5 ± 10.1) as compared to normal breast tissue with MAMCs (14.0 ± 5.8), MD (11.0 ± 4.4) and normal breast tissue without mammographic findings (10.0 ± 2.0). Elevated levels of versican were correlated with higher tumor grade and invasiveness in carcinomas with MD and MAMCs, whereas increased amounts of decorin were associated with in situ carcinomas in MAMCs. Stromal deposition of both proteoglycans was related to higher expression of ERα and PR in tumor cells only in MAMCs. Conclusions: The specific accumulation of versican in breast tissue with high MD and MAMCs only in the presence of malignant transformation and its association with the aggressiveness of the tumor suggests its possible use as molecular marker in non-palpable breast carcinomas.

Microcalcifications taken from 50 systematized mammary excisions were submitted to light microscopic and scanning electron microscope analysis. Microprobe and x-ray diffraction analyses were also performed. Two main types were observed: Type I microcalcifications composed of weddellite crystals. They were observed in benign breast lesions only (11 cases out of 21) or, in lobular carcinomas in situ (L.C.I.S.) of the breast (5 cases out of 6). They were not seen in 3 cases of intraductal carcinoma (I.D.C.) nor in infiltrating (I.C.) carcinomas (20 cases). Type II microcalcifications, non-cristalline in nature, composed of calcium, phosphate, hydroxyapatite or of phosphorus and calcium associated with other elements, were observed in benign lesions (10 cases out of 21) and in all cases of infiltrating carcinomas. The microcalcifications observed on mammography were also found on the radiographs of systematised mammary excisions from the lesion or from its immediate vicinity, but only w...

To examine the feasibility of reconstructing critical size mandibular defects with BMP-7 in a collagen carrier within a pedicled muscle flap in sheep. In 6 adult sheep unilateral mandible was exposed via an extra-oral skin incision in the submandibular region. The overlying periosteum was excised and a 35 mm discontinuity defect created. The sterno-occipitalis muscle was identified in the neck, the clavicular end dissected out, and mobilised to the surgical site. 3.5 mg of rhBMP-7 a recombinant human bone morphogenetic protein derived from a Chinese Hamster ovary cell-line was used. Type-1 bovine collagen derived from diaphyseal bone 1.0 gm was used as a carrier. The BMP-7, type-1 collagen, and carboxymethyl cellulose putty was mixed with normal saline and rolled like a sausage and placed in the muscle flap which inturn rolled like a cigar. The muscle flap was sutured to the bony ends of the surgical defect. A stainless steel reconstruction plate with 3 holes at each end was inserted across the defect over the muscle flap and fixed with screws stabilising the mandible. Assessment was by plain radiography, CT scan, Microradiography and Histology. Radiographically detectable bone formation was first noted at 4 weeks after surgery consistently in all the subjects. Marked mineralization and radiographically detectable calcified tissue formation took place between 8-12 weeks. Microradiography: The unique finding in this study with microradiography was the different stages of mineralisation of the tissue at the surgical site. There were numerous irregular calcific areas where mineralisation was extending from the centre of the foci to the periphery with higher mineralisation at the centre and less mineralisation at the periphery (centripetal).ln general there was a non-homogenous distribution of mineralization. Histology: Histological evaluation of the specimens from the subjects revealed that, the bulk of the defect was filled with bone showing a wide range of maturing bone from woven to lamellar with islands of cartilage and fibrous tissue. This study reliably demonstrated the concept, that a pedicled muscle flap and rhBMP-7 could be utilised successfully for mandibular reconstruction, and this procedure can be used in areas of deficient blood supply, where bone formation might be compromised.

Demineralized dentin matrix (DDM) has been used as a recombinant human bone morphogenetic protein-2 (rhBMP-2) carrier in many clinical trials. To optimize the clinical safety and efficacy of rhBMP-2 with DDM, efforts have been made to improve the delivery of rhBMP-2 by 1) lowering the administered dose, 2) localizing the protein, and 3) prolonging its retention time at the action site as well as the bone forming capacity of the carrier itself. The release profile of rhBMP-2 that is associated with endogenous BMP in dentin has been postulated according to the type of incorporation, which is attributed to the loosened interfibrillar space and nanoporous dentinal tubule pores. Physically adsorbed and modified, physically entrapped rhBMP-2 is sequentially released from the DDM surface during the early stage of implantation. As DDM degradation progresses, the loosened interfibrillar space and enlarged dentinal tubules release the entrapped rhBMP-2. Finally, the endogenous BMP in dentin is released with osteoclastic dentin resorption. According to the postulated release profile, DDM can therefore be used in a controlled manner as a sequential delivery scaffold for rhBMP-2, thus sustaining the rhBMP-2 concentration for a prolonged period due to localization. In addition, we attempted to determine how to lower the rhBMP-2 concentration to 0.2 mg/mL, which is lower than the approved 1.5 mg/mL.

A biodegradable microsphere/scaffold composite based on the synthetic polymer poly(propylene fumarate) (PPF) holds promise as a scaffold for cell growth and sustained delivery vehicle for growth factors for bone regeneration. The objective of the current work was to investigate the in vitro release and in vivo bone forming capacity of this microsphere/scaffold composite containing bone morphogenetic protein-2 (BMP-2) in combination with autologous bone marrow stromal cells (BMSCs) in a goat ectopic implantation model. Three composites consisting of 0, 0.08, or 8 μg BMP-2 per mg of poly(lactic-co-glycolic acid) microspheres, embedded in a porous PPF scaffold, were combined with either plasma (no cells) or culture-expanded BMSCs. PPF scaffolds impregnated with a BMP-2 solution and combined with BMSCs as well as empty PPF scaffolds were also tested. The eight different composites were implanted subcutaneously in the dorsal thoracolumbar area of goats. Incorporation of BMP-2–loaded microspheres in the PPF scaffold resulted in a more sustained in vitro release with a lower burst phase, as compared to BMP-2–impregnated scaffolds. Histological analysis after 9 weeks of implantation showed bone formation in the pores of 11/16 composites containing 8 μg/mg BMP-2–loaded microspheres with no significant difference between composites with or without BMSCs (6/8 and 5/8, respectively). Bone formation was also observed in 1/8 of the BMP-2–impregnated scaffolds. No bone formation was observed in the other conditions. Overall, this study shows the feasibility of bone induction by BMP-2 release from microspheres/scaffold composites.

A novel chitosan-cPGA polyelectrolyte complex hydrogel (C-PGA) has been developed and proven to be an effective dressing for wound healing. The purpose of this study was to evaluate if C-PGA could promote new bone formation in the alveolar socket following tooth extraction. An animal model was proposed using radiography and histomorphology simultaneously to analyze the symmetrical sections of Wistar rats. The upper incisors of Wistar rats were extracted and the extraction sockets were randomly treated with gelatin sponge, neat chitosan, C-PGA, or received no treatment. The extraction sockets of selected rats from each group were evaluated at 1, 2, 4, or 6 wk post-extraction. The results of radiography and histopathology indicated that the extraction sockets treated with C-PGA exhibited lamellar bone formation (6.5%) as early as 2 wk after the extraction was performed. Moreover, the degree of new bone formation was significantly higher (P , 0.05) in the extraction sockets treated with C-PGA at 6 wk post-extraction than that in the other study groups. In this study, we demonstrated that the proposed animal model involving symmetrical sections and simultaneous radiography and histomorphology evaluation is feasible. We also conclude that the novel C-PGA has great potential for new bone formation in the alveolar socket following tooth extraction.

Background: This study aims to investigate: (a) the putative association between the presence of microcalcifications and the expression of both epithelial-to-mesenchymal transition and bone biomarkers, (b) the role of microcalcifications in the breast osteoblast-like cells (BOLCs) formation, and (c) the association between microcalcification composition and breast cancer progression. Methods: We collected 174 biopsies on which we performed immunohistochemical and ultrastructural analysis. In vitro experiments were performed to demonstrate the relationship among microcalcification, BOLCs development, and breast cancer occurrence. Ex vivo investigations demonstrated the significant increase of breast osteoblast-like cells in breast lesions with microcalcifications with respect to those without microcalcifications. Results: In vitro data displayed that in the presence of calcium oxalate and activated monocytes, breast cancer cells undergo epithelial to mesenchymal transition. Also, in ...

A chronic nonunion of a proximal pole fracture of the scaphoid was treated by curettage of the nonunion, single K-wire fixation, and implantation of 50 mg of human bone morphogenetic protein followed by 12 weeks of cast immobilization without any conventional corticocancellous bone grafting or rigid screw fixation. Radiographs showed signs of bony healing by 12 weeks and a magnetic resonance imaging scan 6 years after surgery showed no signs of avascular necrosis. The potential future applications of human bone morphogenetic protein in hand surgery are discussed.

Many bioactive molecules like recombinant human bone morphogenetic protein 2 (rhBMP-2) have been developed for mineralized bone grafts, for which proper scaffolds are necessary to successfully apply the bioactive molecules. In this study, we tested the osteogenic efficacy of rhBMP-2 produced in-house in combination with gelatin sponge as the scaffold carrier in a rabbit radial defect model. The efficacy of the rhBMP-2 was determined by alkaline phosphatase activity assay of C2C12 cells. Two groups of ten rabbits each were treated with rhBMP-2/gelatin sponge, or gelatin sponge only. At 4 weeks, rhBMP-2/gelatin sponge grafts showed more bone regeneration than gelatin sponge grafts, as determined by X-ray radiography, micro-computed tomography, and histological analyses. At 8 weeks, rhBMP-2/gelatin sponge grafts exerted much stronger osteogenic effects. The study demonstrates the improved osteogenic efficacy of the rhBMP-2/gelatin sponge grafts in a rabbit radial bone defect model acting as a bone-inductive material. [BMB Reports 2013; 46(6): 328-333]

Objective The proposed study aimed to evaluate and compare the bone regeneration between commercially available hydroxyapatite–β-tricalcium phosphate (Ossifi; Equinox, the Netherlands), powdered polylactic acid (powdered PLA; Sigma-Aldrich, United States), and three-dimensionally printed PLA (3D-printed PLA; Cubex, SC, United States) using 3D printer (Cube X trio) in an animal model. Materials and Methods Eighteen New Zealand rabbits were divided into three groups with six animals each. Platelet-rich fibrin (PRF) was collected from the venous blood and preserved. Bone defect (4 mm × 2 mm) without disturbing the bone marrow was created and filled with bone graft material (group 1–Ossifi, group 2–powdered PLA, and group 3–3D-printed PLA), over which PRF membranes were placed. The graft material and the barrier were stabilized using resorbable sutures, and all the animals were maintained for 4, 8, and 12 weeks, after which they were euthanized, and bone samples were retrieved. Retrieve...

Calcification is one of the clinical and morphological manifestations of ovarian tumors and it begins at the initial stages of carcinogenesis. Thus, this process can be used for the early diagnostics of some malignant ovarian tumors. We compared the results of ultrasound and histology and found that calcifications of a size less than 200 μm are not detected by ultrasound. These calcified structures are round fragile particles of different sizes. In the EDX (Energy-dispersive X-ray spectroscopy) spectra, the main lines were from Ca and P, and the ratio of these elements corresponds to hydroxyapatite. Thus, we established that hydroxyapatite is the main mineral component of ovarian psammoma bodies and could be used for early diagnostics of ovarian malignant neoplasia.

Inorganic elements are involved in all vital actions of the macro organism: the physiological electrolytic equilibrium and acid-alkali balance control, electron transport, active compound transport, oxidation-reduction reactions and tissue biomineralization [5]. The role of heavy metal salts (elements with an atomic weight exceeding 50) in the pathological mineralization of breast cancer tissue should not be ruled out, considering their ability to enter in covalent bonds with calcium salt molecules. Along with the possibility of their participation in the mineralization progress, The process of pathological biomineralization plays an important role in tumor morphogenesis [1]. It has been established that any calcifications are the result of the opposing action of mineralization activators and inhibitors [2]. The action of klotho protein, matrix gamma-carboxyglutamic acid protein, fibroblast growth factors, pyrophosphates, etc., are described as inhibiting action [3]. Breast cancer tissue biopsies show that bone matrix proteins are excessively expressed under pathologic mineralization, including bone sialoprotein, osteopontin and osteonectin [4].

The aim of this study was to evaluate the efficacy of growth factor loaded collagen membranes on new bone formation during horizontal bone augmentation. Mandibular defects (4 × 4 × 4 mm) were surgically prepared in six male beagle dogs, which were then protected with one of three types of membranes: (1) titanium mesh, (2) titanium reinforced collagen, or (3) rhBMP-2 loaded titanium reinforced collagen. Animals were euthanized 8 and 16 weeks after surgery, and nondecalcified specimens were prepared and histomorphologically investigated to determine the degree of osteogenesis. Data were analyzed with Friedman test. With respect to the degree of osteogenesis at earlier stage (8 weeks after surgery), there was significantly higher new bone ratio in rhBMP-2 loaded membrane group (p &gt; 0.05). However, with respect to the long-term results (16 weeks after surgery), there were no significant differences among the three membranes (p &gt; 0.05). Based on histomorphometric analysis, there we...

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Reviewing the available literature, one could conclude that marrow-derived mesenchymal stem cells (BMSCs) are the 'gold standard' source for bone tissue engineering applications, due to their multilineage differentiation potential and easy accessibility. However, comprehensive studies comparing their osteogenic potential with bone-derived osteoblasts (OBs) to justify the preferred application of BMSCs based on performance are few. To address these shortfalls, in the present study, ovine BMSCs and OBs seeded onto scaffolds were characterized in vitro and transplanted subcutaneously into NOD/SCID mice in combination with and without recombinant human bone morphogenetic protein 7 (rhBMP-7). It was hypothesized that cell origin, ossification type and degree of vascularization and ossification depends on the nature and commitment of transplanted cells and stimulating growth factors, such as rhBMP-7. After retrieval, specimens were analysed by biomechanical testing, µCT analysis, scanning electron microscopy/energy-dispersive X-ray spectroscopy and histo-and immunohistochemistry for osteocalcin, type II collagen and BrdU. The results showed a high degree of cell survival and proliferation ectopically, resulting in active contribution to endochondral osteogenesis. When compared to BMSCs, OBs showed a higher degree of bone deposition while OB-derived bone was of higher maturation. Stimulation with rhBMP-7 increased the rate of bone synthesis for both BMSCs and OBs, additionally promoting neovascularization and osteoclast activity. These results suggest that the origin and commitment of transplanted cells highly influence the type and degree of ossification, that rhBMP-7 represents a powerful adjuvant for bone tissue-engineering applications, and that mature bone is an adequate alternative cell source for bone tissue-engineering applications.

Donepezil is an acetylcholinesterase inhibitor commonly used to treat mild to moderate Alzheimer’s disease. Its use has been associated with increased bone mass in humans and animals. However, the effect of postoperative administration of donepezil on bone healing remains unknown. Therefore, this study aimed to assess the impact of postoperative injection of donepezil on bone healing, titanium-implant osseointegration, and soft tissue healing. Twenty-two Sprague-Dawley rats were randomly assigned to receive a daily dose of either donepezil (0.6 mg/kg) or saline as a control. In each rat, a uni-cortical defect was created in the right tibia metaphysis and a custom-made titanium implant was placed in the left tibiae. After two weeks, rats were euthanized, and their bones were analysed by Micro-CT and histology. The healing of bone defect and implant osseointegration in the rats treated with donepezil were significantly reduced compared to the saline-treated rats. Histomorphometric ana...

In the review article: "Orthopaedic applications of osteogenic protein-1 (BMP-7)" published in International Orthopaedics [5], we presented two patients with resistant non-union who healed after application of a recombinant bone morphogenetic protein-7 (rh-BMP-7). Now we present a very rare patient with a pre-amputation non-union of the distal tibia who was healed with rh-BMP-7 and external fixator by Ilizarov after 14 years of unsuccessful treatment. The morbidity and mortality associated with impaired/delayed fracture healing remain high. About 10% of fractured bones heal slowly (delayed union) or fail to heal (non-union), requiring additional costly medical intervention to repair the fracture [1]. The discovery of bone morphogenetic proteins (BMPs) [4, 5] has increased our understanding of the cascade of events that take place during fracture healing. Several clinical studies demonstrate the capability of these proteins to induce and facilitate this process [2, 3]. Here we report the use of an rh-BMP-7 bound to a collagen carrier (osteogenic protein-1 device) in combination with autologous bone marrow to treat a patient with a resistant, pre-amputation tibial non-union. The patient fell and fractured the distal tibia at age of 49 and was since unsuccessfully treated 11 times over a period of 14 years. Failed treatments included: plate and screws (three times); plate, screws and autologous cancellous bone (three times), gastrocnemius muscle graft (one time), skin graft (one time), conservative immobilisation (two times), and external fixator (one time). The

The utility of recombinant human bone morphogenetic protein-2 (rhBMP-2) in inducing bone formation in fractures of bone is well known. However, the influence of the mechanical environment on the actions of rhBMP-2 on fracture healing is not clear. An experimental model of fractures of the tibia in rabbits was developed and utilized to investigate the role of mechanical environment on rhBMP-2 action. A 1 mm osteotomy gap was stabilized by either a low-or high-stiffness fixator (LSF or HSF, respectively), and local treatment with rhBMP-2 in an absorbable collagen sponge (ACS) was evaluated. The results of the investigation were analysed by both histomorphometry and biomechanics. The LSF caused an increase in mineralized periosteal callus compared to HSF, the rhBMP-2 in ACS accelerated fracture healing only in the LSF group but not in the HSF group. The area of mineralized tissue in interfragmentary callus was determined by fixation stiffness and not by BMP treatment. rhBMP-2 caused higher bone resorption in the endosteal callus during the late stages of fracture healing, but these histological differences did not affect the mechanical properties. Biomechanical evaluation showed only differences at 3 weeks between LSF-rhBMP-2 and LSF-ACS. The bending and torsional properties were higher in the rhBMP-2/ACS group compared to ACS alone at 3 weeks.