Reconstruction of a Unilateral Alveolar Cleft Using a Customized Allogenic Bone Block and Subsequent Dental Implant Placement in an Adult Patient.

Cleft lip and palate is the most common congenital deformity with severe effects on the quality of life of affected patients. The deformity often includes an alveolar cleft (AC). In most cases, osteoplasty will be performed using autogenous bone transplants harvested from the iliac crest. Thus, this treatment represents a highly invasive procedure. With freeze-dried bone allografts (FDBAs) becoming an increasingly accepted alternative to autogenous bone grafting for several indications, their application might also be suitable for AC reconstruction. We present the use of a customized allogenic bone block in a guided bone regeneration procedure for reconstruction of a unilateral AC and the successful insertion of dental implants after a healing period of 6 months. The use of FDBA seems to represent a successful treatment option for AC reconstruction. The allogenic bone block demonstrated high volume stability with ideal integration and revascularization, resulting in functional bone tissue suitable for implantation and esthetic rehabilitation. Nevertheless, further investigations, especially concerning the long-term stability of the augmented bone and dental implants, are needed to draw definite conclusion regarding the performance of allogenic bone blocks in orofacial cleft osteoplasty.

Drug Repurposing: The Anthelmintics Niclosamide and Nitazoxanide Are Potent TMEM16A Antagonists That Fully Bronchodilate Airways.

There is an unmet need in severe asthma where approximately 40% of patients exhibit poor ß-agonist responsiveness, suffer daily symptoms and show frequent exacerbations. Antagonists of the Ca2+-activated Cl- channel, TMEM16A, offers a new mechanism to bronchodilate airways and block the multiple contractiles operating in severe disease. To identify TMEM16A antagonists we screened a library of ∼580,000 compounds. The anthelmintics niclosamide, nitazoxanide, and related compounds were identified as potent TMEM16A antagonists that blocked airway smooth muscle depolarization and contraction. To evaluate whether TMEM16A antagonists resist use- and inflammatory-desensitization pathways limiting ß-agonist action, we tested their efficacy under harsh conditions using maximally contracted airways or airways pretreated with a cytokine cocktail. Stunningly, TMEM16A antagonists fully bronchodilated airways, while the ß-agonist isoproterenol showed only partial effects. Thus, antagonists of TMEM16A and repositioning of niclosamide and nitazoxanide represent an important additional treatment for patients with severe asthma and COPD that is poorly controlled with existing therapies. It is of note that drug repurposing has also attracted wide interest in niclosamide and nitazoxanide as a new treatment for cancer and infectious disease. For the first time we identify TMEM16A as a molecular target for these drugs and thus provide fresh insights into their mechanism for the treatment of these disorders in addition to respiratory disease.

Bilateral maxillary augmentation using CAD/CAM manufactured allogenic bone blocks for restoration of congenitally missing teeth: A case report.

OBJECTIVE: Various biomaterials have been successfully applied in alveolar bone regeneration, however, the reconstruction of extensive osseous defects remains challenging and is often unfeasible with granular grafting materials. Several studies have outlined allogenic bone blocks as valid alternative to autologous block grafting. CLINICAL CONSIDERATIONS: In this report, we demonstrate the regeneration of two large osseous defects in the maxilla with allogenic bone blocks made from human donor bone. The bone blocks were customized using the CAD/CAM technology in order to enable the insertion of four dental implants. CONCLUSIONS: Both blocks perfectly matched the defect geometry, showed limited resorption, led to the formation of sufficient amounts of mineralized bone in both horizontal and vertical dimensions and enabled the installation of implants according to the treatment plan. The implementation of innovative technologies for individualization of allogenic bone blocks simplifies the restoration of complex and extensive osseous defects and poses great benefits for both practitioners and patients. CLINICAL SIGNIFICANCE: The here presented procedure demonstrates the successful regeneration of two extensive osseous defects in a patient suffering from hypodontia using two CAD/CAM manufactured allogenic bone blocks, rendering the procedure far less invasive as compared to guided bone regeneration carried out with autologous transplants. Furthermore, to the best of our knowledge, this is the first case report that radiographically demonstrates the new formation of a cortical bone layer following block grafting with solely cancellous bone blocks.

Treatment of a bilaterally severely resorbed posterior mandible due to early tooth loss by Guided Bone Regeneration using customized allogeneic bone blocks: A case report with 24 months follow-up data.

OBJECTIVE: The use of allogeneic bone grafts for alveolar ridge reconstruction has become an often-discussed alternative to augmentation procedures using autografts. Nevertheless, there still is a lack of literature concerning long-time experiences of allografts for alveolar ridge reconstruction. Especially the results of the use of allogeneic bone blocks for mandible reconstructions need to be further investigated. CLINICAL CONSIDERATIONS: In this case report, we present the use of customized allogenic bone blocks in Guided Bone Regeneration (GBR) procedures for severely deficient mandibulary bones, the implantation after a healing period of 5 month and the 24-month follow-up data. CONCLUSIONS: Customized allogeneic bone blocks seem to be a successful alternative to augmentations with autologous bone blocks reconstructing highly resorbed mandibulary alveolar ridges, showing highest volume stability and no signs of bone resorption after implant loading. CLINICAL SIGNIFICANCE: The application of customized allogeneic bone blocks has shown to be a successful augmentation technique even compared with with autologous bone blocks reconstructing highly resorbed mandibulary alveolar ridges demonstrating no compromises regarding clinical outcome, functionality and esthetics.

Developing the Totality of Evidence for Biosimilars: Regulatory Considerations and Building Confidence for the Healthcare Community.

Biosimilars are highly similar versions of approved branded biologics. Unlike generics, they are not exact replicas of reference products. Minor differences between biosimilars and reference products in some aspects are expected; likewise, biosimilar products will differ from each other. The objective of this review is to discuss the challenges associated with the development and approval of biosimilar products that are unique because of their complex structure and specialized manufacturing processes, which can impact not only efficacy but also immunogenicity and safety. Regulatory guidelines recommend a totality-of-evidence approach focused on stepwise development that involves demonstration of structural similarity and functional equivalence. Structural and functional characteristics of the proposed biosimilar are compared with the reference product; similarity of these functions forms the foundation of the biosimilar development program, including potential animal studies, a human pharmacokinetics/pharmacodynamics equivalence study, and a clinical study to confirm similar efficacy, safety, and immunogenicity. The clinical study should be performed in a sensitive population using appropriate endpoints to allow detection of any clinically meaningful differences between the biosimilar and the reference product if such differences exist. In conclusion, development of biosimilars is focused on the minimization of potential differences between the proposed biosimilar and reference product and the establishment of a robust manufacturing process to consistently produce a high-quality biosimilar product.

Demonstration of Functional Similarity of Proposed Biosimilar ABP 501 to Adalimumab.

BACKGROUND: Due to the complex molecular structure and proprietary manufacturing processes of monoclonal antibodies (mAbs), differences in structure and function may be expected during development of biosimilar mAbs. Important regulatory requirements for approval of biosimilar products involve comprehensive assessments of any potential differences between proposed biosimilars and reference mAbs, including differences in all known mechanisms of action, using sensitive and relevant methods. Any identified structural differences should not result in differences in biofunctional or clinical activity. OBJECTIVE: A comprehensive assessment comparing the Amgen biosimilar candidate ABP 501 with FDA-licensed adalimumab (adalimumab [US]) and EU-authorized adalimumab (adalimumab [EU]) was conducted to demonstrate similarity in biofunctional activity. METHODS: The functional similarity assessment included testing of binding kinetics to soluble tumor necrosis factor α (TNFα) and relative binding to transmembrane TNFα. The neutralization of TNFα-induced caspase activation, TNFα- and lymphotoxin-α (LTα)-induced chemokine production, and cytotoxicity was also tested. Binding to Fc-gamma receptors FcγRIa, FcγRIIa (131H), FcγRIIIa (158V and 158F), and neonatal Fc receptor (FcRn) was compared with the reference mAbs, as was antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity. RESULTS: The data demonstrate that ABP 501 is similar to both adalimumab (US) and adalimumab (EU) with respect to evaluated biofunctional activities. CONCLUSION: Similarity in biofunctional activity is a critical component of the totality of evidence required for demonstration of biosimilarity. The functional similarity demonstrated for ABP 501 comprehensively assesses the known mechanisms of action of adalimumab, supporting the conclusion that ABP 501, adalimumab (US), and adalimumab (EU) are likely to be clinically similar.

Tumor necrosis factor, tumor necrosis factor inhibition, and cancer risk.

OBJECTIVE: Tumor necrosis factor (TNF) is a highly pleiotropic cytokine with multiple activities other than its originally discovered role of tumor necrosis in rodents. TNF is now understood to play a contextual role in driving either tumor elimination or promotion. Using both animal and human data, this review examines the role of TNF in cancer development and the effect of TNF and TNF inhibitors (TNFis) on malignancy risk. RESEARCH DESIGN: A literature review was performed using relevant search terms for TNF and malignancy. RESULTS: Although administration of TNF can cause tumor regression in specific rodent tumor models, human expression polymorphisms suggest that TNF can be a tumor-promoting cytokine, whereas blocking the TNF pathway in a variety of tumor models inhibits tumor growth. In addition to direct effects of TNF on tumors, TNF can variously affect immunity and the tumor microenvironment. Whereas TNF can promote immune surveillance designed to eliminate tumors, it can also drive chronic inflammation, autoimmunity, angiogenesis, and other processes that promote tumor initiation, growth, and spread. Key players in TNF signaling that shape this response include NF-κB and JNK, and malignant-inflammatory cell interactions, each of which may have different responses to TNF signaling. Focusing on rheumatoid arthritis (RA) patients, where clinical experience is most extensive, a review of the clinical literature shows no increased risk of overall malignancy or solid tumors such as breast and lung cancers with exposure to TNFis. Lymphoma rates are not increased with use of TNFis. Conflicting data exist regarding the risks of melanoma and nonmelanoma skin cancer. Data regarding the risk of recurrent malignancy are limited. CONCLUSIONS: Overall, the available data indicate that elevated TNF is a risk factor for cancer, whereas its inhibition in RA patients is not generally associated with an increased cancer risk. In particular, TNF inhibition is not associated with cancers linked to immune suppression. A better understanding of the tumor microenvironment, molecular events underlying specific tumors, and epidemiologic studies of malignancies within specific disease indications should enable more focused pharmacovigilance studies and a better understanding of the potential risks of TNFis.

Native IL-32 is released from intestinal epithelial cells via a non-classical secretory pathway as a membrane-associated protein.

Although IL-32 has been shown to be induced under various pathological conditions, a detailed understanding of native IL-32 intracellular distribution and mechanism of release from cells has not been reported. We examined the expression of IL-32 in the intestinal epithelial cell line HT-29 following TNFα and IFNγ co-stimulation. The subcellular localization of induced IL-32 was associated with the membrane of lipid droplet-like structures and vacuolar structures that co-localized with markers of endosomes and lysosomes. Prolonged co-stimulation resulted in cell death and appearance of IL-32 in the culture medium. IL-32 released from co-stimulated HT-29 cells was found in a detergent-sensitive particulate fraction, and in a step density gradient the IL-32 particulate was buoyant, suggesting association with a membrane-bound vesicle. Upon Triton X-114 partitioning, most of the IL-32 partitioned to the detergent phase, suggesting hydrophobic characteristics. When IL-32-containing vesicles were subjected to protease K treatment, a protease resistant ∼12kDa fragment was generated from ∼24kDa IL-32. We propose that under these conditions, native IL-32 is released via a non-classical secretory route perhaps involving multi-vesicular bodies and exosomes. Demonstration of membrane association for both intracellular and released IL-32 suggests this unique cytokine may have a complex biosynthetic pathway and mechanism of action.

Enhancing antibody Fc heterodimer formation through electrostatic steering effects: applications to bispecific molecules and monovalent IgG.

Naturally occurring IgG antibodies are bivalent and monospecific. Bispecific antibodies having binding specificities for two different antigens can be produced using recombinant technologies and are projected to have broad clinical applications. However, co-expression of multiple light and heavy chains often leads to contaminants and pose purification challenges. In this work, we have modified the CH3 domain interface of the antibody Fc region with selected mutations so that the engineered Fc proteins preferentially form heterodimers. These novel mutations create altered charge polarity across the Fc dimer interface such that coexpression of electrostatically matched Fc chains support favorable attractive interactions thereby promoting desired Fc heterodimer formation, whereas unfavorable repulsive charge interactions suppress unwanted Fc homodimer formation. This new Fc heterodimer format was used to produce bispecific single chain antibody fusions and monovalent IgGs with minimal homodimer contaminants. The strategy proposed here demonstrates the feasibility of robust production of novel Fc-based heterodimeric molecules and hence broadens the scope of bispecific molecules for therapeutic applications.