Bone Morphogenic Protein Research Articles

Sotatercept in pulmonary hypertension and beyond.

Sotatercept binds free activins by mimicking the extracellular domain of the activin receptor type IIA (ACTRIIA). Additional ligands are BMP/TGF-beta, GDF8, GDF11 and BMP10. The binding with activins leads to the inhibition of the signalling pathway and the deactivation of the bone morphogenic protein (BMP) receptor type 2. In this way, sotatercept activates an antiproliferative signalling to the cells of the pulmonary arteries and arterioles with the aim of rebalancing the proliferative and antiproliferative pathway that characterizes the pulmonary arterial hypertension (PAH). Sotatercept is indicated for the treatment of group 1 PAH in combination with drugs that act through the endothelin receptor, nitric oxide or prostacyclin. Its effects, demonstrated in the STELLAR study, are the improvement of exercise capacity and the FC-WHO functional class, together with the reduction of the risk of clinical worsening events. In addition to its antiremodeling effects on the pulmonary circulation, sotatercept has several haematological effects that could suggest its use in the treatment of some blood disorders other than PAH. In this review, we will discuss the effects of the drug on PAH and in parallel provide an in-depth overview of its application in haematological disorders, focusing on clinical and preclinical studies.

Puerarin Promotes Tibial Shaft Fracture Healing in Rats and Its Relationship with BMP-Smad Pathway

Background Fracture healing involves multiple growth factors and the interaction between multiple signaling pathways. Pue (puerarin) is a natural compound with multiple biological activities, but its effect on fracture healing and its relationship with the bone morphogenic protein (BMP)-Smad pathway is not yet clear. Purpose This study intends to assess the effect of puerarin on fracture healing. Methods Seventy rats were used for the study to establish a tibial shaft fracture rat model. The experimental rats were divided into a tibial fracture group, Pue dose group (low, medium, and high), BMP-2 group, Noggin group, Pue + SY-LB-35 group, and Pue + Noggin group through the random number method. We observed the fracture healing through micro-CT and histology, observed the tibial stump tissue with hematoxylin and eosin (HE) staining, and detected the expression of genes related to the BMP-Smad pathway through quantitative real-time polymerase chain reaction (qPCR) and Western blot. Results We found that puerarin can promote tibial shaft fracture healing in rats in a dose-dependent manner; through micro-CT and histological analysis, we observed increased trabecular bone formation during fracture healing in rats treated with puerarin. Callus formation is accelerated, and bone density increases. In addition, puerarin also significantly increased the expression of genes related to the BMP-Smad pathway, including BMP2, Smad1, Smad5, and so on. Conclusion Puerarin can promote tibial shaft fracture healing in rats, and its mechanism is related to upregulating the expression of genes related to the BMP-Smad pathway. It further improves the fracture-healing mechanism and provides candidate drugs for the treatment of tibial shaft fracture healing in clinical medicine.

Exploring Bone Morphogenetic Protein-2 and -4 mRNA Expression and Their Receptor Assessment in a Dynamic In Vitro Model of Vascular Calcification.

Vascular calcification (VC) is a dynamic, tightly regulated process driven by cellular activity and resembling the mechanisms of bone formation, with specific molecules playing pivotal roles in its progression. We aimed to investigate the involvement of the bone morphogenic proteins (BMP-2, BMP-4, BMPR-1a/1b, and BMPR-2) system in this process. Our study used an advanced in vitro model that simulates the biological environment of the vascular wall, assessing the ability of a phosphate mixture to induce the osteoblastic switch in human coronary artery smooth muscle cells (HCASMCs). HCASMCs were grown in mono- and co-culture with human coronary artery endothelial cells (HCAECs) in a double-flow bioreactor (LiveBox2 and IVTech), allowing static and dynamic conditions through a peristaltic pump. The VC was stimulated by incubation in a calcifying medium for 7 days. A BMP system Real-Time PCR was performed at the end of each experiment. In monocultures, BMP-2 expression increased in calcified HCASMCs in static (p = 0.01) and dynamic conditions. BMP-4 and the biological receptors were expressed in all the experimental settings, increasing mainly in dynamic flow conditions. In co-cultures, we observed a marked increase in BMP-2 and BMP-4, BMPR-1a (p = 0.04 and p = 0.01, respectively), and BMPR-2 (p = 0.001) in the calcifying setting mostly in dynamic conditions. The increase in BMP-2/4 in co-culture suggests that these genes might promote the switch towards an osteogenic-like phenotype, data also supported by the rise of both BMPR-1a and BMPR-2. Thus, our findings provide insights into the mechanisms by which dynamic co-culture modulates the BMP system activation in an environment mimicking in vivo VC's cellular and mechanical characteristics.

Evaluation of Biomaterials in Periodontal Regeneration: A Literature Review.

The field of periodontal regeneration focuses on restoring the form and function of periodontal tissues compromised due to diseases affecting the supporting structures of teeth. Biomaterials have emerged as a vital component in periodontal regenerative therapy, offering a variety of properties that enhance cellular interactions, promote healing, and support tissue reconstruction. This review explores current advances in biomaterials for periodontal regeneration, including ceramics, polymers, and composite scaffolds, and their integration with biological agents like growth factors and stem cells. Specifically, biomaterials such as hydroxyapatite and bioactive glass provide scaffolding for cellular adhesion and proliferation, while synthetic and natural polymers offer flexibility and biocompatibility. Growth factors and bone morphogenic proteins (BMP) further support cell differentiation and tissue formation, enhancing clinical outcomes in periodontal defect repair. Moreover, stem cell integration with biomaterials, particularly the periodontal ligament stem cells (PDLSCs) and mesenchymal stem cells (MSCs), shows promise for complex tissue regeneration by stimulating targeted healing responses in periodontal tissue. Although clinical results are encouraging, challenges related to the selection of optimal biocompatible and bioactive materials and standardization of clinical protocols remain. This review examines the potential, limitations, and future directions for biomaterial-based strategies, highlighting the evolving role of these materials in achieving predictable and effective periodontal regeneration.

Discovery of highly potent and ALK2/ALK1 selective kinase inhibitors using DNA-encoded chemistry technology

Activin receptor type 1 (ACVR1; ALK2) and activin receptor like type 1 (ACVRL1; ALK1) are transforming growth factor beta family receptors that integrate extracellular signals of bone morphogenic proteins (BMPs) and activins into Mothers Against Decapentaplegic homolog 1/5 (SMAD1/SMAD5) signaling complexes. Several activating mutations in ALK2 are implicated in fibrodysplasia ossificans progressiva (FOP), diffuse intrinsic pontine gliomas, and ependymomas. The ALK2 R206H mutation is also present in a subset of endometrial tumors, melanomas, non-small lung cancers, and colorectal cancers, and ALK2 expression is elevated in pancreatic cancer. Using DNA-encoded chemistry technology, we screened 3.94 billion unique compounds from our diverse DNA-encoded chemical libraries (DECLs) against the kinase domain of ALK2. Off-DNA synthesis of DECL hits and biochemical validation revealed nanomolar potent ALK2 inhibitors. Further structure-activity relationship studies yielded center for drug discovery (CDD)-2789, a potent [NanoBRET (NB) cell IC50: 0.54 μM] and metabolically stable analog with good pharmacological profile. Crystal structures of ALK2 bound with CDD-2281, CDD-2282, or CDD-2789 show that these inhibitors bind the active site through Van der Waals interactions and solvent-mediated hydrogen bonds. CDD-2789 exhibits high selectivity toward ALK2/ALK1 in KINOMEscan analysis and NB K192 assay. In cell-based studies, ALK2 inhibitors effectively attenuated activin A and BMP-induced Phosphorylated SMAD1/5 activation in fibroblasts from individuals with FOP in a dose-dependent manner. Thus, CDD-2789 is a valuable tool compound for further investigation of the biological functions of ALK2 and ALK1 and the therapeutic potential of specific inhibition of ALK2.

DDDR-27. BMP SIGNALING MODULATES APOPTOTIC AND NON-APOPTOTIC CELL DEATH IN GLIOBLASTOMA

Abstract Recurrence occurs in approximately 90% of glioblastoma (GBM) patients, in part due to the tumors becoming resistant to cell death in response to existing therapies. These therapies often trigger apoptosis; however, apoptosis is only one of several different forms of cell death. We hypothesize that activating non-canonical death mechanisms could provide a back door to killing treatment-resistant GBM cells. To test this hypothesis, we first developed a new imaging-based high-throughput method to directly quantify cell death in 3D models. Using this method, we find patient-derived GBM spheroids to be resistant to the induction of cell death in response to various standard-of-care and experimental therapeutic compounds. To further investigate mechanisms underlying this resistance, we screened a panel of extracellular protein ligands for the modulation of cell death response. We find that bone morphogenic protein (BMP) signaling increases resistance to several apoptosis-inducing compounds, including temozolomide and tyrosine kinase inhibitors. Intriguingly, BMP signaling simultaneously enhanced sensitivity to a novel non-apoptotic cell death pathway triggered by the small molecule CIL56. Efforts are ongoing to further understand how BMP signaling governs apoptotic and non-apoptotic cell death mechanisms. Nevertheless, our results suggest a novel approach to treating GBM, centered upon the modulation of BMP signaling combined with the use of agents that induce non-apoptotic cell death.

Treatment of Bone Defects and Nonunion via Novel Delivery Mechanisms, Growth Factors, and Stem Cells: A Review.

Bone nonunion following a fracture represents a significant global healthcare challenge, with an overall incidence ranging between 2 and 10% of all fractures. The management of nonunion is not only financially prohibitive but often necessitates invasive surgical interventions. This comprehensive manuscript aims to provide an extensive review of the published literature involving growth factors, stem cells, and novel delivery mechanisms for the treatment of fracture nonunion. Key growth factors involved in bone healing have been extensively studied, including bone morphogenic protein (BMP), vascular endothelial growth factor (VEGF), and platelet-derived growth factor. This review includes both preclinical and clinical studies that evaluated the role of growth factors in acute and chronic nonunion. Overall, these studies revealed promising bridging and fracture union rates but also elucidated complications such as heterotopic ossification and inferior mechanical properties associated with chronic nonunion. Stem cells, particularly mesenchymal stem cells (MSCs), are an extensively studied topic in the treatment of nonunion. A literature search identified articles that demonstrated improved healing responses, osteogenic capacity, and vascularization of fractures due to the presence of MSCs. Furthermore, this review addresses novel mechanisms and materials being researched to deliver these growth factors and stem cells to nonunion sites, including natural/synthetic polymers and bioceramics. The specific mechanisms explored in this review include BMP-induced osteoblast differentiation, VEGF-mediated angiogenesis, and the role of MSCs in multilineage differentiation and paracrine signaling. While these therapeutic modalities exhibit substantial preclinical promise in treating fracture nonunion, there remains a need for further research, particularly in chronic nonunion and large animal models. This paper seeks to identify such translational hurdles which must be addressed in order to progress the aforementioned treatments from the lab to the clinical setting.

Treatment of adult spine deformity: A retrospective comparison of bone morphogenic protein and bone marrow aspirate with bone allograft.

This was a retrospective cohort study. ASD patients were stratified by use of intra-operative BMP (BMP +) or not (BMA + I) and surveyed for the development of complications and mechanical failure. Quality of life gained following the procedure was evaluated using quality-adjusted life years (QALYs). Cost was calculated using the PearlDiver database and CMS definitions. Multivariable analyses (ANCOVA) and logistic regression were used to adjust for confounding. 512 patients were included (60% BMP +). At baseline, BMP + patients were older (62.5 vs 60.8years, p < 0.010). Radiographic and quality-of-life metrics did not differ at follow up timepoints (all p > 0.05). BMP use was associated with higher supplemental rod use (OR: 7.0, 1.9 - 26.2, p = 0.004), greater number of levels fused (OR: 1.1, 1.03 - 1.17, p = 0.003) and greater neurological complications (OR: 5.0, 1.3 - 18.7, p = 0.017). Controlling for rod use and levels fused, BMP use was not associated with a lower risk of mechanical complications (OR 0.3, 95% CI: 0.2 - 3.0, p = 0.353), rod breakage (OR: 3.3, 0.6 - 18.7, p = 0.182) or implant failure (OR: 0.3, 0.04 - 1.51). At 2years, the BMP + cohort exhibited higher overall costs ($108,062 vs $95,144, p = 0.002), comparable QALYs (0.163 vs 0.171, p = 0.65) and higher cost per QALY (p = 0.001) at two years. In this analysis, BMP-2 application was not associated with superior outcomes when compared to a less costly biologic alternative (bone marrow aspirate + cancellous bone chips + i-Factor) following ASD surgery. The use of BMP-2 in ASD surgery appears to have reduced cost-efficacy at two years postoperatively.

Derivation of transplantable human thyroid follicular epithelial cells from induced pluripotent stem cells

The production of mature functioning thyroid follicular cells (TFCs) from human induced pluripotent stem cells (iPSCs) is critical for potential novel therapeutic approaches to post-surgical and congenital hypothyroidism. To accomplish this, we developed a novel human iPSC line that expresses fluorophores targeted to the NKX2-1 and PAX8 loci, allowing for the identification and purification of cells destined to become TFCs. Optimizing a sequence of defined, serum-free media to promote stepwise developmental directed differentiation, we found that bone morphogenic protein 4 (BMP4) and fibroblast growth factor 2 (FGF2) stimulated lineage specification into TFCs from multiple iPSC lines. Single-cell RNA sequencing demonstrated that BMP4 withdrawal after lineage specification promoted TFC maturation, with mature TFCs representing the majority of cells present within 1month. After xenotransplantation into athyreotic immunodeficient mice, engrafted cells exhibited thyroid follicular organization with thyroglobulin protein detected in the lumens of NKX2-1-positive follicles. While our iPSC-derived TFCs presented durable expression of thyroid-specific proteins, they were unable to rescue hypothyroidism invivo.

Antiosteoporotic activity of nodakenetin, a coumarin compound from Angelica decursiva, by activation of the Wnt/β-catenin signaling pathway

Angelica decursiva (Umbelliferae) is a medicinal plant widely used to treat colds, coughs and fevers in Korea, Japan, and mainland China. The anti-inflammatory activity of nodakenetin, a furano-coumarin compound from A. decursiva, has been reported, although, the antiosteoporotic activity remains unknown. This study sought to investigate the antiosteoporotic activity and precise mechanism of action of nodakenetin in vitro cell culture and in vivo bone remodeling models. The transcriptional activity of nodakenetin on the Wnt signaling pathway was assessed using the TOPflash/FOPflash assay. The effect of nodakenetin on the osteoblast differentiation was measured using Alizarin red staining and alkaline phosphatase (ALP) activity. Western blotting and real-time RT-PCR were used to assess the effect of nodakenetin on the expression of markers related to Wnt/β-catenin pathway and osteoblast differentiation. The in vivo antiosteoporotic activity of nodakenetin was assessed using an ovariectomized (OVX)-induced bone loss mouse model. Nodakenetin activated the Wnt/β-catenin pathway through regulation of DKK1, β-catenin and other target proteins of the Wnt/β-catenin pathway in HEK293 and MC3T3-E1 cells. Nodakenetin induced the differentiation of MC3T3-E1 cells as shown by enhanced Alizarin red staining and ALP activity. Induction of osteoblast differentiation was related to upregulated expression of bone formation biomarkers such as bone morphogenic proteins and Runx2. Oral administration of nodakenetin in the OVX mouse model effectively protected the deterioration of bone microstructure in OVX mice. Nodakenetin exhibits antiosteoporotic activity in vitro and in vivo through the activation of the Wnt/β-catenin pathway and subsequent induction of osteoblast differentiation.

Perspectives on Sotatercept in Pulmonary Arterial Hypertension.

Sotatercept acts as a type IIA-Fc activin receptor, thereby scavenging free activin from its physiological membrane receptor. Through this type of action, sotaterpect leads to a rebalancing of the proliferation and antiproliferation pathways of pulmonary smooth muscle cells in response to bone morphogenic protein (BMP). Although sotatercept has been approved as the fourth pillar of therapy for group 1 pulmonary arterial hypertension (PAH) in the United States and Europe, several studies are ongoing to broaden the application of the drug to non-Group 1 PAH. We provide an overview of the clinical and preclinical evidence of targeting the activation pathway by sotatercept in the treatment of PAH. We also discuss other potential applications of sotatercept in the context of pulmonary hypertension other than PAH group 1.

Development of a Thermoresponsive Core-Shell Hydrogel for Sequential Delivery of Antibiotics and Growth Factors in Regenerative Endodontics.

Regenerative endodontics requires an innovative delivery system to release antibiotics/growth factors in a sequential trend. This study focuses on developing/characterizing a thermoresponsive core-shell hydrogel designed for targeted drug delivery in endodontics. The core-shell chitosan-alginate microparticles were prepared by electrospraying to deliver bone morphogenic protein-2 for 14 days and transforming growth factor-beta 1 (TGF-β1) for 7-14 days. Methylcellulose (MC) and gelatin were utilized to create the core-shell hydrogel to load a modified triple antibiotic combination (penicillin G/metronidazole/ciprofloxacin (PMC)) and growth factor-loaded microparticles in the shell and the core compartments, respectively. Morphological assessment, core-shell structural analysis, FTIR analysis, rheological analysis, swelling, and degradation rate studies were conducted for characterization. The viability of dental pulp stem cells (DPSCs) upon antibiotic exposure, antibacterial activity, and release studies of PMC and growth factors were investigated. Cellular studies (cell viability, alkaline phosphatase (ALP) activity, osteo/odontoblast gene expression (using Reverse transcription-polymerase chain reaction (RT-PCR)) and in vivo studies (inflammatory response and differentiation potential of the developed hydrogel by subcutaneous implantation in rats via histological examination) were assessed. The hydrogel showed a porous microstructure with interconnected pores. Core-shell structure analysis confirmed the successful extrusion of the MC hydrogel to the surface. FTIR analysis revealed interactions between MC and gelatin. Rheological analysis indicated time-dependent gel formation, supporting thermosensitivity at 37 °C. Swelling occurred rapidly, and degradation reached 62.42% on day 45. Further, antibiotics exhibited no cytotoxicity on DPSCs. Sequential release of antibiotics and growth factors was observed for up to 5 and 14 d, respectively. The hydrogel showed antibacterial activity. DPSCs exhibited increased proliferation, ALP activity, and odontoblast gene expression. In vivo studies showed that the biocompatible drug-loaded hydrogel exhibited more mineralization than the control. The developed core-shell hydrogel containing PMC and growth factor-loaded core-shell microparticles provided a versatile and biocompatible platform for sequential drug delivery in regenerative endodontics. The system demonstrates promising characteristics for dentin regeneration, making it a potential candidate for clinical applications.

HOXD1 inhibits lung adenocarcinoma progression and is regulated by DNA methylation.

The homeobox (HOX) gene family encodes a number of highly conserved transcription factors and serves a crucial role in embryonic development and tumorigenesis. Homeobox D1 (HOXD1) is a member of the HOX family, whose biological functions in lung cancer are currently unclear. The University of Alabama at Birmingham Cancer data analysis Portal of HOXD1 expression patterns demonstrated that HOXD1 was downregulated in lung adenocarcinoma (LUAD) patient samples compared with adjacent normal tissue. Western blotting analysis demonstrated low HOXD1 protein expression levels in lung LUAD cell lines. The Kaplan‑Meier plotter database demonstrated that reduced HOXD1 expression levels in LUAD correlated with poorer overall survival. Meanwhile, an in vitro study showed that HOXD1 overexpression suppressed LUAD cell proliferation, migration and invasion. In a mouse tumor model, upregulated HOXD1 was demonstrated to inhibit tumor growth. In addition, targeted bisulfite sequencing and chromatin immunoprecipitation assays demonstrated that DNA hypermethylation occurred in the promoter region of the HOXD1 gene and was associated with the action of DNA methyltransferases. Moreover, upregulated HOXD1 served as a transcriptional factor and increased the transcriptional expression of bone morphogenic protein (BMP)2 and BMP6. Taken together, the dysregulation of HOXD1 mediated by DNA methylation inhibited the initiation and progression of LUAD by regulating the expression of BMP2/BMP6.

Contemporary Concise Review 2023: Interstitial lung disease.

In this review, we have discussed several important developments in 2023 in Interstitial Lung Disease (ILD). The association of pollution with genetic predispositions increased the risk of Idiopathic Pulmonary Fibrosis (IPF). An interesting comorbidity of malnutrition was not adequately recognized in ILD. Novel genes have been identified in IPF involving predominantly short telomere length and surfactant protein production leading to alveolar epithelial cell dysfunction. Genetics also predicted progression in IPF. Crosstalk between vascular endothelial cells and fibroblasts in IPF mediated by bone morphogenic protein signalling may be important for remodelling of the lung. A novel modality for monitoring of disease included the 4-min gait speed. New treatment modalities include inhaled pirfenidone, efzofitimod, for sarcoidosis, and earlier use of immunosuppression in connective tissue disease-ILD.

Pulmonary Hypertension: Pharmacological and Non-Pharmacological Therapies.

Pulmonary hypertension (PH) is a severe and chronic disease characterized by increased pulmonary vascular resistance and remodeling, often precipitating right-sided heart dysfunction and death. Although the condition is progressive and incurable, current therapies for the disease focus on multiple different drugs and general supportive therapies to manage symptoms and prolong survival, ranging from medications more specific to pulmonary arterial hypertension (PAH) to exercise training. Moreover, there are multiple studies exploring novel experimental drugs and therapies including unique neurostimulation, to help better manage the disease. Here, we provide a narrative review focusing on current PH treatments that target multiple underlying biochemical mechanisms, including imbalances in vasoconstrictor-vasodilator and autonomic nervous system function, inflammation, and bone morphogenic protein (BMP) signaling. We also focus on the potential of novel therapies for managing PH, focusing on multiple types of neurostimulation including acupuncture. Lastly, we also touch upon the disease's different subgroups, clinical presentations and prognosis, diagnostics, demographics, and cost.

Osteogenic effect of alogliptin in chemical-induced bone loss: a tri-modal in silico, in vitro, and in vivo analysis.

To investigate the effects of Alogliptin in chemical-induced post-menopausal osteoporosis. The binding affinity of alogliptin with osteogenic proteins was analysed in silico. The effect of alogliptin on osteogenic proteins and mineralization of osteoblastic cells was evaluated in UMR-106 cells. Further, in vivo anti-osteoporotic activity of alogliptin was evaluated in postmenopausal osteoporosis. Various bone turnover markers were assayed in serum. This followed the analysis of microarchitecture of bone, histology, and immunohistochemistry (IHC) of bone tissue. Docking scores showed that alogliptin has binding affinity for bone alkaline phosphatase (BALP), osteocalcin, and bone morphogenic protein (BMP-2). Alogliptin also enhanced mineralization of osteoblast cells, evidenced with increased ALP, osteocalcin, and BMP-2. Animal studies revealed significant elevation of bone formation markers, bone ALP, osteocalcin and BMP-2, and decreased bone resorption markers, receptor activator of NF-κβ (RANKL), cathepsin K (CTSK), tartrate resistant acid phosphatase (TRAcP5b) in VCD-induced post-menopausal osteoporosis. Micro computed tomography (μCT) analysis and histology of femur bone and lumbar vertebrae demonstrated decrease in trabecular separation and improved bone density. IHC of femur showed reduced DPP4 enzyme. Alogliptin increased mineralization in osteoblast cells. It had beneficial effects also altered bone turnover markers, repaired the trabecular microstructure, improved bone mineral density, and exhibited bone forming capacity targeting DPP-4 enzyme in postmenopausal osteoporosis.

Utilization of Orthobiologics by Foot and Ankle Orthopaedic Surgeons

Category: Basic Sciences/Biologics; Sports Introduction/Purpose: Over the past two decades, there have been significant advancements and increased utilization of orthobiologic technologies. The use of orthobiologic products is often determined by individual surgeons given the limited available literature on efficacy. Currently, there is a paucity of data available evaluating the prevalence of orthobiologics use among foot and ankle orthopaedic surgeons. The primary aim of this study was to investigate orthobiologic utilization among American Orthopaedic Foot &amp; Ankle Society (AOFAS) members. Methods: Members of the AOFAS were sent a survey in 2023 that was designed to assess for usage, motivation for use, and treatment indications in operative and nonoperative settings for the following orthobiologics: leukocyte-poor (PRP-LP), leukocyte-rich (PRP-LR) and leukocyte-unknown (PRP-UNK) platelet rich plasma, bone marrow aspirate concentrate (BMAC), adipose-derived mesenchymal stromal cells, hyaluronic acid, recombinant human bone morphogenic protein (rhBMP-2), recombinant human platelet derived growth factor (rhPDGF-BB), amniotic membrane products, and small cell binding peptide. The survey was completed by 129 respondents representing a balanced cohort in regards to geographic spread, time in practice, and medical practice setting. Results: 92% of respondents utilized ≥1 orthobiologic with no significant differences in utilization based on demographics. Perceived efficacy and support from literature were cited as common reasons to use orthobiologics. Among orthobiologic users, the most common product used was BMAC (74%), followed by rhPDGF-BB (62%) and rhBMP-2 (43%) (Figure 1). The most common indications for BMAC were revision arthrodesis, osteochondral surgery and primary arthrodesis. For rhPDGF-BB and rhBMP, the most common indications were revision and primary arthrodesis. The most common pathology for nonoperative use of PRP-LR and PRP-UNK was tendinopathy versus plantar fasciitis for PRP-LP, whereas the most common operative use of any formulation of PRP was for osteochondral lesions. Among non-users, 70% cited lack of evidence and 50% cited costs in their decision making. Conclusion: Among AOFAS surgeons, there is a high rate of utilization of orthobiologic products. Bone marrow aspirate concentrate was the most common orthobiologic used overall, though there remains significant variability in the usage and treatment indications for each individual orthobiologic. Given that evidence from the literature was frequently cited as a motivation for both users and nonusers for orthobiologics, there remains a need for future studies evaluating the efficacy of orthobiologic products in foot and ankle orthopedics.

Gunshot Injury With Bone Defect of the First Metatarsal Bone A Presentation of 2 Cases Treated With an Iliac Crest Structural Graft, Internal Fixation, and Bone Morphogenic Protein 2.

Gunshot injuries to the foot with segmental bone defects can be challenging to treat. When the vascularity is intact and the soft tissues allows, the goal should be to reconstruct the bony defect. We present 2 cases of a gunshot injury to the foot with a defect of the first metatarsal bone. Both cases were treated, with favorable outcome, with a structural iliac crest graft, internal fixation, and bone morphogenic protein 2.Level of Evidence: V, cases series, technical.

The overexpression of R-spondin 3 affects hair morphogenesis and hair development along with the formation and maturation of the hair follicle stem cells.

Mice hair follicles (HFs) are a valuable model for studying various aspects of hair biology, including morphogenesis, development, and regeneration due to their easily observable phenotype and genetic manipulability. The initiation and progression of hair follicle morphogenesis, as well as the hair follicle cycle, are regulated by various signaling pathways, of which the main role is played by the Wingless-type MMTV integration site family (Wnt) and the Bone Morphogenic Protein (BMP). During the hair follicle cycle, the BMP pathway maintains hair follicle stem cells (HFSCs) in a dormant state while the Wnt pathway activates them for hair growth. Given the pivotal role of the Wnt pathway in hair biology and HFSCs regulation, we investigated the influence of the Wnt modulator - R-spondin 3 (Rspo3), in these processes. For this purpose, we developed a transgenic mice model with the overexpression of Rspo3 (Rspo3GOF) in the whole ectoderm and its derivatives, starting from early morphogenesis. Rspo3GOF mice exhibited a distinct phenotype with sparse hair and visible bald areas, caused by reduced proliferation and increased apoptosis of hair matrix progenitor cells, which resulted in a premature anagen-to-catagen transition with a shortened growth phase and decreased overall length of all hair types. In addition, Rspo3GOF promoted induction of auchene and awl, canonical Wnt-dependent hair type during morphogenesis, but the overall hair amount remained reduced. We also discovered a delay in the pre-bulge formation during morphogenesis and prolonged immaturity of the HFSC population in the bulge region postnatally, which further impaired proper hair regeneration throughout the mice's lifespan. Our data supported that Rspo3 function observed in our model works in HFSCs' formation of pre-bulge during morphogenesis via enhancing activation of the canonical Wnt pathway, whereas in contrast, in the postnatal immature bulge, activation of canonical Wnt signaling was attenuated. In vitro studies on keratinocytes revealed changes in proliferation, migration, and colony formation, highlighting the inhibitory effect of constitutive overexpression of Rspo3 on these cellular processes. Our research provides novel insights into the role of Rspo3 in the regulation of hair morphogenesis and development, along with the formation and maturation of the HFSCs, which affect hair regeneration.

Effect of Adeno-Associated Virus-Transfected Mesenchymal Stem Cells Containing Agents On Bone Formation in Extended İnter-Premaxillary Suture in Rats

Aim: The aim of this study was to investigate the histomorphometric effects of bone marrow-derived mesenchymal stem cells (MSCs) transfected with Adeno Associated Virus (AAV) and containing bone morphogenic protein 7 (Bmp7) or osteoprotegerin (OPG) on bone formation after injection into inter-premaxillary suture in rats with Bmp7 or OPG alone. Materials and methods: In 4 groups (each group n:9), different chemical solutions, namely AAV-Bmp7, AAV-OPG and AAV-Bmp7-OPG and AAV-EGF (control group) were injected into the interpremaxillary suture of rats. Bone volumes (BV), soft tissue volume (STV) and total bone volumes (TBV) of 10 μm serial selection with hemotoxylin and eosin staining were calculated according to the Cavalieri principle. Each point in the region of interest was 1000μm3. Results: The BV for AAV-Bmp7, AAV-OPG, AAV-Bmp7-OPG and AAV-EGF were 46.98±1.50 mm3, 49.40±4.72 mm3, 42.58±2.89 mm3 and 38.82±0.76 mm3, respectively. The STV was 11.53±0.99, 13.31±1.88, 8.00±4.43 and 9.57±1.90 mm3 for Bmp7, OPG, AAV-Bmp7-OPG and AAV-EGF, respectively. TBV was 58.34±2.28 mm3, 63.83±5.17 mm3, 53.74±3.34 mm3 and 48.13±1.54 mm3 for AAV-Bmp7, AAV-OPG, AAV-Bmp7-OPG and AAV-EGF, respectively. The comparison between BV, STV, TBV for AAV-OPG showed a statistically significant difference (p=0.001) compared to AAV-Bmp7 or AAV-Bmp7-OPG and AAV-EGF. Conclusion: During tooth movement and bone remodeling, the ratio of soft and bone tissues is maintained by OPG. Although Bmp7 is not as effective as OPG in bone remodeling, both can reduce the retention time and the risk of recurrence.