Coronary artery disease (CAD) remains a leading cause of global morbidity and mortality despite advances in medical and interventional therapies. Mesenchymal stem cell (MSC) therapy has emerged as a promising regenerative approach for patients with refractory or non-revascularizable CAD. MSCs exhibit unique immunomodulatory, pro-angiogenic, and anti-fibrotic properties, primarily through paracrine mechanisms involving the secretion of cytokines, growth factors, and exosomal microRNAs. Clinical and preclinical studies have demonstrated improvements in myocardial perfusion, left ventricular ejection fraction (LVEF), and functional capacity following MSC-based interventions, particularly in patients with low baseline LVEF and heightened inflammation. Various MSC sources—including bone marrow, adipose tissue, and umbilical cord—offer distinct advantages, while delivery strategies such as intracoronary, intramyocardial, intravenous, and subcutaneous administration impact cell retention and efficacy. Advances in genetic modification, hypoxic preconditioning, and exosome-based therapies aim to enhance MSC survival and therapeutic potency. However, challenges persist regarding cell engraftment, cryopreservation effects, and inter-patient variability. Moving toward precision cell therapy, future approaches may involve stratifying patients by inflammatory status, ischemic burden, and comorbidities to optimize treatment outcomes. MSCs may not yet replace conventional therapies but are increasingly positioned to complement them within a personalized, regenerative framework for CAD management.

Dental and maxillofacial bone grafts may fail to reconstruct large bone deficiencies or otherwise due to insufficient blood supply, infections, or mechanical instabilities. To address this, Bonus Biogroup developed BonoFill, an injectable, autologous bone graft that employs a bone-like mineral scaffold to deliver a three-dimensional multi-cell culture of regenerative cells that support bone regeneration, vascularization, and graft integration. These cells are derived from mesenchymal stromal and other cells obtained from the patient's adipose tissue.

Background & AimBone tissue engineering stands as a principal area of research in mesenchymal cell-based regenerative medicine. The aim is to develop a viable alternative to autologous bone grafting, currently marked by procedural complexities and high failure rates. Bonus Biogroup has been at the forefront of this effort with our innovative product, BonoFill™. This tissue-engineered bone graft utilizes a biodegradable mineral scaffold in an injectable formulation to deliver a three-dimensional culture of hemi-differentiated autologous mesenchymal cells. These cells range from multipotent cells to mature osteoprogenitors, offering capabilities beyond mere bone rebuilding; they support vascularization and graft integration.This abstract presents interim results from our Phase II clinical trials for treating complex maxillofacial bone deficiencies, traditionally treated with autologous bone grafting. All transplantations in this trial have been completed, and the final results will be presented during ISCT 2024. Concurrently, another Phase II clinical trial is ongoing to address critical-sized limb bone defects.Methods, Results & ConclusionIn the Phase II study, subjects with maxillofacial bone deficiencies received BonoFill™ following cyst removal (Fig. 1A) or for sinus augmentation (Fig. 1B). Thirty bone defects were treated with over 14cc of BonoFill™ demonstrating a satisfactory safety profile. Based on 20 out of 30 procedures for subjects who completed the six-month follow-up period, successful bone regeneration was noted in 18/20 treated bones, enabling further reconstructive procedures. The bone height at the graft sites showed an average increase of 9.2±1.2 mm (p<0.0001), resulting in a final average bone height of 14.3±1.3 mm. Notably, this bone augmentation was already apparent three months post-transplantation (Fig. 1C), indicating rapid regeneration and successful graft integration. Biopsies taken six months post-treatment from the graft sites revealed healthy bone formation, abundant vascularization, and no inflammation (Fig. 2). The complete success of BonoFill™, evident in 90% of the cases, along with its safety, marks a formidable solution for complex bone deficiencies. Supported by promising results from a concurrent Phase II trial focusing on critical-sized limb bone defects, the results presented during ISCT 2024 represent a significant milestone in mesenchymal cell-based bone tissue engineering.

Detailed description of several methods mentioned in the manuscript

Supplemental Figures S1-7 supporting figures 1-6. Figure S1: Tumor growth in mice coâ€implanted with gemcitabineâ€educated MSCs and PANC1 cells. Figure S2: Viability and survival of TICs and MSCs in response to chemotherapy. Figure S3: TIC enrichment in pancreatic cancer cells cultured with conditioned medium obtained from gemcitabineâ€educated MSCs. Figure S4: Sphere formation of pancreatic cancer cells in the presence of conditioned medium obtained from gemcitabineâ€educated MSCs. Figure S5: Gemcitabineâ€educated MSCs promote PANC1 tumor growth. Figure S6: TIC enrichment in the presence of recombinant CXCL10. Figure S7: Densitometry analysis of expression of proteins from different signaling pathways.

Fold change in the expression levels of factors secreted by gemcitabine-educated versus untreated MSCs

S31time and sorted at high throughput.Cartilage regeneration is one of the targets for regenerative therapy because of poor regenerative potential.A previous study showed that the human chondrocytes fluorescently labeled with CFSE exhibited a wide variety in fluorescent intensities, indicating heterogeneity in their proliferation rates.The rapid proliferation cell population (low CFSE intensity) showed more matrix production potential than the other cells (high CFSE intensity).To evaluate if LF-GC is applicable to the enrichment of high matrix producing chondrocytes, we analyzed CFSE-stained human chondrocytes on LF-GC.Methods, Results & Conclusion: By defining rapid/slow proliferation cell populations based on CFSE intensity (low 30% population as rapid cells and high 70% population as slow cells), we built a LF-GC classifier based on support vector machine (SVM) with the area under the receiver operating characteristic curve (AUC) of 0.86.With the LF-GC classifier, we enriched the rapid proliferation cell population from 33.6% to 76.8%.After three weeks of culturing the cells as pellets, we measured glycosaminoglycans (GAGs) accumulations to evaluate the extracellular matrix production.The sorted samples accumulated more GAGs compared to the control samples with statistical significance.Here, we demonstrated LF-GC's potential to purify the desired cells without any staining, which suggests that it could be a new effective tool for label-free and selective cell isolation and purification in regenerative medicine.

S21markers, including CRP, ferritin, D-dimer, IL-6, LDH, platelet count, and lymphocyte count, all showed various levels of improvement at day 7 after SBI-101.A comprehensive profiling of 200 exploratory biomarkers and immune cell subsets over timepoints pre-and post-treatment will be presented to characterize the pharmacokinetic and pharmacodynamic effects of SBI-101 on the immune system.Overall, these preliminary results suggest ex vivo MSC therapy carries significant promise and warrants further study in the treatment of patients with severe COVID-19 requiring CRRT.

Mesenchymal stromal cells (MSC) are widely investigated for treating ARDS in Covid-19. Nonetheless, these efforts are overshadowed by studies predating the pandemic that mostly failed to show MSC efficacy in ARDS and recent disappointments with repurposed MSC products. Relying on years of MSC-related experience, Bonus BioGroup developed MesenCure: An enhanced allogeneic MSC therapy for Covid-19, professionalized by a unique combination of culture conditions and optimized in ARDS-relevant models.

INTRODUCTION:Colorectal cancer (CC) is the third most common type of cancer, accounting for 10% of all cancer cases. Adjuvant chemotherapy is recommended in stages II-III CC. Wheatgrass juice (WGJ) from wheat seeds has high nutritional values, may induce synergistic benefits to chemotherapy and may attenuate chemotherapy-related side effects. Extracellular vesicles (EVs) are subcellular membrane blebs. EVs include exosomes (generated in the endosome, in size <150 nm) and microvesicles (shed from the plasma cell membrane) provide information on their parental cells and play a role in intercellular communication. We aimed to elucidate the effects of chemotherapy administration with supportive treatment of WGJ on CC patients' EVs characteristics.METHODS:EVs were isolated from the blood samples of 15 healthy controls (HCs) and 50 CC patients post-surgery, treated by chemotherapy, with or without additional daily WGJ. Blood samples were taken before, during, and at the end of chemotherapy. EVs were characterized by size, concentration, membrane antigens and cytokine content using nanoparticle-tracking analysis, western blot, flow cytometry, and protein array methods.RESULTS:EVs were found to be similar by size and concentration with reduced levels of exosome markers (CD81) on samples at the end of combined treatment (chemotherapy and WGJ). Higher levels of endothelial EVs, which may indicate impairment of the vascular endothelial cells during treatment, were found in CC patients treated by chemotherapy only compared to those with chemotherapy and daily WGJ. Also, EVs thrombogenicity was lower in patients added WGJ compared to patients who had only chemotherapy (levels of tissue factor p = 0.029 and endothelial protein C receptor p = 0.005). Following treatments, levels of vascular endothelial growth factor receptors (VEGFR-1) and the majority of growth-factors/pro-inflammatory cytokines were higher in EVs of patients treated by chemotherapy only than in EVs obtained from patients with the combined treatment.CONCLUSION:Daily consumption of WGJ during chemotherapy may reduce vascular damage and chemotherapy-related thrombogenicity, growth factors and cytokines, as reflected by the characteristics of patient's EVs.

Background & Aim Significant losses of bone tissues that are often too large to heal occur in >10% of all open fractures and present an urgent clinical challenge associated with grave morbidities and huge healthcare costs. Bone autografting, the therapeutic standard in such cases, has a 50% complication rate, with persistent infection or non-union present in 18% of the patients, and further surgery required in 36% of them. Bone tissue engineering technologies have been widely suggested as an alternative to autografts. Such technologies were tested in segmental animal bone defects up to 5 cm long, with success reported mostly for defects under 2.5 cm. This size (or equivalent product volume) is also the upper limit for tissue-engineered bone grafts in clinical development. Considering that much larger defects, with a mean of 5.5 cm, are encountered in the clinic, Bonus is developing autologous tissue-engineered bone grafts for the treatment of defects larger than 2.5 cm. Methods, Results & Conclusion Bonus utilizes proprietary procedures to isolate adipose tissue-derived mesenchymal cells from patients' lipoaspirates, seed them on mineral scaffold particles, and culture them in a specially designed bioreactor (Fig. 1). The expansion of the cells on this scaffold increases their osteo-inducibility, allowing a controlled osteoinduction process that possibly also retains some of these cells' additional regenerative functions. This possibility is exemplified by the higher levels of osteogenic and other genes related to angiogenesis and ECM remodeling expressed by 3D-osteoinduced cells, compared to the 2D ones. After their osteoinduction, the cells and scaffold are washed and formulated into the final product—BonoFill-II. In a preclinical study, BonoFill-II applied to a 3.2 cm defect in a sheep tibia led to a full recovery in 12 weeks (N=7), as well as a full bridging of a 10.5 cm defect (Fig. 2). A phase I/II clinical trial is currently underway testing BonoFill-II in patients with bone defects ≥2.5 cm. So far, we have treated patients with defects up to 8.5 cm long and a history of 2-7 failed prior interventions, with a few representatives that can exemplify this study safety and efficacy (Fig. 3). The performance of BonoFill-II was also tested in Phase I/II trial in patients in need of a bone transplant in the upper maxilla with complete success reported. Overall, the solid underlying science and innovative technologies developed by Bonus position BonoFill-II as a promising alternative to bone autografts. Significant losses of bone tissues that are often too large to heal occur in >10% of all open fractures and present an urgent clinical challenge associated with grave morbidities and huge healthcare costs. Bone autografting, the therapeutic standard in such cases, has a 50% complication rate, with persistent infection or non-union present in 18% of the patients, and further surgery required in 36% of them. Bone tissue engineering technologies have been widely suggested as an alternative to autografts. Such technologies were tested in segmental animal bone defects up to 5 cm long, with success reported mostly for defects under 2.5 cm. This size (or equivalent product volume) is also the upper limit for tissue-engineered bone grafts in clinical development. Considering that much larger defects, with a mean of 5.5 cm, are encountered in the clinic, Bonus is developing autologous tissue-engineered bone grafts for the treatment of defects larger than 2.5 cm. Bonus utilizes proprietary procedures to isolate adipose tissue-derived mesenchymal cells from patients' lipoaspirates, seed them on mineral scaffold particles, and culture them in a specially designed bioreactor (Fig. 1). The expansion of the cells on this scaffold increases their osteo-inducibility, allowing a controlled osteoinduction process that possibly also retains some of these cells' additional regenerative functions. This possibility is exemplified by the higher levels of osteogenic and other genes related to angiogenesis and ECM remodeling expressed by 3D-osteoinduced cells, compared to the 2D ones. After their osteoinduction, the cells and scaffold are washed and formulated into the final product—BonoFill-II. In a preclinical study, BonoFill-II applied to a 3.2 cm defect in a sheep tibia led to a full recovery in 12 weeks (N=7), as well as a full bridging of a 10.5 cm defect (Fig. 2). A phase I/II clinical trial is currently underway testing BonoFill-II in patients with bone defects ≥2.5 cm. So far, we have treated patients with defects up to 8.5 cm long and a history of 2-7 failed prior interventions, with a few representatives that can exemplify this study safety and efficacy (Fig. 3). The performance of BonoFill-II was also tested in Phase I/II trial in patients in need of a bone transplant in the upper maxilla with complete success reported. Overall, the solid underlying science and innovative technologies developed by Bonus position BonoFill-II as a promising alternative to bone autografts.

Nanoghosts (NGs) are nanovesicles reconstructed from the cytoplasmic membranes of mesenchymal stem cells (MSCs). By retaining MSC membranes, the NGs retain the ability of these cells to home in on multiple tumors, laying the foundations, thereby, for the development of a targeted drug delivery platform. The susceptibility of MSCs to functional changes, following their exposure to cytokines or cancer‐derived conditioned‐media (CM), presents the opportunity to modify the NGs by conditioning their source cells. This opportunity is investigated by comparing the membrane protein composition and the tumor uptake of NGs derived from naïve MSCs (N‐NG) against conditioned NGs made from MSCs pre‐treated with conditioned‐media (CM‐NG) or with a mix of the proinflammatory cytokines TNF‐α and IL‐1β (Cyto‐NG). CM‐NGs are found to be more targeted towards immune cells than Cyto‐ or N‐NGs, while Cyto‐NGs are the most tumor‐targeted ones, with similar immune‐targeting capacity as N‐NGs but with a higher affinity towards endothelial cells. Proteomic variations were wider in the CM‐NGs, with exceptionally higher levels of ICAM‐1 compared to N‐ and Cyto‐NGs. From a translational point of view, the data show that the tumor‐targeting ability of the NGs, and possibly that of other MSC‐derived extracellular vesicles, can be enhanced by simple conditioning of their source cells.

The rapid development of biomimetic cell membrane-based nanoparticles is still overshadowed by many practical challenges, one of which is the difficulty to precisely measure the biodistribution of such nanoparticles. Currently, this challenge is mostly addressed using fluorescent techniques with limited sensitivity, or radioactive labeling methods, which rarely account for the nanoparticles themselves, but their payloads instead. Here we report the development of a robust method for the innate radioactive labeling of cells and membrane-based nanoparticles and their consequent sensitive detection and biodistribution measurements. The preclinical potential of this method was demonstrated with Nano-Ghosts (NGs), manufactured from the cytoplasmic membranes of mesenchymal stem cells cultured with radioactively-labeled linoleic acid and achieving a cell labeling efficiency of 36%. Radiolabeling did not affect the physiochemical properties of the NGs, which stably retained their radiolabels. Using radioactivity measurements, we are now able to determine precisely the amount of NGs uptaken by tissues and cells, thereby providing further support to our presumed active NG targeting mechanisms. Biodistribution studies comparing radiolabeled NGs to fluorescently-labeled ones have validated our method and revealed new information, which could not be obtained otherwise, regarding the NGs' unique kinetics and rapid clearance, supporting their excellent safety profiles. The reported approach may be expanded to other membrane-based entities to facilitate and hasten their preclinical development and be used in parallel with other labeling methods to provide different and additional information.