Exosome-Functionalized, Drug-Laden Bone Substitute along with an Antioxidant Herbal Membrane for Bone and Periosteum Regeneration in Bone Sarcoma.

Developing advanced methods for effective bone reconstructive strategies in case of critical bone defects caused by tumor resection, trauma, and other implant-related complications remains a challenging problem in orthopedics. In the clinical management of bone diseases, there is a paradigm shift in using local drugs at the injury site; however, the dead space created during the surgical debridement of necrotic bone and soft tissues (periosteum and underlying muscle) leads to ineffective bone formation, thereby leading to secondary complications, and thus calls for better regenerative approaches. In this study, we have utilized an exosome-functionalized doxorubicin-loaded biodegradable nanocement (NC)-based carrier along with a Cissus quadrangularis (CQ) extract-laden antioxidant herbal membrane for simultaneously managing the periosteum as well as bone formation in the tumor resection model of osteosarcoma. We initially evaluated the efficacy of scaffolds for in vitro mineralization and bone formation. To examine the in vivo effectiveness, we developed a human osteosarcoma cell line (Saos-2)-induced tumor xenograft model with a critical-sized bone defect. The findings revealed that doxorubicin released from NC was successful in killing the tumor cells and was present even after 30 days of implantation. Additionally, the incorporation of exosomes aided the bone formation, resulting in around a 2.6-fold increase in the bone volume compared to the empty group as evaluated by micro-CT. The herbal membrane assisted in the development of periosteum and mineralizing bone callous as validated through histological and immunofluorescence analysis. Thus, our findings describe a one-step biomaterial-based cell-free approach to regenerate bone in osteosarcoma and prevent further fracture due to the complete development of periosteum and lost bone.

Effect of hyperthermia and proton beam radiation as a novel approach in chordoma cells death and its clinical implication to treat chordoma.

PURPOSE: Chordoma is a locally aggressive tumor that most commonly affects the base of the skull/clivus, cervical, and sacral spine. Conventional radiotherapy (RT), cannot be safely increased further to improve disease control due to the risk of toxicity to the surrounding critical structures. Tumor-targeted hyperthermia (HT) combined with Proton Beam Radiation Therapy (PBRT) is known to act as a potent radiosensitizer in cancer control. In this study, we investigated whether PBRT efficacy for chordoma can be enhanced in combination with HT as a radiosensitizer. MATERIAL AND METHODS: Human chordoma cell lines, U-CH2 and Mug-chor1 were treated in vitro with HT followed by PBRT with variable doses. The colony-forming assay was performed, and dose-response was characterized by linear-quadratic model fits. HSP-70 and Brachyury (TBXT) biomarkers for chordoma aggression levels were quantified by western blot analysis. Gene microarray analysis was performed by U133 Arrays. Pathway Analysis was also performed using IPA bioinformatic software. RESULTS: Our findings in both U-CH2 and Mug-Chor1 cell lines demonstrate that hyperthermia followed by PBRT has an enhanced cell killing effect when compared with PBRT-alone (p < .01). Western blot analysis showed HT decreased the expression of Brachyury protein (p < .05), which is considered a biomarker for chordoma tumor aggression. HT with PBRT also exhibited an RT-dose-dependent decrease of Brachyury expression (p < .05). We also observed enhanced HSP-70 expression due to HT, RT, and HT + RT combined in both cell lines. Interestingly, genomic data showed 344 genes expressed by the treatment of HT + RT compared to HT (68 genes) or RT (112 genes) as individual treatment. We also identified activation of death receptor and apoptotic pathway in HT + RT treated cells. CONCLUSION: We found that Hyperthermia (HT) combined with Proton Beam Radiation (PBRT) could significantly increase chordoma cell death by activating the death receptor pathway and apoptosis which has the promise to treat metastatic chordoma.

Exosome-Functionalized Ceramic Bone Substitute Promotes Critical-Sized Bone Defect Repair in Rats.

Ceramic biomaterials are promising alternatives to bone autografts. However, limited bioactivity affects their performance. Therefore, bioactive molecules and cells are often added to enhance their performance. Exosomes have emerged as cell-secreted vesicles, delivering proteins, lipids, and nucleic acids in a paracrine/endocrine fashion. We studied two complementary aspects required for exosome activity/therapy using purified exosomes: first, the intracellular uptake of labeled exosomes and second, the influence of delivered exosomes on cell behavior. Origin-specific differences in the characteristics of purified exosomes, quantification of time-dependent intracellular uptake of PKH-26-labeled exosomes by mesenchymal stem cells (MSCs) and preosteoblasts, and influence on cell behavior were evaluated. Furthermore, exosomes from osteoblasts and MSCs cultured under normal and osteogenic environments were isolated. There is little data available on the concentration and dose of exosomes required for bone regeneration. Therefore, equal amounts of quantified exosomes were implanted in vivo in rat tibia critical defects using a calcium sulfate-nano-hydroxyapatite nanocement (NC) bone filler as the carrier. Bone regeneration was quantified using micro-computed tomography and histology. Along with inducing early maturation and mineral deposition by primary preosteoblasts in vitro, exosome treatment also demonstrated a positive effect on bone mineralization in vivo. Our study concludes that providing a local delivery of exosomes loaded onto a slowly resorbing NC bone filler can provide a potential alternate to autografts as a bone substitute.

Enhanced bone mineralization using hydroxyapatite-based ceramic bone substitute incorporating Withania somnifera extracts.

Withania somnifera (ashwagandha) is used in Indian traditional medicine for its various health benefits. Withaferin-A, a steroidal lactone present in this herb, has shown proteosomal inhibition-based enhancement of bone mineralization. In the present work, chitosan microparticles blended with total methanolic root extract of W. somnifera were incorporated as a porogen in calcium phosphate-based hydroxyapatite bone filler. The controlled release of bioactive molecules enabled enhanced proliferation and differentiation of pre-osteoblasts. Microparticle percentages were optimized to have a minimum effect on the setting time, mechanical strength and degradability of hydroxyapatite bone filler. In vitro cell adhesion, proliferation and differentiation were evaluated to determine the biocompatibility of the composites. On the basis of the desirable results obtained, we provide a preliminary rationale for the use of methanolic extract-blended chitosan microparticle-impregnated calcium phosphate filler for enhanced bone regeneration.

Orthobiologics with phytobioactive cues: A paradigm in bone regeneration.

Bone injuries occur due to various traumatic and disease conditions. Healing of bone injury occurs via a multi-stage intricate process. Body has the potential to rectify most of the bone injuries but some severe traumatic cases with critical size defects may require interventions. Autografts are still considered the "gold standard" for fracture healing but due to limitations associated with it, new alternatives are warranted. The field of orthobiologics has provided novel approaches using scaffolds, bioactive molecules, stem cells for the treatment of bone defects. Phyto-bioactives have been widely used in alternative medicine and folklore practices for curing bone ailments. It is believed that different bioactive constituents in plants work synergistically to give the therapeutic efficacy. Bioactives in plants extracts act upon different signal transduction pathways aiding in bone healing. The present review focuses on the use, chemical composition, mode of delivery, mechanism of action, and possible future strategies of three medicinal plants popularly used in traditional medicine for bone healing: Cissus quadrangularis, Withania somnifera and Tinospora cordifolia. Plants extracts seem to be a natural and non-toxic therapeutic alternative in treating bone injuries. Most of the studies on bone healing for these plants have reported oral administration of the extracts and presented them as a safe alternative without any side effects despite giving higher doses. Forthcoming studies could be directed towards the local delivery of extracts at the defect site. Unification of herbal extracts and orthobiologics could be an interesting direction in the field of bone healing in future. The present review intends to provide a bird's eye view of different strategies used in bone healing, mechanisms involved and future direction of advancements using phytobioactives and orthobiologics.

New insights into bioelectronic medicines: A new approach to tackle diabetic peripheral neuropathy pain in clinics.

Bioelectronic medicines are a newer way to treat and diagnose the diseases associated with biological systems. All vital organs of the body are innervated, commanding brain to regulate the homeostasis functions. Bioelectronic medicines rely on implications of electrical stimulations or signals associated with the nervous system for real-time treatment. Diabetic peripheral neuropathy (DPN) is a most prevalent micro-vascular complication associated with diabetes mellitus. Complex plexus of nerves were affected in this complication with impaired function. Bioelectronic medicines are future hope for effective treatment of DPN.