Damkragh9231

Critical-sized bone defects and nonunions following fracture are common among elderly patients and severely reduce the quality of life. Dysfunctional senescent endothelial and mesenchymal stromal cells (MSCs) inhibit bone defect repair. Here we provide a method to obtain surrogate vascularized juvenile bone by subcutaneous implantation of recombinant human bone morphogenic protein-2 (rhBMP-2)-loaded absorbable gelatin scaffolds. RhBMP-2-induced ossicles showed fewer senenscent MSCs whereas much more type H blood vessels (strongly positive for CD31 and endomucin (Emcn)) and osteoprogenitor cells than native bone (femur and tibiae) even in old mice. Treatment with this juvenile ossicles improved the regenerative capacity in critical-sized cranial defects versus standard treatment in both young and old mice. Furthermore, ossicles with custom size shape were obtained by 3D-printing for irregular bone defects repair. These customizable juvenile ossicles developed in aged individuals provide an alternative to resecting native bone in autologous bone transplantation, with superior regenerative efficacy in elderly patients due to their juvenile phenotype.Besides its broad application in research and biotechnology, genome editing (GE) has great potential for clinical gene therapy, but delivery of GE tools remains a bottleneck. Whereas significant progress has been made in ex vivo GE delivery (e.g., by electroporation), establishment of efficient and safe in vivo delivery systems is still a challenge. Bupivacaine Above and beyond standard vector requirements (safety, minimal/absent toxicity and immunogenicity, sufficient packaging capacity, targeting, straight and low-cost large-scale good manufacturing practice (GMP) production), GE delivery systems ideally use a hit-and-run principle to minimize off-targets as well as display of immunogenic peptides. Since currently used viral vectors do not fulfil all of these requirements, the broad variety of non-viral delivery platforms represents a promising alternative. This review provides a comprehensive analysis of the most relevant aspects of non-viral physical and chemical delivery methods in non-clinical studies and clinical trials, ranging from classic electroporation to advanced drug carriers that can transport GE tools in form of plasmid DNAs (pDNAs), mRNAs, and ribonucleoproteins (RNPs). For comparison, advantages and shortcomings of viral delivery systems are shortly discussed. In summary, we review various delivery approaches and discuss the future perspectives to use drug carriers for in vivo GE in clinical trials.The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated protein 9 (Cas9) system holds great promise for the cancer gene therapy. However, due to complicated signal networks and various compensatory mechanisms in tumors, adjusting a single molecular pathway has limited effects on cancer treatments. Herein, a virus-like nanoparticle (VLN) was reported as a versatile nanoplatform to co-deliver CRISPR/Cas9 system and small molecule drugs for effective malignant cancer treatment. VLN has a core-shell structure, in which small molecule drugs and CRISPR/Cas9 system are loaded in the mesoporous silica nanoparticle (MSN)-based core, which is further encapsulated with a lipid shell. This structure allows VLN maintaining stable during blood circulation. As reaching tumors, VLN releases the CRISPR/Cas9 system and small molecule drugs in response to the reductive microenvironment, resulting in the synergistic regulation of multiple cancer-associated pathways. By loading a single guide RNA (sgRNA) targeting programmed death-ligand 1 and axitinib, VLN achieved to disrupt multiple immunosuppressive pathways and suppress the growth of melanoma in vivo. More importantly, VLN can co-deliver almost any combination of sgRNAs and small molecule drugs to tumors, suggesting the great potential of VLN as a general platform for the development of advanced combination therapies against malignant tumors.Tumor-activatable ultrasmall nanozyme generation is an unprecedented strategy to overcome intrinsically fatal defects of traditional reactive oxygen species (ROS)-based nanoagents for deep tumor penetration, including limited tissue-penetrating depth of external energy, heavy reliance on oxygen and nonspecific toxicity of therapeutic agents. Here, based on the cascade reaction between glucose oxidase (GOx) and ultrasmall peroxidase nanozyme embedded into acid-dissociable zeolitic imidazolate framework-8 (ZIF-8), such a tumor-activatable ultrasmall nanozyme generator is designed for enhanced penetration and deep catalytic therapy. With the aid of mildly acidic tumor microenvironment, the produced gluconic acid from intratumoral glucose can gradually induce the dissociation of ZIF-8 to release ultrasmall peroxidase nanozyme with significant intratumoral penetration. On the other hand, the generated hydrogen peroxide with relatively long-life can be subsequently catalyzed by penetrated peroxidase nanozyme into toxic hydroxyl radicals for deep catalytic therapy. In this way, the well-designed nanoplatform not only can greatly enhance tumor penetration but also directly induce exogenous ROS without oxygen participation and external energy input, thereby thoroughly avoiding the inactivation of traditional ROS-based nanoagents in the extremely hypoxic tumor center and finally resulting in remarkable deep catalytic therapy.Currently, reactive oxygen species (ROS)-induced apoptosis systems have drawn increasing attention in cancer therapy, owing to their specific tumor inhibition ability and great biocompatibility. Herein, we developed a highly dispersed nano-enzyme based on the assembly of natural glucose oxidase (GOD) onto CoFe-layered double hydroxides (CoFe-LDHs) monolayer nanosheets. By virtue of the high dispersion of Fe3+ within the host layer, the CoFe-LDHs nanosheets exhibit a collaborative enhanced Fenton catalytic activity with a rate constant of 3.26 × 10-4 s-1, which is 1-3 orders of magnitude higher than other iron-containing Fenton reaction agents. Subsequently, with a massive H2O2 triggered by GOD, GOD/CoFe-LDHs nanohybrid converts a cascade of glucose into hydroxyl radicals under tumor acid conditions, which is validated by a high maximum velocity (Vmax = 2.23 × 10-6 M) and low Michaelis-Menten constant (KM = 5.40 mM). Through the intracellular catalytic Fenton reaction within the tumor environment, both in vitro and in vivo results demonstrate the excellent antitumor effect of GOD/CoFe-LDHs.