betadex and Osteoporosis

Osteoporosis is a skeletal disorder characterized by a low bone mass and density. Alendronate (Alen), a second-generation bisphosphonate drug, was indicated as the first-line regimen for the treatment of osteoporosis. However, the use of Alen has been limited due to its low bioavailability and gastrointestinal side effects. Herein, Alen-decorated nanoparticles were prepared through ionic cross-linking between poly (lactic-co-glycolic acid), β-cyclodextrin-modified chitosan (PLGA-CS-CD), and Alen-modified alginate (ALG-Alen) for Alen loading and bone-targeted delivery. Alen was selected as a therapeutic drug and a bone-targeting ligand. The nanoparticles have negatively charged surfaces, and sustained release of Alen from the nanoparticles can be observed. Cytotoxicity detected using cell counting kit-8 (CCK-8) assay and lactate dehydrogenase release test on MC3T3 cells showed that the nanoparticles had good cytocompatibility. A hemolysis test showed that the hemolysis ratios of nanoparticles were <5%, indicating that the nanoparticles had no significant hemolysis effect. Moreover, the Alen-decorated nanoparticles exhibited enhanced binding affinity to the hydroxyapatite (HAp) disks compared with that of nanoparticles without Alen modification. Thus, the Alen-decorated nanoparticles might be developed as promising bone-targeted carriers for the treatment of osteoporosis.

Topics: Alendronate; Alginates; Animals; beta-Cyclodextrins; Bone and Bones; Bone Density Conservation Agents; Cell Line; Cell Survival; Delayed-Action Preparations; Drug Carriers; Drug Liberation; Durapatite; Erythrocytes; Goats; Hemolysis; Mice; Nanoparticles; Osteoporosis; Polylactic Acid-Polyglycolic Acid Copolymer

In this study, β-cyclodextrin (β-CD) molecules are used as molecular reservoirs and grafted onto chitosan molecules for calcitriol (VD3) loading, which is a hormonally active metabolite of vitamin D. The resultant molecular complex is co-assembled with an antiosteoporosis drug calcitonin (CT) to form bio-functional multilayer structure on Ti6Al7Nb substrate via layer-by-layer self-assembly, which is capable of releasing VD3 and calcitonin in a sustained manner to modulate osteoblasts, osteoclasts, and macrophages at the bone-implant interface. In vitro results show that the released VD3 and CT individually upregulated the expression of calcium-binding protein (including Calbindin D9k and Calbindin D28k) and BMP2 in osteoblasts in peri-implant regions to stimulate their Ca deposition and differentiation. RAW264.7 cells (a murine macrophage) on the biofunctional implant displayed improved M2 phenotypical differentiation and expression of BMP2 and VEGF genes, but M1 phenotypical differentiation potential and MCF and TRAP gene expression levels are evidently lower. Results from in vivo micro-CT and histological analysis also demonstrate that VD3/CT co-loaded implant can dramatically enhance the bone remodeling under osteoporotic conditions with significantly enhanced interfacial shear strength and improved osseointegration as compared to other groups. The insights in this study offer new avenues for the rational functionalization of titanium implants to effectively repair osteoporotic fractures. STATEMENT OF SIGNIFICANCE: A promising strategy to enhance the recovery rate of osteoporotic fractures is to immobilize antiosteoporotic drugs onto the surface of titanium-based implants. In this study, we grafted beta-cyclodextrin (β-CD) onto chitosan (Chi) molecules to load VD3, which was co-assembled with calcitonin (CT) onto Ti6Al7Nb implants by the layer-by-layer assembly technique. The obtained functional titanium alloy implant (Ti6Al7Nb/LBL/Chi-CD@VD3/ CT) could stably release VD3 and calcitonin agents in a sustained manner. RAW264.7 cells grown on Ti6Al7Nb/LBL/Chi-CD@VD3/CT showed superior M2 phenotypical differentiation efficiency, but lower MCF/TRAP gene expression levels. In vitro and in vivo results showed that the released VD3 and CT individually upregulated the expression of calcium binding proteins and BMP2 in osteoblasts, promoting new bone formation in the peri-implant region.

Topics: Alkaline Phosphatase; Animals; beta-Cyclodextrins; Bone and Bones; Calcitonin; Calcitriol; Cattle; Cell Differentiation; Cell Polarity; Chitosan; Drug Delivery Systems; Gene Expression Regulation; Mice; Osseointegration; Osteoblasts; Osteogenesis; Osteoporosis; Prostheses and Implants; Proton Magnetic Resonance Spectroscopy; Rabbits; Rats; RAW 264.7 Cells; RNA, Messenger; Titanium; Wound Healing

Osteoporosis, a systemic skeletal disease prevalent in elderly women, is associated with post-menopausal estrogen deficiency. Although systemic administration of exogenous estradiol (E2) reduced fragility fractures, the treatment has adverse effects. Localized delivery technologies of E2 could be utilized to circumvent the systemic adverse effects of systemic administration. In this study, a localized E2 delivery system is developed. Mesoporous bioactive glass nanoparticles (MBGNPs) with inherent osteogenic properties are modified with β-cyclodextrin (CD-MBGNPs) to enhance their affinity for E2. To ensure mechanical stability and integrity, E2 loaded CD-MBGNPs are further electrospun with silk fibroin (SF) to produce a nanofibrous mesh (E2@CD-MBGNPs/SF). The incorporation of MBGNPs in SF enhances in vitro apatite formation and sustains the constant release of E2. Moreover, osteoblast proliferation and differentiation markers such as alkaline phosphatase activity, collagen 1 and osteocalcin expression of MC3T3-E1 are augmented in CD-MBGNPs/SF and E2@CD-MBGNPs/SF as compared to SF nanofibers. On the other hand, osteoclast DNA, tartrate resistant acid phosphatase activity and multinucleated cell formation are reduced in E2@CD-MBGNPs/SF as compared to CD-MBGNPs/SF and SF. Hence the presence of CD-MBGNPs in SF stimulates osteoblast function whereas E2 incorporation in CD-MBGNPs/SF reduces osteoclast activity. This is the first report to develop CD-MBGNPs/SF as a localized delivery system for hydrophobic molecules such as estradiol to treat osteoporosis.

Topics: Animals; Apatites; beta-Cyclodextrins; Cell Differentiation; Cell Proliferation; Cells, Cultured; Drug Delivery Systems; Drug Liberation; Estradiol; Fibroins; Mice; Nanofibers; Nanoparticles; Osteoblasts; Osteogenesis; Osteoporosis; Rats

An osteotropic alendronate-beta-cyclodextrin conjugate (ALN-beta-CD) was developed as a bone-targeting delivery system for improved treatment of skeletal diseases. The conjugate shows very strong binding to hydroxyapatite (HA, main component of the skeleton). Its ability in forming molecular inclusion complex with prostaglandin E(1) (PGE(1), a potent bone anabolic agent) was confirmed by phase solubility experiments and differential scanning calorimetry (DSC). In a bilateral rat mandible model, ALN-beta-CD/PGE(1) molecular complex was shown to stimulate strong local bone anabolic reaction. In the control study, ALN-beta-CD itself was also found to be bone anabolic. To investigate this finding, other control groups were studied. The histomorphometry data suggest that ALN-beta-CD itself could generate more new bone at the injection site than its complex with PGE(1). Alendronate (ALN) injection could also cause new bone formation, which locates peripheral to the site of injection. PGE(1), saline or ethanol injections do not have anabolic effect. These findings were also confirmed by micro-CT evaluation of mandibular bones. It is clear that the bone anabolic effect of ALN-beta-CD is independent of mechanical stimuli of the periosteum or ALN injection alone. Further studies are warranted to understand the working mechanism of ALN-beta-CD as a bone anabolic agent.

Topics: Alprostadil; Animals; beta-Cyclodextrins; Bone Regeneration; Calorimetry, Differential Scanning; Disease Models, Animal; Drug Delivery Systems; Minerals; Osteoporosis; Rats; Solubility; Tomography, X-Ray Computed

We examined the usefulness of intranasal (i.n.) administration of a novel osteotropic prodrug of estradiol, estradiol-17beta-succinate-(L-aspartate)6 (E2.17D6), for selective drug delivery to bone. E2.17D6 alone or with 5% 2,6-di-O-methyl-beta-cyclodextrin (DMbetaCD), 5% beta-cyclodextrin (betaCD), or 10% hydroxypropyl cellulose (HPC) as an absorption enhancer was administered to ovariectomized (OVX) mice via the i.n. route. The oral and nasal bioavailability after p.o. or i.n. administration of E2.17D6 (3.7 micromol/kg) in mice amounted to 9.9 and 23.0% of the dose, respectively. The values of nasal bioavailability of E2.17D6 administered with DMbetaCD, betaCD, and HPC were 74.9, 55.8, and 49.1%, respectively. The plasma concentration of E2.17D6 after i.n. administration of E2.17D6-DMbetaCD decreased rapidly to the endogenous level by 6 h, but the concentration in the bone was about 200 times higher than that in plasma, and decreased slowly over a period of about a week. When E2 (total dose 4.4 micromol/kg, i.n., every 3rd day) was administered to OVX mice for 35 d, bone mineral density (BMD), liver weight, and uterus weight increased, whereas E2.17D6-DMbetaCD (total dose 0.44 to 8.8 micromol/kg, i.n., every 7th day) increased only BMD in a dose-dependent manner. In conclusion, intranasally administered E2.17D6-DMbetaCD has a potent antiosteoporotic effect without side effects, and has potential to provide an improved quality of life for patients with osteoporosis.

Topics: Administration, Intranasal; Animals; Aspartic Acid; beta-Cyclodextrins; Bone and Bones; Dose-Response Relationship, Drug; Drug Carriers; Estradiol; Female; Gas Chromatography-Mass Spectrometry; Mice; Mice, Inbred Strains; Molecular Structure; Osteoporosis; Ovariectomy; Prodrugs; Time Factors; Tissue Distribution