Bone targeting nanocarrier-assisted delivery of adenosine to combat osteoporotic bone loss.

Extracellular adenosine has been shown to play a key role in maintaining bone health and could potentially be used to treat bone loss. However, systemic administration of exogenous adenosine to treat bone disorders remains a challenge due to the ubiquitous presence of adenosine receptors in different organs and the short half-life of adenosine in circulation. Towards this, we have developed a bone-targeting nanocarrier and determined its potential for systemic administration of adenosine. The nanocarrier, synthesized via emulsion suspension photopolymerization, is comprised of hyaluronic acid (HA) copolymerized with phenylboronic acid (PBA), a moiety that can form reversible bonds with adenosine. The bone binding affinity of the nanocarrier was achieved by alendronate (Aln) conjugation. Nanocarriers functionalized with the alendronate (Aln-NC) showed a 45% higher accumulation in the mice vertebrae in vivo compared to those lacking alendronate molecules (NCs). Systemic administration of adenosine via bone-targeting nanocarriers (Aln-NC) attenuated bone loss in ovariectomized (OVX) mice. Furthermore, bone tissue of mice treated with adenosine-loaded Aln-NC displayed trabecular bone characteristics comparable to healthy controls as shown by microcomputed tomography, histochemical staining, bone labeling, and mechanical strength. Overall, our results demonstrate the use of a bone-targeting nanocarrier towards systemic administration of adenosine and its application in treating bone degenerative diseases such as osteoporosis.

Dysregulation of ectonucleotidase-mediated extracellular adenosine during postmenopausal bone loss.

Adenosine and its receptors play a key role in bone homeostasis and regeneration. Extracellular adenosine is generated from CD39 and CD73 activity in the cell membrane, through conversion of adenosine triphosphate to adenosine monophosphate (AMP) and AMP to adenosine, respectively. Despite the relevance of CD39/CD73 to bone health, the roles of these enzymes in bona fide skeletal disorders remain unknown. We demonstrate that CD39/CD73 expression and extracellular adenosine levels in the bone marrow are substantially decreased in animals with osteoporotic bone loss. Knockdown of estrogen receptors ESR1 and ESR2 in primary osteoprogenitors and osteoclasts undergoing differentiation showed decreased coexpression of membrane-bound CD39 and CD73 and lower extracellular adenosine. Targeting the adenosine A2B receptor using an agonist attenuated bone loss in ovariectomized mice. Together, these findings suggest a pathological association of purine metabolism with estrogen deficiency and highlight the potential of A2B receptor as a target to treat osteoporosis.

Stimuli-Responsive Supramolecular Hydrogels and Their Applications in Regenerative Medicine.

Supramolecular hydrogels are a class of self-assembled network structures formed via non-covalent interactions of the hydrogelators. These hydrogels capable of responding to external stimuli are considered to be smart materials due to their ability to undergo sol-gel and/or gel-sol transition upon subtle changes in their surroundings. Such stimuli-responsive hydrogels are intriguing biomaterials with applications in tissue engineering, delivery of cells and drugs, modulating tissue environment to promote innate tissue repair, and imaging for medical diagnostics among others. This review summarizes the recent developments in stimuli-responsive supramolecular hydrogels and their potential applications in regenerative medicine. Specifically, various structural aspects of supramolecular hydrogelators involved in self-assembly, the role of external stimuli in tuning/controlling their phase transitions, and how these functions could be harnessed to advance applications in regenerative medicine are focused on. Finally, the key challenges and future prospects for these versatile materials are briefly described.

Oligo(trimethylene carbonate)-poly(ethylene glycol)-oligo(trimethylene carbonate) triblock-based hydrogels for cartilage tissue engineering.

A triblock co-polymer of oligo(trimethylene carbonate)-block-poly(ethylene glycol) 20000-block-oligo(trimethylene carbonate) diacrylate (TMC20) was used as a photo-polymerizable precursor for the encapsulation of primary articular chondrocytes. The efficacy of TMC20 as a biodegradable scaffold for cartilage tissue engineering was compared with non-degradable poly(ethylene glycol) 20000 diacrylate (PEG20) hydrogel. Chondrocytes encapsulated in PEG hydrogels containing oligo(trimethylene carbonate) (OTMC) moieties underwent spontaneous aggregation during in vitro culture, which was not observed in the PEG hydrogel counterparts. The aggregation of cells was found to be dependent on the initial cell density, as well as the mesh size of the hydrogels. Similarly, cell aggregation was also found in biodegradable PEG hydrogels containing caprolactone moieties. The aggregation of cells in TMC20 hydrogels resulted in enhanced cartilage matrix production compared with their PEG20 counterparts over 3 weeks of culture. Taken together, these results indicate that PEG hydrogels containing degradable OTMC moieties promote the aggregation and biosynthetic activity of encapsulated chondrocytes, indicating their potential as scaffolds for the repair of cartilage tissue.

Influence of physical properties of biomaterials on cellular behavior.

PURPOSE: In this study, we evaluated the effect of hydrogel structural properties on proliferation and biosynthesis activity of encapsulated chondrocytes. METHODS: Hydrogels with varying structural and mechanical properties were prepared by photopolymerizing PEGDA precursors having MWs of 3.4 kDa, 6 kDa, 10 kDa, and 20 kDa and were characterized for their swelling ratio, network structure, morphology, and mechanical properties. The effect of hydrogel structural properties on the cellular activity of encapsulated chondrocytes was studied over four weeks. RESULTS: Varying the molecular weight of PEGDA precursors exhibited a significant effect on the structural and mechanical properties of the hydrogels. Large mesh size was found to support cell proliferation. However, extracellular matrix (ECM) accumulation varied with the precursor molecular weight. Both PEGDA 6 kDa and 10 kDa hydrogels supported GAG accumulation, while PEGDA 10 kDa and 20 KDa hydrogels supported collagen accumulation. Chondrocytes cultured in PEGDA 10 kDa hydrogels expressed a relative increase in collagen type II and aggrecan expression while maintaining low collagen type I expression. CONCLUSIONS: Increasing mesh size of the hydrogels resulted in an increase in cellular proliferation exhibiting the strong correlation between mesh size and cell growth, while mesh size had a differential effect on ECM accumulation and expression of cartilage specific markers.

Heparin mimicking polymer promotes myogenic differentiation of muscle progenitor cells.

Heparin and heparan sulfate mediated basic fibroblast growth factor (bFGF) signaling plays an important role in skeletal muscle homeostasis by maintaining a balance between proliferation and differentiation of muscle progenitor cells. In this study we investigate the role of a synthetic mimic of heparin, poly(sodium-4-styrenesulfonate) (PSS), on myogenic differentiation of C2C12 cells. Exogenous supplementation of PSS increased the differentiation of C2C12 cells in a dose-dependent manner, while the formation of multinucleated myotubes exhibited a nonmonotonic dependence with the concentration of PSS. Our results further suggest that one possible mechanism by which PSS promotes myogenic differentiation is by downregulating the mitogen activated extracellular regulated signaling kinase (MAPK/ERK) pathway. The binding ability of PSS to bFGF was found to be comparable to heparin through molecular docking calculations and by native PAGE. Such synthetic heparin mimics could offer a cost-effective alternative to heparin and also reduce the risk associated with batch-to-batch variation and contamination of heparin.

Interconnected macroporous poly(ethylene glycol) cryogels as a cell scaffold for cartilage tissue engineering.

Macroporous networks of poly(ethylene glycol) (PEG) with interconnected pores can be created by cryogelation techniques. In this study, we describe the potential application of such PEG cryogels as scaffolds for cartilage tissue engineering. Three-dimensional macroporous cryogels were evaluated for chondrocyte growth and production of cartilage-specific extracellular matrix (ECM). Seeded primary bovine chondrocytes showed homogeneous distribution throughout the cryogels. DNA content suggests continuous cell proliferation over 4 weeks of in vitro culture. Analysis of the composition of cell-secreted ECM showed a culture-time-dependent increase in the amount of glycosaminoglycan and collagen. The production of ECM by chondrocytes was confirmed using scanning electron microscopy analysis. Further histological and immunohistological analysis of the cell-laden scaffold confirmed the presence of accumulated cartilage-specific ECM within the scaffold. The interconnected macroporous network promoted diffusion of cell-secreted matrix within the cryogels. Our results indicated that interconnected macroporous PEG cryogels successfully supported attachment, viability, proliferation, and biosynthetic activity of seeded chondrocytes.