Yanpeng Liu

Thermoresponsive hydrogels based on a phosphorylated star-shaped copolymer: mimicking the extracellular matrix for in situ bone repair

A type of four-arm star-shaped copolymer (star-PAA(PEA)-PNIPAM), consisting of the thermoresponsive block (poly(N-isopropylacrylamide), PNIPAM) and the phosphorylated functional block O-phosphoethanolamine (PEA) grafted poly(acrylic acid) (PAA(PEA)), is synthesized by atom transfer radical polymerization (ATRP) and subsequent modification. Owing to the unique superiority of the star-shaped structure, star-PAA(PEA)-PNIPAM can transform from the sol to gel state in response to the physiological temperature (37 °C) at a relatively low polymer concentration (>0.5 wt%). In addition, because of the enriched phosphorylated functional groups, the hydrogel formed by star-PAA(PEA)-PNIPAM can mimic the acidic extracellular matrix protein to adsorb calcium ions and mineralize in situ, both in in vitro and in in vivo experiments. Meanwhile, it is favorable for cell adhesion and proliferation due to its appropriate three-dimensional interspace. Thus, the biocompatible star-PAA(PEA)-PNIPAM hydrogel has great potential for bone repair applications.

Bio-inspired peptide decorated dendrimers for a robust antibacterial coating on hydroxyapatite

Bacterial colonization on implanted biomaterials remains a clinically significant problem. In order to achieve relatively long-term antibacterial activity and reduce the incidence of infections associated with the use of biomaterials, a salivary statherin protein (SSP) inspired poly(amidoamine) dendrimer (SSP-PAMAM-NH2) was synthesized and characterized. PAMAM-NH2 has numerous peripheral amino groups, and thus possesses effective antibacterial activity. The SSP bio-inspired peptide sequence DDDEEKC was conjugated to PAMAM-NH2 since it has a strong capability of adsorbing on hydroxyapatite (HA). Moreover, SSP-PAMAM-NH2 is a zwitterionic polymer possessing cationic amino groups and anionic carboxylic groups, thus it can form aggregates by intermolecular electrostatic interactions, thereby promoting its adsorption on HA. Adsorption tests by ATR-IR, UV, QCM-D, and CLSM, all indicated that SSP-PAMAM-NH2 can tightly adsorb on the HA surface. We found that even after being incubated in PBS for 4 weeks, the SSP-PAMAM-NH2 treated HA disks still retained stable antibacterial activity, while the inhibitory impact of PAMAM-NH2 treated disks had disappeared. Animal experiments also demonstrated that SSP-PAMAM-NH2 could significantly reduce infection of HA implanted into the medullary cavity of rats.

Calcitonin-Loaded Thermosensitive Hydrogel for Long-Term Antiosteopenia Therapy

Effective antiosteopenia therapy can be achieved by designing long-term protein/peptide drug delivery systems for bone trabecula restoration. Here we show that a complex of salmon calcitonin and oxidized calcium alginate (sCT-OCA) was prepared and loaded into a thermosensitive copolymer hydrogel for long-term antiosteopenia treatment. The triblock copolymer, poly(d,l-lactic acid-co-glycolic acid)-b-poly(ethylene glycol)-b-poly(d,l-lactic acid-co-glycolic acid) (PLGA-PEG-PLGA) exhibited sol-gel transition at body temperature. The sustained release of sCT from the in situ gelling system was determined by both the degradation of the hydrogel and the decomposition of the sCT-OCA complex. This system showed sustained effects in reducing serum calcium and bone trabecula reconstruction in the treatment of glucocorticoid-induced osteopenia in rats for approximately 30 days after a single subcutaneous injection, which may shed light on antiosteopenia therapy in the future.

Cucurbit[7]-assisted sustained release of human calcitonin from thermosensitive block copolymer hydrogel

Conventional formulations of human calcitonin (hCT), a peptide drug, normally suffer from limited therapeutic efficacy with low stability and short half-life. We have found that the fibrillation of highly amyloidogenic hCT can be inhibited by cucurbit[7] (CB[7]), an amphiphilic small molecule. Meanwhile, a thermogelling copolymer was found to be a suitable candidate for sustained delivery of peptide/protein drugs. Herein, we report a long-term delivery formulation composed of hCT-CB[7] complex and biodegradable thermogel of poly(D,L-lactic acid-co-glycolic acid)-b-poly(ethylene glycol)-b-poly(D,L-lactic acid -co-glycolic acid) (PLGA-PEG-PLGA). A 20wt% PLGA-PEG-PLGA solution exhibited a temperature-sensitive sol-gel transition at 35°C in phosphate buffer solution and slowly degraded over one month at neutral pH. Both the mass fraction of PLGA-PEG-PLGA copolymer and the complexation of hCT-CB[7] moderated the release process. hCT was sustainedly released over three weeks in the 20wt% hydrogel with hCT-CB[7] complex at the ratio of 1:10 or 1:25. Further considering its low cytotoxicity, the delivery system is potential for the clinical application of hCT.

Detecting the Formation and Transformation of Oligomers during Insulin Fibrillation by a Dendrimer Conjugated with Aggregation-Induced Emission Molecule

The fibrillation of protein is harmful and impedes the use of protein drugs. It also relates to various debilitating diseases such as Alzheimers diseases. Thus, investigating the protein fibrillation process is necessary. In this study, poly(amido amine) dendrimers (PAMAM) of generation 3 (G3) and generation 4 (G4) were synthesized and conjugated with 4-aminobiphenyl, an aggregation-induced emission (AIE) moiety, at varied grafting ratios. Among them, one fluorescence probe named G3-biph-3 that was grafted average 3.25 4-aminobiphenyl to the G3, can detect the transformations both from native insulin to oligomers and from oligomers to fibrils. The size difference of native insulin, oligomers, and fibrils was proposed to be the main factor leading to the detection of the above transformations. Different molecular weights of sodium polyacrylate (PAAS) were also applied as a model to interact with G3-biph-3 to further reveal the mechanism. The results indicated that PAMAM with a certain generation and grafted with appropriate AIE groups can detect the oligomer formation and transformation during the insulin fibrillation process.

Bioinspired heptapeptides as functionalized mineralization inducers with enhanced hydroxyapatite affinity

The regeneration of mineral crystals under physiological conditions is an efficient way to repair defects in hard tissues. To achieve robust mineralization on surfaces such as the tooth enamel, an inducer requires strong affinity with the substrates and should be able to induce mineralization. Thus far, most studies used a single molecule containing two components to realize the above functions separately, which might be troublesome to synthesize and purify. In this work, inspired by the statherin in the salivary acquired pellicle, we designed a simple peptide sequence, Asp-Asp-Asp-Glu-Glu-Lys-Cys (peptide-7), to accomplish the dual tasks of adsorption and mineralization on enamel surfaces. We speculate the calcium binding ability of the negatively charged carboxylic acid groups in the peptide itself contributes to the dual functions of peptide-7. In vitro and in vivo experiments demonstrated its excellent repair effect on enamel as compared to fluoride. More importantly, due to the strong affinity between peptides and hydroxyapatite, a compact mineralized crystal layer and a strong adhesion between the regenerated minerals and the bottom substrates were observed, similar to the effect induced by fluoride. This work sheds light on the interaction mechanism between peptide-7 and minerals. In addition, since it is safer than fluoride, peptide-7 may have potential applications in the repair of other hard tissues and the functionalization of biomaterials.

A stimuli-responsive insulin delivery system based on reversible phenylboronate modified cyclodextrin with glucose triggered host-guest interaction

ABSTRACT Injection of insulin is an effective therapy to treat most patients with the type I diabetes and some with type II diabetes. Additionally, the release of insulin under specific conditions has attracted widespread interest. In this study, a smart drug carrier that can release insulin depending on the changes in blood glucose levels was designed. Combining two popular molecules through facile synthetic processes, a drug carrier of reversible phenylboronate group modified cyclodextrin (&bgr;‐CD‐EPDME) was fabricated. The drug carrier is composed of cyclodextrin, which can encapsulate insulin, and phenylboronate, which is sensitive to the cis‐diols in some saccharides. Moreover, &bgr;‐CD‐EPDME can successfully encapsulate insulin and almost completely release insulin in the presence of glucose. The detached phenylboronic acid moiety triggered by glucose can attack the &bgr;‐CD cavity and form a host‐guest complex, which can force out the encapsulated insulin within the cavity. In addition, the insulin released from the &bgr;‐CD‐EPDME@Insulin complex retains its secondary structure, and the drug carrier has been proven to have low cytotoxicity. Thus, this safe and glucose‐responsive drug carrier shows the potential for use in the therapy of diabetes.

Inhibition of the fibrillation of highly amyloidogenic human calcitonin by cucurbit[7]uril with improved bioactivity.

Protein/peptide fibrillation is an important challenge for biotechnological drug development. Salmon calcitonin (sCT) is currently used in the clinical treatment of bone-related diseases such as osteoporosis and hypercalcemia, but it still has the risk of immune responses. Although human calcitonin (hCT) would be a better choice in terms of immunogenicity, it has a strong tendency to irreversibly aggregate in aqueous solutions and form long amyloid fibrils, which significantly reduces its bioavailability and therapeutic potency. Here, we demonstrate that cucurbit[7]uril (CB[7]) can inhibit hCT fibrillation by supramolecular interaction with its aromatic groups (affinity: Phe16 > Tyr12 > Phe19 > Phe22). The hCT-CB[7] complex exhibits low cytotoxicity, even promotes osteoblast proliferation and osteogenic capacity of MC3T3 cells. Meanwhile the hCT-CB[7] complexes shows higher bioactivity compared to hCT in reducing blood calcium levels in rats, and also decreases the immunogenicity of hCT. These results suggest that CB[7] has the potential to improve the therapeutic potency of amyloidogenic protein/peptide drugs such as hCT.