Photoinduced immobilization of 2-methacryloyloxyethyl phosphorylcholine polymers with different molecular architectures on a poly(ether ether ketone) surface.

Poly(ether ether ketone) (PEEK) has seen increasing use in biomedical fields as a replacement for metal implants. Accordingly, the surface functionalities of PEEK are important for the development of medical devices. We have focused on the application of photoinduced reactions in PEEK to immobilize a functional polymer via radical generation on the surface, which can react with hydrocarbon groups. In this study, we used zwitterionic copolymers comprising 2-methacryloyloxyethyl phosphorylcholine (MPC) units and n-butyl methacrylate (BMA) units with various molecular architectures for surface modification. A random copolymer (poly(MPC-co-BMA) (r-PMB)), an AB-type diblock copolymer (di-PMB), and an ABA-type triblock copolymer (tri-PMB) (A segment: poly(BMA); B segment: poly(MPC)) were synthesized with the same monomer compositions. All PMBs were successfully immobilized on the PEEK surface via UV irradiation after the dip-coating process, regardless of their molecular structure. In this reaction, the alkyl group of the BMA unit functioned as a photoreactive site on the PEEK surface. This indicates that the molecular structure differences affect the surface properties. For example, compared to r-PMB and tri-PMB, di-PMB-modified surfaces exhibited an extremely low water contact angle of approximately 10°. The findings of this study demonstrate that this surface functionalization method does not require a low-molecular-weight compound, such as an initiator, and can be applied to the surface of inert PEEK through a simple photoreaction under room temperature, atmospheric pressure, and dry state conditions.

Water-soluble and amphiphilic phospholipid copolymers having 2-methacryloyloxyethyl phosphorylcholine units for the solubilization of bioactive compounds.

We summarize the development and evaluation of new type of phospholipid polymers as a solubilizer for poorly water-soluble compounds. That is, a water-soluble and amphiphilic poly(2-methacryloyloxyethyl phosphorylcholine-random-n-butyl methacrylate) contains 30 mol% hydrophilic 2-methacryloyloxyethyl phosphorylcholine units and its weight-averaged molecular weight is around 5.0 × 104. When the polymer is dissolved in an aqueous medium, a large portion of hydrophobic n-butyl methacrylate units assemble, forming polymer aggregates. To avoid severe biological reactions caused by conventional solubilizers, the phospholipid polymer can be applied for the solubilization of poorly water-soluble bioactive compounds. The polarity inside these polymer aggregate is the same as that of ethanol and n-butanol. Therefore, bioactive compounds, whose solubility is poor in water but good in these alcohols, can be entrapped in the polymer aggregate. The phospholipid polymer can penetrate the cell membrane by molecular diffusion, carrying inside the cell the bioactive compound, without exhibiting significant cytotoxicity. Several animal experiments have revealed that the pharmacological performance of various bioactive compound/phospholipid polymer complexes is excellent. Furthermore, functionalization of the polymer aggregate with biomolecules, such as antibodies and oligonucleotides, can be done, leading to selective capturing of the target molecules. These examples clearly indicate that water-soluble and amphiphilic phospholipid polymer is a candidate for preparing safer formulations and more effective pharmaceutical treatment with several bioactive compounds.

Solubilization of poorly water-soluble compounds using amphiphilic phospholipid polymers with different molecular architectures.

To achieve stable and effective solubilization of poorly water-soluble bioactive compounds, water-soluble and amphiphilic polymers composed of hydrophilic 2-methacryloyloxyethyl phosphorylcholine (MPC) units and hydrophobic n-butyl methacrylate (BMA) units were prepared. MPC polymers having different molecular architectures, such as random-type monomer unit sequences and block-type sequences, formed polymer aggregates when they were dissolved in aqueous media. The structure of the random-type polymer aggregate was loose and flexible. On the other hand, the block-type polymer formed polymeric micelles, which were composed of very stable hydrophobic poly(BMA) cores and hydrophilic poly(MPC) shells. The solubilization of a poorly water-soluble bioactive compound, paclitaxel (PTX), in the polymer aggregates was observed, however, solubilizing efficiency and stability were strongly depended on the polymer architecture; in other words, PTX stayed in the poly(BMA) core of the polymer micelle formed by the block-type polymer even when plasma protein was present in the aqueous medium. On the other hand, when the random-type polymer was used, PTX was transferred from the polymer aggregate to the protein. We conclude that water-soluble and amphiphilic MPC polymers are good candidates as solubilizers for poorly water-soluble bioactive compounds.

The unique hydration state of poly(2-methacryloyloxyethyl phosphorylcholine).

2-Methacryloyloxyethyl phosphorylcholine (MPC) is methacrylate bearing a phosphorylcholine group in the side chain. The phosphorylcholine group generates several unique properties arising from its zwitterionic structure, consisting of a phosphate anion and a trimethylammonium cation. Despite these charged groups, the total electrical charge of the species is zero because of the formation of an inner salt. The polymerization of MPC proceeds both conventional and living radical polymerizations. And, using these method, the corresponding polymer can be obtained efficiently. The product, poly(MPC), is soluble in aqueous media, even if the ionic strength of the solution is high, such as in the presence of 5.0 mol/L NaCl. The polymer does not show any surface active properties, even when the polymer concentration is greater than 1.0 g/dL. Hydration of poly(MPC) mainly occurs by hydrophobic hydration of the three methyl groups in the trimethylammonium group. Thus, this hydration induces an increase in a clathrate cage structure of surrounding water molecules, i.e. an ice-like water state is formed. Because of this unique hydration, poly(MPC) cannot make strong interactions with proteins and cells. Some biomedical applications have used poly(MPC) as a protein and solid-surface modification agents.

Herbal formula Xian-Fang-Huo-Ming-Yin regulates differentiation of lymphocytes and production of pro-inflammatory cytokines in collagen-induced arthritis mice.

BACKGROUND: Xian-Fang-Huo-Ming-Yin (XFHM), a traditional herbal formula, has been used to treat sores and carbuncles for hundreds of years in Asia. Nowadays, its clinical effects in treatment of rheumatoid arthritis (RA) have been validated. In this study, we want to study its possible molecular mechanisms of regulating the differentiation of lymphocytes and production of pro-inflammatory cytokines in collagen-induced arthritis (CIA) mice for RA treatment. METHODS: A high performance liquid chromatography-electrospray ionization/mass spectrometer (HPLC-ESI/MSn) system was used to analyze the constituents of XFHM granules. An arthritics mouse model was induced by collagen and leflunomide (LEF) was used as a positive control medicine. Pathological changes at the metatarsophalangeal joint were studied through Safranin O and immunohistochemical staining. The differentiation of T, B and NK cells was examined by flow cytometry and pro-inflammatory cytokines were assayed using an Inflammation Antibody Array assay. The expression of key molecules of the nuclear factor κB (NF-κB) and Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathways in spleen were studied by western-blot analysis. RESULTS: In our study. 21 different dominant chemical constituents were identified in XFHM. Treatment with XFHM suppressed the pathological changes in arthrosis of CIA. Additionally, XFHM down-regulated the proliferation and differentiation of CD3+ T cells and CD3-CD19+ B cells significantly. However, XFHM had no significant effect on CD3-NK1.1+ NK cells. Further study showed that the production of pro-inflammatory cytokines had been suppressed by inhibiting the activation of NF-κB and JAK/STAT signaling. CONCLUSIONS: XFHM can regulate and maintain the immunologic balance of lymphocytic immunity and inhibit the production of pro-inflammatory cytokines, thus suppressing the pathological changes of RA. Therefore, XFHM may be used as an application of traditional medicine against RA in modern complementary and alternative therapeutics.

[Experimental study of bone morphogenetic protein-2 in noncemented fixation of prosthesis].

OBJECTIVE: To study on the method of combining rhBMP(2) with porous-coated implants, and compared the histological and biomechanical results of 4 types of implants by being inserted into the femur of canine. METHODS: 4 types of implants which are porous-coated implants (PCA), PCA implants combined with BMP, PCA implants coated with hydroxyapatite and HA coated cylindrical implants had been inserted into the femur of 16 canines. After 4, 8, 12 and 24 weeks the femur of the canines with the implants were retrieved. Bone ingrowth and shear strength of the interface was studied and analysed by the means of X-ray, soft X-ray, fluorescence tag, non-decalcification bone ground section, computer-aided image analysis procedure and biomechanical test. RESULTS: X-ray, soft X-ray, fluorescence tag, non-decalcification bone ground section and computer-aided image analysis procedure was used in histological study, and it showed that bony ingrowth into interface more than the other groups, even the maturation of newly formed bone. Non-decalcification bone ground section observation and computer aided image analysis showed the results: the new bone formation ratio of BMP group was 26.58% +/- 4.56% at 4 weeks post-implantation, which was much higher than the other groups, and there was significant difference between BMP group and each of the other group statistically (P < 0.05). Results of biomechanical study using push-out test showed that shear strength of each group appeared to rise with time. Shear strength of BMP group reached a high level at 4 weeks which was 18.94 +/- 5.11Mpa and almost twice of the other group, and there was also statistical difference between BMP group and each of the other group (P < 0.05). And the new bone formation ratio and shear strength of BMP group at 8, 12, 24 weeks post-implantation was still higher than the other group, but there was no statistically difference. CONCLUSIONS: Porous-coated implants combined with rhBMP can enhance bone ingrowth at bone-implant interface especially at the early period post-implantation. Porous-coated implants combined with rhBMP(2) can not only shorten the time of new bone formation and bone ingrowth but also enhance shear strength of bone-implant interface.