[Investigation on radiographic signs of osteoarthrosis in temporomandibular joint with cone beam computed tomography in adolescents].

OBJECTIVE: To investigate the occurrence and radiographic signs of osteoarthrosis of the temporomandibular joints (TMJOA) with cone beam computed tomography (CBCT) in adolescents. METHODS: Individuals with temporomandibular disorders (aged 10-19 years) in the patients group (n=386) and pre-orthodontic patients with malocclusion (aged 10-19 years) in the control group (n=339) were included in the present study. All the patients in both groups had been examined by CBCT. The abnormalities of the condyle were evaluated. The results of radiological findings were compared between the patients group and the controls by using chi-square tests. Inter- and intra-examiner agreements were assessed using Cohen's Kappa coefficient and all statistical analysis was performed using SPSS 13.0. RESULTS: 157 patients in the patients group and 41 subjects in the control group had radiographic signs of TMJOA. The occurrence of OA was significantly higher in the patients group (40.7%, 157/386) than in the controls (12.1%, 41/339), the difference was statistically significant (P<0.05). The occurrence of TMJOA was significantly higher in females (44.6%, 123/276) than in males (30.9%, 34/110) in the patients group (P<0.05) but showed no significant difference in the controls (females: 13.7%, 32/234, and male: 8.6%, 9/105, P>0.05). In addition, the patients group showed significantly higher occurrence of ill-defined cortical bone (31.7%, 65/205) small bony defect and extensive erosion (25.4%, 52/205) than the controls (1.7%, 1/58 and 5.2%, 3/58, respectively, P<0.05), while the subjects in the control group had significantly higher occurrence of flattening and shortening of the condyle (39.7%, 23/58) and sclerosis (39.7%, 23/58) than patients with temporomandibular disorders (6.3%,13/205 and 14.6%, 30/205, respectively, P<0.05). CONCLUSION: TMJOA is not uncommon in adolescent patients with TMD and with malocclusion. Patients in the two study groups had different radiographic signs of OA. The patients with temporomandibular disorders often demonstrate erosive changes, while the pre-orthodontic patients with malocclusion often demonstrate relatively stable radiographic signs.

Biosynthesis and characterization of 3-hydroxyalkanoate terpolyesters with adjustable properties by Aeromonas hydrophila.

Polyhydroxyalkanoates (PHA) terployesters P(3HB-co-3HV-co-3HHx) consisting of 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), and 3-hydroxyhexanoate (3HHx) were produced by wild-type Aeromonas hydrophila 4AK4, its recombinant harboring PHA synthesis genes phaPCJ encoding PHA binding protein phasin, PHA synthase, and enoyl-CoA hydratase, and another its recombinant harboring phaAB encoding beta-ketothiolase and acetoacetyl-CoA reductase, respectively, when grown in lauric acid and/or valerate. The terpolyesters produced by A. hydrophila 4AK4 (phaAB) grown in velarate were found to produce copolymers P(3HB-co-3HV) containing high 3HV fractions with a maximum of 99 mol% 3HV. In terpolyesters, 3HV ranged from 9 to 32 mol% depending on the valerate concentration and strain used. A maximal terpolyester P(3HB-co-3HV-co-3HHx) content in dry cells was 71 wt%. Transmission electron microscopy study of A. hydrophila 4AK4 harboring phaPCJ revealed the full occupation of terpolyester P(3HB-co-3HV-co-HHx) in the cellular spaces. Terpolyesters with various monomer compositions showed changing thermal and mechanical properties. Those with higher 3HV fractions demonstrated an improved property over the lower HV containing ones.

In situ extraction of intracellular L-asparaginase using thermoseparating aqueous two-phase systems.

The feasibility and generic applicability of directly integrating conventional discrete operations of cell disruption by high pressure homogenizer and the product capture by aqueous two-phase extraction (ATPE) system have been demonstrated for the extraction of intracellular L-asparaginase from E. coli. In a side-by-side comparison with the conventional ATPE process, including cell disruption, centrifugal clarification and following ATPE, purification of L-asparaginase via this novel in situ ATPE process yielded a product of L-asparaginase with a higher specific activity of 94.8 U/(mg protein) and a higher yield of 73.3%, both of which in the conventional ATPE process were 78.6 U/(mg protein) and 52.1%, respectively. In the purification of L-asparaginase (pI=4.9), product-debris interactions commonly diminish its recovery. It was demonstrated that immediate extraction of L-asparaginase in ATPE systems when it is released at pH 5.0 during cell disruption effectively increased its recovery in the top phase due to the reduced interaction between L-asparaginase and cell debris and the reduced degradation by contaminated protease. In addition, no clarification step and/or disruptate storage are required in this in situ ATPE, which reduced the number of unit operations and thus shortened the overall process time. This novel process has a good potential for the separation of other intracellular biological products.

Ankylosis of temporomandibular joint caused by psoriatic arthritis: a report of four cases with literature review.

Psoriatic arthritis (PsA) is an inflammatory joint disease associated with psoriasis. PsA is often confused with other diseases such as osteoarthritis and rheumatoid arthritis. PsA involving temporomandibular joints (TMJs) are uncommon: only 19 articles with 43 cases have been documented in the literature. TMJ ankylosis caused by PsA is rare, with only six cases having been reported. The authors present four cases of ankylosis of the TMJ secondary to PsA and review the literature. The findings of this study suggest that more attention should be paid to psoriasis patients with TMJ symptoms and proper treatment should be taken to prevent irreversible TMJ damage.

[Effects of three at-home bleaching agents on enamel structure and structure-related mechanical properties].

OBJECTIVE: To investigate the effects of three differently concentrated at-home bleaching agents on the structure and the structure-related mechanical properties of human enamel. METHODS: Sixty enamel specimens were randomly divided into four groups and treated with 10% carbamide peroxide (CP), 15% CP, 20% CP and distilled water, respectively. The bleaching process was 8 h/day for 14 consecutive days. Baseline and final atomic force microscopy (AFM) surface detection, Raman spectroscopy, attenuated total reflectance-infrared spectroscopy (ATR-IR), microhardness and fracture toughness (FT) measurements were carried out before and after bleaching experiments. RESULTS: CP didn't change the morphology of enamel. Meanwhile, the three bleached groups and the control group had no significant difference in root mean square detection (P = 0.774), ν(2)CO(3)(2-) : ν(1)ν(3)PO(4)(3-) (P = 0.263) and microhardness (P = 0.829). The percentage of relative Raman intensity in the three bleached groups and the control group were (105.74 ± 11.34)%, (104.46 ± 8.83)%, (99.52 ± 9.32)% and (97.62 ± 7.46)%, respectively. There was no significant difference among them (P = 0.062). However, the percentage of laser-induced fluorescence in the three bleached groups and the control group were (20.86 ± 7.23)%, (22.14 ± 7.34)%, (21.10 ± 7.59)% and (100.78 ± 3.70)%, respectively. There was significant difference between either of the bleached groups and the control group (P < 0.001). Moreover, FT declined significantly in the three groups (P = 0.024, P = 0.005, P = 0.013) when compared with the control group. CONCLUSIONS: Under in vitro condition, three differently concentrated at-home bleaching agents wouldn't induce the demineralization and the decline of microhardness on enamel. However, the decrease of FT on enamel seemed to be inevitable after bleaching.

P85, Optison microbubbles and ultrasound cooperate in mediating plasmid DNA transfection in mouse skeletal muscles in vivo.

Pluronic block copolymers, a kind of non-ionic surfactant, also known as poloxamers, and ultrasound-targeted microbubble destruction have been respectively investigated as vectors for gene delivery in vitro and in vivo. However, they are limited for clinical application due to the relatively low transfer efficiency of each individual vector. In the present study, we explored if the combination of P85, a pluronic block copolymer, Optison, a microbubble contrast agent and ultrasound enhances the transfection of plasmid DNA in vivo using mouse skeletal muscle models. Plasmid encoding green fluorescent protein (GFP) was respectively conjugated with 0.05%P85, 10%Optison, or 0.05%P85 plus 10%Optison, and injected into mouse tibialis anterior (TA) muscles with or without ultrasound irradiation (1 MHz, 1 W/cm(2), 2 min and 20% duty cycle). Mice were sacrificed 1 week after injection. The TA muscles were collected and cryo-sectioned into a series of 7 µm slices. To assess the efficiency of plasmid DNA transfection, tissue sections were counterstained with DAPI and scored by counting the number of GFP-positive fibers. Meanwhile the area of damaged muscles was measured based on the tissues stained with hematoxylin and eosin. Both P85 and Optison significantly enhanced the delivery of plasmid DNA in mouse TA skeletal muscles (P<0.01 and P<0.05 respectively, compared to saline control). In combination with Ultrasound irradiation, P85 (P<0.01, compared to P85 alone) but not Optison (P>0.05, compared to Optison alone) exerted a more pronounced effect on the transfection efficiency. Furthermore P85-induced gene delivery was higher than that by Optison regardless of the presence of ultrasound (P<0.01). The highest transfection efficiency was observed when P85, Optison and ultrasound irradiation were administrated together (P<0.01, compared to any other treatment in this study). The area of damaged muscles was enlarged by ultrasound irradiation in the presence of Optison microbubbles (P<0.01, compared to those groups without ultrasound irradiation). These results suggest that P85, microbubbles and ultrasound irradiation synergistically enhance plasmid DNA delivery in mouse skeletal muscles in vivo.

Enhanced gene transduction into skeletal muscle of mice in vivo with pluronic block copolymers and ultrasound exposure.

The pluronic block copolymers are able to enhance the ultrasound-induced gene delivery in vitro. In the present study, the effects of pluronics on the efficiency of gene transfer into skeletal muscle in vivo under sonoporation were investigated. Plasmid DNA encoding green fluorescent protein (GFP) in combination with three different pluronics, F127, L61, and P85, was injected into the tibialis anterior (TA) muscle of mice with and without adjunct ultrasound (1 MHz, 3 W/cm(2) 1 min, 20% duty cycle). Mice were killed 1 week after injection. The TA muscles were removed and snap frozen immediately in isopentane cooled by liquid nitrogen and sections of 7 µm thick were cut. Transfection efficiency was assessed by counting the number of GFP-positive fibers under fluorescence microscopy, and tissue damage by hematoxylin and eosin staining. The results suggested that all three pluronics significantly enhanced transgene expression in skeletal muscle (P < 0.01), especially the P85 showed significantly higher efficiency than the other two pluronics (P < 0.05). Ultrasound synergistically enhanced the gene delivery efficiency with P85 (P < 0.01), but was unable to do so with F127 and L61 groups. In short, P85 displays significantly synergistic effect with ultrasound for enhancing plasmid DNA transduction in skeletal muscle of mice in vivo.

Differentiation of human bone marrow mesenchymal stem cells grown in terpolyesters of 3-hydroxyalkanoates scaffolds into nerve cells.

Polyhydroxyalkanoates, abbreviated as PHA, have been studied for medical applications due to their suitable mechanical properties, blood and tissue tolerance and in vivo biodegradability. As a new member of PHA family, terpolyester of 3-hydroxybutyrate, 3-hydroxyvalerate and 3-hydroxyhexanoate, abbreviated as PHBVHHx, was compared with polylactic acid (PLA), copolyester of 3-hydroxybutyrate and 3-hydroxyhexanoate (PHBHHx) for their respective functions leading to differentiation of human bone marrow mesenchymal stem cell (hBMSC) into nerve cells. Results indicated that 3D scaffolds promoted the differentiation of hBMSC into nerve cells more intensively compared with 2D films. Smaller pore sizes of scaffolds increased differentiation of hBMSC into nerve cells, whereas decreased cell proliferation. PHBVHHx scaffolds with pore sizes of 30-60 microm could be used in nerve tissue engineering for treatment of nerve injury. The above results were supported by scanning electron microscope (SEM) and confocal microscopy observation on attachment and growth of hBMSCs on PLA, PHBHHx and PHBVHHx, and by CCK-8 evaluation of cell proliferation. In addition, expressions of nerve markers nestin, GFAP and beta-III tubulin of nerve cells differentiated from hBMSC grown in PHBVHHx scaffolds were confirmed by real-time PCR.

The improvement of fibroblast growth on hydrophobic biopolyesters by coating with polyhydroxyalkanoate granule binding protein PhaP fused with cell adhesion motif RGD.

Polyhydroxyalkanoates (PHA), a family of biopolyesters, have been studied as tissue engineering biomaterials due to their adjustable mechanical properties, biodegradability and tissue compatibility. Amphiphilic PHA granule binding protein PhaP has been shown to be able to bind to hydrophobic surfaces of polymers, especially PHA, via strong hydrophobic interaction. Genes of PhaP and RGD peptides, which are a cell adhesion motif recognized by many cell surface receptors, were successfully expressed and obtained as a pure fusion protein PhaP-RGD in Escherichia coli DH5α. When films of poly(3-hydroxybutyrate-co-3-hydroxy- hexanoate) (PHBHHx), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and polylactic acid (PLA) were coated with PhaP-RGD, their surface hydrophilicities were all increased compared with their corresponding naked (non-coated) films, respectively. Among the three biopolyesters, PHBHHx demonstrated the strongest affinity to PhaP. In vitro study showed that mouse fibroblasts L929 and mouse embryonic fibroblasts NIH/3T3 attached better and grew faster on all three PhaP-RGD coated films compared with their related behaviors on PhaP coated and non-coated films, respectively. Both fibroblasts attached and grew very well on PhaP-RGD coated PHBHHx, PHBV and PLA, even in their serum-free medium, while the non-coated and PhaP coated biopolyesters poorly supported the cell growth if the two fibroblasts were incubated in their serum free medium. These results indicated that PhaP-RGD could be used as a coating material to improve cell growth on hydrophobic biopolyesters for implant tissue engineering purposes.