TDI-based continuous window compressed spatio-temporal imaging capable of flexible voxels post-interpretation.

To achieve high frame rates and continuous streaming simultaneously, we propose a compressed spatio-temporal imaging framework implemented by combining time-delay-integration sensors and coded exposure. Without additional optical coding elements and subsequent calibration required, this electronic-domain modulation enables a more compact and robust hardware structure, compared to the existing imaging modalities. By exploiting the intra-line charge transfer mechanism, we achieve a super-resolution in both temporal and spatial domains, thus multiplying the frame rate to millions of frames-per-second. In addition, the forward model with post-tunable coefficients, and two reconstruction strategies proposed therefrom, facilitate a flexible voxels post-interpretation. Finally, the effectiveness of the proposed framework is demonstrated by both numerical simulations and proof-of-concept experiments. With the prominent advantages of prolonged time window and flexible voxels post-interpretation, the proposed system will be suitable for imaging random, non-repetitive, or long-term events.

Regulation of IL-17 by lncRNA of IRF-2 in the pearl oyster.

Long noncoding RNAs (lncRNAs), once thought to be nonfunctional, have recently been shown to participate in the multilevel regulation of transcriptional, posttranscriptional and epigenetic modifications and to play important roles in various biological processes, including immune responses. However, the expression and roles of lncRNAs in invertebrates, especially nonmodel organisms, remain poorly understood. In this study, by comparing a transcriptome to the PfIRF-2 genomic structure, we identified lncIRF-2 in the PfIRF-2 genomic intron. The results of the RNA interference (RNAi) and the nucleus grafting experiments indicated that PfIRF-2 might have a negative regulatory effect on lncIRF-2, and PfIRF-2 and lncIRF-2 may have a positive regulatory effect on PfIL-17. Additionally, lncIRF-2, PfIRF-2 and PfIL-17 were involved in responses to the nucleus graft. These results will enhance the knowledge of lncIRF-2, IRF-2, and IL-17 functions in both pearl oysters and other invertebrates.

Analysis and reduction of the TDI CCD charge transfer image shift.

Based on the description of the charge transfer process of a time delay integration charge coupled device (TDI CCD), it is pointed out that the relative displacement of the target image and the transferred charge introduces the charge transfer image shift problem in a line transfer period, which leads to the decrease of the modulation transfer function (MTF) in the scanning direction. Based on the basic definition of MTF, the charge transfer image shift model of the TDI CCD is established. According to the quantitative calculation of this model, the MTF curves of the three-phase TDI CCD are obtained under the condition of different pulse widths of computer interface (CI) signals. The MTF changes with the pulse width of CI signals, and the maximum value at the spatial Nyquist frequency is obtained when the CI signals are equally spaced, which is difficult to achieve in the actual system because of the limitation of computed radiography readout signals. In this paper, we present the improved TDI CCD driving timing, which makes it possible to realize the equal interval distribution of CI signals by adjusting the technology compatibility kit signal. Finally, the experimental platform is built, and the MTF curves are calculated from the obtained target images. The result is consistent with the theoretical model.

Gene cloning and function analysis of cytokine-induced suppressor of cytokine signaling (SOCS) from pearl oyster Pinctada fucata.

Cytokine-induced suppressor of cytokine signaling (SOCS) family acts as a negative regulator of cytokine receptor signaling to control excessive cytokine effects and inhibit a variety of signal transduction pathways, particularly the Janus kinases/signal transducers and activators of transcription (JAK/STAT) pathway. In present study, SOCS-2 homolog (PfSOCS-2) from pearl oyster Pinctada fucata was cloned and its gene has no intron. Multiple sequence alignments and phylogenetic analysis showed that PfSOCS-2 was clustered with other mollusk SOCS-2. LPS or polyI:C challenge and gene expression analysis revealed that PfSOCS-2 involved the innate immune response against bacterial and viral infections and that induction of PfSOCS-2 was varied with the different challenge stimulations. Furthermore, Dual-luciferase reporter assays showed that PfSOCS-2 involved in the regulation of vertebrate target genes containing the IFN-stimulated response element or NF-κB binding site in vitro. These results indicated that SOCS-2 from P. fucata plays a regulatory role against the stimulation.

Molecular characterization of interferon regulatory factor 2 (IRF-2) homolog in pearl oyster Pinctada fucata.

Interferon regulatory factors (IRFs) control many facets of the innate and adaptive immune responses, regulate the development of the immune system itself and involve in reproduction and morphogenesis. In the present study, the IRF-2 homology gene, PfIRF-2 from pearl oyster Pinctada fucata was cloned and its genomic structure and promoter were analyzed. PfIRF-2 encodes a putative protein of 350 amino acids, and contains a highly conserved N-terminal DNA-binding domain and a variable C-terminal regulatory domain. Comparison and phylogenetic analysis revealed that PfIRF-2 shared a relatively higher identity with other mollusk but relatively lower identity with vertebrate IRF-2, and was clustered with IRF-1 subfamily composed of IRF-2 and IRF-1. Furthermore, gene expression analysis revealed that PfIRF-2 involved in the immune response to LPS and poly(I:C) stimulation. Immunofluorescence assay showed that the expressed PfIRF-2 was translocated into the nucleus and dual-luciferase reporter assays indicated that PfIRF-2 could involved and activate interferon signaling or NF-κB signal pathway in HEK293 cells. The study of PfIRF-2 may help better understand the innate immune in mollusk.

[Biomechanical characteristics of hip prosthesis in hip arthroplasty treating elderly patients with Evans I-III intertrochanteric fracture of femur].

OBJECTIVE: To investigate the feasibility of hip arthroplasty in the treatment of elderly patients with Evans I-III intertrochanteric fracture of femur by analyzing its biomechanics characters. METHODS: We solved the CT digital image files with the graphics processing software Mimics at DICOM 3.0 standard, and reconstructed the three-dimensional entity of femur with CAD modeling software Unigraphics. Then the fracture line was defined in the model as the line between the tip of greater trochanter and inferior margin of small trochanter, above which the upper bone was removed. Afterwards the two prosthesises with different stem lengths (120 mm and 170 mm) were implanted into the fracture model respectively as hip arthroplasty with 3 mm bone cement layer between prosthesis and femur, and the bone defect was repatched with 5 mm bone cement layer. A three-dimensional finite element model was established with finite element analysis software ABAQUS 6.5. We formulated different material parameters under the stress condition standing with single leg to build the stress distribution map of the femur prosthesis, and took 5 loci of region of stress concentration to calculate the mean value of stress. RESULTS: The stress distribution maps of the short and long stem length prothesises were similar. And there were two areas of stress concentration, including the upper portion and the lower portion close to the joint of the prosthesis stem, and the stress concentration in the junction part was obviously between the lower portion and the upper area of the small trachanter. The stress reached the first concentration area at the junction and then gradually reached the second concentration area at the interior terminal of the stem. While the stress gradually increased along the lateral prosthesis stem, and reached the stress concentration area at the end. CONCLUSIONS: The stress distribution maps in the femur prosthesises are similar between hip arthroplasty in the treatment of intertrochanteric fracture of femur and the traditional hip arthroplasty surgery. The peak stress values are higher in the long stem prosthesis in the treatment of intertrochanteric fracture of femur than the short type, while they are under the rupture value of the metal.

Molecular cloning, characterization and expression analysis of tumor necrosis factor receptor-associated factor 3 (TRAF3) from pearl oyster Pinctada fucata.

TRAF3 is a highly versatile regulator that negatively regulates JNK and alternative nuclear factor-κB signalling, but positively controls type I interferon production. To investigate TRAF3 function in innate immune responses among invertebrate especially mollusk, we characterized TRAF3 (PfTRAF3) from pearl oyster Pinctada fucata, one of the most important bivalve mollusks for seawater pearl production. PfTRAF3 cDNA is 2261 bp with an open reading frame of 1623 bp encoding a putative protein of 541 amino acids. The deduced PfTRAF3 contains a RING finger domain, two TRAF domains with zinc finger domains and a conserved C-terminal meprin and TRAF homology (MATH) domain. Comparison and phylogenetic analysis revealed that PfTRAF3 from mollusk shared a higher identity with Ciona intestinalis TRAF3 from urochordata, Branchiostoma belcheri TRAF3 from cephalochordate, and even TRAF3 from vertebrate than with insect homologues. Furthermore, gene expression analyses suggested that PfTRAF3 was involved in the immune response to Vibrio alginolyticus.

Molecular cloning and characterization of class I NF-κB transcription factor from pearl oyster (Pinctada fucata).

NF-κB transcription factors play central roles in many important physiological and pathological processes including innate immune responses. Here we report the cloning of an NF-κB transcription factor, PfRelish from pearl oyster Pinctada fucata, one of the most important bivalve mollusks for seawater pearl production. PfRelish full-length cDNA is 3916 bp with an open reading frame of 3558 bp encoding a putative protein of 1186 amino acids. The deduced PfRelish contains a N-terminal RHD, a nucleus localization signal, an IκB-like domain with six ankyrin repeats and a death domain at the C-terminus, which is similar to class I NF-κB transcription factors. Comparison and phylogenetic analysis revealed that class I NF-κBs in mollusks including PfRelish might have most distant relationship to the arthropod Relish. Further expression analysis showed that PfRelish was apparently upregulated after Vibrio alginolyticus injection, which suggested that PfRelish was involved in the immune response to V. alginolyticus.

Theoretical study on the reaction mechanisms of CH3O- with O2(X3Σ(g)-) and O2(a1Δ(g)).

The detailed potential energy surfaces (PESs) of poorly understood ion-molecule reactions of CH(3)O(-) with O(2)(X(3)Σ(g)(-)) and O(2)(a(1)Δ(g)) are accounted for by the density functional theory and ab initio of QCISD and CCSD(T) (single-point) theoretical levels with 6-311++G(d,p) and 6-311++G(3df,2pd) basis sets for the first time. For the reaction of CH(3)O(-) with O(2)(X(3)Σ(g)(-)) ((3)R), it is shown that a hydrogen-bonded complex (3)1 is initially formed on the triplet PES, which is 1.8 kcal/mol above reactants (3)R at the CCSD(T)//QCISD level, from which all the products P(1)-P(8) can be generated. As to the reaction of CH(3)O(-) with O(2)(a(1)Δ(g)) ((1)R), it is found that the two energetically low-lying complexes of (1)1(-31.5 kcal/mol) and (1)2(-24.1 kcal/mol) are initiated on the singlet PES. Starting from them, a total of seven products may be possible, that is, besides P(1), P(2), P(3), P(4), and P(8), which are the same as on the triplet PES, there exist also another two products, P(9) and P(10). For both reactions, taking the thermodynamics and kinetics into consideration, the hydride-transfer species P(1)(CH(2)O + HO(2)(-)) should be the most favorable product followed by P(8)(e + CH(2)O + HO(2)), which is a secondary product of electron-detachment from P(1), and the generation of endothermic P(7)(17.7 kcal/mol) for the reaction of CH(3)O(-) with O(2)(X(3)Σ(g)(-)) is also possible at high temperature, whereas the remaining products are negligible. The measured branching ratio of products for CH(3)O(-) with O(2)(X(3)Σ(g)(-)) by Midey et al. is 0.85:0.15 for P(1) and P(8), and that of CH(3)O(-) with O(2)(a(1)Δ(g)) is 0.52:0.48 with more P(8), which can be rationalized by our theoretical results that P(8) on the triplet PES is 4.9 kcal/mol above (3)R, whereas both P(1) and P(8) on the singlet PES are very low-lying at 45.6 and 25.2 kcal/mol below (1)R energetically. The measured total reaction rate constant of CH(3)O(-) with O(2)(a(1)Δ(g)) is k = 6.9 × 10(-10) cm(3) s(-1) at 300 K, which is larger than that of k = 1.1 × 10(-12) cm(3) s(-1) for the reaction of CH(3)O(-) with O(2)(X(3)Σ(g)(-)). This is understandable because both P(1) and P(8) on the singlet PES can be generated barrierlessly, whereas to give all the products on the triplet PES has to pass the barrier of (3)1(1.8 kcal/mol) at the CCSD(T)//QCISD level. It is expected that the present theoretical study may be helpful for understanding the reaction mechanisms related to CH(3)O(-) and even CH(3)S(-).

Arthroscopic Suture Fixation of Os Acetabuli With Absorbable Suture Anchors-A Double-Pulley Technique.

Os acetabuli is a bone fragment with unknown origin and isolated at the acetabular rim that may be associated with cam-type femoroacetabular impingement. If this bone fragment is too large and threatens the stability of the hip joint after resection, fixation would be recommended. However, conventional rigid fixation with metal screws has some disadvantages. We describe an arthroscopic suture fixation of the Os acetabulum with absorbable anchors penetrating the bone fragment and secured by tying knots in a double-pulley fashion simultaneously. This technique provides a new feasible solution for the fixation of Os acetabuli, avoiding any metal implants and potential damage to the joint.

Theoretical study on the reaction mechanism of NH2(-) with O2 (a1Δg).

A detailed theoretical study of the potential energy surface of poorly understood ion-molecule reaction of NH(2)(-) and O(2) (a(1)Δ(g)) is explored at the density functional theory B3LYP/6-311++G(d,p), ab initio of QCISD/6-311++G(d,p) and CCSD(T)/6-311++G(3df, 2pd) (single-point) theoretical levels for the first time. It is shown that there are six total possible products from P(1) to P(6) on the singlet potential energy surface. Among these, the charge-transfer product P(1) (NH(2) + O(2)(-)) is the most favorable product with predominant abundances, whereas P(4) (NO(-) + H(2)O) and P(2) (HNO + OH(-)) may be the second and third feasible products followed by the almost neglectable P(3) (NO(2)(-) + H(2)), while P(5) (c-NO(2)(-) + H(2)) and P(6) (ONO(-) + H(2)) will not be observed due to their either high barriers or being secondary products. The present theoretical study points out that besides P(1) (NH(2) + O(2)(-)) and P(2) (HNO + OH(-)), P(4) (NO(-) + H(2)O) should be also observed, which is different from the previous experiment study by Anthony Midey et al. in 2008. In addition, almost all of the reaction pathways to products are exothermic and the reaction rate should be very fast since the reaction barriers are very low except for P(5) (c-NO(2)(-) + H(2)) which is in agreement with the measured total reaction rate constant k = 9.0 × 10(-10) cm(3)s(-1) at 300 K in the experiment study. It is expected that the present theoretical study may be helpful for the understanding of the reaction mechanism related to NHX(-), NX(2)(-), PHX(-), and PX(2)(-) (X = H, F, and Cl).

RgBF2(+) complexes (Rg = Ar, Kr, and Xe): the cations with large stabilities.

Rare gas containing cations with general formula [Rg, B, 2F](+) have been investigated theoretically by second-order Mo̸ller-Plesset perturbation, coupled cluster, and complete active space self-consistent field levels of theory with correlation-consistent basis sets. Totally two types of minima, i.e., boron centered C(2) (v) symmetried RgBF(2) (+) (Rg = Ar, Kr, and Xe) which can be viewed as loss of F(-) from FRgBF(2) and linear FRgBF(+) (Rg = Kr and Xe) are obtained at the CCSD(T)∕aug-cc-pVTZ∕SDD and CASSCF(10,8)∕aug-cc-pVTZ∕SDD levels, respectively. It is shown that the RgBF(2) (+) are global minima followed by FRgBF(+) at 170.9 and 142.2 kcal∕mol on the singlet potential-energy surfaces of [Rg, B, 2F](+) (Rg = Kr and Xe) at the CASPT2(10,8) ∕aug-cc-pVTZ∕SDD∕∕CASSCF(10,8)∕aug-cc-pVTZ∕SDD, respectively. The interconversion barrier heights between RgBF(2) (+) and FRgBF(+) (Rg = Kr and Xe) are at least 39 kcal∕mol. In addition, no dissociation transition state associated with RgBF(2) (+) and FRgBF(+) can be found. This suggests that RgBF(2) (+) (Rg = Ar, Kr, and Xe) can exist as both thermodynamically and kinetically stable species, while linear FRgBF(+) (Rg = Kr and Xe) can exist as metastable species compared with the lowest dissociation limit energies just like isoelectronic linear FRgBO and FRgBN(-). From natural bond orbital and atoms-in-molecules calculations, it is found that the positive charge is mainly located on Rg and boron atoms for both types of minima, the Rg-B bonds of ArBF(2) (+), KrBF(2) (+), and XeBF(2) (+) are mostly electrostatic, thus can be viewed as ion-induced dipole interaction; while that of linear FKrBF(+) and FXeBF(+) are covalent in nature. The previous experimental observation of ArBF(2) (+) by Pepi et al. [J. Phys. Chem. B. 110, 4492 (2006)] should correspond to C(2) (v) minimum. The presently predicted spectroscopies of KrBF(2) (+), XeBF(2) (+), FKrBF(+), and FXeBF(+) should be helpful for their experimental identification in the future.

[Transplantation of porcine type I atelocollagen into rabbit cornea].

OBJECTIVE: To investigate corneal wound healing in rabbit after lamellar keratoplasty with porcine type I atelocollagen. METHODS: One hundred Chinese white rabbits, 80 rabbits were randomly divided into collagen materials group (group A) and allograft group (group B), and group A and B were operated for the lamellar keratoplasty in one eye. Rest 20 rabbits' eyes (40 eyes) were used as the source of allograft for group B. After operation, the rabbits' eyes were observed by naked eyes and slit lamp, and recorded the score of the transparency and neovascularization of the cornea. The rabbits were sacrificed at 3rd, 14th day, the 1st, 3rd, and 6th month (each group eight eyes). The corneas were observed by histopathology, and the epithelial cells marker protein K3 were detected by immunohistochemistry. The results were analyzed by Rank sum test. RESULTS: The transparency of the cornea was increased gradually in A and B groups, and reached the peak on the 6th month (group A: χ(2) = 31.250, P = 0.000; group B: χ(2) = 32.566, P = 0.000). The difference had not statistics significance in the comparison of the transparency from the 1st to 6th month (Z = -1.414, 0.000, -0.743;P = 0.157, 1.000, 0.458). The two groups had corneal neovascularization after 7 days, the intensity of corneal neovascularization in A group increased gradually, reached the peak on day 14, then decreased gradually (χ(2) = 20.727, P = 0.001); the intensity of corneal neovascularization in B group increased gradually, reached the peak on day 14, and then decreased gradually (χ(2) = 25.562, P = 0.000). The difference had statistics significance in the comparison of the neovascularization of the cornea on the 6th month between A and B group (Z = -2.070, P = 0.038). Histopathology showed the collagen material nearly disappeared after 1 month of surgery and arrangement of collagen tends to be regular. On the 6th month, arrayed collagen was regular in general, but disordered partly. CONCLUSIONS: Porcine type I atelocollagen can promote corneal cells regeneration. But it needs more study to be a cornea substitute that can completely replace the corneal allograft.

Minimum 7-year Follow-up of A Porous Coated Trabecular Titanium Cup Manufactured with Electron Beam Melting Technique in Primary Total Hip Arthroplasty.

OBJECTIVES: To investigate the cup survivorship, patient satisfaction level, clinical function, and radiographic outcomes of patients who underwent total hip arthroplasty (THA) using electron beam melting (EBM)-produced porous coated titanium cups at mid-term follow up. METHODS: A total of 32 patients (32 hips) from five hospitals in China who underwent primary THA using EBM-produced trabecular titanium cups between May and December 2012 were retrospectively reviewed. The inclusion criteria were: (i) patients who underwent THA with the use of EBM-produced cups with possible 7-year follow up; and (ii) patients with follow-up information, including the cup survivorship, patient satisfaction level, and clinical outcomes such as Harris hip score. The exclusion criteria were: (i) patients with neuropathic diseases; and (ii) patients who underwent THA due to neoplastic disease. Five (15.6%) patients were lost to follow up before the 7-year follow-up and, thus, were excluded; none of these patients died due to disease associated with the THA or had undergone removal of their cups as of our last evaluation. The mean age and body mass index of the patients were 59.37 (range: 38.00-69.00) years and 24.51 (range: 16.50-34.10) kg/m2 , respectively. Thirteen (48.1%) of the patients were female. RESULTS: The average duration of follow-up was 93.48 (range: 89.00-99.00) months. The median Harris hip score improved from 42.00 (interquartile range: 37.00-49.00) to 97.00 (interquartile range: 92.00-97.00) at the latest follow up (P < 0.001). A total of 18 (66.7%) patients rated their satisfaction level as very satisfied, 6 (22.2%) as satisfied, 2 (7.4%) as neutral and 1 (3.7%) as dissatisfied. No intraoperative or postoperative complications were identified. At the latest follow up, all cups were considered to have achieved osteointegration fixation, with three or more of the five signs evident in the most recent X-ray. However, three cups revealed radiolucent lines with a width of less than 1 mm. The median vertical and horizontal distances between the latest postoperative center of rotation relative to the anatomic center of rotation were 2.50 (interquartile range: -3.10, 6.94) mm superiorly and 3.26 (interquartile range: -8.12, 2.38) mm medially, respectively, at the most recent postoperative follow up. Kaplan-Meier survivorship analysis of cups, with the endpoint defined as postoperative radiolucent lines of less than 1 mm in width in at least two zones, reveals that the 8.25-year survival was 96.3% (95% confidence interval: 76.49%-99.47%). CONCLUSION: The mid-term follow-up of patients who underwent primary THA using EBM-produced porous coated titanium cups demonstrated favorable patient satisfaction, good clinical function, excellent survivorship, and adequate biological fixation.

Finite element analysis of cemented prostheses for hip replacement in elderly patients with comminuted intertrochanteric fracture.

BACKGROUND: To determine postoperative stress distribution after cemented arthroplasty in elderly patients with comminuted intertrochanteric fracture, and assist in determining a rational prosthetic stem length. METHODS: A three-dimensional (3D) model of intertrochanteric fracture was established using the Mimics and Unigraphics modeling software, which included the 3D model of comminuted intertrochanteric fracture, two long-stem(#4,#5) and one short stem(#3) prostheses, and the mantle layer of cement. The bone defect of the calcar femorale was replaced with a 5-mm thick cement. Then, the 3D finite element model of those materials was established, boundary conditions of force were imposed, and material parameters were set. Accordingly, a finite element analysis was performed to this model in stress. RESULTS: (1) The stress of the femur in the three-stem replacement prosthesis models increased from proximal end to distal end in the same pattern, while a stress concentration region was found at 5 mm from within the distal tip of the short-stem prosthesis (#3), which had a peak value of 67.85 MPa. However, no stress concentration was found on the long-stem prosthesis model. (2) For the short-stem prosthesis, the stress distribution of the cement-prosthesis interface was significantly concentrated in the distal region around the prosthesis end, in which the peak value of the lateral interface exceeded the fatigue strength of the bone cement. However, the biomechanics for the long prosthesis was better appreciated. CONCLUSION: Long prosthesis stems may theoretically be a better option for comminuted intertrochanteric fractures in elderly patients. However, the application of exceedingly long prosthesis stems would not be a better option. KEY WORDS: Comminuted intertrochanteric fracture, Elderly patients Finite element analysis, Prosthetic replacement.

Intraoperative perfusion-weighted imaging in non-enhanced glioma surgery.

BACKGROUND: The objective of the present study was to observe and analyze the significance of perfusion-weighted imaging for guiding the operation implementation for non-enhanced glioma, and analyze the estimation of the histopathological grade of the non-enhanced glioma and the accuracy of the degree of malignancy degree before surgery. METHODS: Fifty-six patients diagnosed with non-enhanced glioma through conventional magnetic resonance scanning were selected. Before surgery, conventional magnetic resonance scanning and perfusion-weighted imaging were performed on all patients. The property classification was performed with the perfusion-weighted imaging parameters: cerebral blood volume (CBV) and cerebral blood flow (CBF) before surgery. RESULTS: Surgery was performed on the 56 patients. Tumors were excised and processed for histopathological classification and semi-quantitative immunohistochemical analysis of vascular endothelial growth factor (VEGF) levels. Histology was compared after surgery and the classification accuracy rate was analyzed before surgery. Additionally, conventional magnetic resonance scanning and perfusion-weighted imaging were performed on 15 patients during surgery. We compared and analyzed the reference value of perfusion-weighted imaging before and during surgery. Residual diseased tissues were excised; histopathological examination was performed, and semi-quantitative immunohistochemical analysis of VEGF was performed. Regarding maximal magnetic resonance perfusion-weighted imaging measured before surgery, the CBV, CBF, and expression level of VEGF were positively correlated with the pathological grade of tumors. If the CBV and CBF values of the non-enhanced glioma were higher, the grade of malignancy was higher (P<0.01), and the positive expression rate of VEGF was higher (P<0.01). CONCLUSIONS: Magnetic resonance perfusion-weighted imaging can display vessel growth and distribution within non-enhanced gliomas before surgery, and effectively evaluate the histopathological grade and grade of malignancy, and provide accurate guidance for tumor resection during surgery.

Effect of Cross-Linked Hyaluronate Scaffold on Cartilage Repair: An In Vivo Study.

OBJECTIVE: To determine the safety and effectiveness of a cross-linked sodium hyaluronate (CHA) scaffold in cartilage repair. METHODS: Physicochemical properties of the scaffold were determined. The safety and effectiveness of the scaffold for cartilage repair were evaluated in a minipig model of a full-thickness cartilage defect with microfracture surgery. Postoperative observation and hematological examination were used to evaluate the safety of the CHA scaffold implantation. Pathological examination as well as biomechanical testing, including Young's modulus, stress relaxation time, and creep time, were conducted at 6 and 12 months postsurgery to assess the effectiveness of the scaffold for cartilage repair. Furthermore, type II collagen and glycosaminoglycan content were determined to confirm the influence of the scaffold in the damaged cartilage tissue. RESULTS: The results showed that the routine hematological indexes of the experimental animals were within the normal physiological ranges, which confirmed the safety of CHA scaffold implantation. Based on macroscopic observation, it was evident that repair of the defective cartilage in the animal knee joint began during the 6 months postoperation and was gradually enhanced from the central to the surrounding region. The repair smoothness and color of the 12-month cartilage samples from the operation area were better than those of the 6-month samples, and the results for the CHA scaffold implantation group were better than the control group. Greater cell degeneration and degeneration of the adjacent cartilage was found in the implantation group compared with the control group at both 6 and 12 months postoperation, evaluated by O'Driscoll Articular Cartilage Histology Scoring. Implantation with the CHA scaffold matrix promoted cartilage repair and improved its compression capacity. The type II collagen level in the CHA scaffold implantation group tended to be higher than that in the control group at 6 months (2.33 ± 1.50 vs 1.68 ± 0.56) and 12 months postsurgery (3.37 ± 1.70 vs 2.06 ± 0.63). The GAG content in the cartilage of the control group was significantly lower than that of the experimental group (2.17 ± 0.43 vs 3.64 ± 1.17, P = 0.002 at 6 months and 2.27 ± 0.38 vs 4.12 ± 1.02, P = 0.002 at 12 months). Type II collagen and glycosaminoglycan content also demonstrated that CHA was beneficial for the accumulation of both these vital substances in the cartilage tissue. CONCLUSIONS: The CHA scaffold displayed the ability to promote cartilage repair when applied in microfracture surgery, which makes it a promising material for application in the area of cartilage tissue engineering.

Pre-deliver chitosanase to cells: a novel strategy to improve gene expression by endocellular degradation-induced vector unpacking.

A radio-labeled plasmid pTracer/Bsd/LacZ containing LacZ reporter gene was complexed with different molecular weights of chitosans (CS). Mouse myoblast cell line C2C12 was transfected by these chitosan-plasmid DNA complexes, and lipofectamine 2000 was used as control. Forty-eight hours after transfection, the activity of beta-galactosidase and radioactive count of cell lysis were determined. It was found that chitosan, especially low molecular weight species, had a surprising ability to deliver DNA into cells, since the radioactive count of cells transfected by chitosan-DNA complexes was even two times that of cells transfected by lipofectamine 2000. But the beta-galactosidase activity of chitosan/DNA complexes was much lower compared to that of lipofectamine 2000. Chitosanase which could degrade chitosan in specific mode was transported into C2C12 cells by osmotic lysis prior to gene delivery. Then these chitosanase-modified cells were transfected by CS-DNA complexes. The results indicated that beta-galactosidase activity in these cells increased markedly to 425.4 +/- 45.1 U/mg protein, nearly two-fold as that of cells transfected by liposome. This transfection protocol was also applied to 3T3 mouse fibroblast, 2T3 mouse osteoblast and MG63 human osteosarcoma cell lines, and an increased gene expression level was observed without exception. It is thought that the incorporated chitosanase could aid in chitosan degradation, which would promote gene unpacking, consequently increasing gene expression.

Preparation of oxidized glucose-crosslinked N-alkylated chitosan membrane and in vitro studies of pH-sensitive drug delivery behaviour.

Glucose was oxidized to generate a glucose dialdehyde and chitosan (CS) was hydrophobicallly modified with butyl bromide, octyl bromide and dexyl bromide. The analysis of IR and X-ray diffraction results of CS derivatives confirms that the hydrogen bonds and crystallinity were weakened by the incorporation of pendant alkyl. The permeability coefficient P and diffusion coefficient D for model drug vitamin B2 through oxidized glucose-crosslinked alkylated CS membrane were determined under different pH media. The results show that for the same alkylated CS in different pH media, P and D decrease with an increase in pH; for different alkxylated CSs in acidic media, P and D diminish with the increase in the length of alkyl side, which is supposedly originated from the enhancement of hydrophobicity. In alkali medium, P and D show a rising trend with the increase in the length of alkyl chain, which might be related to the loose stacking of network as it occurs to shrink in alkali medium. The preliminary cytotoxicity assay indicates that oxidized glucose-crosslinked alkylate CS membrane is non-toxic in vitro.

Biomimetic surface modification of poly(L-lactic acid) with chitosan and its effects on articular chondrocytes in vitro.

The objective of this study was to investigate the efficiency of two treatments for poly(L-lactic acid) (PLLA) surface modification with chitosan, via entrapment and coupling by using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide. The properties of original PLLA films, chitosan-entrapped and coupled PLLA films were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). The contact angle indicated the change in hydrophilicity and the ESCA data suggested that the modified PLLA films became enriched with nitrogen atoms. The cytocompatibility of modified PLLA films might be improved. Therefore, the attachment and proliferation of bovine articular chondrocyte seeded on modified PLLA films and control one were examined. A whole cell enzyme-linked immunosorbent assay (Cell ELISA) that detects the BrdU incorporation during DNA synthesis and collagen type II secretion was applied to evaluate the chondrocytes on different PLLA films and tissue culture plates. Cell viability was estimated by the MTT assay and cell function were assessed by measuring sulfated glycosaminoglycan secreted by chondrocytes. These results implied that chitosan used to modify PLLA surface through entrapment and coupling could enhance the chondrocyte adhesion, proliferation and function.