Comparisons of Emu Necrotic Femoral Head Micro Structure Repaired in Two Different Methods.

OBJECTIVE: To compare emu necrotic femoral head micro structure repaired in two different methods. METHODS: Fifteen adult emus were divided into 3 groups (all n=5), and the right femoral head was selected to research. The first group was the control group; in the second group, femoral head necrosis was made by cryogen with liquid nitrogen; and in the third group, femoral head necrosis was made by local pure ethanol injection. Right femurs were taken for micro CT examination,then femoral head micro structures were compared among these three groups. RESULTS: No infection or unexpected death was found in all groups. Compared with normal group, necrotic femoral heads in cryogen group showed that bone mineral density significantly reduced after repaire (P=0.015), trabecular space significantly reduced (P=0.001), bone volume fraction significantly enlarged (P=0.036), bone surface/volume fraction (P=0.032) and trabecular numbers (P=0.002) significantly enlarged; trabecular thickness showed no significant difference (P=0.060). Compared with control group, necrotic femoral heads in ethanol group showed that bone mineral density significantly enlarged after repaire (P=0.001), trabecular thickness (P=0.003) and bone surface/volume fraction (P=0.022) significantly enlarged, trabecular space (P=0.001) and bone volume fraction (P=0.001) significantly reduced; the trabecular numbers showed no significant difference (P=0.143). Compared with ethanol group, necrotic femoral heads in cryogen group showed significant lower bone mineral density after repair (P=0.001), significantly lower bone volume fraction (P=0.001), significantly lower trabecular thickness (P=0.001), significantly higher bone surface/volume fraction (P=0.022) and higher trabecular numbers (P=0.003); the trabecular space showed no significant difference (P=0.398). CONCLUSION: Different repair methods make reconstructed femoral head weight bearing area have different bone structure and bone mineral density, along with different bone trabecular quality.

[Study of animal model of osteonecrosis induced by local ethanol injection in emu].

OBJECTIVE: To establish a new animal model of osteonecrosis of the femoral head by local ethanol injection in emu. METHODS: Eight milliliter ethanol was injected slowly to the operated femoral head with customized probe in twenty adult male emus. Postoperatively, hip magnetic resonance imaging was performed at 1, 4, 8, 12 weeks. After emus were sacrificed, the femurs were collected for micro-computed tomography and histological analysis. RESULTS: No emu demonstrated signs of infection or died unexpectedly. Magnetic resonance imaging examination showed broad edema at proximal femur at 1(th) week, and the edema decreased with time, till local edema at femoral head at the 12(th) week. Histological images showed human-like osteonecrotic changes with active bone repair. There were significant differences in trabecular structure and bone mineral density between the operated and intact femoral heads. No collapse was found 6 months after the operation. CONCLUSIONS: This emu model of femoral head osteonecrosis by local ethanol injection can progress to early stage osteonecrosis. The different repair methods may have certain correlation with the results of osteonecrosis of the femoral heads.

Basic research and clinical applications of bisphosphonates in bone disease: what have we learned over the last 40 years?

It is now 40 years since bisphosphonates (BPs) were first used in the clinic. So, it is timely to provide a brief review of what we have learned about these agents in bone disease. BPs are bone-specific and have been classified into two major groups on the basis of their distinct molecular modes of action: amino-BPs and non-amino-BPs. The amino-BPs are more potent and they inhibit farnesyl pyrophosphate synthase (FPPS), a key enzyme of the mavalonate/cholesterol biosynthetic pathway, while the non-amino-BPs inhibit osteoclast activity, by incorporation into non-hydrolyzable analogs of ATP. Both amino-BPs and non-amino-BPs can protect osteoblasts and osteocytes against apoptosis. The BPs are widely used in the clinic to treat various diseases characterized by excessive bone resorption, including osteoporosis, myeloma, bone metastasis, Legg-Perthes disease, malignant hyperparathyroidism, and other conditions featuring bone fragility. This review provides insights into some of the adverse effects of BPs, such as gastric irritation, osteonecrosis of the jaw, atypical femoral fractures, esophageal cancer, atrial fibrillation, and ocular inflammation. In conclusion, this review covers the biochemical and molecular mechanisms of action of BPs in bone, particularly the discovery that BPs have direct anti-apoptotic effects on osteoblasts and osteocytes, and the current situation of BP use in the clinic.

Reconstruction of the medial patellofemoral ligament using hamstring tendon graft with different methods: a biomechanical study.

OBJECTIVE: To measure the tensile strength of the normal medial patellofemoral ligament (MPFL), and evaluate the biomechanics of different fixation methods of the hamstring tendon graft on the patella. METHODS: Eight fresh cadaver knees were prepared by isolating the patella, leaving only the MPFL as its attachment to the medial condyle of femur. The MPFL was reconstructed by three different methods: four-suture fixation, anchors-single suture fixation, and anchors-double suture fixation. The tensile strength and the elongation of the normal MPFL and the tendon grafts were measured. RESULTS: The tensile strength of the four-suture fixation group (234.86±49.02 N) was stronger than that of the normal MPFL (146.91±25.30 N, P=0.0014) and the anchors-single suture group (159.17±49.07 N, P=0.0077), while weaker than that of the anchors-double suture group (314.74±78.46 N, P=0.0052) CONCLUSIONS: With regard to the tensile strength, the four-suture fixation method is reliable for clinical use. Compared with the anchor-suture method, the four-suture fixation method which has no specific implants is more economical, convenient and efficient.

[Combined effects of NEL-like type 1 gene and zoledronate in preventing collapse of the femoral head].

OBJECTIVE: To determine if combined therapy consisting of NEL-like type 1 gene (NELL-1) and zoledronate can prevent the collapse of the femoral head and stimulate the new bone formation in an animal model of osteonecrosis. METHODS: Ischemic osteonecrosis was surgically induced in 24 SD rats, whicih were equally randomly divided into three groups: combination group, treated with both NELL-1 and zoledronate; sham operation group; and placebo group, treated with normal saline solution. The animals were killed 5 weeks after surgery. Radiography, MicroCT, histology, and immunohistochemistry were performed to analyze the results. RESULTS: Morphologically, the femoral head was at good shape in the combination group, while mildly flattened femoral head was seen in the placebo group. No heterotopic ossifications were observed in each group. MicroCT assessment showed significantly higher total and bone mineral volume in the combination group than in the placebo group (P<0.01), whereas no such significant difference was found when compared with the sham operation group(P>0.05). Histological assessment showed more active osteoblast activity and reduced osteoclast activity in the combination group compared with placebo group. CONCLUSION: A combination of NELL-1 and zoledronate can decrease the femoral head deformity while stimulating bone formation in a traumatic rat osteonecrois model, showing a potential to reverse the osteonecrosis.

The volatile flavor and the antioxidant properties of a novel chrysanthemum rice wine during natural aging.

Chrysanthemum morifolium cv. Fubaiju, a traditional tea in southern China with high nutritional and health functions was used in this study. Optimized production conditions of a novel chrysanthemum rice wine (FRW) were obtained by the Box-Behnken design response surface experiment. FRW with best sensory quality was developed with 0.68% chrysanthemum, 0.79% Jiuqu and 0.81:1 liquid-to-solid ratio. Compared with rice wine (RW) control, the total phenolic and flavonoid contents, as well as antioxidant activity of the FRW increased significantly. GC-MS analysis showed that more flavor compounds including alcohols, aldehydes, acids, and esters were detected in FRW. During the aging process, it was found that the antioxidant substances, the antioxidant activity and the flavor substances decreased, with the wine body tending to be homogenized. After 6 months of storage, overall sensory quality of FRW was more harmonious, with special nectar taste, which dramatically improved the flavor characteristics and functionality compared with traditional RW.

Effect and mechanism of zoledronate on prevention of collapse in osteonecrosis of the femoral head.

OBJECTIVE: To observe the effect and mechanism of zoledronate on prevention of collapse in an animal model of osteonecrosis. METHODS: Ischemic osteonecrosis was surgically induced in 16 SD rats (which were further divided into zoledronate group and placebo group); another 8 rats were used as sham surgery group (n=8). The animals were killed 5 weeks after surgery. Radiographic, Micro-CT, histological, and immunohistochemical assessments were performed. RESULTS: Radiographic assessment showed better preservation of the femoral head shape in the zoledronate group than in the placebo group but not significantly different from the sham surgery group. Micro-CT assessment showed higher total volume, bone volume, and total mineralized content in the zoledronate group(all P0.05). Compared with the placebo group, the zoledronate group had reduced osteoclast and osteoblast activity, as confirmed by histological examinations. CONCLUSION: Zoledronate can decrease the femoral head deformity by reducing the osteoclast activity while suppressing new bone and vessels formation in a rat model of traumatic osteonecrosis, and therefore may delay the collapse of femoral head.

[In vitro cartilage tissue engineering with cartilage extracellular matrix-derived porous scaffolds and bone marrow mesenchymal stem cells].

OBJECTIVE: To develop a novel cartilage ECM-derived porous scaffold (CEDPS) and investigate the attachment, proliferation and distribution of bone marrow mesenchymal stem cells (BMSCs) cultured in vitro within the scaffolds. METHODS: Cartilage microfilaments were prepared after pulverization and gradient centrifugation and prepared into suspension after acellularization treatment. The scaffolds were examined by histological staining, scanning electron microscope (SEM), biochemical and biomechanical analysis. After labeling with PKH26, the canine BMSCs were seeded onto the scaffolds. The attachment, proliferation and differentiation of cells were observed by inverted fluorescent microscope and SEM. RESULTS: On histology, most extracellular matrices were retained in the scaffold after the removal of cell fragments. Safranin O staining and immunofluorescence examination with collagen II antibodies provided positive results. Biochemical analysis showed that the collagen content was (708.2 ± 44.7) µg/mg, glycosaminoglycan (254.7 ± 25.9) µg/mg and DNA (0.021 ± 0.007) µg/mg. Mechanical testing showed the compression moduli (E) were (1.226 ± 0.288) and (0.052 ± 0.007) MPa under dry and wet conditions respectively. Inverted fluorescent microscope and SEM showed moderate cell adhesion, chondrocyte-like morphology and matrix synthesis around cells. CONCLUSION: The CEDPS retains most extracellular matrices after a thorough decellularization so as to possess an excellent microstructure with ideal biomechanical characteristics and a good biocompatibility. Thus it is a suitable candidate as an alternative cell-carrier for cartilage tissue engineering. Chondrogenic BMSCs and CEDPS may be used to construct cartilage-like tissue in vitro.

[A new animal model of osteonecrosis induced by focal alternative cooling and heating insults].

OBJECTIVE: To establish a new animal model of osteonecrosis of the femoral head(ONFH) with improved consistency and incidence of femoral head collapse for studies on the mechanism of osteonecrosis. and on the assessment of treatment effectiveness. METHODS: Twenty adult male emus were used. Guide instrumentation was constructed to position the customized probe just articularly and at the proximal part of the femoral head. An alternating focal liquid nitrogen freezing and radiofrequency heating was applied. At 2, 4, 8, 12 and 16 weeks after surgery, hip magnetic resonance imaging (MRI) was performed. Before the emus were sacrificed, barium sulfate was infused to lower extremities for microangiography. The femoral samples were scanned by micro-computed tomography (Micro-CT) and evaluated histologically. RESULTS: No bird demonstrated signs of infection or died unexpectedly. Hip MRI showed changes massive edema at the 4th week, increasingly localized abnormal signals at the 8th'" week, and femoral head collapse at the 12'h week. Micro-CT scans and histological images at the 16th week showed human-like osteonecrotic changes with impaired local blood supply. Bone mineral density of the collapsed head was (380. 31 + 28. 12) mg/cm3 and trabecular spaces were (0. 86 ±0.32) mm; both were significantly lower than those in the control side, which were (415.75 41.28) mg/cm3 and (1. 17 ± 0. 17) mm, respectively (P < 0. 05). Bone volume fraction of the collapsed head was(47.28 ± 17. 14)% and trabecular thickness was (506. 17 ± 220. 58) p.m; both were significantly higher than those at control side, which were (30. 92 ± 4. 01)% and (325. 50 ±44. 53) pm, respectively (P <0. 05). The microangiography at the 16th week showed that vessel volume fraction was (0. 315 ± 0. 055)% , which was significantly higher than the collapsed side [ (0. 142 ± 0. 059)% ] (P <0. 05). CONCLUSIONS: The emu model of fem-oral head osteonecrosis was successfully established using focal alternating cooling and heating insults. The models, with improved consistency and incidence of femoral head collapse, can be used in studies on the mechanism of osteonecrosis and on the assessment of treatment effectiveness.

Bioinformatic analysis of cytokine expression in the proximal and distal nerve stumps after peripheral nerve injury.

In our previous study, we investigated the dynamic expression of cytokines in the distal nerve stumps after peripheral nerve injury using microarray analysis, which can characterize the dynamic expression of proteins. In the present study, we used a rat model of right sciatic nerve transection to examine changes in the expression of cytokines at 1, 7, 14 and 28 days after injury using protein microarray analysis. Interleukins were increased in the distal nerve stumps at 1-14 days post nerve transection. However, growth factors and growth factor-related proteins were mainly upregulated in the proximal nerve stumps. The P-values of the inflammatory response, apoptotic response and cell-cell adhesion in the distal stumps were higher than those in the proximal nerve stumps, but the opposite was observed for angiogenesis. The number of cytokines related to axons in the distal stumps was greater than that in the proximal stumps, while the percentage of cytokines related to axons in the distal stumps was lower than that in the proximal nerve stumps. Visualization of the results revealed the specific expression patterns and differences in cytokines in and between the proximal and distal nerve stumps. Our findings offer potential therapeutic targets and should help advance the development of clinical treatments for peripheral nerve injury. Approval for animal use in this study was obtained from the Animal Ethics Committee of the Chinese PLA General Hospital on September 7, 2016 (approval No. 2016-x9-07).

A novel tissue engineered nerve graft constructed with autologous vein and nerve microtissue repairs a long-segment sciatic nerve defect.

Veins are easy to obtain, have low immunogenicity, and induce a relatively weak inflammatory response. Therefore, veins have the potential to be used as conduits for nerve regeneration. However, because of the presence of venous valves and the great elasticity of the venous wall, the vein is not conducive to nerve regeneration. In this study, a novel tissue engineered nerve graft was constructed by combining normal dissected nerve microtissue with an autologous vein graft for repairing 10-mm peripheral nerve defects in rats. Compared with rats given the vein graft alone, rats given the tissue engineered nerve graft had an improved sciatic static index, and a higher amplitude and shorter latency of compound muscle action potentials. Furthermore, rats implanted with the microtissue graft had a higher density and thickness of myelinated nerve fibers and reduced gastrocnemius muscle atrophy compared with rats implanted with the vein alone. However, the tissue engineered nerve graft had a lower ability to repair the defect than autogenous nerve transplantation. In summary, although the tissue engineered nerve graft constructed with autologous vein and nerve microtissue is not as effective as autologous nerve transplantation for repairing long-segment sciatic nerve defects, it may nonetheless have therapeutic potential for the clinical repair of long sciatic nerve defects. This study was approved by the Experimental Animal Ethics Committee of Chinese PLA General Hospital (approval No. 2016-x9-07) on September 7, 2016.

[Biomechanical evaluation of tendon graft fixation at the tibial site in anterior cruciate ligament reconstruction with Intrafix and bioabsorbable interference screw].

OBJECTIVE: The fixation strength of the hamstring tendon graft on the tibial side is considered the weak point in anterior cruciate ligament (ACL) reconstruction. This work tested the hypotheses that some of these devices will resist graft slippage under loads better than others, and that some will have higher ultimate strength than others. METHODS: Fourteen fresh frozen human cadaver knees underwent fixation of the hamstring tendon harvested from the knee to be used as graft material to reconstruct the ACL and were divided into two equal groups based on the fixation methods: bioabsorbable interference screw group and Intrafix group. A MTS 858 MiniBionix II testing machine was used to carry out tensile testing under an axial load parallel to the tibial tunnel with a velocity of 10 mm/min. Ultimate failure load, displacement of 100N, displacement of 400N, stiffness and mode of failure were recorded respectively. RESULTS: The maximum load for the Intrafix fixation group was (719.094+/-160.478) kgxmxs(-2), significantly higher than that of the bioabsorbable interference screw fixation group [(476.640+/-64.226) kgxmxs(-2), P<0.05]. The displacement levels of 100 kgxmxs(-2) and 400 kgxmxs(-2) for the Intrafix fixation group was (1.025+/-0.326) mm and (4.728+/-1.992) mm respectively, not significantly different from those of the bioabsorbable interference screw fixation [(1.335+/-0.539) mm and (7.564+/-4.307) mm respectively, t=0.2173, 0.0944, both P>0.05], and the stiffness of the Intrafix fixation group was (96.770+/-36.848) kgxm(-1)xs(-2) not significantly different from that of the bioabsorbable interference screw fixation group [(63.976+/-31.003 kgxm(-1)xs(-2)), t=0.0967, P>0.05]. After randomized t-test of significance, the displacement levels of 100N and 400N of these 2 groups were not significantly different. CONCLUSION: Both tibial site fixation with bioabsorbable interference screw and that with Intrafix in ACL reconstruction with 4-stranded hamstring tendon grafts can meet daily physiological demand. Tibial site fixation with Intrafix shows a higher failure load.

Association between glaucoma and the risk of Alzheimer's disease: A systematic review of observational studies.

To address inconsistency as well as investigate the relationship between glaucoma and the risk of Alzheimer's disease (AD). We systematically conducted this meta-analysis based on observational studies published up to 15 January 2018, identified from PubMed and Web of Science. Two team members independently extracted the data and assessed the quality of each included study. Summary relative risk (RR) and 95% confidence intervals (CIs) were calculated using a random-effects model. Eight observational studies with 6870 AD cases were included. The majority of these studies (n = 6) were graded as low risk according to the Newcastle-Ottawa Quality Assessment Scale. Individuals diagnosed with glaucoma, compared to those who were not, had an increased risk of AD (RR = 1.52; 95% CI: 1.41-1.63; I2  = 97%, p < 0.001). A significant finding was also observed for primary open-angle glaucoma (RR = 1.52; 95% CI: 1.41-1.63; I2  = 97%, p < 0.001). However, when stratified by study design, only the case-control studies (RR = 1.08; 95% CI: 0.89-1.31; I2  = 37.3%, p = 0.207) yielded significant results, while the cohort studies did not (RR = 1.08; 95% CI: 0.89-1.31; I2  = 97.7%, p < 0.001). Of note, our meta-regression analysis suggested that study design might be a source of heterogeneity (p = 0.009). Additionally, a significantly positive association was observed when the analyses were restricted to Asia (RR = 2.03; 95% CI: 1.02-4.07). There was no significant publication bias in these analyses. Recent evidence suggests that glaucoma may increase the risk of AD. Additional cohort studies are needed to confirm these findings and to have improved knowledge on the true nature of this association.

[Research of the peak force of pullout about different thoracic hemi-epiphyseal compression staples in animal].

OBJECTIVE: To evaluate the initial dislocation resistance of the pullout force of different material or different type of goats thoracic hemi-epiphyseal compression staples. METHODS: According to the measuring anatomic data of thoracic vertebra (T6 approximately T11) specimens in goats (age 2 approximately 3 months), we designed stainless steel staple whose original distances between two opened prone were 10 mm and three kinds of shape memory alloy (SMA) staples whose original distances between two opened prone were respectively 5.0, 6.5 and 8.0 mm. Four specimens were used and each was cut into three functional spinal units (FSU) of T6 - T7, T8 - T9 and T10 - T11, then FSU were redistributed. Stainless steel staples and SMA staples were implanted into both sides of FSU. There were four groups: group I was stainless steel staples. group II was SMA of 5.0 mm. group III was SMA of 6.5 mm. group IV was SMA of 8.0 mm. The peak force of pullout in each FSU was measured on 858 biomechanical test machine. RESULTS: The peak force of group I was (20.62 +/- 9.15) N, group II was (74.18 +/- 8.81) N, group III was (51.28 +/- 5.44) N, group IV was (39.13 +/- 7.54) N. It is found that there were statistically different between all groups (P < 0.05). The peak force values of group I were lowest than others. CONCLUSIONS: Differences of the initial dislocation resistance in different materials. SMA staples exhibit higher pullout force. Meanwhile, the pullout forces were also different between SMA staples, the smaller original distance the higher peak force of pullout.

Extracellular matrix from human umbilical cord-derived mesenchymal stem cells as a scaffold for peripheral nerve regeneration.

The extracellular matrix, which includes collagens, laminin, or fibronectin, plays an important role in peripheral nerve regeneration. Recently, a Schwann cell-derived extracellular matrix with classical biomaterial was used to mimic the neural niche. However, extensive clinical use of Schwann cells remains limited because of the limited origin, loss of an autologous nerve, and extended in vitro culture times. In the present study, human umbilical cord-derived mesenchymal stem cells (hUCMSCs), which are easily accessible and more proliferative than Schwann cells, were used to prepare an extracellular matrix. We identified the morphology and function of hUCMSCs and investigated their effect on peripheral nerve regeneration. Compared with a non-coated dish tissue culture, the hUCMSC-derived extracellular matrix enhanced Schwann cell proliferation, upregulated gene and protein expression levels of brain-derived neurotrophic factor, glial cell-derived neurotrophic factor, and vascular endothelial growth factor in Schwann cells, and enhanced neurite outgrowth from dorsal root ganglion neurons. These findings suggest that the hUCMSC-derived extracellular matrix promotes peripheral nerve repair and can be used as a basis for the rational design of engineered neural niches.

[The clinical application and biomechanical study of reconstruction of anterior and posterior cruciate ligament with hamstring tendons knot implant fixation].

OBJECTIVE: To investigate the feasibility of reconstruction of anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL) with hamstring tendons knot implant fixation. METHODS: Fifty-two cases of ACL and PCL old injury were reconstructed under the arthroscopy with double bundles hamstring tendons knot implant fixated in the bottle-necked femoral tunnel. The tibia side were fixated by tendon weave suture cross tied a knot at the bone bridge of tibia. In this group reconstruction of ACL was in 25, PCL and ACL at the same time reconstruction in 15, PCL in 12. The failure test and displacement evaluation were used to study the biomechanics of reconstruction of ACL by hamstring tendon knot implant fixation (n = 13) in porcine knees. The control group were reconstructed with bone-patellar tendon-bone B-PT-B and interference screw (n = 11). The tibia side tendon weave suture immobility by cross tied a knot at the tibia bone bridge (n = 7) and interference screw in the tibia tunnel (n = 8). RESULTS: Forty-nine cases were followed up, average 14.6 months. The results of Lanchman test was negative in 46 cases, positive in 3 cases. Preoperative Lysholm score was 56.7, and postoperative was 92.8. According to the knee joint effective evaluate standard, 46 were excellent and 3 good. The maximal pull-out force of hamstring tendons knot implanted fixation had been greater than B-PT-B fixation by interference screw. There were similar displacement in 100 N and 400 N load between hamstring tendons knot implant fixation and B-PT-B (P > 0.05). Peak force test maximal displacement evaluation and failure energy absorption measurement showed that hamstring tendons knot implant fixation was significantly greater than B-PT-B (P < 0.01). The failure test and displacement evaluation of tendon fixation by weave suture knot at the tibia bone bridge cross tie were greater than B-PT-B fixation by interference screw. CONCLUSION: The methods of reconstruct of ACL and PCL by hamstring tendons knot implant fixation are feasible. The advantages include: it is biological fixation, not machine fixation; it may benefit for tendons with bone tunnel healing and decrease medical cost. Decreasing or eliminating displacement is helpful to prevent relaxation after reconstruction of ACL and PCL.

[Experimental study on preparation of decellularized artery vascular graft matrix and explantation of carotid artery allografts].

OBJECTIVE: To set up a new process to access the preparation of decellularized artery grafts. And to evaluate the feasibility of decellularized artery allografts was evaluated. METHODS: This study compared the effects of four extraction chemicals [1% t-octyl-phenoxypolyethoxyethanol (Triton X-100), 1% tri (n-butyl) phosphate (TnBP), and 1% sodium dodecyl sulfate (SDS) and trypsin (0.125, 0.25%) on thoracic artery vascular for 24 h (except trypsin for 2 h). At the base of it, a four-step process, including hypotonic, hypertonic solutions and combining with 1% Triton X-100 and 1% SDS detergents, were performed in rabbit thoracic artery vascular. Histological examination, tensile tests and expanding-burst tests were done on the samples. The decellularized carotid artery allografts were transplanted in other rabbits. RESULTS: Treatment with 1% SDS or 1% Triton X-100 for 24 h could remove most cells with retention of near normal structure. A four-step process could remove all cells with the extracellular matrix well conserved. The pulling mechanical properties and burst pressure of decellularized carotid artery were similar to the control. The decellularized carotid artery allografts (diameter of 2 mm) were patent at explanting up to 2 months. CONCLUSIONS: The acellular artery vascular graft matrix is well prepared with four-step process including detergents, such as TritonX-100, SDS without compromising the graft structure or mechanical properties significantly. The carotid artery allografts (diameter of 2 mm) decellularized by the process are patent at explanting up to 2 months.

[The preparation, structure evaluation and preliminary application of biomimetic biphasic calcium phosphate scaffold].

OBJECTIVE: To fabricate biomimetic biphasic calcium phosphate BCP ceramic scaffolds using three-dimensional (3D) gel-lamination technology and evaluated their structure with 3D parameters and related method. METHODS: Series two-dimensional images of femoral head's specimen of dogs were obtained by micro-computed tomography (Micro-CT). According to these images, porous biomimetic biphasic calcium phosphate (BCP) ceramic scaffolds with oriented trabecular structure were fabricated by three-dimensional (3D) gel-lamination technology. And then, the three-dimensional structure of the scaffolds were reconstructed by computer according to Micro-CT images of these scaffolds and evaluated by three-dimensional parameters. These parameters included bone volume fraction (BVF, BV/TV), bone surface/bone volume (BS/BV) ratio, trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular spacing (Tb.Sp) and structure model index (SMI). The biomechanical properties and biocompatibility of these scaffolds were also evaluated in the study. Six scaffolds, which were combined with BMCs (bone mesenchymal cells, BMCs), were planted into the bone defect of six dogs' femoral head respectively. RESULTS: There was no significant difference between trabecular samples and BCP scaffolds in BV/TV, Tb.Th, Tb.N, and Tb.Pf (P > 0.05). The trabecular system of the scaffold, which had some orientation, represented plate-like model. With a micro-porous porosity of 62%, the average compressive modulus and ultimate strength along the axis of the scaffolds reached (464.0 +/- 36.0) MPa and (5.6 +/- 0.8) MPa respectively. The results of animal test indicated that the trabeculae of these scaffolds were covered by a layer of new bone after 10 weeks of operation. CONCLUSION: Porous BCP scaffolds have been produced with oriented microarchitectural features designed to facilitate vascular invasion and cellular attachment and with initial mechanical properties comparable to those of trabecular bone.

[Cytokine expression in murine cornea tissue during recurrent herpetic stromal keratitis].

OBJECTIVE: To investigate the expression of cytokine in CD(4)(+) T helper type 1 (Th1) and T helper type 2 (Th2) cells in murine cornea during recurrent herpetic stromal keratitis (HSK) and its relationship with the HSK. METHODS: Seven weeks after corneal inoculation with HSV-1, the eyes of latently infected BALB/c mice were irradiated by UV-B and examined for the signs of inflammation and viral reactivation. The corneas of six mice with recurrent stromal disease and six controls were examined to detect the content of Th1 type cytokines (IFN-gamma and IL-12) and Th2 type cytokines (IL-4 and IL-10) on days 0, 3, 7, 10, 14, 21 and 28 after irradiation. RESULTS: Peak of the inflammation process of HSK, manifested by stromal opacification, occurred 7-14 days after viral reactivation in latently infected mice. In semi-quantitative RT-PCR analyses, IFN-gamma, IL-12 p40, IL-10 and IL-4 mRNA were expressed before and during the clinical course of recurrent HSK. The highest expression of IFN-gamma and IL-10 was revealed during the occurrence and development of corneal inflammation (3 -14 days after the reactivation of virus) with a decline at the recovering process (after 14 days). The expression of IFN-gamma and IL-10 was co-related with the degree of corneal opacification. Large amounts of IL-12 p40 mRNA were detected in the cornea 3 days after the virus reactivation and maintained over the whole course of observation. IL-4 levels increased at 3-14 days after the reactivation of virus and declined after day 14. CONCLUSIONS: Th1 and Th2 cytokines present together throughout the course of the inflammatory process accompanying recurrent corneal HSV-1 infection, which is likely associated with a memory response to viral antigens. The expression of IL-10 mRNA parallels with the expression of IFN-gamma and is co-related with the degree of corneal inflammation. The damage and repair of corneal tissue in recurrent HSK may depend upon the balance between the destructive cytokines and protective cytokines at the site of virus reactivation.

MicroRNAs' Involvement in Osteoarthritis and the Prospects for Treatments.

Osteoarthritis (OA) is a chronic disease and its etiology is complex. With increasing OA incidence, more and more people are facing heavy financial and social burdens from the disease. Genetics-related aspects of OA pathogenesis are not well understood. Recent reports have examined the molecular mechanisms and genes related to OA. It has been realized that genetic changes in articular cartilage and bone may contribute to OA's development. Osteoclasts, osteoblasts, osteocytes, and chondrocytes in joints must express appropriate genes to achieve tissue homeostasis, and errors in this can cause OA. MicroRNAs (miRNAs) are small noncoding RNAs that have been discovered to be overarching regulators of gene expression. Their ability to repress many target genes and their target-binding specificity indicate a complex network of interactions, which is still being defined. Many studies have focused on the role of miRNAs in bone and cartilage and have identified numbers of miRNAs that play important roles in regulating bone and cartilage homeostasis. Those miRNAs may also be involved in the pathology of OA, which is the focus of this review. Future studies on the role of miRNAs in OA will provide important clues leading to a better understanding of the mechanism(s) of OA and, more particularly, to the development of therapeutic targets for OA.