Zoledronic acid and teriparatide have a complementary therapeutic effect on aseptic loosening in a rabbit model.

BACKGROUND: Revisions are mainly caused by wear debris-induced aseptic loosening. How to effectively suppress debris-induced periprosthetic osteolysis has become an urgent problem. Both zoledronic acid and teriparatide can increase the bone mass around prostheses and increase the stability of prostheses. A hypothesis was proposed: the combination of the two drugs may have a better treatment effect than the use of either drug alone. METHODS: We created a rabbit model to study the effect and mechanism of the combination of zoledronic acid and teriparatide in the treatment of aseptic loosening. Thirty-two adult male New Zealand white rabbits were selected and treated with TKA surgery, and a titanium rod prosthesis coated evenly with micrometre-sized titanium debris was implanted into the right femoral medullary cavity. All rabbits were randomized into four groups (control group = 8, zoledronic acid group = 8, teriparatide group = 8, and zoledronic acid + teriparatide group = 8). All the animals were sacrificed in the 12th week, and X-ray analyses, H&E staining, Goldner-Masson trichrome staining, von Kossa staining, and RT-PCR and Western blotting of the mRNA and protein of OCN, OPG, RANKL and TRAP5b in the interface membrane tissues around the prostheses were immediately carried out. RESULTS: The results shown that both zoledronic acid and teriparatide could inhibit debris-induced peri-prosthetic osteolysis and promote new bone formation. Zoledronic acid was more capable of inhibiting osteoclast activation and peri-prosthetic osteolysis, while teriparatide was more capable of promoting osteoblast function and peri-prosthetic bone integration. CONCLUSION: This research confirmed that the combination of zoledronic acid and teriparatide could prevent and treat aseptic loosening of the prosthesis more effectively. However, the safety of this combination and the feasibility of long-term application have not been ensured, and the clinical application requires further experiments and clinical research support.

Early Patellofemoral Function of Medial Pivot Prostheses Compared with Posterior-Stabilized Prostheses for Unilateral Total Knee Arthroplasty.

OBJECTIVE: To systematically evaluate the patellofemoral joint design of medial pivot prosthesis, which incorporates a variety of "patella-friendly" design features, by comparing clinical and radiographic results with another prosthesis. METHODS: All consecutive patients who underwent unilateral total knee arthroplasty (TKA) with medial pivot prosthesis (Group MP, 126 cases) between September 2016 and April 2018 were enrolled in this retrospective study. For each patient reviewed, a control patient was matched, according to age, gender, side, body mass index (BMI), preoperative range of motion (ROM), and operating period, who had received primary unilateral TKA with a conventional posterior-stabilized prosthesis at the same period as the study group (Group PS, 126 cases). All patients underwent at least 1-year follow-up. At the preoperative and final follow-up periods, data on the Knee Society Score (KSS) score, WOMAC score, Kujala score, and ROM were collected. Merchant views were taken with the knee flexion at 30°, 60°, and 90° to measure patella shift and tilt. Preoperative posterior condylar angle (PCA) was also measured. Postoperative complications, including anterior knee pain, maltracking, patellar clunk or crepitus (PCC), were evaluated. RESULTS: There were no significant differences in the demographics or clinical characteristics between the two groups. No statistically significant difference was identified in the KSS total score, including knee score and function score, or in the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score between the two groups after the operation. We found statistically significant differences in the postoperative Kujala scores and the ROMs between the two groups. The mean Kujala score in group MP was better than in group PS (MP 77.16 ± 3.80 vs PS 75.97 ± 4.06, P < 0.05), while the ROM in group PS was significantly higher than in group MP (MP 122.24° ± 4.45° vs PS 123.78° ± 6.05°, P < 0.05). Simultaneously, the preoperative/postoperative Kujala score improvement in group MP was observed to be significantly larger than in group PS (MP 27.82 ± 5.31 vs PS 26.17 ± 4.89, P < 0.05), but the average ROM improvement in group PS was significantly greater than in group MP (MP 19.00° ±9.90° vs PS 21.57° ± 9.62°). In the 90° Merchant view, the mean patella tilt of group MP was statistically smaller than that of group PS (MP 4.21° ± 1.62° vs PS 4.74° ± 1.95°, P < 0.05), and the average patella tilt change in group MP was significantly greater than in group PS (MP -3.8° ± 1.43° vs PS -3.23° ± 1.33°, P < 0.05). Preoperative PCA did not show significant differences between the two groups. Two cases of PCC and three cases of anterior knee pain were noted in group MP, and nine cases and six cases, respectively, were observed in group PS. The incidence of PCC was significantly lower in group MP (1.6% vs 7.1%, P < 0.05). There was no significant difference in follow-up time between the two groups. CONCLUSION: The medial pivot prosthesis could achieve satisfactory outcomes with better patellofemoral performance attributed to its "patella-friendly" design characteristics compared to the conventional posterior-stabilized prosthesis.

Carboxymethyl chitosan reduces inflammation and promotes osteogenesis in a rabbit knee replacement model.

BACKGROUND: The major causes of failure after total knee arthroplasty (TKA) include prosthesis loosening and infection. This study aimed to investigate the role of carboxymethyl chitosan (CMC) in knee arthroplasty. METHODS: A total of 20 New Zealand white rabbits that were divided into two groups (10 in the control group and 10 in the chitosan group) were included in the study. They underwent TKA surgery, and all were implanted with titanium rod prostheses; the prosthesis in the chitosan group was coated with CMC. After 12 weeks, all rabbits were euthanized, and the following analyses of some specific surface membrane tissues around the prosthesis were performed: X-ray analysis; micro-computed tomography scan; haematoxylin and eosin, Van Gieson, and Von Kossa staining; reverse transcription polymerase chain reaction; and Western Blotting. RESULTS: The result of CCK8 test showed CMC can promote cell proliferation and increase cell viability. Radiological result showed better amount of bone deposits and more bone formation in the chitosan group. HE staining result showed CMC reduces inflammation around the prosthesis. The VG and Von Kossa staining results showed CMC can promote bone deposition around prosthesis. And according to the results of PCR and WB, the OCN content was higher in the chitosan group, while the MMPs content was lower. The chitosan group has an increased OPG/RANKL ratio than the control group. CONCLUSION: CMC can effectively inhibit the inflammatory response around the prosthesis and osteoclast activation and promote osteogenesis by interfering with the osteoprotegerin/receptor activator of nuclear factor kappa-Β ligand/receptor activator of nuclear factor kappa-Β signalling pathway.

Comparison of Postoperative Effects between Medial Pivot Prosthesis and Posterior Stabilized Prosthesis.

OBJECTIVE: To compare the postoperative inflammation and pain response between medial pivot (MP) and posterior stabilized (PS) prostheses among total knee arthroplasty (TKA) patients. METHODS: A prospective cohort study was conducted from January 2019 to May 2019 at the Affiliated Hospital of Qingdao University. The study included patients diagnosed with stage III or IV Kellgren-Lawrence knee osteoarthritis (KOA) who had failed conservative treatment, had undergone no previous knee surgeries, had varus substantial deformities (11°-20° deviation), and had received their first unilateral TKA. A total of 109 patients who underwent PS prosthesis TKA and 98 patients who underwent MP prosthesis TKA were continuously enrolled. Inflammation biomarkers, such as leukocyte (white blood cells), erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP), together with hemoglobin (Hb), the visual analog pain score (VAS) and range of motion (ROM) were compared between the two groups. The Student t-test was applied to analyze continuous parameters, and the χ2 -test was used for categorical parameters. The linear mixed model was used for the repeated measurement data from the follow-up visits. Multivariate backward logistic and linear regression models were used to determine the factors potentially influencing prostheses and VAS scores. RESULTS: All these enrolled patients were followed up at 2, 4, 7, and 30 days after TKA. There were no significant differences between the PS group and the MP group in body mass index (BMI), gender, laterality, usage of nonsteroidal anti-inflammatory drugs (NSAIDs) and opioids, and drain tube extubation time (P > 0.05). Compared with the PS group, the MP group were older (67.5 years vs 65.4 years), and had a higher mid-vastus approach rate (67.3% vs 26.6%), a shorter tourniquet duration (68.3 ± 10.2 h vs 73.9 ± 11.2 h), a larger prosthetic pad (10.8 ± 1.2 mm vs 10.4 ± 1.2 mm), and a lower drain tube diversion volume (187.6 ± 119.3 mL vs 234.0 ± 155.7 mL). In the linear mixed model, MP prostheses had less CRP and ESR elevation and less Hb decrease than PS prostheses (P for group × time < 0.001). There were no significant differences in the changing trends between MP and PS prostheses by time for VAS scores and ROM. In the multivariate logistic regression model, MP prostheses showed significant differences compared with PS prostheses in treatment approach (odds ratio [OR] = 3.371, 95% confidence interval [CI]: 1.953-7.127; P < 0.001), ultrasound treatment start time (OR = 2.669, 95% CI: 1.385-5.141; P = 0.003), and tourniquet duration (OR = 0.954, 95% CI: 0.925-0.984; P = 0.003). Higher VAS scores on the second day postoperatively were related to high VAS scores preoperatively, use of opioids, high drain tube diversion, long tourniquet duration, and long drain tube extubation (P < 0.05), respectively. CONCLUSION: The MP prostheses showed potential advantages compared with PS prostheses in TKA in inflammatory responses.

Esmolol inhibits inflammation and apoptosis in the intestinal tissue via the overexpression of NF-κB-p65 in the early stage sepsis rats.

BACKGROUND/AIMS: Accumulating evidence reveals esmolol could protect the gut mucosa through the regulation of immune response and inflammation in patients with sepsis. However, its underlying mechanism is not fully understood. MATERIALS AND METHODS: Diamine oxidase (DAO), intestinal fatty acid-binding protein (I-FABP), interleukin (IL)-6, and IL-10 in the plasma of rats were detected by ELISA assay. Western blotting was utilized to measure the expression levels of NF-kappa B-p65, Bcl-2, and cleaved caspase-3 in the intestinal tissues. The survival analysis was performed in each group. RESULTS: The plasma levels of DAO and IL-10 levels were increased, whereas that of I-FABP and IL-6 were decreased in the sepsis rats after esmolol treatment, indicating that after the esmolol treatment, the intestinal inflammation and damages were remarkably reduced as compared to those in the normal saline treated sepsis rats. NF-κB-p65 and Bcl-2 were highly expressed, but cleaved caspase-3 showed lower expression in the esmolol treated groups. However, at the same time, we observed contrasting results in the normal saline treated group. Western blotting data indicated that the esmolol treatment inhibited the inflammation and apoptosis in the intestinal tissue due to the overexpression of NF-κB-p65 in the celiac sepsis rats. The survival analysis results indicate that the esmolol infusion should be used in the early stages sepsis rats. CONCLUSION: Esmolol can suppress inflammation and apoptosis in the intestinal tissue via the overexpression of NF-kappa B-p65 in the early stage sepsis rats. kappa BEarly-stage use of esmolol might be an ideal treatment method for sepsis.

Hip survival rate in the patients with avascular necrosis of femoral head after transtrochanteric rotational osteotomy: a systematic review and meta-analysis.

BACKGROUND: The clinical outcome of transtrochanteric rotational osteotomy (TRO) for osteonecrosis of the femoral head (ONFH) remains controversial, and the promising clinical results of several Japanese studies could not be reproduced in American and European studies. Trying to solve controversies on TRO for ONFH rising from apparently conflicting studies, a meta-analysis was conducted to assess the 5- and 10-year hip survival rates (with conversion to artificial joint replacement and radiographic failure as endpoints) after TRO. METHODS: All eligible studies were searched in seven comprehensive databases including PubMed, Web of Science, Embase, Cochrane Library, VIP Database, China Knowledge Resource Integrated Database, and Wan Fang Database prior to June 2019. The outcomes evaluated were 5- and 10-year hip survival rates after TRO. The odds ratio and risk difference for the non-comparative binary data with the 95% confidence intervals (CIs) were calculated for each outcome. The included studies were assessed for methodologic bias and potential reasons for heterogeneity were explored. RESULTS: Nineteen studies of TRO for ONFH were eligible for this meta-analysis according to inclusion criteria. Based on the previous report, two calculation methods (Methods 1 and 2) were adopted in this meta-analysis. Furthermore, we performed a sub-group analysis of the 5- and 10-year hip survival rates (Method 1) after TRO for ONFH: Asian sub-population and non-Asian sub-population. Taking conversion to artificial joint replacement as the endpoint, 5- and 10-year hip survival rates (Method 1) after TRO for ONFH in the Asian population were 0.86 (95% CI = 0.82-0.89) and 0.72 (95% CI = 0.65-0.78), respectively, and 5- and 10-year hip survival rates after TRO for ONFH in the non-Asian population were 0.55 (95% CI = 0.43-0.67) and 0.42 (95% CI = 0.28-0.55), respectively. The 5- and 10-year hip survival rates (Method 2) after TRO for ONFH were 0.90 (95% CI = 0.79-0.95) and 0.89 (95% CI = 0.81-0.94), respectively. Taking radiographic failure as the endpoint, 5- and 10-year hip survival rates after TRO for ONFH were 0.70 (95% CI = 0.64-0.76) and 0.53 (95% CI = 0.46-0.61), respectively. CONCLUSIONS: The 5- and 10-year hip survival rates after TRO for ONFH were satisfactory in the Asian population, and were acceptable in the non-Asian population despite high early failure rates.

Micrometer-Sized Titanium Particles Induce Aseptic Loosening in Rabbit Knee.

Wear debris induced aseptic loosening is the leading cause of total knee arthroplasty (TKA) failure. The complex mechanism of aseptic loosening has been a major issue for introducing effective prevention and treatment methods, so a simplified yet efficient rabbit model was established to address this concern with the use of micrometer-sized titanium particles. 20 New Zealand white rabbits were selected and divided into two groups (control = 10, study = 10). A TKA surgery was then performed for each of them, with implantation of a titanium rod prosthesis which was coated evenly with micrometer-sized titanium in the study group and nothing in the control group, into right femoral medullary cavity. After 12 weeks, all the animals were euthanized and X-ray analyses, H&E staining, Goldner Masson trichrome staining, Von Kossa staining, PCR, and Western blotting of some specific mRNAs and proteins in the interface membrane tissues around the prosthesis were carried out. The implantation of a titanium rod prosthesis coated with 20 µm titanium particles into the femoral medullary cavity of rabbits caused continuous titanium particle stimulation around the prosthesis, effectively inducing osteolysis and aseptic loosening. Titanium particle-induced macrophages produce multiple inflammatory factors able to activate osteoclast differentiation through the OPG/RANKL/RANK signaling pathway, resulting in osteolysis while suppressing the function of osteoblasts and reducing bone ingrowth around the prosthesis. This model simulated the implantation and loosening process of an artificial prosthesis, which is an ideal etiological model to study the aseptic prosthetic loosening.

[Clinical efficiency and activity range of stiff knee by total knee arthroplasty].

OBJECTIVE: To observe clinical effect and influencing factor of total knee arthroplasty (TKA) for the treatment of stiff knee. METHODS: From January 2010 to October 2014, 20 patients(25 knees) with stiff knee were treated with TKA. Among them, including 2 males(3 knees) and 18 females(22 knees), aged from 55 to 78 years old with an average of(64.5± 4.9) years old, the courses of disease ranged from 5 to 21 years with an average 8.3 years. Preoperative and postoperative HSS (hospital for special surgery knee score) score, activity range and complications were observed and compared. RESULTS: All patients were followed up from 12 to 69 months with an average of 35.3 months. Ten patients occurred complications after operation. HSS score was improved from 32.36±12.31 preoperatively to 80.70±18.52 postoperatively, and had statistical difference between two groups;7 knees obtained excellent results, 15 knees good and 3 knees moderate. Activity range was improved from(39.4±5.3)°preoperatively to (92.5±11.2)° at the latest follow up. CONCLUSIONS: Total knee arthroplasty for stiffness knees is feasible and could obtain satisfied activity range and function.

F-box genes: Genome-wide expansion, evolution and their contribution to pollen growth in pear (Pyrus bretschneideri).

F-box gene family, as one of the largest gene families in plants, plays crucial roles in regulating plant development, reproduction, cellular protein degradation and responses to biotic and abiotic stresses. However, comprehensive analysis of the F-box gene family in pear (Pyrus bretschneideri Rehd.) and other Rosaceae species has not been reported yet. Herein, we identified a total of 226 full-length F-box genes in pear for the first time. And these genes were further divided into various subgroups based on specific domains and phylogenetic analysis. Intriguingly, we observed that whole-genome duplication and dispersed duplication have a major contribution to F-box family expansion. Furthermore, the dynamic evolution for different modes of gene duplication was dissected. Interestingly, we found that dispersed and tandem duplicate have been evolving at a high rate. In addition, we found that F-box genes exhibited functional specificity based on GO analysis, and most of the F-box genes were significantly enriched in the protein binding (GO: 0005515) term, supporting that F-box genes might play a critical role for gene regulation in pear. Transcriptome and digital expression profiles revealed that F-box genes are involved in the development of multiple pear tissues. Overall, these results will set stage for elaborating the biological role of F-box genes in pear and other plants.

Astragaloside IV suppresses inflammatory mediator production in synoviocytes and collagen­induced arthritic rats.

The aim of the current study was to investigate the effects of Astragaloside­IV (AS­IV) on inflammatory mediators in synoviocytes and collagen­induced arthritic rats. Synoviocytes were stimulated with lipopolysaccharides (LPS) and Sprague­Dawley rats were injected with type II collagen. AS­IV was administered to the LPS­stimulated synoviocytes and collagen­induced arthritis (CIA) rats. The inflammation of LPS­stimulated synoviocytes and CIA rats was assessed using enzyme­linked immunosorbent assays and western blotting. Using Cell Counting Kit­8 analysis, it was demonstrated that AS­IV (5, 20 and 50 mg/ml) inhibited the LPS­stimulated synoviocytes proliferation in a dose­dependent manner. AS­IV significantly inhibited the LPS­stimulated inflammatory response, as indicated by the expression levels of TNF­α, IL­1ß, IL­6 and IL­8. In addition, treatment with AS­IV significantly reduced the LPS­stimulated cyclooxygenase (COX)­1, COX­2, high mobility group box 1 protein (HMGB1) and intercellular adhesion molecule 1 overexpression, and intranuclear nuclear factor (NF)­κBp65 subunit accumulation and activation of c­Jun N­terminal kinase (JNK)1/2 and p38. Similar to the protective effects of AS­IV on LPS­stimulated synoviocytes, AS­IV treatment significantly reduced the expression levels of tumor necrosis factor α, interleukin (IL)­1ß, IL­6 and IL­8 expression levels, and attenuated intranuclear NF­κBp65 subunit accumulation and overexpression of COX­2 and inducible nitric oxide synthase in CIA rats. In conclusion, the results of the present study demonstrated for the first time, to the best of our knowledge, that AS­IV protects synoviocytes against LPS­ and collagen­induced inflammatory responses through inhibition of the HMGB1­dependent JNK1/2­ and p38­activated NF­κB/COX­2 pathway.

Effect of the human insulin-like growth factor 1 gene transfection to human umbilical cord blood mesenchymal stem cells.

OBJECTIVE: To observe the effect of transfecting the gene human insulin-like growth factor (hIGF)-1 into human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) via non-viral vector. METHODS: This study was performed in the Affiliated Hospital of Qingdao University, Qingdao, China from June 2012 to May 2013. Twelve hUCB samples were harvested, and isolated in lymphocyte separation medium, and then cultured. Surface antigen expression in MSCs was detected by flow cytometry. Recombinant plasmid pIRES2-enhanced green fluorescent protein (EGFP)-hIGF-1 was transfected into MSCs by X-treme GENE HP DNA transfection reagent. Then, EGFP was observed with reverse fluorescent microscope at different time points. Enzyme-linked immunosorbent assay was used to determine the hIGF-1 protein concentration in supernatants. Immunofluorescence microscopy and reverse transcription polymerase chain reaction were used to detect the expression of hIGF-1 in the hUCB-MSCs. Expression of type II collagen was detected by immunohistochemistry staining. RESULTS: Transfection efficiency was 28.74 +/- 7.31%. The cluster of differentiation (CD)90, CD105, and CD146 expression increased CD34, CD45, and anti-HLA-DR expression decreased. Results of immunofluorescence microscopy and RT-PCR confirmed expression of the hIGF-1 gene. The hIGF-1 protein concentration in the supernatants showed a peak level at 34.63 +/- 1.61 ng/ml 48 hours after transfection. Immunohistochemical analysis of transfected hUCB-MSCs proved that type II collagen could be expressed positively. CONCLUSION: Human IGF-1 gene can be transfected into hUCB-MSCs, and expressed at a high level with subsequent expression of type II collagen.

Impact of high altitude on the hepatic fatty acid oxidation and synthesis in rats.

High altitude (HA) affects energy metabolism. The impact of acute and chronic HA acclimatization on the major metabolic pathways is still controversial. In this study, we aimed to unveil the impact of HA on the key enzymes involved in the fatty acid (FA) metabolism in liver. Rats were exposed to an altitude of 4300 m for 30 days and the expressions of two key proteins involved in FA ß-oxidation (carnitine palmitoyl transferase I, CPT-I; and peroxisome proliferator-activated receptor alpha, PPARα), two proteins involved in FA synthesis (acetyl CoA carboxylase-1, ACC-1; and AMP-activated protein kinase, AMPK), as well as the total ketone body in the liver and the plasma FFAs were examined. Rats without HA exposure were used as controls. We observed that the acute exposure of rats to HA (3 days) led to a significant increase in the expressions of CPT-I and PPARα and in the total hepatic ketone body. Longer exposure (15 days) caused a marked decrease in the expression of CPT-I and PPARα. By 30 days after HA exposure, the expression levels of CPT-I and PPARα returned to the control level. The hepatic ACC-1 level showed a significant increase in rats exposed to HA for 1 and 3 days. In contrast, the hepatic level of AMPK showed a significant reduction throughout the experimental period. Plasma FFA concentrations did not show any significant changes following HA exposure. Thus, increased hepatic FA oxidation and synthesis in the early phase of HA exposure may be among the important mechanisms for the rats to respond to the hypoxic stress in order to acclimatize themselves to the stressful environments.

Protective effects of BML-111 on cerulein-induced acute pancreatitis-associated lung injury via activation of Nrf2/ARE signaling pathway.

The aim of this study was to investigate whether BML-111 can exert protective effects on cerulein-induced acute pancreatitis-associated lung injury (APALI) via activation of nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant responsive element (ARE) signaling pathway. Severe acute pancreatitis (SAP) was established by intraperitoneal injection of cerulein (50 µg/kg) seven times at hourly intervals and Escherichia coli lipopolysaccharide (10 mg/kg) once after the last dose of cerulein immediately. BML-111 (1 mg/kg) was administered 1 h before the first injection of cerulein. Samples were taken at 3, 6, 12, and 24 h after the last injection. Pathologic lesions of the pancreas and lung tissues as well as the levels of serum amylase were analyzed; Myeloperoxidase (MPO), malondialdehyde (MDA), superoxide dismutase (SOD), Nrf2, heme oxygenase-1 (HO-1), and NAD(P)H: quinone oxidoreductase-1 (NQO1) of lung tissue were determined. The findings revealed that the injuries of pancreas and lung were typically induced by cerulein. The administration of BML-111 reduced the levels of serum amylase, lung MPO, lung MDA, the wet-to-dry weight ratio, and the pathology injury scores of the lung and pancreas, which increased in the SAP group. The expressions of Nrf2, HO-1, NQO1, and activity of SOD in lung tissue increased in the BML-111 group compared with those in the SAP group. This study indicates that BML-111 may play a critical protective role in APALI induced by cerulein. The underlying mechanisms of protective role may be attributable to its antioxidant effects through the activation of Nrf2/ARE pathway.

BML-111, a lipoxin receptor agonist, ameliorates 'two-hit'-induced acute pancreatitis-associated lung injury in mice by the upregulation of heme oxygenase-1.

The objective of this study is to investigate the effects of BML-111 on acute pancreatitis-associated lung injury (APALI) induced by cerulein with subsequent an LPS administration in mice and its possible mechanisms. One hundred and twenty-eight mice were randomly allocated to four groups, namely the APALI group, the BML-111 pretreatment group, the BM-111 control group, and the control group. The 'two-hit' mice APALI model was established by intraperitoneal injection of cerulein 7 times at hourly intervals and Escherichia coli lipopolysaccharide (LPS) once after the last dose of cerulein immediately. The samples were taken at 3, 6, 12, and 24 h after the last injection. Serum levels of amylase, TNF-a, IL-1ß and IL-10, were determined. Histological score of the pancreas and lung, the wet/dry weight ratio, and heme oxygenase-1 (HO-1) expression in the lung were also evaluated. BML-111 pretreatment significantly reduced the serum levels of amylase, TNF-α, IL-1ß, the wet/dry weight ratio of lung, and the pathology injury scores of pancreas and lung, and the serum levels of IL-10 were markedly increased. The severity of pancreatic and lung histology were also significantly improved by the administration of BML-111, and the expressions of HO-1 in lung tissues also increased in the BML-111 group compared with those in the APALI group. In conclusion, BML-111 exerts protective effects on APALI induced by cerulein and LPS. In addition to its anti-inflammatory effects, the beneficial effects may also be due to the upregulation of HO-1 expression in the lung tissues.

Repair of large osteochondral defects with mix-mosaicplasty in a goat model.

Osteochondral defects in weight-bearing regions must be repaired with cartilage and subchondral bone support simultaneously, as well as the integration between the 2, particularly in young, active patients. In this study, a new method called mix-mosaicplasty was used to reconstruct large osteochondral defects (6-mm diameter) in the weight-bearing region of the femoral condyle of goats. Two periosteum-bone plugs and 1 osteochondral plug harvested from the proximal tibia and intertrochlea groove were assembled to fill the defects in a mosaic mode. The goats were euthanized 16 weeks postoperatively, and the result of the repair process was assessed using macroscopy, morphologic analysis, electron microscope observation, glycosaminoglycan assay, and magnetic resonance imaging. Sixteen weeks postoperatively, the superficial surface of the defective region was covered with regenerated cartilage, and the periosteum-bone plugs were combined with each other. However, cleavage between cartilage plugs was noted. The donor site, which was filled with periosteum-bone plugs, was regenerated with fibrocartilage-like tissue. The repaired tissue was composed of small chondrocyte-like cells arranged tightly within an evenly distributed extracellular matrix containing type II collagen. Cells of the regenerated tissue in periosteum-bone plugs were smaller and distributed more densely. Electron microscopy demonstrated regular matrix fibers and abundant organelles within the repaired tissue. No significant differences of glycosaminoglycan content were observed between reconstructed tissue and normal hyaline cartilage. Magnetic resonance imaging revealed the healing process between plugs other than the control group. The new technique of mix-mosaicplasty can reconstruct full-thickness osteochondral compound defects in the weight-bearing region of the femoral condyle.

Reconstruct large osteochondral defects of the knee with hIGF-1 gene enhanced Mosaicplasty.

OBJECTIVE: To investigate a compound technique including gene therapy, injectable tissue engineering and Mosaicplasty to reconstruct large osteochondral defect. METHODS: Plasmid vector containing hIGF-1 cDNA was created and transfected into BMSCs in vitro with FuGene6. After gene expression determination, cells were mixed with calcium alginate gel. Osteochondral defects were created on the femoral condyle of goats in a diameter of 6mm. Osteochondral plugs were harvested from the intertrochlea groove and pressed into the recipient sites in a mosaic mode. Gene modified BMSCs-scaffold complex was applied to fill the residual defects. Control groups were also set up. At 4 and 16 weeks, specimens were investigated in gross and under microscopy, electromicroscopy and MRI detection. RESULTS: hIGF-I gene was expressed effectively with the peak concentration at 34.75 ng/ml. Subchondral bone and cartilage were integrated well in gene enhanced Mosaicplasty group. The reconstructed tissue filled up the gaps between columns, which appeared better than other groups. The regenerated cartilage was integrated with neighbor tightly in regular arrange. Extracellular matrix distributed evenly and deeply stained by alcian blue. Quantitative histologic assessments showed higher score in gene enhanced Mosaicplasty group. Glycosaminoglycan assay revealed no difference between groups involving Mosaicplasty. MRI analysis demonstrated the healing process between the subchondral bone other than control groups. CONCLUSIONS: hIGF-I gene enhanced tissue engineering can modify the outcome of Mosaicplasty to reconstruct large osteochondral defects in weight-bearing region.

Protective effect of exogenous IGF-I on the intestinalmucosal barrier in rats with severe acute pancreatitis.

BACKGROUND: Severe acute pancreatitis (SAP) can result in intestinal mucosal barrier (IMB) dysfunction. This study was undertaken to demonstrate the effect of IGF-I on the intestinal mucosal barrier in rats with SAP and its possible mechanisms. METHODS: Seventy-two male Wistar rats were randomly divided into three groups: a sham operation (SO group, n=24), a SAP group not treated with IGF-I (SAP group, n=24), and a SAP group treated with IGF-I (IGF-I group, n=24). SAP was induced in the rats by injecting 5.0% sodium taurocholate into the biliary-pancreatic duct. The SO rats were given an infusion of normal saline instead. The rats in the IGF-I group underwent the SAP procedure and were given a subcutaneous injection of IGF-I at 30 minutes before the operation and at 3 hours after the operation. Eight rats in each group were sacrificed at 6, 12 and 24 hours after operation. Apoptosis of mucosal cells in the small intestine was determined by TUNEL. The levels of endotoxin and DAO and serum amylase were also measured. Pathologic changes in the small intestine were monitored. Changes of bax and bcl-2 mRNA expression in the small intestine were determined by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The levels of serum amylase were lower in the IGF-I group than in the SAP group at all three time points (P<0.05). The levels of endotoxin in the IGF-I group were higher than those in the SAP group at 6 hours, but lower in the IGF-I group than in the SAP group at 12 and 24 hours (P<0.05). The levels of diamine oxidase were higher in the IGF-I group at 6 hours but lower than those in the SAP group at 12 and 24 hours. The pathological score of the small intestine was lower in the IGF-I group than in the SAP group, and the difference was statistically significant at 12 and 24 hours. The pathologic changes observed under electron microscopy were better in the IGF-I group than those in the SAP group. The apoptosis index of intestinal epithelial cells was significantly decreased in the IGF-I group compared with the SAP group. Compared with the SO group, the mRNA expression levels of bax were increased at each time point in the SAP group, and were significantly decreased in the IGF-I group as compared with the SAP group at each time point (P<0.05). The expression levels of bcl-2 were weak and not different between the SO group and the SAP group (P>0.05). They were significantly increased in the IGF-I group versus the SO and SAP groups (P<0.05). The ratio of bax and bcl-2 mRNA expression levels at each time point in the SAP group were significantly higher than those in the SO group, but they were obviously decreased in the IGF-I group. CONCLUSIONS: Exogenous IGF-I seems to protect mucosal cells in the small intestine against SAP-induced apoptosis and could alleviate SAP-induced injury of the intestinal mucosa. The underlying mechanisms include enhanced mRNA expression of bcl-2 and inhibition of bax mRNA expression.

[Repair full-thickness meniscal defects with injectable tissue engineering technique].

OBJECTIVE: To investigate the effectiveness of injectable tissue engineering to repair full-thickness meniscal defects. METHODS: From June 2008 to February 2009 full-thickness of meniscal defects were created in the anterior corner of goats, which with no blood supply, in a diameter of 2 mm. Then bone marrow stem cells (BMSCs) was mixed with injectable calcium alginate gel to fill the defects. Other groups include the calcium alginate gel and empty group were served as control groups. At different time points, the animals were sacrificed and macroscopy, microscopy determination, electroscopy and MRI detection were performed to assess the outcomes of repairing. RESULTS: The meniscal defects had been filled thoroughly in 16 weeks after operation with white, tough and elastic repair tissue similar to normal meniscal fibrocartilage in the tissue engineering groups. The repair tissue was mainly fibrochondrocytes in line with the calcium alginate fiber. Thick matrix secreted by the cells crammed the space between fibers. The view under electroscopy demonstrated that the microstructure of the repair tissue was normal and cells were in a fibrocartilage phenotype. CONCLUSION: The full-thickness meniscal defects in regions without blood supply can be reconstructed effectively with injectable tissue engineering.

Tendon healing in vivo and in vitro: neutralizing antibody to TGF-ß improves range of motion after flexor tendon repair.

Adhesion formation between the flexor tendon and its surrounding fibro-osseous sheath results in a decreased postoperative range of motion (ROM) in the hand. Transforming growth factor-beta (TGF-ß) is a key cytokine in the pathogenesis of tissue fibrosis. In this study, the effects of TGF-ß1 neutralizing antibody were investigated in vitro and in vivo. In the in vitro investigation, primary cell cultures from rabbit flexor tendon sheath, epitenon, and endotenon were established and each was supplemented with TGF-ß along with increasing doses of TGF-ß1 neutralizing antibody. Collagen I production was measured with enzyme-linked immunosorbent assay. In the in vivo study, rabbit zone-II flexor tendons were transected and then immediately repaired. Transforming growth factor-ß1 neutralizing antibody or phosphate-buffered saline solution (control) was added to the repair sites, and the forepaws were tested for ROM and repair strength at 8 weeks postoperatively. Transforming growth factor-ß1 neutralizing antibody reduced TGF-ß upregulated collagen production. Intraoperative application of TGF-ß1 neutralizing antibody significantly improved the ROM of the operatively treated digits. The effect on breaking strength of the tendon repair was inconclusive.

[The efficacy of open-wedge high tibial osteotomy for varus knee].

OBJECTIVE: To introduce the biplanar opening high tibial osteotomy with rigid fixation for adult varus knee. METHODS: Twelve patients with varus knee and degeneration of medial compartment received the biplanar opening high tibial osteotomy between June 2001 and July 2008. The pre-operative deformity was about 11.5 degrees without osteoarthritis changes in other compartments. The average range of motion was more than 90 degrees . One of the patients had ruptures of LCL and ACL, who received osteotomy after reconstruction of the ligaments. The main symptom before the operation was pain in medial compartment. The range of motion, Lysholm score and subjective satisfactory examinations were assessed before and after the osteotomy. RESULTS: All of the osteotomy sites were healed at twelve to sixteen weeks after operation. No complications such as plate broken or injury of nerve or blood vessel had occurred. The mean correct angle was 9.5 degrees . No degenerative changes had developed in other compartments of the knee. The mechanical axis of the lower extremities was maintained during the follow-up. The overall satisfactory rate was 83.3%. Statistically significant changes exist in the Lysholm score and varus degree. CONCLUSION: The open-wedge high tibial osteotomy is suitable for the symptomatic genu varum in younger patients with good short-term and mid-term results.