Loop technique for anterior cruciate ligament reconstruction combined with anterolateral structure reinforcement: technical description and clinical results.

PURPOSE: We describe a surgical technique for ACL reconstruction combined with anterolateral structure reinforcement and report early clinical follow-up results. METHODS: The semitendinosus and gracilis tendons are braided into 5 strands and the ACL femoral tunnel and tibial tunnel are created. The graft is passed through the tunnel with the use of a traction suture and the tibial end is fixed with absorbable interference screws at 30° of knee flexion. The ACL graft traction suture is used as an anterolateral reconstruction structure to pass through the proximal exit of the ACL femoral tunnel and then through the depth of the iliotibial bundle to the anterior to Gerdy's tubercle, a bony tunnel is created from the anterior to Gerdy's tubercle to the goose foot, and the traction suture is passed through this bony tunnel to form a Loop structure at 20° of knee flexion. Between March 2021 and May 2022 IKDC score, Lysholm score, and Tegner score were performed preoperatively and 6-12 months postoperatively in 24 consecutive patients who met the indications for this procedure and underwent surgery. The patient's maximum flexion angle, the circumference of the thigh, and the stress X-ray between the operated and healthy knee were measured. RESULTS: Patients showed significant improvement in IKDC score, Lysholm score and Tegner score at a mean follow-up of 7 months postoperatively compared to preoperatively. No significant increase in anterior tibial displacement was found between the patient's operated side and the healthy side. CONCLUSION: The Loop technique ACLR combined with ALSA can be used in patients with an ACL tear combined with a high degree of positive pivot shift. The patient's subjective perception was significantly improved from the preoperative period and knee stability was restored. LEVEL OF EVIDENCE: IV, therapeutic study.

Substantive molecular and histological changes within the meniscus with tears.

BACKGROUND: The meniscus plays a vital role in the normal biomechanics of the knee. However, it is not well studied at the molecular level. The purpose of this study was to determine whether molecular and pathological changes in the meniscal tissue vary depending on the presence or absence of meniscal and/or anterior cruciate ligament tear (ACL). METHODS: Six normal menisci (group A), seven simple torn menisci (group B) and seven torn menisci with concomitant anterior cruciate ligament tears (group C) were collected. We observed the pathological changes in the menisci and used real-time polymerase chain reaction along with immunohistochemistry and in situ hybridisation to examine the levels of ACAN, ADAMTS5, COL10A1, CEBPß, MMP13 and miR-381-3p, miR-455-3p, miR-193b-3p, miR-92a-3p, respectively. Patients were scored preoperatively and postoperatively using the Lysholm Knee Scoring Scale and International Knee Documentation Committee Subjective Knee Evaluation Form. RESULTS: Compared with group A, the expression levels of ADAMTS5, COL10A1, CEBPß, and MMP13 and all the miRNAs were increased while ACAN was down-regulated in groups B and C. Additionally, the gene expression and miRNA levels were higher in group C than that in group B, except for ACAN, which was lower. Several fibrochondrocytes strongly expressed ADAMTS5, CEBPß, and MMP13 in groups B and C and had high levels of miR-381-3p and miR-455-3p than that in group A. Postoperative Lysholm and IKDC scores were higher in group B than in group C. CONCLUSIONS: Our findings suggest that the meniscus tended to degenerate after it was injured, especially when combined with a torn ACL. The miRNAs investigated in this study might also contribute to meniscus degeneration. Patients with a combined injury patterns might have relatively worse joint function.

MicroRNA-193b-3p regulates matrix metalloproteinase 19 expression in interleukin-1ß-induced human chondrocytes.

Micro(mi)RNAs are small, non-coding RNA molecules known to play a significant role in osteoarthritis (OA) initiation and development, and similar to matrix metalloproteinases (MMPs), they participate in cartilage degeneration and cleave multiple extracellular matrices. The aim of this study was to determine whether the expression of MMP-19 in interleukin (IL)-1ß-induced human chondrocytes is directly regulated by miR-193b-3p. Expression levels of miR-193b-3p and MMP-19 in normal and osteoarthritis (OA) human cartilage, and interleukin-1 ß (IL-1ß)-induced human chondrocytes were determined by real-time polymerase chain reaction. Additionally, expression level of MMP-19 in IL-1ß-induced human chondrocytes was estimated by Western blotting and immunohistochemistry analyses. The effect of miR-193b-3p on MMP-19 expression was evaluated using transient transfection of normal human chondrocytes with miR-193b-3p mimic or its antisense inhibitor (miR-193b-3p inhibitor), and siMMP-19. The putative binding site of miR-193b-3p in the 3'-untranslated region (UTR) of MMP-19 mRNA was validated by luciferase reporter assay. miR-193b-3p expression was reduced in OA cartilage compared to that in normal chondrocytes, while the opposite was observed for MMP-19. Upregulation of MMP-19 expression was correlated with downregulation of miR-193b-3p in IL-1ß-stimulated normal chondrocytes. Increase in miR-193b-3p levels was associated with silencing of MMP-19. Overexpression of miR-193b-3p suppressed the activity of the reporter construct containing the 3'-UTR of human MMP-19 mRNA and inhibited the IL-1ß-induced expression of MMP-19 and iNOS in chondrocytes, while treatment with miR-193b-3p inhibitor enhanced MMP-19 expression. MiR-193b-3p is an important regulator of MMP-19 in human chondrocytes and may relieve the inflammatory response in OA.

tRNA-derived fragment 3031B regulates human anterior cruciate ligament cell proliferation and survival by targeting RELA.

tRNA-derived fragments (tRFs) are novel short noncoding RNAs that play pivotal roles in cell proliferation and survival. However, knowledge of the biological roles of tRFs in anterior cruciate ligament (ACL) cells is limited. Here, we intended to investigate the function of tRF-3031B in ACL cell. We used the tRF and tiRNA array to analyze tRF and tiRNA expression profiles in osteoarthritis (OA) ACL cells and normal ACL cells, and qRT-PCR and fluorescence in situ hybridization (FISH) were used to determine tRF-3031B expression. The results showed that tRF-3031B was expressed at low levels in OA ACL and Interleukin-1ß (IL-1ß) treated ACL cells. We found that RELA was the target of tRF-3031B. When ACL cells were transfected with tRF-3031B mimics, RELA expression was suppressed, whereas transfection with tRF-3031B inhibitors had the opposite effect. The rescue and dual-luciferase reporter assays showed that tRF-3031B silenced the RELA expression by binding to its untranslated region (3'-UTR). Hence, this study showed the novel function of tRF-3031B in regulating ACL cell proliferation and survival by targeting RELA, and these findings may offer a new direction for the study of ACL degeneration and pathophysiological of OA.

miRNA expression profile during osteogenic differentiation of human adipose-derived stem cells.

Human adipose-derived stem cells (hADSC) are capable of differentiating into an osteogenic lineage. It is believed that microRNAs (miRNAs) play important roles in regulating this osteogenic differentiation of human adipose-derived cells, although its molecular mechanism remains unclear. We investigated the miRNA expression profile during osteogenic differentiation of hADSCs, and assessed the roles of involved miRNAs during the osteogenic differentiation. We obtained and cultured human adipose-derived stems cells from donors who underwent elective liposuction or other abdominal surgery at our institution. miRNA expression profiles pre- and post-osteogenic induction were obtained using microarray essay, and differently expressed miRNAs were verified using quantitative real-time polymerase chain reaction (qRT-PCR). The expression of osteogenic proteins was detected using an enzyme-linked immunosorbent assay. Putative targets of the miRNAs were predicted using online software MiRanda, TargetScan, and miRBase. Eight miRNAs were found differently expressed pre- and post-osteogenic induction, among which four miRNAs (miR-17, miR-20a, miR-20b, and miR-106a) were up-regulated and four miRNAs (miR-31, miR-125a-5p, miR-125b, and miR-193a) were down-regulated. qRT-PCR analysis further confirmed the results. Predicted target genes of the differentially expressed miRNAs based on the overlap from three public prediction algorithms: MiRanda, TargetScan, and miRBase Target have the known functions of regulating stem cell osteogenic differentiation, self-renewal, signal transduction, and cell cycle control. We identified a group of miRNAs that may play important roles in regulating hADSC cell differentiation toward an osteoblast lineage. Further study of these miRNAs may elucidate the mechanism of hADSC differentiation into adipose tissue, and thus provide basis for tissue engineering.

The influence of body mass index on life quality and clinical improvement after total hip arthroplasty.

BACKGROUND: The effect of body mass index (BMI) on the outcome of total hip arthroplasty (THA) remains controversial. The purpose of this study was to examine whether revision rate and postoperative outcomes following THA were influenced by BMI. MATERIALS AND METHODS: We retrospectively evaluated 714 patients (751 hips) who underwent primary THA in our department between March 1991 and April 2006. They were followed prospectively for 5-20 years with 24 deaths (24 hips) and 33 losses (34 hips). Patients were separated into three groups based on BMI: underweight, normal and obese groups. A survival analysis was performed using revision as the endpoint, and a case-matched study that was matched for age, gender, and laterality was designed; outcomes were assessed with the Harris Hip score, 36-item short-form health survey, complication rate and radiological examination. RESULTS: The preoperative scores were lower for the obese group, and the postoperative scores were higher for the normal group. Patients in the obese group obtained the greatest overall improvement in clinical scores from admission to the last follow-up. We found a significantly higher complication rate in the obese group and underweight group. It appears that being underweight was associated with an increased dislocation rate, and obese patients were more likely to have osteolysis, deep vein embolism, and pulmonary thrombosis. The log rank test for survival showed no significant differences among the three groups. CONCLUSIONS: Abnormal BMI does not prevent functional rehabilitation after THA; however, patients with abnormal BMI have to face higher complication rates and poorer clinical outcomes following this operation.

Hyaluronan size alters chondrogenesis of mesenchymal stem cells cultured on tricalcium phosphate-collagen-hyaluronan scaffolds.

Hyaluronan (HA) provides a favorable environment for chondrogenesis of bone marrow mesenchymal stem cells (BMSCs). A previous report from our group indicated that addition of HA increases the chondro-inductive capacity of scaffolds. Therefore, this study aimed to investigate whether the Mw of the HA could affect chondrogenesis of BMSCs seeded on TCP-COL-HA scaffolds. Human BMSCs (hBMSCs) and rabbit BMSCs (rBMSCs) were isolated and expanded. TCP-COL scaffolds and TCP-COL-HA scaffolds with two different HA Mws were assessed for their capacity to induce cartilage regeneration from hBMSCs in vitro and in vivo. The results showed that about 96.96% of hBMSCs expressed CD44. Moreover, Hyal-1 and chondrogenic marker genes expressions were increased in hMSCs seeded on TCP-COL-HA scaffolds, and blocking the HA-CD44 interaction with an anti-CD44 antibody reduced the expression levels of Hyal-1 and chondrogenic marker genes. Additionally, TCP-COL-HA scaffolds with 2000 kDa Mw showed greater induction of BMSC chondrogenesis induction compared with those with 80 kDa Mw. Similar results were observed in an ectopic implantation nude mouse model. In a rabbit osteochondral defect repair model, rBMSCs seeded on TCP-COL-HA scaffolds with 2000 kDa Mw showed greater cartilage regeneration than those seeded with 80 kDa Mw. In addition, hBMSC-seeded TCP-COL-HA scaffolds with 2000 kDa Mw showed a significantly higher mechanical strength than those with 80 kDa Mw. Collectively, these results indicate that the Mw of HA could affect chondrogenesis of BMSCs seeded on TCP-COL-HA scaffolds. The TCP-COL-HA scaffolds might be used as allogenic off the shelf products in cartilage tissue engineering in future.

tRNA-derived fragment TRF365 regulates the metabolism of anterior cruciate ligament cells by targeting IKBKB.

tRNA-derived fragments (tRFs) are new noncoding RNAs, and recent studies have shown that tRNAs and tRFs have important functions in cell metabolism via posttranscriptional regulation of gene expression. However, whether tRFs regulate cellular metabolism of the anterior cruciate ligament (ACL) remains elusive. The aim of this study was to investigate the role and action mechanism of tRFs in ACL cell metabolism. A tRF array was used to determine tRF expression profiles in different human ACL cells, and quantitative real-time polymerase chain reaction and fluorescence in situ hybridisation were used to determine TRF365 expression. ACL cells were transfected with a TRF365 mimic or a TRF365 inhibitor to determine whether TRF365 regulates IKBKB expression. A rescue experiment and dual-luciferase reporter assay were conducted to determine whether the 3'-untranslated region (UTR) of IKBKB has a TRF365-binding site. TRF365 was weakly expressed in osteoarthritis (OA) ACL and interleukin-1ß-treated ACL cells. IKBKB was highly expressed in OA ACL and interleukin-1ß-treated ACL cells; transfection with the TRF365 mimic suppressed IKBKB expression, whereas transfection with the TRF365 inhibitor had the opposite effect. A dual-luciferase reporter assay showed that TRF365 silenced the expression of IKBKB by binding to its 3'-UTR. Thus, TRF365 regulates the metabolism of ACL cells by targeting IKBKB. In summary, TRF365 may provide a new direction for the study of ACL degeneration and on the pathophysiological process of OA.

[Clinical analysis of revision after primary hip replacement in the early stage].

OBJECTIVES: To analyze the reason of revisions no more than 5 years after primary hip replacement, and to discuss the methods how to prevent and manage. METHODS: Retrospectively review 11 cases with revision no more than 5 years after primary total hip replacement from January 2002 to June 2007. The reasons for revision were as follows: 2 cases were recurrent dislocation due to malposition of acetabular prosthesis; 5 cases were loosening of acetabular prosthesis; 1 case was abrasion of the native acetabulum by bipolar femoral head; 2 cases were periprosthetic femoral fractures and 1 case was periprosthetic infection. The average follow-up time was 36 months. Each patient was assessed according to Harris hip score. The revision procedures including liner only, acetabular prosthesis only, or both acetabular prosthesis and femoral prosthesis depending on the reasons for revision, two-stage revision was performed on 1 case with periprosthetic infection. RESULTS: The average of Harris hip score was increased from 46 (28 to 62) preoperatively to 86 (75 to 96) at follow up. The complication occurred in 2 cases: one was postoperative haematoma formation who was performed further surgery for clearance of haematoma, another was slight instability of the hip joint who was accepted skin traction for 3 weeks. CONCLUSIONS: The main reason for revision after primary total hip replacement is related to uncorrected insert of acetabular prosthesis. Improving surgical technique of insert of acetabular prosthesis is important in primary total hip replacement.

[Comparison of hemostatic effect and safety in primary unilateral total hip arthroplasty receiving different anticoagulants after anti-fibrinolysis with tranexamic acid].

OBJECTIVE: To compare the hemostatic effect and safety in primary unilateral total hip arthroplasty (THA) receiving nadroparin calcium, enoxaparin sodium, rivaroxaban, or apixaban after anti-fibrinolysis with tranexamic acid (TXA) and explore the best anticoagulant. METHODS: A retrospective analysis was conducted on 184 patients who underwent the primary unilateral THA between January 2014 and December 2018, administrated 15 mg/kg TXA before surgery and received nadroparin calcium, enoxaparin sodium, rivaroxaban, or apixaban. The patients were divided into four groups based on the different anticoagulants: 46 patients received nadroparin calcium; 45 patients received enoxaparin sodium; 47 patients received rivaroxaban; the other 46 patients received apixaban. There was no significant difference in age, gender, body mass, body mass index, the types of hip joint diseases, complications, anesthesia mode, operation time, and preoperative laboratory indexes (hemoglobin, hematocrit, platelet, prothrombin time, activated partial prothrombin time, blood volume) ( P>0.05). Perioperative blood data (total blood loss, hidden blood loss, dominant blood loss, postoperative drainage volume, maximum loss of hemoglobin, and blood transfusion rate) and complications (incision, bleeding, and thrombosis) were recorded and compared between groups. RESULTS: There was no significant difference in total blood loss, hidden blood loss, dominant blood loss, postoperative drainage volume, maximum loss of hemoglobin, and blood transfusion rate between groups ( P>0.05). The comparison of postoperative complications showed that 1 case (2.1%) of redness and swelling of incision occurred in the rivaroxaban group, and 1 case (2.2%) of the other 3 groups each had poor incision healing. No incision infection, fat liquefaction, or other incision complications occurred in the 4 groups. There was no significant difference in incision complication between groups ( P>0.05). There were 2 cases (4.3%) bleeding events (1 case of right inguinal hematoma and 1 case of subcutaneous ecchymosis in front of left leg) in the nadroparin calcium group, while no bleeding event occurred in the other 3 groups, which had no significant difference in bleeding complication between groups ( χ 2=5.612, P=0.132). There was 1 case (2.2%) of intermuscular vein thrombosis of the lower extremity in the nadroparin calcium group and no case in the other 3 groups, which had no significant difference between groups ( χ 2=2.789, P=0.425). Neither deep venous thrombosis nor pulmonary embolism occurred in any group. CONCLUSION: No significant difference in the hemostatic effect and incidences of complications for patients underwent primary unilateral THA receiving nadroparin calcium, enoxaparin sodium, rivaroxaban, or apixaban after anti-fibrinolysis with TXA. One of the four anticoagulants can be selected to prevent thrombosis after anti-fibrinolysis with TXA, which has certain safety.

[Analysis of hemostatic effect and safety of tranexamic acid in primary simultaneous bilateral total hip arthroplasty].

OBJECTIVE: To explore the effect of tranexamic acid (TXA) on the transfusion rate, dominant blood loss, and postoperative complications in simultaneous bilateral total hip arthroplasty (SBTHA). METHODS: A clinical data of 72 patients who underwent the primary SBTHA between January 2010 and December 2018 was retrospectively analyzed. A single dose of 15 mg/kg TXA was administered intravenously before 5-10 minutes of operation in 48 patients of trial group and 24 patients were not treated with TXA in the control group. There was no significant difference between the two groups ( P>0.05) in the gender, age, body mass index, the type of disease, American Society of Anesthesiologists (ASA) grading, comorbidity, and preoperative hospital stay, hemoglobin, hematocrit, platelet count, coagulation function tests. The operation time, intraoperative blood loss, and postoperative transfusion rate, dominant blood loss, complication, and hospital stay were recorded and compared between the two groups. RESULTS: The median operation time of the trial group was 208.0 minutes, and that of the control group was 202.5 minutes, with no significant difference ( Z=-1.046, P=0.295). Postoperative transfusion was performed in 26 patients (54.2%) in the trial group and 21 patients (87.5%) in the control group, and the difference of transfusion rate between the two groups was significant ( χ 2 =7.843, P=0.005). However, there was no significant difference in the amount of transfused suspended red blood cells and plasma between the two groups ( P>0.05). The median intraoperative blood loss was 550 mL in the trial group and 600 mL in the control group, with no significant difference ( Z=-1.378, P=0.168). The postoperative drainage volume and median dominant blood loss in the trial group were (542±269) and 1 050 mL, respectively, which were significantly lower than those in the control group [(710±316) and 1 270 mL] ( P<0.05). There was 1 case of skin tension blisters around the incision, 1 case of lower limb numbness and muscle strength loss, and 1 case of lacunar cerebral infarction in the trial group, while in the control group, there was 1 case of skin ecchymosis around the incision and 1 case of bilateral lower limb numbness and muscle strength loss, which showed no significant difference in the incidences of complications ( P>0.05). No pulmonary embolism or deep venous thrombosis was found in the two groups. The median postoperative hospital stay and median total hospital stay were 9.0 and 13.0 days in the trial group, while 9.0 and 13.0 days in the control group, respectively, with no significant difference ( P>0.05). CONCLUSION: For patients who are treated with the primary SBTHA, TXA can reduce transfusion rate and perioperative dominant blood loss, and has a good hemostatic effect without increasing complications of incision, pulmonary embolism, deep venous thrombosis, and hospital stay. Therefore, TXA is relative safe.

Effect of 3 different anticoagulants on hidden blood loss during total hip arthroplasty after tranexamic acid.

Comparison of different anticoagulants in blood management and complications with tranexamic acid (TXA) in total hip arthroplasty (THA) is unclear. Our aim was to compare the efficacy and safety among receiving nadroparin calcium, enoxaparin sodium or rivaroxaban after TXA in THA.150 patients undergoing primary unilateral THA were received 15 mg/kg intravenous TXA (IV-TXA) before skin incision, followed by 1 of nadroparin calcium (Group A), enoxaparin sodium (Group B), or rivaroxaban (Group C) randomly during hospitalization. The primary outcome was hidden blood loss (HBL). Other outcomes such as the maximum hemoglobin (Hb) drop, total blood loss (TBL), the volume of drainage, transfusion rate, length of hospital stay (LOS), and complications were also compared.There were no statistically significant differences in HBL, the maximum hemoglobin (Hb) drop, transfusion rate, and complications among 3 groups. LOS was significantly higher for patients in Group B than Group A (P = .026). Neither deep venous thrombosis (DVT) nor pulmonary embolism (PE) occurred in any group.There were no differences in efficacy and safety in patients undergoing THA receiving nadroparin calcium, enoxaparin sodium, or rivaroxaban after anti-fibrinolysis with TXA.

Primary non-Hodgkin's lymphoma of the spine with neurologic compression treated by radiotherapy and chemotherapy alone or combined with surgical decompression.

Neurologic compression is a disastrous consequence for the patients with primary non-Hodgkin's lymphoma (NHL) of the spine, and such a condition has not been carefully taken into account in the treatment guidelines. The aim of this study was to compare the effect of radiotherapy and chemotherapy alone or combined with surgical decompression on primary NHL of the spine with neurologic compression. Sixteen patients with primary NHL in the vertebrae of the spine were treated between 1994 and 2006. Thirteen patients had neurologic compression. The neurologic deficits in 11 patients involved soft tissue extension from the vertebral tumors and 3 had vertebral fractures with motor signs and 5 had radicular pain. Five patients were treated by radiotherapy and chemotherapy alone while 8 were combined with surgical decompression. The decompression operation for tumors resulted in neurologic recovery in 6 patients. Five patients were not operated on but three received emergent radiotherapy before chemotherapy, 4 of whom had complete recovery in their neurologic symptoms. Of all patients, 3 relapsed. At average follow-up of 61.5 months (range 2-156 months), 4 patients had died after an average interval of 23.3 months from treatment (range 3-71 months). The 5-year overall survival rate was 82% with 60% for the patients in the surgical group, 100% for the patients in the non-surgical group. There was no difference between the groups (chi(2)=3.559, P=0.059). The 5-year overall survival was 100% for the 8 patients who completed CHOP chemotherapy and radiotherapy. It appears that optimum treatment in these patients depends on the cause of the neurologic deficits, whereas the survival is not influenced by the surgical or non-surgical treatment. The results suggest that chemotherapy and radiotherapy alone is the ideal treatment for these patients whose neurologic compression was only due to soft tissue extension. The authors emphasize the importance of chemotherapy and radiotherapy followed by surgical decompression depending on individual priorities in the indications for operation on primary NHL of spine with neurologic compression.

Interferonalpha enhances etoposide-induced apoptosis in human osteosarcoma U2OS cells by a p53-dependent pathway.

Interferonalpha (IFNalpha) induces cell cycle arrest and triggers apoptosis and chemosensitivity. But the mechanism of IFNalpha in regulating chemosensitivity has not been fully understood. To study whether IFNalpha affected chemosensitivity of osteosarcoma cells, we treated p53-wild U2OS cells and p53-mutant MG63 cells with IFNalpha and etoposide, alone or in combination, and then examined growth inhibition, cell cycle arrest and apoptosis. IFNalpha enhanced etoposide-induced growth inhibition and apoptosis in p53-wild U2OS cells but not p53-mutant MG63 cells in a dose- and time-dependent manner. Etoposide-induced G2/M phase arrest was also enhanced by IFNalpha. The enhanced apoptosis was associated with the accumulation of transcriptionally active p53 accompanied with increased Bax and Mdm2, as well as decreased Bcl-2. IFNalpha also activated caspases-3, -8 and -9 protein kinases and PARP cleavage in response to etoposide in U2OS cells. Moreover, the combination-induced cytotoxicity and PARP cleavage were significantly reduced by caspase pan inhibitor and p53 siRNA. Thus we conclude that IFNalpha enhances etoposide-induced apoptosis in human osteosarcoma U2OS cells by a p53-dependent and caspase-activation pathway. The proper combination of IFNalpha and conventional chemotherapeutic agents may be a rational strategy for the treatment of human osteosarcoma with functional p53.

MicroRNA-193b-3p regulates chondrogenesis and chondrocyte metabolism by targeting HDAC3.

Histone deacetylase 3 (HDAC3) plays a pivotal role in the repression of cartilage-specific gene expression in human chondrocytes. The aim of this study was to determine whether microRNA-193b-3p (miR-193b-3p) regulates the expression of HDAC3 during chondrogenesis and chondrocyte metabolism. Methods: miR-193b-3p expression was assessed in a human mesenchymal stem cell (hMSC) model of chondrogenesis, in interleukin-1ß (IL-1ß)-treated primary human chondrocytes (PHCs), and in non-degraded and degraded cartilage. hMSCs and PHCs were transfected with miR-193b-3p or its antisense inhibitor. A direct interaction between miR-193b-3p and its putative binding site in the 3'-untranslated region (3'-UTR) of HDAC3 mRNA was confirmed by performing luciferase reporter assays. Chondrocytes were transfected with miR-193b-3p before performing a chromatin immunoprecipitation assay with an anti-acetylated histone H3 antibody. To investigate miR-193b-3p-transfected PHCs in vivo, they were seeded in tricalcium phosphate-collagen-hyaluronate (TCP-COL-HA) scaffolds, which were then implanted in nude mice. In addition, plasma exosomal miR-193b-3p in samples from normal controls and patients with osteoarthritis (OA) were measured. Results: miR-193b-3p expression was elevated in chondrogenic and hypertrophic hMSCs, while expression was significantly reduced in degraded cartilage compared to non-degraded cartilage. In addition, miR-193b-3p suppressed the activity of reporter constructs containing the 3'-UTR of HDAC3, inhibited HDAC3 expression, and promoted histone H3 acetylation in the COL2A1, AGGRECAN, COMP, and SOX9 promoters. Treatment with the HDAC inhibitor trichostatin A (TSA) increased cartilage-specific gene expression and enhanced hMSCs chondrogenesis. TSA also increased AGGRECAN expression and decreased MMP13 expression in IL-1ß-treated PHCs. Further, 8 weeks after implanting PHC-seeded TCP-COL-HA scaffolds subcutaneously in nude mice, we found that miR-193b overexpression strongly enhanced in vivo cartilage formation compared to that found under control conditions. We also found that patients with OA had lower plasma exosomal miR-193b levels than control subjects. Conclusions: These findings indicate that miR-193b-3p directly targets HDAC3, promotes H3 acetylation, and regulates hMSC chondrogenesis and metabolism in PHCs.

Better Prognosis of Senile Patients with Intertrochanteric Femoral Fracture by Treatment with Open Reduction Internal Fixation than by Hip Arthroplasty.

PURPOSE: To compare the postoperative survival and mortality rates in intertrochanteric femoral fracture (IFF) patients who underwent either open reduction internal fixation (ORIF) or hip arthroplasty. METHODS: Clinical data from senior patients who had IFF and underwent ORIF or hip arthroplasty were analyzed retrospectively. Survival curves were compared between groups with Kaplan-Meier method and log-rank test. Significant independent prognostic factors were identified by Cox multivariate regression analysis. RESULTS: All patients recovered fully post-surgery. Although 31 patients died during the follow-up period (ORIF, mean 45.4 months; arthroplasty, mean 51.6 months), mortality rate did not differ significantly between the groups. The 1-yr and 2-yr survival rate estimates for the ORIF group were 92.2%, and 86%, respectively; they were 85% and 74% for the arthroplasty group. Average survival lengths for ORIF and arthroplasty groups were 88 and 67 months, respectively. The effect of surgical approaches on survival differed significantly (log-rank test c2 = 6.402, p = 0.011). Multivariate Cox regression model indicated that surgical choice (p = 0.036) was a significant independent risk factor for the prognosis of senile IFF, even with adjustment for age (p = 0.002). CONCLUSION: The overall postoperative prognosis was superior in senile IFF patients treated with ORIF.

P14ARF sensitizes human osteosarcoma cells to cisplatin-induced apoptosis in a p53-independent manner.

The tumor suppressor p14ARF, encoded by the INK4a/ARF locus, is often disrupted in human cancers, p14ARF triggers cell cycle arrest and sensitizes cells to apoptosis in the presence of collateral signals. To investigate the role of p14ARF in chemotherapeutic drugs-induced apoptosis, p14ARF was overexpressed by stable transfection in human osteosarcoma cell lines, U2OS (p53-wt/p14ARF-null) and MG63 (p53-mt/p14ARF-null). The results showed that ectopic p14ARF sensitized both cell lines to cisplatin-induced cytotoxicity and apoptosis. This sensitization of cisplatin-induced apoptosis was associated with upregulation of p53, Bax and p21 in U2OS cells. Conversely, such a result was not observed in MG63 cells. Moreover, the sensitization of cisplatin-induced cytotoxicity in U2OS cells was unaltered by p53 siRNA. Together, we show here p14ARF sensitizes human osteosarcoma cells to cisplatin-induced apoptosis in a p53-independent manner. Proper combinations of p14ARF gene transfer and conventional chemotherapy may be a valuable strategy in human osteosarcoma treatment.

Biological character of human adipose-derived adult stem cells and influence of donor age on cell replication in culture.

To investigate the biological character of human adipose-derived adult stem cells (hADAS cells) when cultured in vitro and the relationship between hADAS cell's replication activity and the donor's age factor, and to assess the stem cells as a new source for tissue engineering. hADAS cells are isolated from human adipose tissue of different age groups (from adolescents to olds: <20 years old, 21-40 years old, 41-60 years old and >61 years old groups). The protein markers (CD29, CD34, CD44, CD45, CD49d, HLA-DR, CD106) of hADAS cells were detected by flow cytometry (FCM) to identify the stem cell, and the cell cycle was examined for P20 hADAS cells to evaluate the safety of the subculture in vitro. The generative activity of hADAS cells in different age groups was also examined by MTT method. The formula "TD = t x log2/logNt - logN0" was used to get the time doubling (TD) of the cells. The results showed that the cells kept heredity stabilization by chromosome analysis for at least 20 passages. The TD of these cells increased progressively by ageing, and the TD of the <20 years old group was lower than that of the >61 years old group (statistical analysis of variance (ANOVA), P=0.002, P<0.05). These findings suggested that a higher level of hADAS cells replication activity was found in the younger donators, and they represent novel and valuable seed cells for studies of tissue engineering.

[EXPRESSIONS OF CARTILAGE DEGENERATIVE RELATED GENES AND microRNAs IN TORN MENISCUS].

OBJECTIVE: To investigate the expressions of cartilage degenerative related genes in meniscus, and to evaluate the potential effect of meniscal damage on cartilage degeneration, and to analyze the relationship between microRNAs (miRNAs) expression and cartilage degeneration. METHODS: Meniscal tissue was collected from 5 patients undergoing partial meniscectomy between September 2012 and October 2013 (experimental group), and normally meniscal tissue without tearing from amputees was used as controls (control group). Pathological changes of menisci were observed; and real-time fluorescent quatitative PCR was performed to examine the relative expression levels of cartilage degenerative related genes and miRNAs: Aggrecan (ACAN), type X collagen (COL10A1), matrix metalloproteinases 13 (MMP-13), CCAAT enhancer binding protein ß (CEBP-ß), a disintegrin and metalloproteinase with thrombospondinmotif 5 (ADAMTS-5), miR-193b, miR-92a, and miR-455-3p in meniscus. RESULTS: There were varying degrees of degenerative pathological changes in torn meniscus of experimental group. Compared with normal meniscus of control group, the expression of ACAN was decreased, while the expressions of COL10A1, CEBP-ß, ADAMTS-5, and MMP-13 were increased in torn meniscus of experimental group; and significant difference was found (P < 0.05) except ACAN and MMP-13 (P > 0.05). The expressions of miR-92a, miR-455-3p, and miR-193b in torn meniscus of experimental group were significantly higher than those in normal meniscus of control group (P < 0.05). CONCLUSION: Meniscal tissue has the intrinsic tendency of degeration after meniscus tear. The torn meniscus has greater stimulative impact on cartilage degeneration than normally morphological meniscus without tearing. The cartilage degenerative related miRNAs, including miR-193b, miR-92a, and miR-455-3p may contribute to the up-regulation of osteoarthritis.

MiR-193b regulates early chondrogenesis by inhibiting the TGF-beta2 signaling pathway.

Cartilage generation and degradation are regulated by miRNAs. Our previous study has shown altered expression of miR-193b in chondrogenic human adipose-derived mesenchymal stem cells (hADSCs). In the current study, we investigated the role of miR-193b in chondrogenesis and cartilage degradation. Luciferase reporter assays showed that miR-193b targeted seed sequences of the TGFB2 and TGFBR3 3'-UTRs. MiR-193b suppressed the expression of early chondrogenic markers in chondrogenic ATDC5 cells, and TNF-alpha expression in IL-1b-induced PMCs. In conclusion, MiR-193b may inhibit early chondrogenesis by targeting TGFB2 and TGFBR3, and may regulate inflammation by repressing TNF-alpha expression in inflamed chondrocytes.