Immune response and effect of adenovirus-mediated human BMP-2 gene transfer on the repair of segmental tibial bone defects in goats

BACKGROUND: Tissue-engineered bone may be used for filling bone defects. There are, however, no reports on this technique used in large animals. METHODS: We evaluated the effectiveness of, and immune response in repairing diaphyseal bone defects by gene transfer using bone morphogenetic proteins (BMPs). We used adenovirus-mediated human BMP-2 (Adv-hBMP-2) gene-transduced bone marrow stromal cells (BMSCs) to repair 2.1-cm segmental tibial bone defects in goats (group I, n = 7). An Adv-ssgal-transduced BMSC group (group II, n = 5), a non-transduced BMSC group (group III, n = 5), and an untreated group (group IV, n = 2) were used as controls. Self-secreted extracellular matrix was used as cellular carrier. RESULTS: Radiographic and histomorphometric examination demonstrated more callus in the bone defects of group I compared to other groups.Week 24 after implantation, the defect healing rates of groups I, II, III, and IV were 6/7, 1/5, 2/5, and 0/2, respectively. The maximum compressive strength of new tissue in the bone defects of group I was higher than those of groups II and III. Temporary cellular and persistent humoral immune responses against adenovirus were detected after hBMP-2 gene transfer. INTERPRETATION: We found that Adv-hBMP-2 genetransduced BMSCs had superior osteoinductivity in the repair of tibial bone defects in goats, but it could cause temporary cellular and persistent humoral immune responses against adenovirus.     

腺病毒介导的人骨形成蛋白2基因治疗的免疫学研究

目的评价机体对腺病毒介导的人骨形成蛋白2(adenovirusmediatedhumanbonemorphogeneticprotein2,Ad-hBMP-2)基因治疗的免疫学反应。方法崇明山羊12只,手术截去右侧胫骨中段2.1cm,制备胫骨干缺损模型,随机分为2组。将Ad-hBMP-2转染的山羊骨髓间充质干细胞(marrowmesenchymalstemcells,MSCs,转染组,n=7)及未转染的MSCs(未转染组,n=5)分别植入骨缺损内。于术后4、8、16及24周摄X线片检查骨缺损愈合情况,并对治疗前后机体对腺病毒的细胞和体液免疫反应进行检测。结果X线片示,转染组4～8周,骨缺损内有连续性骨痂形成;24周,有6侧骨缺损完全愈合,部分骨髓腔再通。未转染组4～8周,骨缺损内骨痂形成较少,与骨端界限清楚;24周,仅2侧骨缺损愈合。淋巴细胞与MSCs混合培养示淋巴细胞刺激指数(stimulationindex,SI)于植入后14d升高,转染组4.213±1.278,未转染组-0.310±0.147,且差异有统计学意义(P<0.05);28d后下降,转染组2.544±0.957,未转染组3.104±0.644,差异无统计学意义(P>0.05)。治疗后14、28、49和120d,转染组的血浆腺病毒中和抗体滴度分别为2.359±0.226、2.297±0.200、2.214±0.215和2.297±0.210,未转染组分别为-0.175±0.335、-0.419±0.171、0±0.171和0.874±0.524,两组各时间点差异均有统计学意义(P<0.05)。结论腺病毒介导的基因治疗可引起机体对腺病毒的细胞及体液免疫反应,从而逐渐消除腺病毒基因和相关蛋白的影响。     

HA复合rhBMP-2转染的BMSCs对羊胫骨延长骨愈合的影响

目的 评价HA复合rhBMP-2的腺病毒(adenovirus mediated rhBMP-2,Adv-rhBMP-2)转染的羊BMSCs对骨痂延长骨愈合的影响. 方法 成年山羊19只,雌雄不限,体重15～20 kg.于每只羊髂嵴上穿刺抽取骨髓10 mL,常规传代培养BMSCs.取第3代BMSCs,以感染复数200转染腺病毒.取0.25%胰蛋白酶消化转染后3 d的细胞1×108个,与HA充分混匀制备Adv-rhBMP-2/BMSCs/HA.建立山羊右侧胫骨延长模型,术后立即于截骨部位注射自体细胞复合物.按术后注入物质不同随机分成4组A组为Adv-rhBMP-2/BMSCs/HA组(n=6),B组为Adv-rhBMP-2/BMSCs组(n=5),C组为Adv-β-gal/BMSCs/HA组(n=4),D组为空白组(n=4).术后第7天开始行胫骨延长,速度1 mm/d,共延长4周.术后5、8、12周摄X线片观察骨痂生长情况,12周处死动物,取标本分别行骨密度检测、生物力学测定、组织学观察和骨形态计量学分析. 结果 X线片检查示,术后5、8周A、B组骨痂生成量明显多于C、D组,X线片定量评分A、B组明显高于C、D组,差异均有统计学意义(P＜0.05);12周各组均在骨延长部位形成连续骨痂.骨密度测定术后12周A、B、C、D组延长部位骨矿物质含量分别为(4.175 ±1.921)、(2.600 ±0.638)、(2.425±0.826)和(1.175 ±0.574)g,骨矿物质密度分别为(0.612 ±0.196)、(0.630 ±0.159)、(0.450 ±0.166)和(0.266 ±0.113)g/cm2,A、B组显著高于C、D组(P＜0.05).生物力学测定A、B、C、D组最大载荷分别为(490.20 ±155.08)、(350.59±80.48)、(221.95±68.79)和(150.65±92.29)N,弹性模量为(178.24±105.80)、(105.88 ±27.09)、(81.18±48.67)和(50.35±47.64)Mpa,各指标A组显著高于C、D组(P＜0.05).组织学观察见A组骨延长处大量新生骨组织,骨小梁多为纵行网状排列.骨形态计量分析示A、B、C、D组新骨体积分别为72.35%±5.68%、67.58%±7.42%、49.63%±4.87%和38.87%±2.35%,A组新生骨生成量明显比D组多(P＜0.05). 结论 HA复合rhBMP-2修饰的BMSCs制备的组织工程骨可促进羊胫骨延长骨愈合.     

Repairing of goat Tibial Bone Defects with BMP-2 Gene–Modified Tissue-Engineered Bone

Bone defects larger than a critical size are major challenges in orthopedic medicine. We combined tissue-engineered bone and gene therapy to provide osteoprogenitor cells, osteoinductive factors, and osteoconductive carrier for ideal bone regeneration in critical-sized bone defects. Goat diaphyseal bone defects were repaired with tissue and genetically engineered bone implants, composed of biphasic calcined bone (BCB) and autologous bone marrow derived mesenchymal stem cells (BMSC) transduced with human bone morphogenetic protein-2 (hBMP-2). Twenty six goats with tibial bone defects were divided into groups receiving implants by using a combination of BCB and BMSCs with or without the hBMP-2 gene. In eight goats that were treated with BCB that contained hBMP-2 transduced BMSC, five had complete healing and three showed partial healing. Goats in other experimental groups had only slight or no healing. Furthermore, the area and biochemical strength of the callus in the bone defects were significantly better in animals treated with genetically engineered implants. We concluded that the combination of genetic and tissue engineering provides an innovative way for treating critical-sized bone defects.     

In vitro osteogenic differentiation and in vivo bone-forming capacity of human isogenic jaw periosteal cells and bone marrow stromal cells

OBJECTIVE: To compare the in vitro osteogenic differentiation and in vivo ectopic bone forming capacity of human bone marrow stromal cells (BMSCs) and jaw periosteal cells (JPCs), and to identify molecular predictors of their osteogenic capacity. SUMMARY BACKGROUND DATA: JPC could be an appealing alternative to BMSC for the engineering of cell-based osteoinductive grafts because of the relatively easy access to tissue with minimal morbidity. However, the extent of osteogenic capacity of JPC has not yet been established or compared with that of BMSC. METHODS: BMSCs and JPCs from the same donors (N = 9), expanded for 2 passages, were cultured for 3 weeks in osteogenic medium either in monolayers (Model I) or within 3-dimensional porous ceramic scaffolds, following embedding in fibrin gel (Model II). Cell-fibrin-ceramic constructs were also implanted ectopically in nude mice for 8 weeks (Model III). Cell differentiation in vitro was assessed biochemically and by real-time RT-PCR. Bone formation in vivo was quantified by computerized histomorphometry. RESULTS: JPCs had lower alkaline phosphatase activity, deposited smaller amounts of calcium (Model I), and expressed lower mRNA levels of bone sialoprotein, osteopontin, and osterix (Models I and II) than BMSCs. JPCs produced ectopic bone tissue at lower frequency and amounts (Model III) than BMSCs. Bone sialoprotein, osteopontin, and osterix mRNA levels by BMSCs or JPCs in Model II were markedly higher than in Model I and significantly more expressed by cells that generated bone tissue in Model III. CONCLUSIONS: Our data indicate that JPCs, although displaying features of osteogenic cells, would not be as reliable as BMSCs for cell-based bone tissue engineering, and suggest that expression of osteoblast-related markers in vitro could be used to predict whether cells would be osteoinductive in vivo.     

自体移植的大鼠骨髓间充质干细胞在脊髓损伤处的存活及分化

目的观察自体移植的骨髓间充质干细胞(BMSC)在脊髓损伤(SCI)处的存活及向神经细胞分化情况。方法将36只雄性SD大鼠随机分为PBS注射组(PBS组)和BMSC自体移植组(BMSC组)。BMSC组大鼠在术前均进行自体BMSC分离培养。2组大鼠均采用改良Allen撞击装置制备T9水平的SCI模型,PBS组于损伤处注射PBS,BMSC组注射第4代自体BMSC。于移植后2、3、5周采用Basso-Beattie-Bresnahan(BBB)评分法进行运动功能评分;移植后2、3、5周取损伤部位脊髓,采用Hoechst33342染色法观察移植细胞存活情况,同时行免疫荧光化学染色检测自体移植的BMSC中微管相关蛋白2(MAP-2)及胶质纤维酸性蛋白(GFAP)的表达情况。结果移植后2、3、5周,BMSC组大鼠BBB评分均高于PBS组,差异有统计学意义(P<0.05)。移植后2、3、5周,BMSC组大鼠损伤处脊髓有不同数量Hoechst33342标记细胞存活,其中2周时细胞存活较多,5周时细胞存活较少;移植后2、3周,未检测到移植细胞表达MAP-2及GFAP,5周时检测到部分移植细胞表达MAP-2和GFAP。结论SCI后立即进行移植的自体BMSC可在损伤处存活,部分存活细胞可向神经元和星形胶质细胞方向分化,促进大鼠后肢运动功能的恢复。     

富集骨髓基质干细胞复合多孔β-磷酸三钙促进骨缺损修复的实验研究

目的 评估富集骨髓基质干细胞(BMSCs)复合β-磷酸三钙(β-TCP)在促进骨缺损修复中的作用. 方法 抽取12只崇明山羊骨髓血,用梯度离心法分别将其中的BMSCs富集、分离后与β-TCP复合.12只山羊双侧股骨内髁钻孔,制备骨缺损模型.所有山羊左侧股骨内髁缺损通过富集BMSCs/β-TCP进行局部填充治疗(实验组),其中10只羊右侧股骨内髁缺损以单纯β-TCP颗粒填充治疗(对照组),2只羊右侧股骨内髁缺损旷置(空白组),评价富集效率.术后16周处死取材,将所得标本进行影像学观察和形态学计量,实验组和对照组之间进行比较. 结果 BMSCs富集后有核细胞数和碱件磷酸酶染色呈阳性的细胞集落形成单位(CFUs/ALP+)数均有显著提高,差异有统计学意义(P<0.05).X线侧位片和μ-CT形态计量分析显示实验组新生骨长入率(70.27%±8.47%)显著高于对照组(21.40%±5.77%),差异有统计学意义(t=11.97,P<0.05).X线侧位片、CT平扫与三维重建以及μ-CT图像均可见实验组疗效优于对照组. 结论 在促进骨缺损修复中,富集BMSCs/β-TCP的疗效优于单纯β-TCP颗粒,有良好的应用前景.     

Massive bone reconstruction with heat‐treated bone graft loaded autologous bone marrow‐derived stromal cells and β‐tricalcium phosphate composites in canine models

Bone marrow‐derived stromal cells (BMSCs) contain mesenchymal stem cells that are capable of forming various mesenchymal tissues. We hypothesized that BMSCs and β‐tricalcium phosphate (β‐TCP) composites would promote the remodeling of large‐sized autologous devitalized bone grafts; therefore, the aim of this study was to evaluate the effects of the composites on the remodeling of autologous devitalized bone grafts. Autologous BMSCs cultured in culture medium containing dexamethasone (10^?7 M) were loaded into porous β‐TCP granules under low‐pressure. Theses BMSC/TCP composites were put into the bone marrow cavity of autologous heat‐treated bone (femoral diaphysis, 65‐mm long, 100°C, 30 min) and put back to the harvest site. In the contralateral side, β‐TCP without BMSC were used in the same manner as the opposite side as the control. Treatment with the BMSC/TCP composites resulted in a significant increase in thickness, bone mineral density, and matured bone volume of the cortical bone at the center of the graft compared to the control. Histological analysis showed matured regenerated bone in the BMSC loaded group. These results indicate that BMSC/TCP composites facilitated bone regeneration and maturation at the graft site of large‐sized devitalized bone. This method could potentially be applied for clinical use in the reconstruction of large bone defects such as those associated with bone tumors. © 2013 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 31:1308–1316, 2013

     

Lycium barbarum polysaccharide protects against osteonecrosis of femoral head via regulating Runx2 expression

BackgroundOsteonecrosis of femoral head (ONFH) is a pathological state caused by lack of blood supply in femoral head. This study aimed to explore the function of Lycium barbarum polysaccharide (LBP), an antioxidant agent extracted from L. barbarum, on ONFH.MethodsOsteonecrosis rat model was generated using lipopolysaccharide (LPS) and methylprednisolone followed by examination of body weight, blood glucose, morphology, and BMSC osteoblast differentiation. The effect and underlying mechanism of LBP on the proliferation, apoptosis, and osteoblast differentiation of BMSC were determined with or without LPS or hypoxia treatment using CCK-8. Alizarin Red S staining, flow cytometry, and western blot, respectively.ResultLBP could protect against glucocorticoid-induced ONFH in rats, resulting in improved sparse trabecular bone, empty lacunae and bone cell coagulation. Moreover, LBP promoted the proliferation and osteoblast differentiation of bone mesenchymal-derived stem cells (BMSCs) in a dose-dependent manner. Furthermore, LBP enhanced osteoblast differentiation of BMSCs under hypoxia condition. Mechanistically, we found that LBP treatment enhanced Runx2 and ALP expression in BMSCs. LBP restored the expression of Runx2 and ALP under hypoxia, suggesting that LBP might be involved in regulating Runx2/ALP expression and contributed to osteoblast differentiation. Knockdown of Runx2 significantly inhibited BMSCs proliferation, while LBP treatment did not rescue the osteoblast differentiation ability of BMSCs with Runx2 knockdown.ConclusionOur findings suggested that LBP protects against ONFH via regulating Runx2 expression, which could be utilized to treat patients suffering ONFH.     

慢病毒介导骨形成蛋白-2基因转染骨髓间质干细胞修复大鼠颅骨骨缺损

目的 观察携带人骨形成蛋白-2(hBMP-2)基因的慢病毒(lentivirus)转染骨髓间质干细胞构建的组织工程化骨对大鼠颅骨极量骨缺损的修复效果. 方法 300～350g雄性 SD大鼠取股骨骨髓体外培养扩增骨髓间质干细胞(BMSC),以lentivirus载体介导hBMP-2基因转染BMSC,诱导其向成骨细胞分化.将细胞接种于经纤维连接蛋白修饰的β-磷酸三钙(β-TCP),共同培养10d.30只大鼠均造成颅骨极量缺损(直径8mm)模型,分为Lev- BMP2转染细胞+β-TCP组(A组:n=10);BMSC +β-TCP组(B组:n=10);β-TCP组(C组:n=10)于第4、8、12、20周从影像学及组织学角度观察比较颅骨缺损修复情况.结果 无论X线摄片及组织切片均表明,A组颅骨骨缺损修复优于同期的B组和C组.在各时间点A组和B组在植骨区均有新骨形成,而C组仅见边缘区成骨.术后20周新生骨小梁相对体积(TBV,%)A组为(60.83±11.49),B组为(40.37±9.02),C组为(18.37±6.02),差异有统计学意义(P<0.01). 结论 Lev- BMP2转染BMSC复合β-TCP构建的组织工程化骨对大鼠颅骨极量骨缺损有较好的修复效果.     

New nasal fracture classification for patients with silicone implants

Abstract

Aesthetic nasal augmentation has increased in popularity among Asian populations, and nasal bone fracture is the most common type of facial bone fracture. In Asia, the frequency of nasal bone fractures is also increasing among patients who have undergone silicone augmentation rhinoplasty. The increasing prevalence of this injury presents a challenge to the surgeon. Thirty-six patients who had previously undergone augmentation rhinoplasty with silicone implant presented with nasal bone fracture from June 2007 through December 2011. The patients were grouped into three categories: patients with fractures in the high level (type I), patients with fractures in the low level (type II), and patients with fractures throughout the entire nasal bone, from base to top (type III). The largest group comprised patients with type I fractures (n = 24, 67%), followed by type II (n = 4, 11%), and Type III (n = 8, 22%) fractures. Symptoms and surgical outcomes for nasal bone fractures may be different in patients with silicone implants. A novel classification system for nasal bone fractures is required, as is a new approach to diagnosis and treatment.     

Post-operative hip centre restoration and migration after impaction bone grafting in revision and complex primary hip arthroplasty

Introduction/objectives

Although impaction grafting proved efficacy in the reconstruction of acetabular defects in primary and revision hip arthroplasty, its role in large segmental defects is still debatable. Our objective is to determine hip centre restoration and last follow-up migration after acetabular reconstruction with impaction grafting in different types of acetabular defects.

Methods

This is a single-centre retrospective radiographic study of (107) total hip arthroplasty (42 primary and 65 revision) in (104) patients using impaction grafting. The available radiographs were examined for normal, preoperative, immediate postoperative, and last follow-up vertical (Y) and horizontal (X) hip centre. Maximum acetabular defect distance (MADD), presence, and size of the mesh were recorded.

Results

In type I and II AAOS defects, the post-operative hip centre was not significantly different from the normal hip centre on the contralateral healthy side. In type III defects, there was a significant variation between the normal hip centre and the post-operative hip centre (P value 0.034 and 0.001 for Y and X, respectively). At 44-month follow-up of 36 hips, 31 (86%) hips migrated. The mean migration ± SD was 5.72 ± 3.7, 2, 4.15 ± 1.2, and 11.26 ± 3.9 mm for types I, II, and III, respectively (P value 0.211). Hips with MADD > 15 mm, especially with large mesh sizes migrate significantly more (P value = 0.042, 0.037, and 0.039, respectively).

Conclusion

Hip centre restoration was better, and migration was less for type I and II AAOS rather than for type III. Other options for reconstruction should be considered.

     

Effects of Platelet‐Rich Plasma and Bone Marrow Mesenchymal Stem Cells on Meniscal Repair in the White‐White Zone of the Meniscus

ObjectiveTo investigate the role of autologous platelet‐rich plasma (PRP) on the repair of meniscal white‐white zone injury through promoting the proliferation of canine bone marrow‐derived mesenchymal stem cells (BMSCs).MethodsA total of 24 beagle dogs were selected to construct meniscal white‐white zone injury models in both lateral knee joints. All subjects were divided into four groups: control, BMSCs, PRP, and PRP + BMSCs. Immunohistochemistry was applied in the expression detection of type I and type II collagens. HE staining and methylene blue staining were performed to observe the injury of cartilage of lateral femoral condyle in each group. ELISA was used to detect the osteopontin (OPN) content in cartilage of lateral femoral condyle. HE staining and magnetic resonance imaging (MRI) were used to observe the healing of meniscus in each group. Outcome measures include the expression of OPN in the synovial fluid of knee joint, the expression of type I collagen and type II collagen, the healing of meniscus injury, and the damage degree of lateral femoral condyle cartilage.ResultsCompared with the control group, the expressions of type I and type II collagens were enhanced in the PRP group and the PRP + BMSCs group. Compared with 1 week before modeling, the expression of OPN was elevated in the control group and the BMSCs group at 3 weeks after modeling. There were no significant differences in the above indicators between the PRP group and the PRP + BMSCs group. According to MRI and pathological section after HE staining, meniscal healing in the PRP group and the PRP + BMSCs group was significantly improved as compared to that of the control group and the BMSCs group (all P < 0.05), and there was no significant difference between the PRP group and the PRP + BMSCs group (P > 0.05). All subjects were divided into the non‐healing group and the healing group in accordance with the HE staining results in previous experiment. The injury of cartilage of lateral femoral condyle was significantly heavier in the non‐healing group than that in the healing group.ConclusionThe application of PRP alone or in combination with BMSCs could promote the clinical healing rate of meniscal white‐white zone injury.     

Influence of decortication of the recipient graft bed on graft integration and tissue neoformation in the graft-recipient bed interface

The objective of the present study was to assess the influence of decortication of the posterior elements of the vertebra (recipient bed) and the nature of the bone graft (cortical or cancellous bone) on graft integration and bone, cartilage and fiber neoformation in the interface between the vertebral recipient bed and the bone graft. Seventy-two male Wistar rats were divided into four experimental groups according to the presence or absence of decortication of the posterior vertebral elements and the use of a cortical or cancellous bone graft. Group I—the posterior elements were decorticated and cancellous bone used. Group II—the posterior elements were decorticated and cortical graft was used. Group III—the posterior elements were not decorticated and cancellous graft was used. Group IV—the posterior elements were not decorticated and cortical graft was used. The animals were killed 3, 6 and 9 weeks after surgery and the interface between the posterior elements and the bone graft was subjected to histomorphometric evaluation. Mean percent neoformed bone was 40.8% in group I (decortication and cancellous graft), 39.13% in group II (decortication and cortical graft), 6.13% in group III (non-decorticated and cancellous graft), and 9.27% in group IV (non-decorticated and cortical graft) for animals killed at 3 weeks (P = 0.0005). For animals killed at 6 weeks, the mean percent was 38.53% for group I, 40.40% for group II, 10.27% for group III, and 7.6% for group IV (P = 0.0005), and for animals killed at 9 weeks, the mean was 25.93% for group I, 30.6% for group II, 16.4% for group III, and 18.73% for group IV (P = 0.0026). The mean percent neoformed cartilage tissue was 8.36% for group I, 7.46% for group II, 11.1% for group III, and 9.13% for group IV for the animals killed at 3 weeks (P = 0.6544); 6.6% for group I, 8.07% for group, 7.47% for group III and 6.13% for group IV (P = 0.4889) for animals killed at 6 weeks, and 3.13% for group I, 4.06% for group II, 10.53% for group III and 12.07% for group IV (P = 0.0006) for animals killed at 9 weeks. Mean percent neoformed fibrous tissue was 11% for group I, 6.13% for group II, 26.27% for group III and 21.87% for group IV for animals killed at 3 weeks (P = 0.0008); 7.67% for group I, 7.1% for group II, 9.8% for group III and 10.4% for group IV (P = 0.7880) for animals killed at 6 weeks, and 3.73% for group I, 4.4% for group II, 6.67% for group III and 6.8% for group IV (P = 0.0214) for animals killed at 9 weeks. The statistically significant differences in percent tissue formation were related to decortication of the posterior elements. The use of a cortical or cancellous graft did not influence tissue neoformation. Ossification in the interface of the recipient graft bed was of the intramembranous type in the decorticated animals and endochondral type in the non-decorticated animals.     

Cell-mediated immune response is better preserved by laparoscopy than laparotomy

BACKGROUND: This study compares the effects of carbon dioxide pneumoperitoneum versus laparotomy on cellular-mediated immune response in a murine model. METHODS: Sixty-eight female C3H/He mice were sensitized to keyhole limpet hemocyanin (KLH) and to a mouse mammary carcinoma cell line (MC2) before surgery. Animals were randomized into 4 groups: group I, anesthesia (control); group II, pneumoperitoneum with carbon dioxide; group III, extraperitoneal wound; group IV, laparotomy. All animals were challenged subsequently with KLH and MC2 tumor cells. Delayed-type hypersensitivity skin reaction (DTH) to KLH was measured on postoperative days (PODs) 1, 2, 4, and 5. Tumor growth was assessed weekly as an indicator of postoperative cellular immune response. RESULTS: Compared with preoperative values, postoperative DTH skin reactions were significantly less for all PODs in groups III and IV (P < .05), on POD 1 and 4 in group II (P < .05) and POD 4 for group I (P < .05). Group IV showed significantly fewer DTH skin reactions for all PODs compared with groups I and II (P < .05) and all PODs except on day 2 compared with group III (P < .05). Tumor growth was significantly increased at postoperative week 2 (n = 3/17 mice) and 3 (n = 4/17 mice) in group IV, when compared with groups I and II (P < .05). CONCLUSIONS: Cellular immunity is preserved after carbon dioxide pneumoperitoneum compared with extraperitoneal incisions and laparotomy as measured by DTH and the ability to reject an immunogenictumor.     

Nonmodular Tapered Fluted Titanium Stems Osseointegrate Reliably at Short Term in Revision THAs

Background

The ideal femoral component for revision THA is undecided. Cylindrical nonmodular stems have been associated with stress shielding, whereas junctional fractures have been reported with tapered fluted modular titanium stems. We have used a tapered fluted nonmodular titanium femoral component (Wagner Self-locking [SL] femoral stem) to mitigate this risk. This component has been used extensively in Europe by its designer surgeons, but to our knowledge, it has not been studied in North America. Added to this, the design of the component has changed since early reports.

Questions/purposes

We asked: (1) Does the Wagner SL stem have low rates of rerevision and other complications at a minimum 2 years after surgery? (2) Is the Wagner SL stem associated with high levels of patient function and pain relief at a minimum 2 years after surgery? (3) Does the Wagner SL stem have low rates of subsidence at a minimum 2 years after surgery? (4) Is the Wagner SL stem associated with proximal femoral bone remodeling at a minimum 2 years after surgery?

Method

Between May 2011 and December 2012, we performed 198 femoral revisions, of which 104 (53%) were performed using the Wagner SL femoral stem; during that period, our institution gradually shifted toward increasing use of these stems for all but the most severe revisions, in which modular fluted stems and proximal femoral replacements still are used on an occasional basis. Median followup in this retrospective study was 32 months (range, 24–46 months), and one patient was lost to followup before the 2-year minimum. The femoral deformities in this series were Paprosky Type I (10 hips), Paprosky Type II (26), Paprosky Type IIIA (52), Paprosky Type IIIB (nine), and Paprosky Type IV (two). Functional assessment was performed using the Oxford Hip Score (OHS), WOMAC, SF-12, and the University of California Los Angeles (UCLA) activity score. All complications and cases of revision were documented. All patients had radiographs performed within 1 year of the latest followup. These were assessed by two surgeons for signs of proximal femoral bone remodeling and subsidence.

Results

Complete preoperative scores were available for 98 patients (98 of 104; 94%). The mean OHS preoperatively and at final followup were 39 (SD, 15) and 87 (SD, 19), respectively (p < 0.001; mean difference, 48; 95% CI, 43–53). Average WOMAC scores were 44 (SD, 15) and 87 (SD, 20), respectively (p < 0.001; mean difference, 43; 95% CI, 38–48). At final followup, signs of restoration of proximal femoral bone stock was noted in 45 of 103 hips (44%). Six (six of 104; 6%) patients had subsidence of 10 mm to 15 mm. In the remainder (98 of 104; 94%), the mean subsidence was 2 mm (range, 0–9 mm). One revision was performed for loosening associated with infection.

Conclusions

The Wagner SL stem is a viable option for patients with Paprosky Types II and III defects undergoing revision THA. This component provides high levels of patient function with low revision rates and low rates of subsidence during the early postoperative phase. They provide a viable alternative to modular components for treatment of Types II and III defects without the risk of junctional fractures. They can be used for very selected Type IV defects, however this extent of bone loss is most easily addressed with other techniques such as a proximal femoral replacement.

Level of Evidence

Level IV, therapeutic study

     

The effect of fibrin clot and C vitamin on the surgical treatment of Achilles tendon injury in the rat model✰

BackgroundThis study aimed to determine the histological, biochemical, and biomechanical efficacy of fibrin clot and vitamin C in the healing of Achilles tendon ruptures (ATR) in a rat model.Methods52 adult Wistar-Albino rats (300–450 g) were used in the study. 12 rats were divided into four groups as Monitor (Group I), Control (Group II), Fibrin Clot (Group III), Fibrin Clot with vitamin C (Group IV). Four rats were used for fibrin clot preparation. Fibroblast Growth Factor (FGF) and Vascular Endothelial Growth Factor (VEGF) were measured on the 3rd, 7th, 14th, and 21st days. Four rats were sacrificed on the 21st day from each group for histological evaluation. The rest of the rats were sacrificed at 42nd day, half for biomechanical and a half for histological evaluation.ResultsThe 42nd-day HSS score of group IV was significantly lower than those of group I, group II and group III (p = 0.036, p = 0.019, and p = 0.036, respectively). Group IV showed a significantly higher Maximum force N value than those of group I, group II and group III (p = 0.034, p = 0.034 and, p = 0.025, respectively). The blood FGF and VEGF levels of group III and group IV on the 3rd, 7th, 14th, and 21st days were higher than those of group I and group II (p < 0.05).ConclusionFibrin clot and vitamin C produced a stronger tendon structure in terms of biomechanics while providing histological and biochemically better quality tendon healing in the surgical treatment of ATR. This model can be used to accelerate high-quality tendon healing after ATR.Level of EvidenceLevel II, experimental study.     

不同比例骨软骨镶嵌成形术联合基因增强组织工程方法治疗骨软骨缺损

目的：研究骨软骨镶嵌成形术与联合基因增强组织工程方法治疗急性骨软骨缺损,并观察在不同比例下的组织修复情况,以期发现两者间的最佳组合。方法：对携带hTGF-β1的重组腺病毒转染BMSCs（hTGF-β1转染组）,与采用Adv-βgal转染BMSCs（Adv-βgal转染组）及未转染BMSCs（未转染空白对照组）进行WesternBlot检测hTGF-β1、Ⅱ型胶原及Aggrecan表达。雄性6月龄崇明山羊18只,体重22～25kg,取自体骨髓进行BMSCs分离及培养,传至第3代。每只动物双膝股骨内髁进行实验,分为AR、AL、BR、BL、CR、CL6组。AR为骨软骨移植覆盖面积为44.44%单纯组,AL为44.44%联合组,BR为33.33%单纯组,BL为33.33%联合组,CR为22.22%单纯组,CL为22.22%联合组。于双膝股骨内髁负重区采用骨钻制备直径为9mm,深为3mm的骨软骨缺损后,单纯组采用骨软骨镶嵌成形术的自体骨软骨柱修复,联合组同时将转染hTGF-β1的BMSCs藻酸钠混合液注入空隙,加入氯化钙产生凝胶。术后24周取材,行大体及组织学观察,并参照O＇Driscoll,KeeleyandSalter组织形态学评分标准进行评分,行免疫组织化学及透射电镜观察。结果：hTGF-β1转染组细胞hTGF-β1、Ⅱ型胶原及Aggrecan表达均强于Adv-βgal转染组及未转染空白对照组。大体观察,组织学染色以及透射电镜检查显示各组的缺损有不同程度的修复。33.33%联合组、44.44%单纯组、44.44%联合组的整体修复效果差别无统计学意义,33.33%单纯组、22.22%单纯组、22.22%联合组的整体修复效果差于前3组。结论：骨软骨镶嵌成形术联合基因增强组织工程的方法可以有效修复骨软骨缺损。随着自体骨软骨移植覆盖缺损面积的减少,骨软骨镶嵌成形术联合基因增强组织工程方法的修复优越性可以得到更好的体现。