OBJECTIVE: To observe the effects of hyaluronic acid (HA) and basic fibroblast growth factor (bFGF) on the proliferation of the cells from medial collateral ligament (MCL) and anterior cruciate ligament (ACL) cells. METHODS: The MCL cells and ACL cells of mature New Zealand white rabbit were cultured, while HA, bFGF or HA and bFGF were added to the cell culture media, the cellular proliferation was assayed by MTT method. RESULTS: HA only had no effect on the preoliferation of ACL cells, but had a small stimulatory effect on the proliferation of MCL cells. The addition of 1 ng/ml bFGF enhanced the proliferation of both MCL and ACL cells significantly, and this enhancement was maximal in the concentration of 50 ng/ml. However, the enhancement of proliferation of MCL and ACL cells could be achieved when the combination of HA in concentration of 100 micrograms/ml and bFGF in concentration of 100 ng/ml. CONCLUSION: It is evident that bFGF can enhance the proliferation of the ligament cells. HA can maintain the normal growth of ACL cells with no effect on the proliferation of the cells, while HA has a small stimulatory effect on the proliferation of MCL cells. However, when bFGF is coordinated with HA, more improvement of cellular proliferation can be achieved. HA can be used as a potential carrier for bFGF to enhance the healing of ligamentous tissue injuries.
ObjectiveTo investigate the effect of transforming growth factorβ1 (TGF-β1) and basic fibroblast growth factor 1 (bFGF-1) on the cellular activities, prol iferation, and expressions of ligament-specific mRNA and proteins in bone marrow mesenchymal stem cells (BMSCs) and ligament fibroblasts (LFs) after directly co-cultured. MethodsBMSCs from 3-month-old Sprague Dawley rats were isolated and cultured using intensity gradient centrifugation. LFs were isolated using collagenase. The cells at passage 3 were divided into 6 groups: non-induced BMSCs group (group A), non-induced LFs group (group B), non-induced co-cultured BMSCs and LFs group (group C), induced BMSCs group (group D), induced LFs group (group E), and induced co-cultured BMSCs and LFs group (group F). The cellular activities and prol iferation were examined by inverted contrast microscope and MTT; the concentrations of collagen type Ⅰ and type Ⅲ were determined by ELISA; and mRNA expressions of collagen types I andⅢ, fibronectin, tenascin C, and matrix metalloproteinase 2 (MMP-2) were measured by real-time fluorescent quantitative PCR. ResultsA single cell layer formed in the co-cultured cells under inverted contrast microscope. Group F had fastest cell fusion ( > 90%). The MTT result indicated that group F showed the highest absorbance (A) value, followed by group D, and group B showed the lowest A value at 9 days after culture, showing significant difference (P < 0.05). Moreover, the result of ELISA showed that group F had the highest concentration of collagen type Ⅰ and type Ⅲ (P < 0.05); the concentration of collagen type Ⅲ in group E was significantly higher than that in group D (P < 0.05), but no significant difference was found in the concentration of collagen type Ⅰ between 2 groups (P > 0.05). The ratios of collagen type Ⅰ to type Ⅲ were 1.17, 1.19, 1.10, 1.25, 1.17, and 1.18 in groups A-F; group D was higher than the other groups. The real-time fluorescent quantitative PCR results revealed that the mRNA expressions of collagen type Ⅰ and type Ⅲ and fibronectin were highest in group F; the expression of tenascin C was highest in group D; the expression of MMP-2 was highest in group E; and all differencs were significant (P < 0.05). ConclusionDirectly co-cultured BMSCs and LFs induced by TGF-β1 and bFGF-1 have higher cellular activities, proliferation, and expressions of ligament-specific mRNA and protein, which can be used as a potential source for ligament tissue engineering.
Objective To study the effectiveness of anterior cruciate l igament (ACL) reconstruction using autologous periosteum wrapping tendon allograft by comparing with using simple tendon allograft. Methods Between March 2008 and November 2008, 68 patients with ACL injury were treated, who were in accordance with the inclusion criteria. They were divided into 2 groups randomly according to different treatment methods: ACL was reconstructed with autologous periosteum wrapping tendon allograft in 31 patients (test group) and with simple tendon allograft (control group) in 37 patients. There was no significant difference in gender, age, disease duration, the cause of injury, and functional score preoperatively between 2 groups (P gt; 0.05). Anatomic single-bundle ACL reconstruction was performed in 2 groups. Results Little exudation at tibial tunnel incision was found in 1 case respectively in both groups at 2 weeks after operation and was cured by dressing change and antibiotics. The other incisions healed by first intention. The patients were followed up 24-29 months (mean, 26 months) in the test group and 24-32 months (mean, 27 months) in the control group. CT showed bone tunnel enlargement in both groups at 2 years after operation, but the rate of the tunnel enlargement was less inthe test group (5/31, 16.1%) than in the control group (14/37, 37.8%), showing significant difference (χ2=3.948, P=0.047). At 2 years after operation, the results of Lachman test and pivot shift test were negative in 23 cases (74.2%) and 25 cases (80.6%) of the test group, and in 26 cases (70.3%) and 30 cases (81.1%) of the control group, respectively. KT-1000 examination showed the displacement of the test group [(1.74 ± 0.88) mm] was less than that of the control group [(2.36 ± 0.83) mm], showing significant difference (t= —2.979, P=0.004). There was no significant difference in Lysholm score, Hospital for Special Surgery (HSS) score, Tegner score, and International Knee Documentation Committee (IKDC) score between 2 groups at 2 years after operation (P gt; 0.05). Conclusion Compared with simple tendon allograft, ACL reconstruction with autologous periosteum wrapping tendon allograft can improve tendon-bone heal ing, and decrease the rate of bone tunnel enlargement, so it has good short-term outcome.
Objective To investigate effectiveness of applying the Bone Morphingbased image-free computer-assisted system for the ligament balancing managementin the total knee arthroplasty (TKA). Methods Between November 2002 and June 2003, twenty-one posterior stabilized total knee prostheses (Ceraver, France) were implanted in 21 patients using the Bone Morphing based image-free Ceravision system.This cohort included 5 men and 16 women with an average age of 72.4 years, two undergoing high tibial osteotomy and 1 undergoing distal femoral osteotomy before. The preoperative deviation was measured by the full-length AP X-rays. The knees were in varus deviation in 14 patients and in valgus deviation in 7 patients, with an average of 2.36°(varus 13°-valgus 13°). The frontal X-rays ofthe knee were assessed, the mean value of the varus force-stress test was 8.47°(varus 2°-varus 20°), and the mean value of the valgus forcestress test was 3.63°(varus 7°-valgus 12°). Results With the Ceravisionrecorded data, the intraoperative alignment was assessed, the mean lower limb axis was 3.33°(varus 12°-valgus 10°),and compared with the preoperative data, the difference was significant (Plt;0.05); the mean value of the varus force-stress test was 6.47°(varus 0°-varus 24°), the mean value of the valgus force-stress test was 4.32°(varus 8°- valgus 15°), and compared with the preoperative data, the difference was significant (Plt;0.05). The post-prosthetic alignment on Ceravision with a deviation of 0.175°(varus 2°- valgus 3°) was compared with the postoperative alignment by the full-length AP X-rays, with a deviation of 0.3°(varus 3.5°-valgus 1.5°), the difference wasn’t significant(Pgt;0.05).The clinical check-up performed 3 months after operation showed that the average range of movement (ROM) was 115°(105-130°), the mean frontal laxity was 0.27 mm(0.2-0.5 mm). The femoral and tibial components were implanted in the satisfactory 3 dimensional position without ligament imbalance in all the patients, andthere were no instability or patella complications.Conclusion Utilization of the Bone Morphing based image-free computer-assisted system can achieve an accurate component 3 dimensional alignment, optimal bone resection, optimal control of surgical decision in releasing the soft tissues, rotating the femoral component to gain an extension/flexion rectangular gap, and managing theligament balancing so as to achieve a satisfactory initial clinical outcome. This system can be routinely used in the TKA.
To research the operative method and the cl inical efficacy of repairing and reconstructing tendon and l igament with allograft tendon. Methods From September 2000 to May 2007, 164 cases with tendon and l igament injuries were treated, including 116 males and 48 females aged 21-47 years old (average 31.5 years old). There were 126 cases of anterior cruciate l igament injury, 18 cases of complete acromioclavicular dislocation, 10 cases of old dislocation of radialhead, 4 cases of Achilles tendon rupture, 2 cases of tibial is anterior muscle rupture, 2 cases of patellar tendon rupture, and 2 cases of rectus femoris rupture. Time interval between injury and hospital admission was 4-345 days (average 75 days). Allograft tendon 10-26 cm in length with suture anchor or absorbable interference screw was used to reconstruct the l igament and tendon. Results All wounds healed by first intention, except one case of rectus femoris rupture. All patients were followed for 10-36 months (average 21 months). The international knee documentation commitee and the Lysholm score of patients with anterior cruciate l igament injury 12 months after operation were significantly higher than that of before operation (P lt; 0.01). According to the Lazzcano and Karlsson score standard, 13 cases of complete acromioclavicular dislocation at 10-12 months after operation were graded as excellent and 5 cases were good. According to Arner Lindholm score standard, 3 cases of Achilles tendon rupture at 8-16 months after operation were graded as excellent and 1 case was good. For the patients with tibial is anterior muscle rupture, at 10-17 months after operation, the limitation of dorsal extension in ankle joint was 5°, and the muscle strength in the anterior tibial is muscle was decreased. For the patients with patellar tendon rupture, one completely restored the motion range of the knee joint 14 months after operation, the other had knee extension l imitation of 10° at 13 months after operation. For the patients with rectus femoris rupture, one had 15° of extension limitation at 18 months after operation, the other suffered limitation of both extension and genuflex at 12 months after operation. According to the Broberg-Morrey score standard, 7 caseswith old dislocation of radial head were graded as excellent and 3 cases were good at 3-36 months after operation. Conclusion Allograft tendon is a good material for repairing and reconstructing tendon and ligament injuries, but attention should be paid to postoperatively early exercise under appropriate protection and early management of local rejection.
ObjectiveTo compare the clinical efficacy between deep medial collateral ligament (dMCL) repair and conservative treatment for complete MCL rupture. MethodsBetween August 2009 and December 2013, 36 patients with grade 3 MCL rupture underwent superior MCL (sMCL) reconstruction with tibial Inlay technique. Of 36 cases, 19 received dMCL repair (repair group), and 17 received conservative treatment (conservation group) after sMCL reconstruction. There was no significant difference in gender, age, knee sides, type of injury, disease duration and preoperative medial joint opening, knee Lysholm scores, and International Knee Documentation Committee (IKDC) score between 2 groups (P > 0.05). The Lysholm and IKDC scores, medial joint opening, range of motion (ROM), visual analogue scale (VAS) scores, and complications were used to assess the knee joint function. ResultsAll patients achieved primary incision healing without acute postoperative complications of incision infection and deep vein thrombosis in the lower limb. The patients were followed up 28-65 months (mean, 46.3 months) in the repair group, and 26-69 months (mean, 45.9 months) in the conservation group. No knee stiffness, vascular or nerve injury, and knee joint infection occurred in 2 groups. All the patients recovered medial stability at 2 years postoperatively. At 2 years after operation, no significant difference was shown in knee ROM between 2 groups (t=0.26, P=0.80); the VAS score of the repair group was significantly lower than that of the conservation group (t=5.22, P=0.00); medial joint opening, IKDC score, and Lysholm score were significantly improved when compared with preoperative ones in 2 groups (P < 0.05), but no significant difference was found between 2 groups (P>0.05). ConclusionWhether or not additional dMCL repair is performed can recover medial stability after sMCL reconstruction. However, the additional dMCL repair is better in relieving medial knee pain than the conservative treatment.
ObjectiveTo evaluate the effectiveness of semitendinous and gracilis transfer for the treatment of medial collateral ligament (MCL) injury caused by total knee arthroplasty (TKA). MethodsBetween March 2009 and May 2014, 11 patients (11 knees) with MCL injuries caused by primary TKA were treated by semitendinous and gracilis transfer in primary TKA (injury group). Another 18 patients (21 knees) without MCL injury were included as the control group. There was no significant difference in gender, age, injury sides, disease duration, body mass index, knee varus deformity, and preoperative Knee Society Score (KSS) between 2 groups (P>0.05), with comparability. KSS score was used to evaluate the function after operation. ResultsPrimary healing of incision was obtained in all patients, and no complications of joint instability and pain occurred. The follow-up time was 6-29 months in injury group and was 7-34 months in control group. At last follow-up, the KSS clinical score and functional score were significantly increased to 89.82±3.76 and 89.54±3.50 in the injury group (P<0.05) and were significantly increased to 90.19±3.39 and 90.00±3.53 in the control group (P<0.05) respectively, but no significant difference was shown between 2 groups (t=0.158, P=0.877; t=0.820, P=0.432). X-ray films showed no prosthetic loosening or subsidence during follow-up. ConclusionThe semitendinous and gracilis transfer is reliable for the treatment of MCL injury caused by TKA. The insertions of semitendinous tendon and gracilis are close to that of the knee MCL, which can effectively improve knee function.
ObjectiveTo discuss whether human amniotic mesenchymal stem cells (hAMSCs) possesses the characteristic of mesenchymal stem cells, and could differentiate into ligament cells in vitro after induction. MethodsThe hAMSCs were separated through enzyme digestion, and the phenotypic characteristics of hAMSCs were tested through flow cytometry. The cells at passage 3 were cultured with L-DMEM/F12 medium containing transforming growth factor β1 (TGF-β1)+basic fibroblast growth factor (bFGF) (group A), containing hyaluronic acid (HA) (group B), containing TGF-β1+bFGF+HA (group C), and simple L-DMEM/F12 medium (group D) as control group. The morphology changes of cells in each group were observed by inverted phase contrast microscope at 21 days after induction; the cellular activities and proliferation were examined by sulforhodamine (SRB) colorimetric method; and specific mRNA and protein expressions of ligament including collagen type I, collagen type III, and tenascin C (TNC) were measured by real-time fluorescence quantitative PCR and immunohistochemical staining. ResultsThe flow cytometry result indicated that hAMSCs expressed mesenchymal stem cell phenotype. After 21 days of induction, the cells in groups A, B, and C grew like spindle-shaped fibroblasts under inverted phase contrast microscope, and cells showed single shape, obvious directivity, and compact arrangement in group C. The SRB result indicated that the cells in each group reached the peak of growth curve at 6 days; the cellular activities of groups A, B, and C were significantly higher than that of group D at 6 days after induction. Also, the immunohistochemical staining results showed that no expressions of TNC were detected in 4 groups at 7 days; expressions of collagen type I in groups A, B, and C were significantly higher than that in group D at 7, 14, and 21 days (P<0.001); the expressions of collagen type III in groups A, B, and C were significantly higher than that in group D at 14 and 21 days (P<0.001). There was an increasing tendency with time in collagen type I of group B, in collagen type III and TNC of groups A and C, showing significant difference among different time points (P<0.001). The real-time fluorescence quantitative PCR results revealed that the mRNA expressions of collagen type I and TNC in group C were significantly higher than those in groups A and B (P<0.05), and the mRNA expression of collagen type III in group B were significantly higher than that in groups A and C at 21 days (P<0.05). The mRNA expressions of collagen type I and TNC in groups A and C and mRNA expression of collagen type III in group C had an increasing tendency with time, showing significant difference among different time points (P<0.001). ConclusionThe hAMSCs possesses the characteristics of mesenchymal stem cells and excellent proliferation capacity. After in vitro induction, the expressions of ligament specific genes can be up-regulated and the synthesis of ligament specific proteins can be also strengthened. As a result, it can be used as one of ligament tissue engineering seed cell sources.
Objective To investigate the radiographic and clinical outcomes of dorsal intercarpal ligament capsulodesis (DILC) procedure for chronic static scapholunate dissociation. Methods Between January 2008 and January 2011, 12 patients with chronic static scapholunate dissociation were treated with DILC. Of 12 cases, 10 were male and 2 were female with an average age of 42 years (range, 20-66 years). All injuries were caused by falling. The interval from injury to surgery was 3-19 months (mean, 8 months). Physical examination at admission showed wrist tenderness and limited range of motion (ROM). Radiological examination showed that scapholunate gap was greater than 3 mm on posteroanterior view, and scapholunate angle more than 60° on the lateral view. Before operation, the grip strength was (25.4±8.2) kg; the wrist ROM was (56.7±11.5)° in flexion and (52.0±15.2)° in extension; visual analogue scale (VAS) score was 6.3±1.4; and disabilities of arm, shoulder & hand (DASH) score was 39.5±7.4. According to Garcia-Elias staging criteria, all cases were rated as stage 4, indicating that the scapholunate interosseous ligament was completely injured and reduction could easily be obtained. Eight patients had wrist instability. Results Primary healing of incision was achieved, no complication was found. All patients were followed up 13-34 months (mean, 15.9 months). During surgery, all deformities were corrected completely, but 7 patients (58%) recurred at 1 month after Kirschner wire removal. Compared with preoperative ones, the scapholunate gap, scapholunate angle, radiolunate angle, lunocapitate angle, and wrist height ratio at 1 month after Kirschner wire removal and last follow-up showed no significant difference (P > 0.05); the wrist flexion and extension ROM were significantly decreaed to (46.8±7.2)° and (42.0±9.0)° at last follow-up (P < 0.05); the grip strength was significantly increased to (32.7±9.6) kg at last follow-up (P < 0.05); VAS score and DASH score were improved to 1.7±1.0 and 8.1±8.7 (P < 0.05). Conclusion Carpal collapse will recur in short time after DILC. DILC is not the best way to treat chronic static scapholunate dissociation.
Objective To discuss the effectiveness of operation technique for antero-medial rotatory instability (AMRI) of the knee joint caused by motorcycle. Methods Between June 2007 and December 2009, 32 cases of AMRI caused by motorcycle were treated. There were 28 males and 4 females with an average age of 35.5 years (range, 20-50 years). The interval between injury and surgery was 5-10 days (mean, 7 days). The anterior cruciate ligament (ACL) was injured at the attachment point of the condyles crest; the medial collateral ligament (MCL) was injured at central site in 19 cases, at medial condyles of femur in 10 cases, and at medial condyles of tibia in 3 cases, which were all closed injuries. The bone avulsion of condyles crest was fixed by steel wire and MCL was repaired. Results Red swelling and a little effusion occurred at the incision in 1 case, and the other incisions healed by first intention. Traumatic arthritis of the knee occured in 5 cases. Thirty-two cases were followed up 16-22 months (mean, 18.5 months). The X-ray examination showed that the fracture union time was 5-8 weeks (mean, 6 weeks) after operation. At last follow-up, the extension of knee joint was 0° and the flexion of the knee joint was 110-170° (mean, 155°). According to the synthetic evaluating standard of International Knee Documentation Committee, 24 cases were rated as A level, 6 cases as B, 1 case as C, and 1 case as D at last follow-up. Lysholm knee score was 85.93 ± 3.76 at last follow-up, which was significantly higher (t=53.785, P=0.000) than preoperative score 37.54 ± 3.43. Conclusion In patients with AMRI caused by motorcycle, steel wire is used to fix the bone avulsion of condyles crest and MCL should be repaired simultaneously as far as possible. And associating with the early postoperative functional exercise, the short-term effectiveness is satisfactory, but long-term effectiveness still need further follow-up observation.