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.
The study aimed to evaluate the safety and function of poly(lactic-acid-co-ε-caprolactone) (PLCL)/fibrinogen nanofibers (P/F-Ns), and provide theoretical basis for the clinical application. The surface morphology, mechanical properties, the hydrophilicity and the fibrinogen content of P/F-Ns were tested by scanning electron microscope, the material testing machine, the contact angle meter and the microplate reader, respectively. The cell adhesion, proliferation and ligament remodeling genes expression of Hig-82 cells on P/F-Ns were conducted through cell counting kit-8 (CCK-8) and real-time quantitative PCR analyses, respectively. The results showed that with the increase of the fibrinogen content, the pore sizes and hydrophilicity of three P/F-Ns increased, but the mechanical properties decreased. Cell adhesion and proliferation tests showed that P/F-N-2 held the best ability to promote cell adhesion and proliferation. The ligament remodeling genes expressions of Hig-82 cells on P/F-N-1, P/F-N-2 and P/F-N-3 were all up-regulated compared to P/F-N-0 on days 3 and 7. All the three P/F-Ns containing fibrinogen (P/F-N-1, P/F-N-2 and P/F-N-3) had better biocompatibility compared to P/F-N-0, and could be efficiently applied to the reconstruction of anterior cruciate ligament.
Objective To investigate the method and short-term effectiveness of arthroscopic reconstruction of anterior cruciate l igament (ACL) using RetroButton-allogeneic tendon-interference screw. Methods Between June 2009 and October 2009, 23 patients with ACL rupture were treated by arthroscopic reconstruction with RetroButton-allogeneic tendon-interference screw. There were 15 males and 8 females with an average age of 32.5 years (range, 19-46 years), including 17 left knees and 6 right knees. The injury causes were sport trauma (13 cases), traffic accident (8 cases), and fall ing injury(2 cases). There were 7 acute cases (lt; 6 weeks) and 16 chronic cases (gt; 6 weeks). Among the cases, 11 cases compl icated by medial meniscus injury, 6 by lateral meniscus injury, 3 by the both injuries, and 5 by articular cartilage injury. All cases had no injuries of posterior cruciate l igament, medial or lateral collateral l igament, or posterolateral structure. The time from injury to operation ranged from 3 weeks to 32 months. Lysholm and International Knee Documentation Committee (IKDC) scores were used for subjective evaluation, while Lachman test and KT-1000 measurement for objective evaluation. Results All wounds healed by first intention. The symptoms of unstable knee were improved obviously. No high fever, infection, or immunologic rejection were observed. Refractoriness synovitis and joint effusion occurred in 1 case after operation, and was improved after articular cavity flushing for 7 times within 3 weeks. All cases were followed up 10-17 months (mean, 14.7 months). There were significant differences in Lysholm score, IKDC score, Lachman test, and KT-1000 measurement between pre-operation and last follow-up (P lt; 0.05). Conclusion Arthroscopic reconstruction of ACL with RetroButton-allogeneic tendon-interferencescrew is simple and safe, and its short-term effectiveness is satisfactory.
Objective To investigate the cl inical outcome of treating dorsal wrist gangl ion with an improved surgical strategy by excising the gangl ion completely along their stalk and repairing the dorsal carpal l igaments under brachial anesthesia. Methods From March 2005 to January 2007, 34 patients with dorsal wrist gangl ion were treated and studied retrospectively. There were 14 males and 20 females, aged 25-65 years (43 years on average). The left sides were involved in 22 cases and right sides in 12 cases. Thirteen cases of relapse received excision for 1 to 4 times under local anesthesia, with amean period of 17 months (14 days to 7 years) from excision to recurrence. Twenty-one patients were first attack cases with a mean period of 11 months (15 days to 8 years) from diagnosis to excision. The size of the gangl ion ranged from 1.5 cm × 1.2 cm to 4.5 cm × 4.0 cm. Now, each surgical process was performed under brachial anesthesia, and a pneumathode tourniquet was used. In 6 patients, the stalks of gangl ion did not invade the carpal l igaments, and gangl ion was removed completely without immobil ization after operation. In 28 patients, the stalks of gangl ion invaded the carpal l igaments, gangl ion was excised completely along its stalk to the dorsal carpal structure; the l igaments were sutured directly in 16 cases and were repaired with adjacent tissue such as the wall of sheathing canal of extensor tendon in 12 cases. The wrists were immobil ised for 3 weeks. Results Primary wound heal ing was achieved in all incisions. All patients were followed up for 26-36 months with an average of 31.5 months. Only 2 cases (5.9%) recurred. The range of motion of the wrist remained normal and the symptom of the dorsal wrist was rel ieved sl ightly. Patients’ satisfaction score ranged from 60 to 100, with an average of 83.8. Conclusion The gangl ion should be excised completely together with defect repair of dorsal carpal l igament under brachial anesthesia and the wrist immobil ised for 3 weeks, the recurrence rate will be reduced greatly.
Objective To investigate the method and curative effect of malposed-suture hanging and fastening method of double-lariat lock catch knot in repairing origin of medial collateral l igament (MCL) rupture. Methods From February 2008 to February 2009, 36 patients with acute MCL rupture were treated with malposed-suture hanging and fastening method of double-lariat lock catch knot. There were 21 males and 15 females with an average age of 40 years (range, 17-58 years),including 19 left knees and 17 right knees. Repture was caused by traffic accident in 5 cases, by fall ing in 11 cases, by kicking in 3 cases, by crush in 4 cases, and by sprain in 13 cases. The X-ray films of double knees at stress state showed the medial joint space of affected knee joint increased 6.5-13.5 mm (11.2 mm on average) when compared with that of normal knee joint. The time from injury to operation was 36 hours to 8 days (3.5 days on average). Results All wounds healed by first intention. Thirty-one cases were followed up 12-20 months (15 months on average). No compl ication of wound infection, deep venous thrombosis, and l igament rerupture occurred. The medial joint space of affected knee joint increased 1.5-5.6 mm (3.5 mm on average) when compared with that of normal knee joint. According to Lysholm assessment standard, the results were excellent in 20 cases and good in 11 cases, the excellent and good rate was 100%. Conclusion Repair of origin of MCL with malposedsuture hanging and fastening method of double-lariat lock catch knot has advantages of less injury, rel iable fixation, and rapid recovery of knee stabil ity.
ObjectiveTo explore the feasibility of Lisfranc ligament reconstruction with autogenous tendon through biomechanical testing. MethodsTwelve fresh-frozen cadaveric lower limbs were prepared three sequential testing conditions:intact Lisfranc ligament (intact group), disrupted Lisfranc ligament (disrupted group), and Lisfranc ligament reconstruction (reconstruction group). Under fixing on the Bose mechanical test machine, three models were given 0-600 N axial loading in the neutral position and the plantar flexion of 30° according to the speed of 10 N/s, every 100 N load with a 1-minute interval. The medial cuneiform (C1) and the second metatarsal (M2) base displacement and the foot transverse arch height were recorded under different loads. ResultsIn the neutral position and the plantar flexion of 30°, C1-M2 displacement and foot transverse arch height showed an increasing trend with increased load under 0-600 N axial loading. There were significant differences in C1-M2 displacement variation in 2 positions among groups (P＜0.05). In disrupted group, the C1-M2 displacement variation in neutral position was significantly lower than that in plantar flexion of 30° (t=7.392,P=0.000). In the neutral position, the foot transverse arch height variation in the disrupted group and the reconstruction group was significantly higher than that in the intact group (P＜0.05), but there was no significant difference between the disrupted group and reconstruction group (P>0.05). ConclusionLisfranc ligament reconstruction with autogenous tendon can reduce the C1-M2 displacement variation and stabilize Lisfranc joint to a certain degree. Reconstruction of both dorsal ligament and Lisfranc ligament will not improve the buffering capacity. The C1-M2 displacement variation in the plantar flexion of 30° is more obvious than that in neutral position, so it is helpful to improve clinical diagnosis of occult Lisfranc damage.
Objective To introduce the research advance of the ligament injury and ligament healing.Methods Recent original articlesrelated to such aspects of ligament were reviewed extensively.Results The ligament properties would be influenced when the situations of the biochemistry and biomechanics had changed. Injuries to ligaments induce a healing response that is characterized by scar formation. Graft could not recovery the ultrastructure, anatomy and biomechanics of the normal ligament.Conclusion The healing ligament is weaker than normal one, and the graft could not reconstruct normal ligament at present.
Objective To evaluate the feasibility of the anterior cruciate ligament (ACL) reconstruction with 6 strands of hamstring tendons enveloped by periosteum. Methods Between April 2008 and April 2009, 34 patients with ACL injury were treated, ACL of whom was reconstructed with 6 strands of hamstring tendons enveloping of periosteum and double Rigidfix fixation. There were 30 males and 4 females, aged 19-54 years with an average of 29.4 years. The causes of injury included sport in 19 cases, traffic accident in 8 cases, falling from height in 5 cases, and other in 2 cases. The locations were left knee in 19 cases and right knee in 15 cases. The disease duration was 3 weeks to 18 months (median, 9.4 months). The results of Lachman test and anterior drawer test were positive. The Lysholm knee score was 61.5 ± 3.6. MRI examination revealed ACL rupture in 26 cases and ACL injury in 8 cases. Results All incisions healed by first intention, and no early complication occurred. Twenty-eight cases were followed up 12-32 months (mean, 16.1 months). The result of Lachman test was negative at 12 months after operation; in all patients, knee extension reached 0°, and flexion reached 120-150° (mean, 132.5°). The AP and lateral X-ray films and MRI showed no bone tunnel expansion. At last follow-up the therapeutic effect evaluation was excellent in 25 cases, good in 1 case, and fair in 2 cases; the excellent and good rate was 92.9%. The postoperative Lysholm score was 91.0 ± 3.2, showing significant difference when compared with preoperative score (t=32.78, P=0.00). Conclusion Six strands of hamstring tendons can ensure sufficient tensile strength, and use of the double Rigidfix absorbable screw makes fixation more reliable. Facing outside suture of periosteal flap can promote tendon-bone healing, so it is a good method of ACL reconstruction.
ObjectiveTo investigate the effectiveness of one-stage arthroscopic reconstruction and strict immobilization for 6 weeks for treatment of knee dislocation. MethodBetween August 2010 and May 2013, 22 cases (22 knees) of knee dislocation were treated with one-stage reconstruction and strict immobilization for 6 weeks. There were 15 males and 7 females, aged 21-54 years (mean, 31.5 years). The left knee and right knee were involved in 8 cases and 14 cases respectively. The disease causes were traffic accident in 12 cases, falling from height in 6 cases, and sports injury in 4 cases. The time between injury and operation was less than 2 weeks in 6 cases, 2-3 weeks in 10 cases, and more than 3 weeks in 6 cases. The results of anterior drawer test, posterior drawer test, and Lachman test were positive in all patients. The posterior displacement of the tibia was more than 10 mm. The results of valgus stress test and varus stress test were positive in 13 cases and 11 cases respectively. The preoperative knee range of motion was (58.2±28.4) °, Lysholm score was 39.7±4.6. All patients had anterior cruciate ligament rupture and posterior cruciate ligament rupture; combined injuries included medial collateral ligament rupture in 11 cases, lateral collateral ligament rupture in 9 cases, both medial and lateral collateral ligament rupture in 2 cases, femoral condylar avulsion fracture in 2 cases, and meniscus injury in 7 cases. No nerve or blood vessel injury was observed. ResultsAll cases obtained primary healing of incision without infection. All the patients were followed up 12-48 months (mean, 27.8 months). At 12 months after operation, the results of the anterior drawer test, posterior drawer test, Lachman test, valgus stress test, and varus stress test were all negative; the knee range of motion increased was significantly to (121.3±7.9) °(t=30.061, P=0.000) ; Lysholm score was 87.2±6.1, showing significant difference when compared with preoperative score (t=24.642, P=0.000) . ConclusionsA combination of arthroscopic one-stage reconstruction and strict immobilization for treatment of knee dislocation is a safe and effective method, good stability and joint function can be achieved.