ObjectiveTo summarize the application status and progress of the strategies to augment tendon-to-bone healing. MethodsThe present researches focused on augmentation of tendon-to-bone healing were extensively reviewed. ResultsThe present strategies to augment healing of tendon-to-bone by enhancing the location environment, and increasing the cell numbers and relative growth factor. The mainly strategies include using calcium phosphate materials, biocompatible scaffolds and glue, growth factors, cell matrix, platelet-rich plasma, and periosteum. Although periosteum have been used in clinical and got some possitive effects, the others still not be used in clinical and needs further studies. ConclusionThere are many strategies to enhance the ability of tendon-to-bone healing, which got some positive results, but results of studies were varied. Thus, further fundamental research and clinical studies are required to achieve the best effects.
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 evaluate the surgical procedure and short-term effectiveness of one-stage repair and reconstruction of knee dislocation with multiple ligament injuries (KDMLI). Methods Between September 2010 and April 2014, 9 cases (9 knees) of KDMLI were treated. There were 7 males and 2 females with an average age of 42 years (range, 27-57 years). Injury was caused by traffic accident in 3 cases, heavy-weight crushing in 3 cases, sports sprain in 2 cases, and falling from height in 1 case. The average time from injury to operation was 11 days (range, 3-19 days). The results of posterior drawer test and Lachman test were positive in all patients. The results of varus stress testing were three-degree positive in 4 cases, and the results of valgus stress testing were three-degree positive in 6 cases. The Lysholm score of knee was 27.2±6.3; the International Knee Documentation Committee (IKDC) score was 29.7±6.5; and the range of motion (ROM) was (52.6±12.8)°. All patients suffered from posterior cruciate ligament (PCL) injury and femoral avulsion injury of anterior cruciate ligament (ACL). Combined injuries included medial collateral ligament (MCL) injury in 4 cases (medial meniscus injury in 1 case), lateral collateral ligament (LCL) injury in 2 cases, and MCL and LCL injuries in 2 cases (medial meniscus and lateral meniscus injuries in 1 case). Autologous harmstring tendon was used to reconstruct PCL under arthroscopy combined with limited open in situ suture for repair of femoral avulsion injury of ACL, and repair of MCL, LCL, and other injury in one-stage operation. Results All incisions healed by first intention. Joint effusion of knee occurred in 1 case and was cured after removal of fluid combined with pressure bandage. All patients were followed up 12-36 months with an average of 22 months. At last follow-up, the result of posterior drawer test was negative in all patients. The results of Lachman test were one-degree positive in 2 cases; the result of varus stress testing was one-degree positive in 1 case; the results of valgus stress testing were one-degree positive in 2 cases; and flexion dysfunction of the knee was observed in 1 case. The Lysholm score of knee was 87.3±6.6; the IKDC score was 88.9±6.8; and the ROM was (121.7±12.3)°, all showing significant differences when compared with preoperative ones (t=44.246, P=0.000; t=37.903, P=0.000; t=19.894, P=0.000). Conclusion For KDMLI, one-stage repair and reconstruction using autologous harmstring tendon to reconst ruct PCL under arthroscopy combined with limited open in situ suture repair of femoral avulsion injury of ACL, and repair MCL, LCL, and other injury has such advantages as minimal invasiveness, reliable fixation, less complications, and fast recovery, which can significantly improve the stability, ROM, and function of knee and obtain good short-term effectiveness.
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.