ObjectiveTo investigate the effectiveness of percutaneous compression plate (PCCP) fixation for femoral neck fracture.MethodsA clinical data of 100 patients with femoral neck fractures who were treated with internal fixation were analyzed retrospectively. The fractures were fixed with the cannulated screws (CS) in 55 patients (CS group) and with the PCCP in 45 patients (PCCP group). There was no significant difference in gender, age, the cause of injury, the fracture type, complications, and disease duration between the two groups (P>0.05). The quality of fracture reduction, bone resorption, screw slipping, femoral neck shortening, complications (nonunion, failure of fixation, and osteonecrosis of femoral head), and functional recovery of hip (Harris score) were compared between the two groups.ResultsAll incisions healed by first intention. All patients were followed up 24-56 months, with an average of 30.7 months. The quality of fracture reduction was excellent in 26 cases, good in 18 cases, fair in 9 cases, and poor in 2 cases in CS group and excellent in 21 cases, good in 17 cases, fair in 4 cases, and poor in 3 cases in PCCP group, showing no significant difference between the two groups (Z=−0.283, P=0.773). The incidence of nonunion in PCCP group was significantly lower than that in CS group (P=0.046), and the fracture healing time in PCCP group was shorter than that in CS group (t=2.155, P=0.034). There was no significant difference in the incidences of bone resorption, screw slipping, femoral neck shortening, failure of fixation, and osteonecrosis of femoral head between the two groups (P>0.05). The overall complication rates were 27.27% (15/55) in CS group and 8.89% (4/45) in PCCP group, showing significant difference (χ2=5.435, P=0.020). The Harris score in PCCP group at 6 months after operation was significantly higher than that in CS group (t=−2.073, P=0.041). However, there was no significant difference in the Harris score at 12, 18, and 24 months after operation between the two groups (P>0.05).ConclusionStable sliding compression of PCCP is benefit for the femoral neck fracture healing, especially shortening union.
Objective To investigate and compare the difference between two implants of reconstructing anterior cruciate l igament (ACL) for the early heal ing of implants tunnel interface in terms of biological mechanism. Methods Fiftyfive adult New Zealand rabbits weighing 2.0-2.8 kg were selected. Patellar l igament with tibia-bone block was obtained fromthe left knee joint serving as donor site, right knee joint served as the recipient site of autograft for ACL reconstruction. Thebone block end of implant was bone-bone interface heal ing model, while the l igament end was tendon-bone interface heal ing model. The general condition of rabbits was observed after operation, the gross observation and histology observation were conducted at 2, 4 and 8 weeks after operation (n=5), and biomechanics examination was conducted at 4 and 8 weeks after operation (n=20). Results Rabbits behaved normally after operation. The gross observation indicated that ACL had complete continuity and moderate tension during experiment. Histology observation: most part of bone-bone interface was connected by fibrous tissue, while the tendon-bone interface was mainly filled by granulation tissue 2 weeks after operation; most part of bone-bone interface was bone union, and there were osteogenesis reaction and large quantity of fibroblasts in the tendonbone interface 4 weeks after operation; complete bone union was evident in bone-bone interface, and the appearance of Sharpey fibers and the formation of indirect insertion occured in part of tendon-bone interface 8 weeks after operation. Biomechanics observation: the pull-out rate for tendon-bone interface and bone-bone interface 4 weeks after operation was 85% and 15%, respectively; while it was 95% and 5% 8 weeks after operation, respectively; indicating there was a significant difference between two groups (P lt; 0.001). Conclusion In the early stage after ACL reconstruction, bone-bone interface is better than tendonbone interface in terms of intensity and speed of heal ing.
ObjectiveTo investigate the effect of hamstring tendon transfected with adenovirus-mediated transforming growth factor β1 (AdTGF-β1) genes on the histomorphology of tendon-bone interface healing after anterior cruciate ligament (ACL) reconstruction in rabbits. MethodsAdTGF-β1 and AdGFP were diluted to 5×108 PFU/mL with DMEM. Forty-eight New Zealand white rabbits were divided into 3 groups randomly (n=16), weighing 1.6-2.5 kg for ACL reconstruction with hamstring tendon autograft. Hamstring tendon was cultured and transfected with AdTGF-β1 (group A) and AdGFP (group B) for 12 hours before ACL reconstruction, and was cultured with DMEM in group C. After 12 hours of transfection, the expression of green fluorescence was observed in groups A and B under fluorescence microscopy; TGF-β1 protein level was detected by ELISA in group A. At 2, 4, 8, and 12 weeks after operation, the specimens were harvested for HE and Masson staining; the number of fibroblasts was counted, and the Buark grading was used to evaluate tendon-bone interface healing. ResultsGreen fluorescence was observed after 12 hours of transfection in groups A and B. TGF-β1 protein level reached (221.0±12.2) ng/mL at 12 hours in group A. The histological observation showed that few fibroblasts and collagen fibers were found, and Sharpey fibers appeared in group A; regular Sharpey fibers were seen in the interface, and integrity interface in some areas at 12 weeks. But fibroblasts of groups B and C were less than those of group A, with loose tendon-bone interface; no integrity interface was observed at 12 weeks. The number of fibroblasts and Buark grading of group A were significantly higher than those of groups B and C (P<0.05), but no significant difference was found between groups B and C (P>0.05). ConclusionHamstring tendon transfected with AdTGF-β1 gene can promote the healing of tendon-bone interface after ACL reconstruction.
Objective To investigate the effect of autologous osteochondral tissue and periosteum transplantation on tendon-bone healing of rotator cuff in rabbits. Methods Twenty-four male New Zealand white rabbits were randomly divided into autologous osteochondral tissue and periosteum transplantation group (experimental group, n=12) and simple suture group (control group, n=12). Both groups were subjected to acute supraspinatus tendon injury and repaired with corresponding techniques. At 4, 8, and 12 weeks after operation, 4 specimens from each group were taken from the right shoulder joint for histological examination (HE staining, Masson staining, and Safranin O-fast green staining), and the left shoulder was subjected to biomechanical tests (maximum tensile load and stiffness). Results Both groups of animals survived until the completion of the experiment after operation. At 4 weeks after operation, both groups showed less collagen fibers and disorder at the tendon-bone junction. At 8 weeks, both groups showed reduced inflammation at the tendon-bone junction, with more organized and denser collagen fibers and chondrocytes. The experimental group showed better results than the control group. At 12 weeks, the experimental group showed typical tendon-bone transition structure, with increased generation of collagen fibers and chondrocytes, and the larger cartilage staining area. Both groups showed an increase in maximum tensile load and stiffness over time (P<0.05). The stiffness at 4 weeks and the maximum tensile load at 4, 8, and 12 weeks in the experimental group were superior to control group, and the differences were significant (P<0.05). There was no significant difference in stiffness at 8, 12 weeks between the two groups (P>0.05). Conclusion Autologous osteochondral tissue and periosteum transplantation can effectively promote the fiber and cartilage regeneration at the tendon-bone junction of rotator cuff and improve the biomechanical effect of shoulder joint in rabbits.
Objective To explore the effect of chitosan (CS) hydrogel loaded with tendon-derived stem cells (TDSCs; hereinafter referred to as TDSCs/CS hydrogel) on tendon-to-bone healing after rotator cuff repair in rabbits. Methods TDSCs were isolated from the rotator cuff tissue of 3 adult New Zealand white rabbits by Henderson step-by-step enzymatic digestion method and identified by multidirectional differentiation and flow cytometry. The 3rd generation TDSCs were encapsulated in CS to construct TDSCs/CS hydrogel. The cell counting kit 8 (CCK-8) assay was used to detect the proliferation of TDSCs in the hydrogel after 1-5 days of culture in vitro, and cell compatibility of TDSCs/CS hydrogel was evaluated by using TDSCs alone as control. Another 36 adult New Zealand white rabbits were randomly divided into 3 groups (n=12): rotator cuff repair group (control group), rotator cuff repair+CS hydrogel injection group (CS group), and rotator cuff repair+TDSCs/CS hydrogel injection group (TDSCs/CS group). After establishing the rotator cuff repair models, the corresponding hydrogel was injected into the tendon-to-bone interface in the CS group and TDSCs/CS group, and no other treatment was performed in the control group. The general condition of the animals was observed after operation. At 4 and 8 weeks, real-time quantitative PCR (qPCR) was used to detect the relative expressions of tendon forming related genes (tenomodulin, scleraxis), chondrogenesis related genes (aggrecan, sex determining region Y-related high mobility group-box gene 9), and osteogenesis related genes (alkaline phosphatase, Runt-related transcription factor 2) at the tendon-to-bone interface. At 8 weeks, HE and Masson staining were used to observe the histological changes, and the biomechanical test was used to evaluate the ultimate load and the failure site of the repaired rotator cuff to evaluate the tendon-to-bone healing and biomechanical properties. Results CCK-8 assay showed that the CS hydrogel could promote the proliferation of TDSCs (P<0.05). qPCR results showed that the expressions of tendon-to-bone interface related genes were significantly higher in the TDSCs/CS group than in the CS group and control group at 4 and 8 weeks after operation (P<0.05). Moreover, the expressions of tendon-to-bone interface related genes at 8 weeks after operation were significantly higher than those at 4 weeks after operation in the TDSCs/CS group (P<0.05). Histological staining showed the clear cartilage tissue and dense and orderly collagen formation at the tendon-to-bone interface in the TDSCs/CS group. The results of semi-quantitative analysis showed that compared with the control group, the number of cells, the proportion of collagen fiber orientation, and the histological score in the TDSCs/CS group increased, the vascularity decreased, showing significant differences (P<0.05); compared with the CS group, the proportion of collagen fiber orientation and the histological score in the TDSCs/CS group significantly increased (P<0.05), while there was no significant difference in the number of cells and vascularity (P>0.05). All samples in biomechanical testing failed at the repair site during the testing process. The ultimate load of the TDSCs/CS group was significantly higher than that of the control group (P<0.05), but there was no significant difference compared to the CS group (P>0.05). Conclusion TDSCs/CS hydrogel can induce cartilage regeneration to promote rotator cuff tendon-to-bone healing.
Objective To investigate whether the Runx2 gene can induce the differentiation of human amniotic mesenchymal stem cells (hAMSCs) to ligament fibroblasts in vitro and promote the tendon-bone healing in rabbits. Methods hAMSCs were isolated from the placentas voluntarily donated from healthy parturients and passaged, and then identified by flow cytometric identification. Adenoviral vectors carrying Runx2 gene (Ad-Runx2) and empty vector adenovirus (Ad-NC) were constructed and viral titer assay; then, the 3rd generation hAMSCs were transfected with Ad-Runx2 (Ad-Runx2 group) or Ad-NC (Ad-NC group). The real-time fluorescence quantitative PCR and Western blot were used to detect Runx2 gene and protein expression to verify the effectiveness of Ad-Runx2 transfection of hAMSCs; and at 3 and 7 days after transfection, real-time fluorescence quantitative PCR was further used to detect the expressions of ligament fibroblast-related genes [vascular endothelial growth factor (VEGF), collagen type Ⅰ, Fibronectin, and Tenascin-C]. The hAMSCs were used as a blank control group. The hAMSCs, hAMSCs transfected with Ad-NC, and hAMSCs were mixed with Matrigel according to the ratio of 1 : 1 and 1 : 2 to construct the cell-scaffold compound. Cell proliferation was detected by cell counting kit 8 (CCK-8) assay, and the corresponding cell-scaffold compound with better proliferation were taken for subsequent animal experiments. Twelve New Zealand white rabbits were randomly divided into 4 groups of sham operation group (Sham group), anterior cruciate ligament reconstruction group (ACLR group), anterior cruciate ligament reconstruction+hAMSCs transfected with Ad-NC-scaffold compound group (Ad-NC group), and anterior cruciate ligament reconstruction+hAMSCs transfected with Ad-Runx2-scaffold compound group (Ad-Runx2 group), with 3 rabbits in each group. After preparing the ACL reconstruction model, the Ad-NC group and the Ad-Runx2 group injected the optimal hAMSCs-Matrigel compunds into the bone channel correspondingly. The samples were taken for gross, histological (HE staining and sirius red staining), and immunofluorescence staining observation at 1 month after operation to evaluate the inflammatory cell infiltration as well as collagen and Tenascin-C content in the ligament tissues. ResultsFlow cytometric identification of the isolated cells conformed to the phenotypic characteristics of MSCs. The Runx2 gene was successfully transfected into hAMSCs. Compared with the Ad-NC group, the relative expressions of VEGF and collagen type Ⅰ genes in the Ad-Runx2 group significantly increased at 3 and 7 days after transfection (P<0.05), Fibronectin significantly increased at 3 days (P<0.05), and Tenascin-C significantly increased at 3 days and decreased at 7 days (P<0.05). CCK-8 detection showed that there was no significant difference (P>0.05) in the cell proliferation between groups and between different time points after mixed culture of two ratios. So the cell-scaffold compound constructed in the ratio of 1∶1 was selected for subsequent experiments. Animal experiments showed that at 1 month after operation, the continuity of the grafted tendon was complete in all groups; HE staining showed that the tissue repair in the Ad-Runx2 group was better and there were fewer inflammatory cells when compared with the ACLR group and the Ad-NC group; sirius red staining and immunofluorescence staining showed that the Ad-Runx2 group had more collagen typeⅠ and Ⅲ fibers, tending to form a normal ACL structure. However, the fluorescence intensity of Tenascin-C protein was weakening when compared to the ACLR and Ad-NC groups. Conclusion Runx2 gene transfection of hAMSCs induces directed differentiation to ligament fibroblasts and promotes tendon-bone healing in reconstructed anterior cruciate ligament in rabbits.
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 observe the effect and significance of autologous fibrin clot on tendon-bone healing after anterior cruciate ligament (ACL) reconstruction.MethodsBetween October 2014 and January 2016, 34 patients (34 knees) with ACL injury were enrolled in the study. During ACL reconstruction, autologous fibrin clot was used in 17 cases (trial group) and was not used in 17 cases (control group). The anterior drawer test, Lachman test, and axial displa-cement test were positive in 2 groups before operation. There was no significant difference in gender, age, causes of injury, injury side, disease cause, and preoperative knee joint activity, Lysholm score, and American Hospital for Special Surgery (HSS) score between 2 groups (P>0.05), with comparable. The results of anterior drawer test, Lachman test, and axial displacement test were recorded and compared between 2 groups after operation. The knee joint activity, Lysholm score, and HSS score were used to evaluate the knee function recovery at 6, 24, and 48 weeks after operation; the graft signal intensity, graft signal to noise ratio, bone tunnel expansion, and graft tendon-bone node T2 value were measured.ResultsAll patients were followed up 48 weeks. Surgical incision healed at stage I. No joint infection and joint adhesion occurred. The drawer test, Lachman test, and axial shift test were negative in 2 groups. At 6, 24, and 48 weeks after operation, the Lysholm score of trial group was significantly higher than that of control group (P<0.05); there was no significant difference in knee joint activity between 2 groups (P>0.05). The HSS score of trial group was significantly higher than that of control group at 24 and 48 weeks (P<0.05), but no significant difference was found at 6 weeks (P>0.05). MRI measu-rement showed that there was significant difference in graft signal intensity, bone tunnel expansion, and graft signal to noise ratio between 2 groups at 6, 24, and 48 weeks after operation (P<0.05). There was no significant difference in graft tendon-bone node T2 value between 2 groups (P>0.05) at 48 weeks after operation, but difference was significant at 6 and 24 weeks (P<0.05).ConclusionAutologous fibrin clot can effectively enhance graft revascularization, and accelerate the process of tendon-bone healing after ACL reconstruction.
ObjectiveTo review the bioactive strategies that enhance tendon graft healing after anterior cruciate ligament reconstruction (ACLR), and to provide insights for improving the therapeutic outcomes of ACLR. Methods The domestic and foreign literature related to the bioactive strategies for promoting the healing of tendon grafts after ACLR was extensively reviewed and summarized. ResultsAt present, there are several kinds of bioactive materials related to tendon graft healing after ACLR: growth factors, cells, biodegradable implants/tissue derivatives. By constructing a complex interface simulating the matrix, environment, and regulatory factors required for the growth of native anterior cruciate ligament (ACL), the growth of transplanted tendons is regulated at different levels, thus promoting the healing of tendon grafts. Although the effectiveness of ACLR has been significantly improved in most studies, most of them are still limited to the early stage of animal experiments, and there is still a long way to go from the real clinical promotion. In addition, limited by the current preparation technology, the bionics of the interface still stays at the micron and millimeter level, and tends to be morphological bionics, and the research on the signal mechanism pathway is still insufficient.ConclusionWith the further study of ACL anatomy, development, and the improvement of preparation technology, the research of bioactive strategies to promote the healing of tendon grafts after ACLR is expected to be further promoted.
ObjectiveTo assess the effect of microfracture and biomimetic hydrogel scaffold on tendon-to-bone healing in a rabbit rotator cuff tear model.MethodsGelatin and methacrylic anhydride were used to synthesize gelatin methacryloyl (GelMA). Then the GelMA were treated with ultraviolet rays and vacuum freeze-drying method to obtain a biomimetic hydrogel scaffold. The morphology of the scaffold was observed by gross observation and scanning electron microscope. Degradation of the scaffold was determined at different time points. Twenty-four adult New Zealand rabbits, weighting 2.8-3.5 kg and male or female, were surgically created the bilateral acute rotator cuff tear models. One shoulder was treated with microfractures on the footprint and transosseous suture (control group, n=24). The other shoulder was treated with the same way, except for putting the scaffold on the footprint before transosseous suture (experimental group, n=24). The general conditions of rabbits were observed postoperatively. Tendon-to-bone healing was evaluated by gross observation, Micro-CT, HE staining, and bio-mechanical testing at 4 and 8 weeks after operation.ResultsThe scaffold was white and has a porous structure with pore size of 31.7-89.9 μm, which degraded slowly in PBS solution. The degradation rate was about 95% at 18 days. All the rabbits survived to the completion of the experiment. Micro-CT showed that there was no obvious defect and re-tear at the tendon-to-bone interface in both groups. No difference was found in bone mineral density (BMD), tissue mineral density (TMD), and bone volume/total volume (BV/TV) between the two groups at 4 and 8 weeks postoperatively (P>0.05). HE staining showed that the fibrous scar tissue was the main component at the tendon-to-bone interface in the control group at 4 and 8 weeks postoperatively; the disorderly arranged mineralized cartilage and fibrocartilage formation were observed at the tendon-to-bone interface in the experimental group at 4 weeks, and the orderly arranged cartilage formation was observed at 8 weeks. Besides, the tendon maturation scores of the experimental group were significantly higher than those of the control group at 4 and 8 weeks (P<0.05). There was no significant difference in the ultimate load to failure and stiffness between the two groups at 4 weeks (P>0.05); the ultimate load to failure at 8 weeks was significantly higher in the experiment group than in the control group (t=4.162, P=0.009), and no significant difference was found in stiffness between the two groups at 8 weeks (t=2.286, P=0.071).ConclusionCompared with microfracture alone, microfracture combined with biomimetic hydrogel scaffold can enhance tendon-to-bone healing and improve the ultimate load to failure in rabbits.