Modified arthroscopic Brostr?m procedure for the treatment of chronic ankle instability combined with multiple ligament laxity / 中华创伤杂志

Objective:To investigate the effectiveness of modified arthroscopic Brostr?m procedure for the treatment of chronic ankle instability combined with multiple ligament laxity.Methods:A retrospective case series study was used to analyze the clinical data of 26 patients with chronic ankle instability combined with multiple ligament laxity treated at Union Hospital, Tongji Medical College of Huazhong University of Science and Technology from January 2016 to December 2020, including 10 males and 16 females; aged 18-48 years [(27.5±7.1)years]. All patients underwent arthroscopic repair of the anterior talofibular ligament (ATFL) by the modified Brostr?m procedure. Healing of surgical incisions was observed after operation. The change of talus tilt angle for ankle stability evaluation, the American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score for ankle function evaluation, and the visual analogue score (VAS) for pain evaluation were assessed before operation, at 3 months postoperatively and at the last follow-up. Complications were observed.Results:All patients were followed up for 18-47 months [(25.3±8.5)months]. All surgical incisions were healed at stage I. The talus tilt angle was decreased from preoperative (15.6±4.7)° to (4.1±1.3)° and (3.5±0.9)° at 3 months postoperatively and at the last follow-up (all P<0.01). The AOFAS ankle-hindfoot score was improved from preoperative (65.8±14.5)points to (86.5±5.6)points and (93.4±4.2)points at 3 months postoperatively and at the final follow-up (all P<0.01). The VAS was decreased from preoperative 3.0 (2.0, 4.0)points to 1.5 (0.0, 2.0)points and 1.0 (0.0, 1.2)points at 3 months postoperatively and at the last follow-up (all P<0.01). Significantly higher AOFAS ankle-hindfoot score and lower VAS were found at the final follow-up when compared with the scores at 3 months postoperatively (all P<0.05). One patient developed superficial peroneal nerve injury, which was recovered spontaneously without special treatment. Conclusion:For chronic ankle instability combined with multiple ligament laxity, the modified arthroscopic Brostr?m procedure has advantages of improved ankle stability, good ankle function recovery, obvious pain relief and less postoperative complications.

Arthroscopic medullary decompression combined with platelet-rich plasma therapy for bone marrow edema of the talus / 中华创伤杂志

Objective:To report the efficacy of arthroscopic medullary decompression combined with platelet-rich plasma (PRP) in the treatment of bone marrow edema of the talus.Methods:A retrospective case series study was used to analyze the clinical data of 17 patients with bone marrow edema of the talus admitted to Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 2018 to July 2020. There were 11 males and 6 females, with the age range of 15-56 years [(45.7±4.3)years]. All patients were subjected to arthroscopic medullary decompression combined with the administration of PRP. Operation time and wound healing were recorded. Maximum area of bone marrow edema was measured by MRI preoperatively and at 6 and 12 months postoperatively. Ankle range of motion (ROM), visual analog score (VAS) and American Association of Foot and Ankle Surgery (AOFAS) ankle-hindfoot score were measured preoperatively and at 6 and 12 months postoperatively. Complications were also detected.Results:All patients were followed up for 12-41 months [(16.7±2.1)months]. Operation time was 45.2-68.5 minutes [(53.4±12.4)minutes]. All wounds were healed at stage I. The maximum area of bone marrow edema decreased from (28.2±6.9)mm 2 preoperatively to (16.3±5.7)mm 2 at 6 months postoperatively and to (7.1±1.7)mm 2 at 12 months postoperatively (all P<0.01). Ankle ROM increased from (52.2±8.9)° preoperatively to (72.3±3.1)° at 6 months postoperatively and to (83.1±2.8)° at 12 months postoperatively (all P<0.01). VAS decreased from (8.2±0.6)points preoperatively to (6.5±0.4)points at 6 months postoperatively and to (3.1±0.8)points at 12 months postoperatively (all P<0.01). AOFAS ankle-hindfoot score increased from (32.4±4.8)points preoperatively to (54.4±6.5)points at 6 months postoperatively and to (88.7±4.3)points at 12 months postoperatively (all P<0.01). There were significant differences in maximum area of bone marrow edema of the talus, ankle ROM, VAS and AOFAS ankle-hindfoot score at 12 months postoperatively when compared with those at 6 months postoperatively (all P<0.01). One patient showed the symptom of localized skin numbness postoperatively, and improved with nerve nutrition therapy. Conclusion:Arthroscopic medullary decompression combined with PRP therapy for bone marrow edema of the talus presents good short-term clinical benefits in terms of reduced extent of bone marrow edema, improved ankle ROM, attenuated pain, improved ankle joint function and few postoperative complications.

Comparison of arthroscopy-assisted reduction and internal fixation versus open reduction and internal fixation for tibial plateau fractures / 中华创伤骨科杂志

Objective:To compare the efficacy between arthroscopy-assisted reduction and internal fixation (ARIF) versus open reduction and internal fixation (ORIF) in the treatment of tibial plateau fractures.Methods:A retrospective analysis was done of the 75 patients with tibial plateau fracture who had been treated by ARIF or ORIF at Department of Orthopaedics, Union Hospital Affiliated to Tongji Medical Collage from January 2016 to August 2018. They were 58 men and 17 women, aged from 20 to 54 years (average, 47 years). The left side was affected in 42 cases and the right side in 33. By the Schatzker classification, there were 23 cases of type Ⅰ, 49 cases of type Ⅱ and 3 cases of type Ⅲ. Of them, 40 were treated by ARIF and 35 by ORIF. The 2 groups were compared in terms of operation time, incision length, intraoperative blood loss, hospital stay, postoperative complications and the Hospital for Special Surgery (HSS) scores 12 months after operation.Results:There was no statistically significant difference between the 2 groups in the preoperative general data, showing the 2 groups were comparable ( P>0.05). The patients were followed up for 12 to 15 months (average, 13.5 months) after operation. The wounds in the 75 patients healed at one stage with no complications like neurovascular lesions. All the fractures healed within 6 months after operation. Compared with the ORIF group, the ARIF group had significantly longer operation time (58.3 min ± 4.2 min versus 48.4 min ± 5.2 min), a significantly shorter incision (4.3 cm ± 0.9 cm versus 6.2 cm ± 0.8 cm), and significantly less intraoperative blood loss (60.8 mL ± 4.5 mL versus 72.8 mL ± 6.5 mL) ( P<0.05). There was no significant difference between the 2 groups in hospital stay (5.1 d ± 0.6 d versus 5.5 d ± 1.6 d) ( P>0.05). Fifteen patients in the ARIF group and 5 in the ORIF group were complicated with soft tissue injury, showing a statistically significant difference ( P<0.05). The excellent and good rate by HSS scores was 100% (40/40) for the ARIF group and 85% (34/40) for the ORIF group, showing no significant difference ( P>0.05). Conclusions:In the treatment of tibial plateau fractures of Schatzker types Ⅰ-Ⅲ, both ARIF and ORIF may result in good efficacy. However, ARIF can evaluate and treat the complicated soft tissue injuries to the knee joint more precisely, showing advantages of shorter operation time, a smaller incision and less blood loss.

MRI presentation and pathogenesis of tennis legs / 中华放射学杂志

Objective To evaluate the imaging features of MRI of tennis legs and to explore the pathogenesis of tennis legs. Methods A retrospective analysis was made on the MRI images of 38 patients with tennis legs which met the criteria and were clinically diagnosed in our hospital from May 2014 to June 2018. All patients underwent non?enhanced MRI. Coronal T1WI、T2WI fast spin echo (TSE) and transverse proton density weighted imaging (PDWI) were performed. The signs of fluid collection between gastrocnemius muscle (GM) and soleus muscle (SM),muscle and tendon injuries, superficial vein dilatation of calf were observed and recorded. Results Coronal T1WI, T2WI TSE and transverse PDWI sequences showed 30 (75.0%) places fluid collection (hematoma or effusion) between medial head of the gastrocnemius muscle (MCM) and SM, 11 (27.5%) places fluid collection (hematoma or effusion) between lateral head of gastrocnemius muscle(LGM)and SM,7 (17.5%) places fluid collection (hematoma or effusion) in MGM and 2 (5.0%) placesin SM. There were 17 (42.5%) places that hematoma or effusion spread around the fascia of the lower leg. The diameter and thickness of hematoma or effusion are about 1.7-22.3 cm and 0.2-3.5 cm, respectively. Rupture of the GM was seen in 37 (92.5%) places,including 37 places rupture of the MGM at the myotendinous junction, 15 places rupture of the LGM at the myotendinous junction, 24 places tendonrupture of MGM,3 places tendon rupture of MGM and LGM,and 2 places tendon rupture of LGM. The maximum diameter of tendon rupture was 1.2-27.0 mm. The muscle rupture of MGM was seen in one place, and muscle rupture of MGM and LGM was seen in one place at the same time. Rupture of the SM was seen in 15 (37.5%) places, including 15 places rupture of the SM at the myotendinous junction, 2 places muscle rupture of SM, 6 places tendonrupture of SM. The maximum diameter of tendon rupture was 2.5-14.9 mm. Rupture of plantaris tendon (PT) was seen in 4 (10.0%) places. Superficial vein dilatation was seen in 3 (7.5%) places. Conclusion This study shows that the rupture of the MGM at the myotendinous junction and the tendon is the main responsibility of tennis leg.

Seeding fresh bone marrow aspirate directly on scaffolds to construct tissue-engineered ligament / 中国组织工程研究

BACKGROUND:There are several reports about the application of fresh bone marrow aspirate being injected directly to repair partial ligament injury, but the application about fresh bone marrow aspirate directly being planted on scaffolds to build tissue-engineered ligament is rarely mentioned. OBJECTIVE:To evaluate the feasibility of applying fresh bone marrow aspirate planted directly on scaffolds to construct tissue-engineered ligament METHODS:We constructed fibroin fiber/smal intestinal submucosa composite scaffold, then planting fresh bone marrow directly to built bone marrow seeding group and planting seed cel s (bone marrow mesenchymal stem cel s) on the scaffold to built cel seeding group. The control group had no treatment. After that, we detected the density of cel adhesion, cel proliferation ability and extracel ular matrix secretion. Then, the composite in the bone marrow seeding group was implanted into the broken anterior cruciate ligament in rabbits, and material biocompatibility in vivo was evaluated after 12 weeks. RESULTS AND CONCLUSION:After 4 hours of incubation, bone marrow seeding group was significantly higher than the cel seeding group in cel adhesion density and proliferation rate (P<0.05). Bone marrow seeding group and cel seeding group showed higher type I, III col agen secretion compared with the control group (P<0.05), but the col agen secretion of bone marrow seeding group and cel seeding group showed no significant difference. Composite cel scaffold implantation in vivo did not cause fatal immune rejection and severe inflammatory reaction, and no significant ligament regeneration and vascularization occurred. These findings indicate that fresh bone marrow aspirate can be seeded directly on scaffolds to construct tissue-engineered ligament, and the short-term biocompatibility in vivo is good.

Mobilization efficiency of granulocyte colony-stimulating factor and stem cell factor to bone marrow mononuclear cells and mechanisms / 华中科技大学学报（医学）（英德文版）

The mobilization efficiency of granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF) to bone marrow mononuclear cells (MNCs) in mice was observed, and the changes of CXCL12/CXCR4 signal were detected in order to find out the mobilization mechanism of stem cells. Kunming mice were randomly divided into two groups. The mice in treatment group were subjected to subcutaneous injection of G-CSF at a dose of 100 mug/kg and SCF at a dose of 25 mug/kg every day for 5 days, and those in control group were given isodose physiological saline. The MNCs were separated, counted and cultured, and the colony-forming unit-fibroblast (CFU-F) was evaluated. CD34(+)CXCR4(+) MNCs were sorted by flow cytometry. The expression of CXCL12 protein in bone marrow extracellular fluid was detected by ELISA, and that of CXCL12 mRNA in bone marrow was measured by RT-PCR. The results showed that the counts of MNCs in peripheral blood and bone marrow were increased after administration of G-CSF/SCF (P<0.01). The factors had a dramatic effect on the expansion capability of CFU-F (P<0.05). Flow cytometric of bone marrow MNCs surface markers revealed that CD34(+)CXCR4(+) cells accounted for 44.6%+/-8.7% of the total CD34(+) MNCs. Moreover, G-CSF/SCF treatment induced a decrease in bone marrow CXCL12 mRNA that closely mirrored the fall in CXCL12 protein. In this study, it is evidenced that G-CSF/SCF can effectively induce MNCs mobilization by disrupting the balance of CXCL12/CXCR4 signaling pathway in the bone marrow and down-regulating the interaction of CXCL12/CXCR4.

Effect of RGD-modified silk material on the adhesion and proliferation of bone marrow-derived mesenchymal stem cells / 华中科技大学学报（医学）（英德文版）

In order to investigate the effect of Arg-Gly-Asp (RGD) peptide-modified silk biomaterial on the adhesion and proliferation of bone marrow-derived mesenchymal stem cells (MSCs), MSCs of third generation were seeded onto the surface of RGD-decorated silk (silk-RGD group), silk alone (silk group) or tissue culture plate (TCP group). After incubation for 4 or 12 h, MSCs were examined quantitatively by using precipitation method for cell attachment. The cell proliferation, which was defined as cell density, was compared among the three groups after culture for 1, 2, 3, and 4 days. Cell skeleton, which was labeled fluorescently, was observed under laser confocal microscope after 24 h of culture. The results showed that cell adhesion rate in silk-RGD group was higher than in silk group (P0.05). There were no significant differences in the cell proliferation among the three groups at different time points (P>0.05 for all). Laser confocal microscopy revealed that in silk-RGD group, MSCs, strongly fluorescently stained, spread fully, with stress fibers clearly seen, while in silk group, actin filaments were sparsely aligned and less stress fibers were found. It was concluded that RGD peptide could improve the adhesion of MSCs to the silk scaffold, but had no impact on the proliferation of the cells.

Mechanical strength and in vitro degradation of a silk scaffold for tissue-engineered ligaments / 中国组织工程研究

BACKGROUND: Presently, the biomaterial used in ligament tissue engineering such as collagen protein, polylactic acid, polyglycolic acid, small intestinal submucosa, glycan and nanomaterial are characterized by rapid degradation, resulting in inflammatory reaction after applying in host.OBJECTIVE: To investigate mechanical strength and in vitro degradation of silk scaffold and explore the reaction to macrophages.DESIGN: Controlled experiment.SETTING: Experiments were performed at the Department of Orthopaedics, Union Hospital, Huazhong University of Science and Technology from September 2004 to January 2005.MATERIALS: White raw Bombyx mori silkworm fibers of size 20/22 (according to the manufacturer) were obtained from the market. Bundles of 30 parallel fibers were prepared for a bundle of scaffold, which was put into fervens 5g/L Na2CO3for degumming. Ratio of Na2CO3 solution (Ml) to raw silk (g) was 1000.METHODS: In vitro degradation: 8cm long silk scaffold was weighed after drying. Subsequently, the silk scaffold was separately dipped into phosphate buffer saline (PBS) and 1.0g/L collagenase prepared with PBS. Twelve weeks later, silk scaffold was weighed to calculate weight loss rate. Simultaneously, tensile test was performed to detect the ultimate tensile strength (UTS) of samples. Culture of monocyte strain RAW264.7:2×108L-1 macrophage suspension (1mL) were separately added in a silk scaffold group, a control group and a lipopolysaccharide (LPS) group. At days 1 and 7, cell supernatant was collected from each group. Tumor necrosis factor-α(TNF-α) levels were measured by enzyme linked immunosorbent assay (ELISA).MAIN OUTCOME MEASURES: ① Changes in weight loss rate and UTS of the silk matrices after incubated with collagenase and the PBS. ②TNF-αlevels in the supernatant of each groups at days 1 and 7.RESULTS: Mass of silk matrices reduced by over 50% after incubated with collagenase for 8 weeks, but no change was found in PBS. UTS decreased by over 50% 8 weeks after incubated with collagenase, but no change was detected in PBS. At days 1 and 7, TNF-α levels in the supernatant was less in the silk scaffold group; TNF-α levels in the supernatant was significantly higher in the LPS group than in the silk scaffold group (P<0.01), but no significant difference in TNF-α levels was measured between the silk scaffold group and the control group (P>0.05).CONCLUSION: After 12-weeks degradation, silk scaffold still has good mechanical properties. Macrophages possess immunological inertia at days 1 and 7 after inoculated with macrophages.

Protective Effect of Niacinamide on interleukin-1β-induced Annulus Fibrosus Type Ⅱ Collagen Degeneration in vitro / 华中科技大学学报（医学）（英德文版）

The protective effect of niacinamide on interleukin-1β (IL-1β)-induced annulus fibrosus (AF) type Ⅱ collagen degeneration in vitro and the mechanism were investigated. Chiba's intervertebrai disc (IVD) culture models in rabbits were established and 48 IVDs from 12 adult Japanese white rabbits were randomly divided into 4 groups: normal control group, niacinamide-treated group, type Ⅱ collagen degneration group (IL-1β) and treatment group (niacinamide+IL-1β). After culture for one week, AFs were collected for inducible nitric oxide synthase (iNOS), cysteine containing aspartate specific protease-3 (Caspase-3) and type Ⅱ collagen immunohistochemical examination, and type Ⅱ collagen reverse transcription polymerase chain reaction (RT-PCR). The results showed that rate of iNOS positive staining AF cells in the 4 groups was 17.6%, 10.9%, 73.9% and 19.3% respectively. The positive rate in treatment group was significantly lower than in the type Ⅱ collagen degeneration group (P＜0.01). Rate of Caspase-3 positive staining AF cells in the 4 groups was 3.4%, 4.2%, 17.6% and 10.3% respectively. The positive rate in treatment group was lower than in the type Ⅱ collagen degeneration group (P＜0.01). Type Ⅱ collagen staining demonstrated that lamellar structure and continuity of collagen in treatment group was better reversed than in the degeneration group. RT-PCR revealed that the expression of type Ⅱ collagen in treatment group was significantly stronger than that in type Ⅱ collagen degeneration group (P＜0.01). It was concluded that niacinamide could effectively inhibit IL-1β stimulated increase of iNOS and Caspase-3 in AF, and alleviate IL-1β-caused destruction and synthesis inhibition of type Ⅱ collagen. Niacinamide is of potential for clinical treatment of IVD degeneration.

Protective effect of niacinamide on interleukin-1beta-induced annulus fibrosus type II collagen degeneration in vitro / 华中科技大学学报（医学）（英德文版）

The protective effect of niacinamide on interleukin-1beta (IL-1beta)-induced annulus fibrosus (AF) type II collagen degeneration in vitro and the mechanism were investigated. Chiba's intervertebral disc (IVD) culture models in rabbits were established and 48 IVDs from 12 adult Japanese white rabbits were randomly divided into 4 groups: normal control group, niacinamide-treated group, type II collagen degneration group (IL-1beta) and treatment group (niacinamide+IL-1beta). After culture for one week, AFs were collected for inducible nitric oxide synthase (iNOS), cysteine containing aspartate specific protease-3 (Caspase-3) and type II collagen immunohistochemical examination, and type II collagen reverse transcription polymerase chain reaction (RT-PCR). The results showed that rate of iNOS positive staining AF cells in the 4 groups was 17.6%, 10.9%, 73.9% and 19.3% respectively. The positive rate in treatment group was significantly lower than in the type II collagen degeneration group (P<0.01). Rate of Caspase-3 positive staining AF cells in the 4 groups was 3.4%, 4.2%, 17.6% and 10.3% respectively. The positive rate in treatment group was lower than in the type II collagen degeneration group (P<0.01). Type II collagen staining demonstrated that lamellar structure and continuity of collagen in treatment group was better reversed than in the degeneration group. RT-PCR revealed that the expression of type II collagen in treatment group was significantly stronger than that in type II collagen degeneration group (P<0.01). It was concluded that niacinamide could effectively inhibit IL-1beta stimulated increase of iNOS and Caspase-3 in AF, and alleviate IL-1beta-caused destruction and synthesis inhibition of type II collagen. Niacinamide is of potential for clinical treatment of IVD degeneration.

Cryopreserved allogenic osteochondral pillars transplantation for cartilage defects / 中国组织工程研究

0.05), and the total score were similar in the two groups. Type Ⅱ collagen in the two groups was strongly positive by immunohistochemistry staining. CONCLUSION: Cryopreserved allogenic osteochondral pillars transplantation can repair small full-thickness articular cartilage defects. The chondrocytes are alive in short time, and they can secret cartilage matrix without obvious rejection. It has similar efficacy in histology with autogenic osteochondral pillar transplantation.

Biomechanical properties of compound ligament with small intestinal submucosa wrapping silk fibers / 中国组织工程研究

0.05).CONCLUSION:The excellent mechanical properties of composite ligament can meet the mechanical requirements of appropriate ligament tissue engineering scaffolds.

The use of calcaneal anatomic plate in arthroscopically-assisted open reduction and internal fixation of intra-articular calcaneal fractures / 华中科技大学学报（医学）（英德文版）

To discuss and evaluate the method and effect of using calcaneal anatomic plate in treatment of intra-articular fractures of the calcaneus with assistant of arthroscope, 86 intra-articular fractures of the calcaneus in 78 patients were reduced by open reduction, and rigid fixation was made with calcaneal anatomic plate under assistant of arthroscope. The average follow-up duration was 18 months (range 12-30 months). The effect of treatment was evaluated according to AOFAS and X-ray before and after operation. The results showed that 86 patients have obtained satisfactory reduction according to X-ray, and there was significant difference before and after operation (P < 0. 01), the total excellent and fine rate was 91.86%. Treating intra-articular fractures of the calcaneus with calcaneal anatomic plate under arthroscope may provide more chance to achieve anatomical reconstruction, which can lead to satisfied recovery of function and few complication.

Up-regulation of Niacinamide in Intervertebral Disc Aggrecan in vitro / 华中科技大学学报（医学）（英德文版）

The regulatory effects of niacinamide (Nia) on intervertebral disc (IVD) aggrecan in vitro was investigated. Chiba's 10 ng/mL interleukin-1 (IL-1)-induced rabbit IVD degeneration model in vitro was established. 0.5, 0.25 and 0.05 mg/mL Nia was added to normal and degenerated IVDs for intervention. On the first and second week after intervention, safranin O-fast green staining intensity and glycosaminoglycan (GS) content were measured. The expression of aggrecan core protein was detected by RT-PCR. The results showed: (1) After treatment with 0.5 mg/mL Nia for one week, the GS content in nucleus pulposus (NP) was increased by 44.8 % as compared with control group (P＜0.01); The GS content in IL-1 induction groups was increased with the increase of Nia concentrations: After treatment with 0.5 mg/mL for one week, the GS content in NP was increased by 68.3 % as compared with control group (P＜0.01). After two weeks, GS content in NP and fibrous rings was still higher than in control group at the same period (P＜0.01)and untreated group (P＜0.01). (2) Safranin O-fast green staining revealed that with the increase of Nia concentrations, staining density in NP and fibrous rings was increased and histological structure damage to IVDs by IL-1β was alleviated. (3) RT-PCR showed that the expression of core protein gene in IL-1β-induced degenerated IVDS was increased with the increase of Nia concentrations.It was concluded that under conditions in vitro, Nia could up-regulate the expression of aggrecan in IVDs and protect IVDs from IL-1β-induced degeneration at least partially, which offers a potential choice for IVD degeneration clinical therapy.

The Use of Calcaneal Anatomic Plate in Arthroscopically-assisted Open Reduction and Internal Fixation of Intra-articular Calcaneal Fractures / 华中科技大学学报（医学）（英德文版）

To discuss and evaluate the method and effect of using calcaneal anatomic plate in treatment of intra-articular fractures of the calcaneus with assistant of arthroscope, 86 intra-articular fractures of the calcaneus in 78 patients were reduced by open reduction, and rigid fixation was made with calcaneal anatomic plate under assistant of arthroscope. The average follow-up duration was 18 months (range 12-30 months). The effect of treatment was evaluated according to AOFAS and X-ray before and after operation. The results showed that 86 patients have obtained satisfactory reduction according to X-ray, and there was significant difference before and after operation (P＜0.01), the total excellent and fine rate was 91.86 %. Treating intra-articular fractures of the calcaneus with calcaneal anatomic plate under arthroscope may provide more chance to achieve anatomical reconstruction, which can lead to satisfied recovery of function and few complication.

Treatment of bone defect with osteoblasts transfected by transforming growth factor-beta 1 gene in combination with biomimetic biodegradable polymer scaffolds / 中国组织工程研究

BACKGROUND: How to deal with bone defect is a big problem to surgeons. In recent years, the development in the technology of molecular biology and tissue engineering provides broad prospect for the clinical treatment of bone defect, which is one of the important study directions in department of orthopedics. The transforming growth factor-beta 1(TGF-β1),one of the important factors in bone formation, plays an important role in bone metabolism and recovery.OBJECTIVE: To observe the therapeutic effects of bone defects with osteoblasts transfected . By TGF-β1 combining with biomimetic biodegradable polymer scaffolds.DESIGN: Randomized controlled trial.SETTING: Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Siderophilin, trypsin, 3H-proline and sirius red, etc.MATERIALS: The experiment was completed in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from March to August in 2003. Twenty healthy adult Sprague-Dawley(SD) rats of SPF grade, weighing 200-250 g, were provided by the Experimental Animal Center of Tongji Medical College.METHODS: The osteoblasts transfected by TGF-β1 gene, combining with poly-DL-lactic acid scaffolds modified with poly-L-lysine, were transplanted into rat tibia defect. Radiographs and histological analysis were performed to evaluate the repair effects.MAIN OUTCOME MEASURES: The X-ray evaluation and histology observation were performed at the 4th and 8th weeks after the operation.RESULTS: Totally 20 SD rats were included in result analysis without one rat missing. ①In the experiment group, X-ray image indicated callus formation, while histology observation showed osteoid tissue and new bone formation, and osteoblasts attached to the surface of the materials after 4 weeks. Eight weeks later, the defect was essentially repaired, and the bone density of new bone was similar to that of the autogenous bone. ②In the control group, there was no formation of callus and osteoid tissue, and few osteoblasts attached to the surface of the materials, and a lot of lymphocytes infiltrated and blood capillary grew in the lacune of materials after 4 weeks. Eight weeks later, the imbedded materials were substituted mostly by fibrous tissue.CONCLUSION: The ideal repair effect of bone defect can be obtained through the combination of molecular biology with tissue engineering.

Expression of rat transforming growth factor beta 1 gene in the transfected osteoblasts / 中国组织工程研究

BACKGROUND: The cells of biomaterial compound cytokine gene or compound transfected cytokine gene that are transplanted into the area of bone defect can promote bone repair.OBJECTIVE: To investigate the feasibility of gene therapy for bone defect after rat transforming growth factor (TGF)β1 gene is transfected into osteoblasts.DESIGN: A controlled and observational experiment.SETTING: Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Five newborn Sprague-Dawley rats of either gender were included.METHODS: The experiment was conducted in the laboratory of Orthopedic Department, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, from February to September 2000.Rat osteoblasts were transfected with pcDNA3-TGF-β1 plasmid by lipofectamine mediated gene transfer, and the plasmid pcDNA transfected cells were set as control group. The transient expression of TGF-β1 was detected by strept avidin-biotin-peroxidase complex (SABC) method and in situ hybridization detection 24 hours later. The cells transfected by G418 for 2 weeks were detected with SABC to investigate the stable expression of TGF-β1 gene.MAIN OUTCOME MEASURES: SABC method and in situ hybridization detection were applied to detect gene expression.of pcDNA3-TGF-β1 transfected osteoblasts and in situ hybridization detection: After the osteoblasts were transfected for 24 hours, cytoplast was full of brown granules and there were no brown granules in the cytoplast of blank carrier transfected cells, indicating that TGF-β1 mRNA was signifiG418: transfected cells still had high expression of TGF-β1 after 2-week G418 screening.CONCLUSION: Osteoblasts can express cytokine immediately with high effect using gene transfection technique. The stable and high expression is presented after TGF-β1 gene is transfected into osteoblasts at the moment of transfection and after 2-week screening, proving the feasibility of gene therapy for bone defect when cytokine gene is transfected into osteoblasts.

Gene-enhanced tissue engineering: applications in osteoinduction using cultured mesenchymal stem cells transduced with the bFGF gene / 生物医学工程学杂志

To investigate the effect of basic fibroblast growth factor(bFGF) gene transfection on the proliferation and differentiation of mesenchymal stem cells (MSCs) and to provide basis for accelerating bone defect repairing using gene-enhanced tissue engineering technology, Rabbit periosteum-derived MSCs were transfected with the full-length rat bFGF cDNA in vitro. The transient and stable gene expression of bFGF were determined by immunohistochemistry. The proliferation and the synthesis alkaline phosphatase (ALP) and osteocalcin(OC) of the transfected MSCs were also examined. The results showed that bFGF cDNA could be transferred into osteoblasts and expressed stably at least 4 weeks. The proliferation and OC content of genetically modified MSCs were increased significantly, whereas the ALP activity remained no change. In conclusion, transfer of gene encoding bFGF to MSCs increases its proliferation and osteogenesis property. Based on the successful conjunction of the existing techniques of tissue engineering with the novel possibilities offered by modern gene transfer technology, an innovative concept, molecular tissue engineering, was put forward for the first time. As a new branch of tissue engineering, it represents both a new area and an important trend in tissue engineering research.

Expression of rhBMP-7 gene in transduced bone marrow derived stromal cells / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To explore the possibility of expression of exogenous gene in transduced bone marrow derived stromal cells (BMSCs).</p><p><b>METHODS</b>The marker gene, pbLacZ, was transferred into cultured BMSCs and the expression of transduced gene by X-gal staining was examined. Then plasmid pcDNA3-rhBMP7 was delivered to cultured BMSCs. Through immunohistochemical staining and RT-PCR assay, the expression of rhBMP7 gene was detected.</p><p><b>RESULTS</b>The exogenous gene could be expressed efficiently in transduced BMSCs.</p><p><b>CONCLUSION</b>The present study provided a theoretical basis to gene therapy on the problems of bone and cartilage tissue.</p>

Expression of transforming growth factor beta 1 in mesenchymal stem cells: potential utility in molecular tissue engineering for osteochondral repair / 华中科技大学学报（医学）（英德文版）

The feasibility of using gene therapy to treat full-thickness articular cartilage defects was investigated with respect to the transfection and expression of exogenous transforming growth factor (TGF)-beta 1 genes in bone marrow-derived mesenchymal stem cells (MSCs) in vitro. The full-length rat TGF-beta 1 cDNA was transfected to MSCs mediated by lipofectamine and then selected with G418, a synthetic neomycin analog. The transient and stable expression of TGF-beta 1 by MSCs was detected by using immunohistochemical staining. The lipofectamine-mediated gene therapy efficiently transfected MSCs in vitro with the TGF-beta 1 gene causing a marked up-regulation in TGF-beta 1 expression as compared with the vector-transfected control groups, and the increased expression persisted for at least 4 weeks after selected with G418. It was suggested that bone marrow-derived MSCs were susceptible to in vitro lipofectamine mediated TGF-beta 1 gene transfer and that transgene expression persisted for at least 4 weeks. Having successfully combined the existing techniques of tissue engineering with the novel possibilities offered by modern gene transfer technology, an innovative concept, i.e. molecular tissue engineering, are put forward for the first time. As a new branch of tissue engineering, it represents both a new area and an important trend in research. Using this technique, we have a new powerful tool with which: (1) to modify the functional biology of articular tissue repair along defined pathways of growth and differentiation and (2) to affect a better repair of full-thickness articular cartilage defects that occur as a result of injury and osteoarthritis.