Objective To study the effects of platelet-rich plasma(PRP) on the proliferation and osteogenetic activity of the marrow mesenchymal stem cells(MSCs) cultured in vitro to elucidate the cellular and molecularmechanism by which PRP accelerates bone repair.Methods The human MSCs were cultured in vitro and randomly divided into the experimental group(n=9) and control group(n=9). In the experimental group, the MSCs were interfused with PRP(10 μl/ml culture media). The proliferation ability of the cells was tested by flow cytometry and MTT, the osteogenetic activity by alkaline phosphatase(ALP) measuring and tetracycline fluorometry, and cbfal mRNA expression by reverse transcriptPCR.Results PRP could stimulate the MSCs proliferation. The flow cytometry assay showed that the MSCs proportion of S period of the experimental group significantly increased 14.5±0.4 in comparison with that of the control group 7.2±0.5 (P＜0.01) after 24 hours. MTT value showed that MSCs proliferatedto platform period earlier in the experimental group than in the control group. There was a significant increase in ALP activity of the experimental group 7.79±1.98,9.51±2.31and 14.03±3.02 when compared with that of the control group 2.06±0.77,2.84±0.82 and 2.58±0.84 after 3, 6 and 9 days(P＜0.05). The number of mineral nodes increased. Reverse transcript-PCR showed that the expression of cbfal mRNA were elevated gradually at 2,4 and 8 hours after interfused with PRP.Conclusion The effect of PRP on accelerating bone repair is related to its effects on stimulating the proliferation of MSCs, increasing the cbfal expression and promoting the osteogenetic activity.
Objective To review the lately new progress of fish collagen as biomedical materials, and then analyze feasibility and risk management of its application as a substitute of collagen originated from mammals in clinical practice. Methods Based on extensive research on new application and investigation of fish collagen, the paper was prepared to bring comprehensive analysis of its research and application status, and then several key points were focused on. Results Fish collagen has been proved to be a novel collagen of rich source, low risk of virus transmission, low biological risk, less religious barrier, and high biocompatibility. Fish collagen has promising prospect when applied in clinical practice as novel collagen especially as a substitute of collagen derived from mammals. However, very few related translational medicine research of fish collagen has been reported up to now in China. Conclusion As a novel potential substitute of collagen source derived from mammals, fish collagen is concerned to be clinical feasible and necessary in translational medicine. However, massive applied basic researches should be focused on in the further investigations.
ObjectiveTo assess the mid-term effectiveness of arthroscopic surgery in treatment of femoroacetabular impingement (FAI).MethodsBetween July 2014 and December 2015, 131 patients (132 hips) with FAI were enrolled in this study. There were 68 males and 63 females with an average age of 42.5 years (range, 17-68 years). The FAI was located at left hip in 72 cases, right hip in 58 cases, and bilateral hips in 1 case. The mean disease duration was 35 months (range, 3-120 months). Under arthroscopy, the femoral neck osteoplasty and/or acetabular rim trimming were performed, then the torn labrum was repaired. At the same time, the iliopsoas tendon was divided and the torn round ligament was debrided. The effectiveness was assessed by the visual analogue scale (VAS) and the Harris hip score (HHS).ResultsAfter operation, the perineum nerve palsy and numbness of dorsal feet occurred in 2 cases and 3 cases, respectively. There was no other complication. All patients were followed up 20-36 months (mean, 27.7 months). There were significant differences in the VAS scores and HHS scores between pre-operation, post-operative 3 months, and the last follow-up (P<0.05). Imaging examination showed that joint interspace of the involved hip was normal, and there was no sign of impingement and osteoarthritis in the femoral neck or acetabulum.ConclusionArthroscopic labral repair and osteoplasty is an effective and safe management for FAI in the mid-term follow-up.
Objective To review and analyze the properties, products, and appl ications of chitosan so as to explore the key molecular structure parameters which can affect the properties and appl ications significantly, and to reveal the relationship between molecular structures and properties so as to provide reference for further development of chitosan industryand scientific research. Methods Based on the collection and analysis of related l iterature, patents and medical productsderived from chitosan, as well as the author’s experiences in research and development, evaluation and standardization of chitosan, the paper was prepared to bring more attentions into the correlativity between structure and properties of chitosan. Results Potential risks in cl inical appl ication of chitosan-based preparations were seriously proposed in addition to a scientific review and analysis on relationships between chitosan structure and properties, as well as the present situations of developments and appl ications of chitosan. Conclusion The molecular structure is the crucial factor that can bring not only positive but also passive effects to the properties and appl ications of chitosan, especially for highly purified chitosan, molecular weight, and deacetylation degree are the most important parameters that should be focused more attention on.
【Abstract】 Objective To establ ish a animal model of osteonecrosis of femoral head in canine l ike human.Methods The thermal field of canine’s femoral head was three-dimensionally analyzed with fluent 6.2 software so that the best cryosurgery patent could be designed to maximize the osteonecrosis and minimize extra surgery trauma with the cryosurgery system invented by Shanghai Jiaotong University. Liquid nitrogen was pressurized to 0.5 MPa, poured into femoral head for 6.5 minutes, rewarming to 2 for 5 minutes and then repoured into it again for another 6.5 minutes. Ten three-foot canines were conducted as the animal models of osteonecrosis of femoral head according to the method above. At the end of followup,the results were reviewed by radiologic and pathologic check. Two dogs were conducted as control group. Results In the experimental group, one of the ten canines was testified to occur osteonecrosis of femoral head after one week pathologically, cell death and vessel breakage of cavitas medullaris in the femoral head was obvious under microscope; in other nine canines beingstill under follow-up, five with three-month follow-up at least progressed to the collapse of femoral head l ike human (Ficat III). In control group, no osteonecrosis was found. Conclusion Cryosurgery for osteonecrosis of the femoral head in three-foot canine model may become a method to establ ish the animal model of osteonecrosis of femoral head l ike human.
To evaluate the cl inical results of less invasive stabil ization system (LISS) for femur supercondylar and intercondylar fractures. Methods From March 2004 to November 2005, 47 patients with 49 intercondylar and supercondylar fractures were treated. Of all the patients, there were 34 males and 13 females with an average of 39.7 years (range 19-56 years). The locations were left side in 21 cases and right side in 28 cases. Fracture was caused by traffic accident in 31 cases, fall ing in 8 cases, violence in 6 cases and others in 2 cases. Forty-nine fractures included 14 intercondylar fractures, 21supercondylar fractures and 14 intercondylar and supercondylar fractures; 32 closed fractures and 17 open fractures. According to the AO typing, there were 6 type 33-A1, 8 type 33-A2 , 10 type 33-A3, 7 type 33-C1, 3 type 33-C2 and 15 type 33-C3. The disease course was 30 minutes to 6 days. Articular surface reduction was first performed, then the LISS plate was inserted via two incisions and locking screws were used later. Results The average operation time was 126 minutes (range 48-248 minutes). The blood loss was 180 mL(range 60-1 200 mL). The average follow-up time was 18.6 months (range 12-23 months). There were 4 patients with AP angular deformity and 5 patients with lateral angular deformity (range 2-5°). External rotation deformity was presented in 2 patients. There were no plate breakage, screw loosen and fixation failure. Average bone union time was 5.6 months (range 3-8 months) without infection case. Six cases were treated with il iac bone transplantation for delayed union. Conclusion LISS is one kind of effective treatment to femoral intercondylar and supercondylar fractures.
Objective To investigate the effect of various concentration of platelet-rich plasma (PRP) on osteogenic differentiation of rabbit skeletal muscle-derived stem cells (SMSCs) cultured in vitro. Methods Blood drawn from the central ear arteries of 9 one-year-old New Zealand white rabbits weighing 2.5-3.0 kg (male and female) was used to prepare PRP (Landesberg method). Full blood count and platelet count in PRP were tested. Soleus muscle of right hindl imb in rabbit was obtained and used to culture SMSCs in vitro. The cells at passage 3 were randomly divided into different groups: the experimental groups in which the cells were treated by conditioned culture media with various concentrations of autologousPRP (6.25%, 12.50%, 25.00%, 50.00%), and the control group in which the cells were treated with the media without PRP. At different time points after intervention, osteogenetic activity of the cells was detected by ALP staining observation, ALP activity detection was conducted, al izarin red staining for calcium nodules and immunofluorescence staining for osteocalcin were performed, and core binding factor α1 (Cbfα1) of osteogenic gene expression was tested by RT-PCR. Results The full blood PRP count and the platelet count in PRP was (3.06 ± 0.46) × 105/μL and (18.08 ± 2.10) × 105/μL, respectively. ALP staining: the cells in all the experimental groups were positive for the staining with many black sediment particles in cytoplasm; the cells in the control group were negative staining. ALP activity: all the experimental groups were higher than the control group (P lt; 0.05), the experimental group at 12.50% was superior to other experimental groups at each time point (P lt; 0.05). Al izarin red staining: at 14 days after culture, orange-red calcium nodules were evident in all the experimental groups; no orange-red calcium nodules were observed in the control group with a mineral ization rate of zero; there were significant difference between the experimental groups and the control group in terms of mineral ization rate (P lt; 0.05), the experimental group at 12.50% had a higher mineral ization rate than other experimental groups (P lt; 0.05). Immunofluorescence staining for osteocalcin: at 7 days after culture, the experimental groups were positive for the staining with yellow fluorescence in cytoplasm, and the result of the control group was negative. RT-PCR detection: no obvious changes of the gene expression were noted at 4, 12, and 24 hoursafter culture in the control group; the gene expression in all the experimental groups was significant superior to that of control group, especially at 12 hours, and the expression in the experimental group at 12.50% was the highest. Conclusion PRP can obviously promote the osteogenic differentiation of SMSCs cultured in vitro in a concentration-dependent manner, and the 12.50% is proved to be the ideal concentration.
Objective To review the research and application progress of bioactive glass in bone repair. Methods The recently published literature concerning bioactive glass in bone repair was reviewed and summarized. Results Bioactive glass can classified different types, such as bioactive glass particulate, bioactive glass scaffold, bioactive glass coating, injectable bioactive glass cement, and bioactive glass delivery system. Bioactive glass has been well studied in the field of bone repair due to its excellent biological properties. Also, the remarkable progress has been made in various aspects. Conclusion Bioactive glass is a reliable material of bone repair and will play an even more important role in the future.
Objective To investigate the possibility of constructing eukaryoticexpression vector for human angiopoietin 1（hAng-1），transfecting it to bonemarrow mesenchymal stem cells (MSCs) so as to repair bone defect. Methods The eukaryotic expression vector pcDNA3-hAng-1 was constructed by recombinant DNA technique, transfected into MSCs by liposome DOTAP, and selected with G418. The hAng-1 expression of mRNA and protein was detected by reverse transcript-PCR and Western Blot. Results After the recombinant eukaryotic expressionvector for hAng-1 was digested with Xho-I and BamH-I, electrophoresis revealed 1.4 kb fragment for hAng-1 gene and 5.4 kb fragment for pcDNA3 vector. In the transfected MSCs, the mRNA and protein expression of hAng-1 gene were detected with reverse transcriptPCR and Western Blot. Conclusion The constructed eukaryotic expression vector hAng-1 could be expressed in the transfected MSCs, thus to provide the basis for bone repair with tissue engineering.
Objective To describe the disease characteristics of osteonecrosis of the femoral head (ONFH) in patients with systemic lupus erythematosus (SLE) who experiencing prolonged glucocorticoid (GC) exposure. Methods Between January 2016 and June 2019, 449 SLE patients meeting the criteria were recruited from multiple centers. Hip MRI examinations were performed during screening and regular follow-up to determine the occurrence of ONFH. The cohort was divided into ONFH and non-ONFH groups, and the differences in demographic baseline characteristics, general clinical characteristics, GC medication information, combined medication, and hip clinical features were compared and comprehensively described. ResultsThe age at SLE diagnosis was 29.8 (23.2, 40.9) years, with 93.1% (418 cases) being female. The duration of GC exposure was 5.3 (2.0, 10.5) years, and the cumulative incidence of SLE-ONFH was 9.1%. Significant differences (P<0.05) between ONFH and non-ONFH groups were observed in the following clinical characteristics: ① Demographic baseline characteristics: ONFH group had a higher proportion of patients with body mass index (BMI)<20 kg/m2 compared to non-ONFH group. ② General clinical characteristics: ONFH group showed a higher proportion of patients with cutaneous and renal manifestations, positive antiphospholipid antibodies (aPLs) and anticardiolipin antibodies, severe SLE patients [baseline SLE Disease Activity Index 2000 (SLEDAI-2K) score ≥15], and secondary hypertension. Fasting blood glucose in ONFH group was also higher. ③ GC medication information: ONFH group had higher initial intravenous GC exposure rates, duration, cumulative doses, higher cumulative GC doses in the first month and the first 3 months, higher average daily doses in the first 3 months, and higher proportions of average daily doses ≥15.0 mg/d and ≥30.0 mg/d, as well as higher full-course average daily doses and proportion of full-course daily doses ≥30.0 mg/d compared to non-ONFH group. ④ Combined medications: ONFH group had a significantly higher rate of antiplatelet drug use than non-ONFH group. ⑤ Hip clinical features: ONFH group had a higher proportion of hip discomfort or pain and a higher incidence of hip joint effusion before MRI screening than non-ONFH group. Conclusion The incidence of ONFH after GC exposure in China’s SLE population remains high (9.1%), with short-term (first 3 months), medium-to-high dose (average daily dose ≥15 mg/d) GC being closely associated with ONFH. Severe SLE, low BMI, certain clinical phenotypes, positive aPLs, and secondary hypertension may also be related to ONFH.