Objective To observe the main biological characteristics and chondrogenesis potency of bone marrow -derived stromal cells(MSCs) after cytokinesinduction or gene modification in vitro. Methods MSCs from an adult New Zealand white rabbit were isolated and cultivated, and then MSCs were divided into the common medium group(Group A, 15%FBS in DMEM), the induced group by cytokines (Group B), the transfected group(Group C)with adenovirus-hepatocyte growth factor transgene (adHGF). The medium of group B consisted of transforming growth factor-β1(TGF-β1,10 ng/ml), basic fibroblast growth factor(bFGF,25 ng/ml) addexamethasone (DEX,10-7mol/L) with 15%FBS in DMEM. Cartilage slices wereobtained from femoral condyles and patellar grove in the same rabbit. The minced cartilage was digested in Ⅱ collagenase (3 mg/ml) to obtain chondrocytes(Group D). The change of cell appearance, proliferation capacity, glycosaminoglycans(GAG), immunohistochemical staining for type Ⅰ, Ⅱ collagen were observed during the 5th passage MSCs and MSCs after induction or gene modification. Expression of mRNA for type Ⅰ and Ⅱ collagen was detected by RT-PCR. Results Primary MSCs proliferated as shortspindle shape, while the 5th MSCs showed longspindle shape. Positive stain of type Ⅰ collagen could be found in groups A, B and C, while positivestain of type Ⅱ collagen was shown in groups B and D. The content of GAG in group B was higher than that in group A, but there was no significant difference between them（Pgt;0.05）, and there was significant difference between groups A and D（Plt;0.05）. No significant difference was noted in groups A,B and C on proliferation by MTT（Pgt;0.05）,except that of at the fourth day after transfection between groups A and C（Plt;0.05）. RT-PCR demonstrated that MSCs always had higher levelsof mRNA type Ⅰ collagen in groups A, B and C. The expression of mRNA type Ⅱ collagen was identified in groups B and D, and only low levels of mRNA type Ⅱ collagen in group C. Conclusion The above results indicate MSCs have a natural tendency of osteogenic differentiation in vitro culture, and also demonstrate the chondrogenic potency with the technique of cytokines induction or gene modification after passage. MSCs can be transfected efficiently being seed cells in tissue engineered bone or cartilage to accept target genes such as adHGF, and have a higher levels of expression in vitro, which lasted 4 weeks at least.
Objective To compare the clinical efficacy of glucosamine hydrochloride and Chinese traditional medicine of angelicae pubescentis and loranthi decoction in the treatment of knee osteoarthritis. Methods We included 142 patients with mild-to-moderate knee osteoarthritis treated between January 2014 and July 2015. The patients were randomly divided into treatment group and control group. The 72 patients in the treatment group received glucosamine hydrochloride, while the other 70 patients in the control group took oral Chinese medicine of angelicae pubescentis and loranthi decoction. The treatment course was one month. We observed the clinical curative effect of both the two groups. Results After the treatment, the difference in Visual Analogy Score (VAS) and Severity Index of Osteoarthritis (ISOA) in the two groups were significant compared with those before the treatment (P ＜ 0.05) . There were significant differences between the two groups in terms of VAS pain score, ISOA and treatment effectiveness (P ＜ 0.05) . Conclusion Glucosamine hydrochloride can obviously relieve knee osteoarthritis symptoms and improve knee function, which has a better curative effect than the traditional Chinese medicine of angelicae pubescentis and loranthi decoction.
Objective To compare the characteristics of gelatin microspheres crossl inked by glutaraldehyde (GA) or geni pin (GP). Methods Gelatin microspheres, prepared by the improved emulsified cold-condensation method, were crossl inked by GP and GA, respectively. After being dispersed in PBS, two kinds of microspheres with 60% degree of cross l inking were compared in terms of morphology, swell ing and degrading properties. rhBMP-2 were loaded into the GP and GAmicrospheres, and the encapsulation rate, drug loading and releasing capacity were measured; 100%, 50% and 25% leaching l iquid of GP and GA microspheres were respectively cultured with rat osteoblast (DMEM group as the control), and cell prol iferation was measured by MTT method to grade the cell cytotoxicity. Results GP and GA microspheres were both spherical with the diameters of (78 ± 18) μm and (65 ± 10) μm, and there were no difference between both microspheres in drug loading and encapsulation rate. But, GP microspheres, with long degrading period (28 days) compared to GA microspheres (21 days), had better dispersibil ity, and swell ing rate (89.0% ± 4.8%), the percentage of cumulative drug releasing at 10 days (78.80% ± 4.96%) were both lower than GA microsphere (118.0% ± 7.6%, 90.50% ± 5.12%). The percentages of drug loading of GP and GA were (921 ± 73) and (965 ± 62) ng/g, and the encapsulation rates were 88.5% ± 2.1% and 89.7% ± 1.8%; showing no significant difference (P gt; 0.05). The cell cytotoxicity of 100%, 50% and 25% leaching l iquid of GP microspheres was all at the level I, but leaching l iquid of GA microspheres with corresponding concentration were at the levels of III, III and II. Conclusion GP crossl inked gelatin microspheres are superior to GA crossl inked gelatin microspheres and can be widely used in tissue engineering field.
Objective To compare the effect of small intestinal submucosa(SIS)and polypropylene mesh(PPM) on repairing abdominal wall defects in rats, and toprobe into the feasibility of using SIS to repair the abdominal wall defects. Methods 100 SD rats(50 males and 50 females)were randomly divided into 2 groups(n=50). Their weight ranged from 200 to 250 g.Full thickness abdominal wall defects (2 cm×2 cm) were created by surgery and were repaired with SIS and PPM respectively. At different postoperative time (1st, 2nd, 4th, 8th and 12th week), animals were sacrificed to make histological observation. The tensile strengthand the development of adhesions were measured and observed. Results 95 animals survived and were healthy after surgery. No inflammatory response and obvious immunoreaction were observed in both groups. One week after operation, the tensile strengthof abdominal wall in SIS group (204.30±5.13 mmHg) was lower than that in PPMgroup(240.0±10.0 mmHg) at 1st week(P＜0.05),and there were no difference at 4th, 8th, 12th week. Adhesions were more marked in PPM group thanthat in SIS group(P＜0.05). Conclusion Both SIS and PPM are histologically compatible when used in rats and can maintain sufficient tensile strength. SIS is superior to PPM in regards to tissue compatibility and adhesion formation.
Objective To compare the effect of two different operations on treating severely comminuted intercondylar fracture. Methods From December 2001 to October 2003, 20 cases of severely comminuted intercondylar fracture were operated. Of the 20 cases, 7(group 1) were treated with closed reduction and retrograde intramedullary nailing through arthroscope, 13(group2) were treated with open reduction and retrograde intramedullary nailing without arthroscope. Follow-ups were conducted after operation. Results All cases of fracture were recovered. Swelling in group 1 was alleviated more obviously than that in group 2. In group 1, all knees could flex to 120° during 6th to 9th weeks after the operation. In group 2, only 4 could flex 110°.Conclusion Retrograde intramedullary nailing through arthroscope proves to be less invasive and more effective in treating heavily comminuted intercondylar fracture.
Objective To compare the effects of flap delay and vascular endothelial growth factor (VEGF) on the viability of the rat dorsal flap. Methods Thirty rats were divided into 3 groups: saline group, flap delay group and VEGF group. The rats in flap delay group underwent flap delay by keeping bipedicle untouched, and the cranial pedicle was cut 7 days later. The rats in VEGF group were given VEGF solution locally when the flaps were elevated in the operation. The ratsin saline group were given saline solution in the same way. Five days after thesingle pedicle flaps were performed, the flap survival rate was measured. Theflap tissues were collected to measure and analyze the microvascular density, diameter and sectional area by immunochemical method. Results The flap survival rate of flap delay group was similar to that of VEGF group andthere is no statistically significant difference(Pgt;0.05). The vascular diameter of flap delay group was much larger than that of saline group and VEGF group, showing statistically significant difference (Plt;0.05). The vascular density of VEGF group was much higher than that of saline group and flap delay group, showing statistically significant difference (Plt;0.05). The vascular sectional area of flap delay group was similar to that of VEGF group(Pgt;0.05). Conclusion The change in the flap after flap delayis manifested as obvious dilatation of microvessels, while the change in the flap after the injection of VEGF is manifested as obvious vascular proliferation. Both flap delay and VEGF can increase the vascular sectional area and the viability of the flap, but the mechanism is different.
Objective To verify the effect of palatoplasty with or without velopharyngeal muscular reconstruction on the velar movement through the lateral radiography. Methods From October 1988 to October 2000, 62 patients with cleft palate and velopharyngeal insufficiency were treated. Of them, 32 were repaired by velopharyngeal muscular reconstruction (group A) and 30 by modified von Langenbeck’s procedure (group B). The lateral radiographs and cephalometric analysis were taken. The velopharyngeal closure, velar extensibility, the angle changes of velar elevation, the distance changes of velar levator eminence based on anatomy (LEA) to velopharyngeal closure line (VCL), the varieties in LEA, the comparison of LEA and velar levator eminence based on phonation (LEP), the distance comparison between posterior nasal spine (PNS) to LEA and LEP through soft palate line (SPL) were measured. Results During the phonation, group A was significantly greater than group B in the velopharyngeal closure rate(Plt;0.01)， the velar extensibility(Plt;0.05) and the location comparison between LEA and LEP(Plt;0.01); group B was significantly greater than group A in velar elevation angle (Plt;0.05), the varieties in LEA(Plt;0.05). In velar rest position, the distance of LEA to VCL was greater inB group than in A group(Plt;0.01). There was significant difference in thedistance comparison between LEA and LEP(Plt;0.05), difference between LEA and LEP(Plt;0.01) and the distance PNS-SPL-LEA and PNSSPL-LEP(Plt;0.05) within group B; contrary to the results within group A(P＞0.05). Conclusion The velopharyngeal muscular reconstruction in palatoplasty can result in a near normalizationof anatomic measurement of velar levator muscles and improve the velar functionand velopharyngeal competence. Repositioning of velar muscles in a more anatomic correct, transverse position is more important to improve the velar length andaccordant velar movement with velopharyngeal muscles in functional palatoplasty. The velar angle change and velar elevation during phonation are not determinative factors for velopharyngeal competence.
Objective To compare the effect and coverage of bacteriostasis of chitosan and sodium hyaluronate. Methods Each of the five bacteria, Proteus mirabilis, Escherichia coli, Candida albicans, Pseudomonas aeruginosa, Staphylococcus aureus, was cultivated for 33 tubes of broth culture. Leaving three tubes each group as control group, ploidy diluted concentration of high relative molecular weight chitosan, low relative molecular weight chitosan and sodium hyaluronate were added respectively in the broth culture. All the tubes were cultivated for 18 hours at 37 ℃ with homeothermia. Then the growth of bacteria was observed. ResultsThe minimal inhibitory concentrations (MIC) of high relative molecular weight chitosan were : Proteus mirabilis 0.031%, Escherichia coli 0.063%, Candida albicans 0.063%, Pseudomonas aerugionosa 0.063%, Staphylococcus aureus 0.063%; and the MIC of low relative molecular weight chitosan were: Proteus mirabilis 0.125%, Escherichia coli 0.025%, Candida albicans 0.25%, Pseudomonas aeruginosa 0.25%, Staphylococcus aureus 0.125%; bacteria grew well in each tube of sodium hyaluronate group and control group. Conclusion The above results show that sodium hyaluronate has no bacteriostasis, while chitosan has bacteriostasison broad spectrum and high relative molecular weight chitosan has ber effect.
Objective To compare the outcomes of two operative methods, the anterior decompression in subsection and the anterior decompression in one section, which were used to treat multilevel cervical spondylotic myelopathy (CSM). Methods Data of multilevel CSM undergoing the anterior decompression in subsection (33 cases, the subsection group) and the anterior decompression in one section (19 cases, the one section group) from July 1999 to January 2004 were retrospectively analyzed. The- incidence of perioperative complications and the rate of fusion were evaluated by the postoperative X-ray and MRI examinations, and improvement of the neurological function was evaluated by the JOA score.Results The incidence of perioperative complications was 36.8%in the one section group, mainly including immigration of the plate and grafts,which was settled by the revision surgery; while the incidence of perioperative compilcations was 12.1% in the subsection group, mainly including the immigration of the titanium mesh. There was a significant difference between the two groups (Plt;0.05). 84.2％ of the patients in the one section group and 81.8％ of the patients in the subsection group developed bony fusion by the end of the follow-up (9-31 mon, averaged 112 mon), and there was no significant differencebetween the two groups (Pgt;0.05). According to the JOA score, the ratio of the improvement in the neurological function was 70.4% in the subsection group and 64.4% in the one section group. There was no significant difference between the two groups (Pgt;0.05). Conclusion The anterior decompression in subsection is more rational for the surgical treatment on the multilevel CSM than the anterior decompression in one section. It can provide an equal decompressive effect but a more stable local mechanical environment right after the surgery and can maintain it well, which is critical for the bony fusion.
To compare the effectiveness of microdiscectomy and macrodiscectomy on the single-level lumbar disc protrusion (LDP). Methods Between November 2002 and October 2005, 241 patients with LDP underwent 2 surgical procedures: microdiscectomy (group A, 93 cases) and macrodiscectomy (group B, 148 cases). All patients had singlelevel LDP. In group A, there were 51 males and 42 females with an average age of 32.3 yeares (range, 18-47 years); there were 23cases of protrusion, 52 cases of prolapse, and 18 cases of sequestration with an average disease duration of 8.5 months (range, 1-18 months), including 8 cases at L2,3 level, 11 cases at L3,4 level, 35 cases at L4,5 level, and 39 cases at L5, S1 level. In group B, there were 81 males and 67 females with an average age of 31.8 years (range, 16-50 years); there were 37 cases of protrusion, 85 cases of prolapse, and 26 cases of sequestration with an average disease duration of 9.3 months (range, 1-20 months), including 9 cases at L2,3 level, 15 cases at L3,4 level, 63 cases at L4,5 level, and 61 cases at L5, S1 level. There was no significant difference in age, sex, segment level, type, or disease duration between 2 groups (P gt; 0.05). Results Immediate back and sciatic pain rel ief was achieved in 225 (93.4%) patients after operation. The satisfactory rates were 91.4% in group A and 87.8% in group B at 1 week after operation, showing no significant difference (P gt; 0.05). The length of incision, amount of bleeding, amount of drainage, and hospital ization time in group A were significantly fewer than those in group B (P lt; 0.05); while the operative time in group A was longer than that in group B, but showing no significant difference (P gt; 0.05). Dural laceration occurred in 4 cases of groupA and 5 cases of group B, superficial infections of incision occurred in 5 cases of group B and intervertebral space nfections occurred in 4 cases of group B, and epidural hematoma occurred in 1 case of group A. The perioperative compl ication rate (5.4%, 5/93) in group A was significantly lower (P lt; 0.05) than that in group B (9.5%, 14/148). LDP recurred in 4 cases (4.3%) of group A and in 9 cases (6.1%) of group B postoperatively, showing no significant difference (P gt; 0.05); of them, 11 cases received second operation and 2 cases were treated conservatively. All cases were followed up 36-77 months (mean, 51.4 months). There were significant differences in visual analog scale (VAS) and Oswestry disabil ity index (ODI) between 2 groups at the last follow-up and preoperation (P gt; 0.05), but there was significant difference in VAS at 1 week postoperatively between 2 groups (P lt; 0.05). VAS and ODI were obviously improved at 1 week and last follow-up when compared with preoperation (P lt; 0.05). There was no significant difference in the improvement rates of VAS and ODI between 2 groups at last follow-up (P gt; 0.05). According to cl inical evaluation of Modified Macnab criteria, the excellent and good rate was 90.3% in group A and 86.5% in group B at final follow-up (P gt; 0.05). Conclusion Both macrodiscectomy and microdiscectomy are effective for LDP, furthermore microdiscectomy is less invasive than macrodiscectomy. Microdiscectomy is recommended to treat single-level LDP.