Objective To explore the relationship of the limited resource of the autologous bone marrow mesenchymal stem cells (MSCs) in articularcavity to the treatment results of full-thickness articular cartilage defect, and to investigate whether the extrogenous sodium hyaluronate(SH) promotes the migration of MSCs cultured in vitro tothe articular defect in vivo. Methods Sixty-six Japan rabbits were made the model of the full-thickness articular cartilage defect (5 mm width and 4 mm depth).The autologous MSCs were extracted from the rabbit femur, cultured in vitro, labeledby Brdu, and injected into the injured articular cavity with or without SH. Theexperiment was divided into 4 groups; group A (MSCs and SH, n=15); group B (MSCs, n=15); group C (SH, n=18); and group D (non-treatment, n=18). The morphologic observation was made by HE staining, Mallory staining and immunohistochemical staining after 5 weeks, 8 weeks and 12 weeks of operation. Results There were significant differences in the thickness of repairing tissue between group A and group B(Plt;0.01); but there were no significant differences between group A and group C, and between group B and group D(P＞0.05). Thehistological observation showed that the main repairing tissue was fibrocartilage in group A and fiber tissue in group B. Conclusion MSCs cultured in vitro and injected into the articular cavity can not improve the treatment results of the articular cartilage defect. Extrogenous SH has effect on repairing cartilage defect. The extrogenous SH has no effect on the chemotaxis of the MSCs, and on the collection of MSCs into the joint defect.
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 investigate the effect of sodium hyaluronate (SH) intra-articular injection on the treatment of knee osteoarthritis (OA), andto compare the contents of free radicals and inflammatory factors in joint fluids of pre-and pro-treatment as to explore the treatment mechanism of SH. Methods Ninety-two patients (111 knees) with mild(51)，moderate(35) and serious(25) knee OA were treated with intra-articular injections of SH (20 mg once a week for 5 weeks). According to Lysholm scoring, clinical signs such aspain, swelling,and the ability to walk, squat, run, go upstairs and downstairs were assessed before and after the treatment, and the contents of nitric oxide (NO), superoxide dismutase(SOD), malonic dialdehyde(MDA) and IL-1β、TNF-α in joint fluids from the OA joints before 1st,2nd, and 5th injection and 3 months after each injection were observed. Results All cases were followed up for 3 months. The improvements in the signs and function of knees were excellent in 42 knees, good in 38 knees, fair in 21 knees and poor in 10 knees, with 72.1% excellent and good results. The lighter the illness was, the better the improvement was: the rate of the excellent and good was 92.1% in mild group, 68.6% in moderate group and 42.9% in serious group. The contents of oxygen free radicals and IL-1β、TNF-α of the patients with mild and moderate OA decreasedmarkedly after being treated with SH（Plt;0.05）, but these decreased lightlyin serious OA group（Pgt;0.05）. SH had mild effect on the contents of NO. Three months after treatment, only in mild OA group the contents of NO significantly decreased（Plt;0.05）, and no significant change in moderate and serious groups was observed（Pgt;0.05）. Conclusion SH intraarticular injection has a positive effect on the relief of clinical symptoms and on the improvement of articular function of knee OA. The therapeutical effect of SH on OA is achieved possibly by decreasing the contents of free radicals especially oxygen free radicals and inflammatory factors in joint fluids.
OBJECTIVE To review the physiological function of sodium hyaluronate in joints and its clinical applications. METHODS Many literatures were reviewed and analysed on therapeutic mechanism and the application foreground of sodium hyaluronate. RESULTS Extrinsic sodium hyaluronate plays an important role in improving synovial fluid and protecting cartilages as well as suppressing inflammation, so it is used in the treatment of joint diseases such as knee osteoarthritis, rheumatoid arthritis or temporomandibular osteoarthritis. CONCLUSION Sodium hyaluronate possesses a good applied prospect in joint diseases.
Objective To investigate the effects of sodium hyaluronate solution on the proliferation and differentiation of myoblasts. Methods The 3rd subculture myoblasts from muscle of infant SD rat were cultured in four growth media, in which the concentrations of sodium hyaluronate were 0.05% (group A) , 0.1%( group B), 0.2% (group C)and 0 (group D, control group), respectively. The proliferation rate of myoblasts in each medium was observed through growth curves by means of count and MTT. At the same time, the subculture myoblasts were cultured in differentiated media in which the concentrations of sodium hyaluronate were 0 and 0.1%. The capacity of fusion of myoblasts was compared between two kinds of differentiated media. Results There were the nearly same proliferation curse in Groups A, B and C: increasing by logarithm at 2 days and reaching peak value at 4 days. The myoblasts in Group D increased slowly: increasing by logarithm at 3 days, doubling at 5 days and reaching peak value at 6 days. MTT has the similar curse to counting. The myoblast proliferation of Group B was more than that of the other groups. The peak value of myoblast fusion was 35% at 6 days in common differentiated media; slowly reached 11.7% at 7 days in the differentiated media in which the concentrations of sodiumhyaluronate was 0.1%.Conclusion Sodium hyaluronate at certain concentration can be a decent media for myoblasts, it can accelerate proliferation and differentiation of myoblasts.
Objective To investigate whether the implanted myoblasts with the soluble carriers can improve the repairing efficiency for theseverelycryodamaged tibialis anterior muscles. Methods The skeletal myoblasts were isolated from the newborn SD rats by the use of the enzyme digestion. They were purified and serially subcultivated; the subcultivated myoblasts of the 3rd generation were marked with BrdU. The severelycryodamaged tibialis anterior muscle models were established from 84 SD rats aged 5 months. They were randomly divided into 4 groups, including Group A1 (the implanted myoblasts with the carriersF12 containing 0.1% sodium hyaluronate), Group A2 (the implanted myoblasts, with the carriersF12 that did not contain 0.1% sodiumhyaluronate), Group B1 (the implanted carrier solution containing 0.1% sodium hyaluronate, but with no myoblasts), and Group B2 (with no carrier solution or myoblasts). Six rats were killed at the following time points: at 2, 5 and 9 days,and 2, 4, 8 and 12 weeks after operation; the immunohistochemical and the Mallory staining studies were performed for an evaluation on the repairing efficiencyfor the severelycryodamaged tibialis anterior muscles. By the imaging analysis, the number of the survived cells in each group was compared at 2 days, and the area ratio of the collagen fiber in each group was also compared at 8 weeks. Results The BrdU immunohistochemical staining showed that the number of the remaining implanted cells was significantly greater in Groups A1 than in Group A2, the migrating area of the myoblasts was greater, the distribution of the cells was more uniform, the cell differentiating potential was undestroyed, the repairing efficiency for the severelycryodamaged tibialis anterior muscles was significantly improved. There was no bluestained nucleus at each time point in Group B. The Mallory staining showed that the fibrous degeneration inthe tissue repairing process was significantly inhibited in Groups A1, A2 and B1; the inhibition was most obvious in Group A1, and next in Group A2. The imaging analysis indicated that at 2 days after operation, the number of the survived cells was significantly-greater in Group A1 than in Group A2 (Plt;0.05). At 8 weeks after operation, the collagen fiber was the least in Group A1, less in Group A2, more in Group B1,and the most in Group B2 (Plt;0.05). Conclusion The implanted myoblasts can significantly improve the repairing efficiency for the severelycryodamaged muscle tissues, and the implanted carrier solution containing 0.1% sodium hyaluronate can improve the implanting efficiency for the myoblasts.
OBJECTIVE To investigate the therapeutic effect of percutaneous lumbar discectomy (PLD) combined with sodium hyaluronate (SH) injection in the treatment of lumbar intervertebral disc herniation. METHODS Forty-eight patients suffered from lumbar disc herniation were divided into two groups and treated by PLD combined with SH injection into epidural cavity (treatment group) or single PLD (control group) respectively. All patients were followed up for 24 months. The therapeutic effects in both groups were assessed and compared according to Macnab’s criterion. RESULTS The patients in the treatment group got much more significant improvement than those in the control group, with shorter therapeutic course and more safety. CONCLUSION PLD combined with SH injection into epidural cavity is more effective and safety in the treatment of lumbar disc herniation than of pure PLD.
OBJECTIVE To investigate the effects of intra-articular injection of sodium hyaluronate in post-operation treatment of the knee. METHODS From January 1998 to February 2001, 4 ml of sodium hyaluronate injection was injected into the knee joint of the 134 cases at the end of arthroscope operation, or the 91 cases undergoing open operation of the knee at the time when the drain tube was removed (treatment group). Five days after operation, the hydrarthrosis was removed and 2 ml of sodium hyaluronate was injected into the knee joint. According to the patient’s condition, injection of sodium hyaluronate was performed once a week for several weeks. Clinical evaluation was made by evaluating pain visual analog scale (VAS) and painless range of movement (ROM) of the joint at every definite point of time. The 85 patients in control group used nothing at the same time. RESULTS The VAS score of patients in the treatment group was significant lower than that of the control group. The period to the maximal painless ROM of the joint was 6 days in the treatment group after open operation, while 9 days in the control group. CONCLUSION Sodium hyaluronate appears effective in relieving post-operation pain of the knee joint.
ObjectiveTo fabricate an injectable composite bone substitute with hyaluronic acid (HA) and calcium sulfate and to evaluate the biocompatibility and effect of the composite on cell proliferation, osteogenic differentiation in vitro and osteogenic capability in vivo. MethodsCalcium sulfate powder was mixed with HA solution, cross-linked HA solution, and phosphate buffer solution (PBS) in a ratio of 2∶1 (W/V) to get composites of CA+HA, CA+HAC, and CA. The standard extracts from above 3 materials were prepared according to ISO10993-5, and were used to culture mouse MC3T3-E1 cells. The composite biocompatibility and cell proliferation in different concentrations of extract were tested with cell counting kit-8 (CCK-8). The cells were cultured with standard medium as a control. The optimal concentration was selected for osteogenic differentiation test, and ELISA Kit was used to determine the alkaline phosphatase (ALP), collagen type I (COL-I), and osteocalcin (OCN). The femoral condylar bone defect was made on New Zealand white rabbits and repaired with CA+HA, CA+HAC, and CA. Micro-CT was done to evaluate new bone formation with bone volume/tissue volume (BV/TV) ratio at 6 and 12 weeks. HE staining was used to observe bone formation. ResultsCA+HA and CA+HAC were better in injectability and stability in PBS than CA. The biocompatibility test showed that absorbance (A) value of CA group was significantly lower than that of control group (P<0.05) at 6, 12, and 24 hours after culture, but no significant difference was found inA values between CA+HA group or CA+HAC group and control group (P>0.05). The proliferation test showed 25% and 50% extract of all 3 materials had significantly higherA value than control group (P<0.05). For 75% and 100% extract, only CA+HA group had significantly higherA value than control group (P<0.05). And 50% extract was selected for osteogenic differentiation test. At 14 and 21 days, ALP, COL-I and OCN concentrations of CA+HA group and CA+HAC group were significantly higher than those of CA group and control group (P<0.05). Micro-CT results showed higher BV/TV in CA+HA group and CA+HAC group than CA group at 6 and 12 weeks (P<0.05), but no significant difference was found between CA+HA group and CA+HAC group (P>0.05). HE staining revealed that a little bone tissue was seen in CA+HA group and CA+HAC group, but there was no bone formation in CA group at 6 weeks; more streak bone tissue in CA+HA group and CA+HAC group than CA group at 12 weeks. ConclusionComposites prepared with calcium sulfate and HA or with cross-linked HA are stable, injectable, and biocompatible. The materials have excellent effect on proliferation and differentiation of mouse MC3T3-E1 cells. They also show good osteogenic capability in vivo. So it is a potential bone substitutes for bone defective diseases.
Objective To determine the effectiveness of sodium hyaluronate (SHA) in preventing intraperitoneal (IP) adhesion. Methods Thirty-eight rats were randomly divided into A,B,C groups, normal saline, 6% Dextran-40 or SHA were applied on the present serosal injury respectively, during operation. Biopsy was taken on the 14th postoperative day.Results There were statistically significant differences in the extent of adhesion among three groups (Ｐ＜0.01). Mild inflammatory changes and less fibrous proliferation were found in group C by microscopy and decreased production of collagen (by fibroblast) and active mesothelial cells proliferation were observed in group C under electron microscope. Conclusion SHA appeares to reduce the extent of postoperative IP adhesion, which is more satisfactory than Dextran-40.