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 explore the ability of insulin-like growth factor-Ⅰ (IGF-Ⅰ) and hyaluracan acid in prompting chondrogenesis of engineering cartilage tissue.Methods Human articular chondrocytes were isolatedand cultured in DMEM plus 10% fetal bovine serum. They were divided into three groups：hyaluracan acid+chondrocytes + IGF-Ⅰ group(IGF-Ⅰ group), hyaluracan acid+chondrocytes group(cell group), hyaluracan acid group(control group). The ability of chondrogenesis was investigated by HE and toluidine blue staining, human collagen Ⅱ immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR).Results Both cell group and IGF-Ⅰ group could develop into cartilage tissue in the sixth week while control group could not. The number of cartilage lacuna in IGF-Ⅰ group were more than that in cell group. Human collagen Ⅱ immunohistochemistry showed that there were ber positive cell in IGF-Ⅰ group than in cell group, collagen Ⅱ mRNA expression was more higher and collagen Ⅰ mRNA expression was lower in IGF-Ⅰ group than in cell group. Conclusion Insulin growth factorⅠ can prompt chondrogenesis of engineering cartilage tissue and ameliorate the quality of engineering cartilage tissue in vitro.
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.
Objective To explore the efficacy and safety of bioactive material(combest) which is combinated with bioglass and hyaluronan on burn wound healing．Methods From March to September 2006, 20 patients were treated; including 16 males and 4 females, aging 18-58 years(40 years on average)．The wounds were classified as deep degrees Ⅱ in 7 cases, granulated wounds in 9 cases and graft site wounds in 4 cases. Twenty wounds in one side were repaired with Combest as the test group and 20 wounds in the other side with blank cream as the control group. The wounds in size ranged from 2.0 cm×1.5 cm to 40.0 cm×20.0 cm. The wound healing rate was observed, and the blood test and the indices of hepatic and renal function were determined on the 1st, 3 rd,6 th, 11 th, 16 th and 21 st days of treatment．Results Wound healed within 3 weeks in 11 cases of the test group (3 cases on the 11 st day, 4 on the 16 th day, and 4 on the 21 st day) , but no wound healing was observed within 3 weeks in the control group. The healing size accounted for 2/3 of wounds in 18 cases of the test group and in 1 case of the control group. The excellent and good rates were 95%(18 cases and 1 case) in the test group and 50% (1 case and 9 cases) in the control group, showing significant difference (Plt;0.01). For all patients, no obvious changes were found in the blood test and hepatic or renal function indices. Conclusion Combest combinated with bioglass and hyaluronan is beneficial to the proliferation of the granulation and wound healing with good safety.
Objective To review the recent advances of cross-linking reagent for producing hyaluronic acid(HA) derivative so as to provide more advice for thedevelopment of HA reagent. Methods Recent original articles related to the species, characteristic, cross-linking methodology and mechanism of the cross-linking reagent to producing HA derivative were summarized and systematically analyzed. Results The derivatives after special kinds of reagents modification would remain their own good biocompatibility and change their original rheololgical characterization and obtain relative long organism residence time. Conclusion Development of hyaluronic derivatives may widen their medical application.
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 effects of sodium hyaluronate in supplementary treatment of comminuted fracture of ankle. METHODS Thirty-seven patients suffered from comminuted fracture of ankle were operated for restoration by routing methods, and received 2 ml of sodium hyaluronate injection intra-articularly before the closure of incision. The ankle was fixed and given the second intra-articular injection on the 3rd day after operation. Then, the patients were given sodium hyaluronate injection intra-articularly at a week intervals till the paste was removed after 4 weeks. All patients were followed up. The clinical results were evaluated by measuring the symptoms of pain, and the function of walking and other daily living activities. RESULTS All the patients were followed up for 6 to 27 months, among them, 30 patients were cured completely without any symptoms, the ankle function for walking and daily living activities was normal, 6 patients felt pain with violent activity or walking exceeding 1 km, one patient suffered from comminuted fracture with compressed depression was not improved due to his ankle being not restored properly. CONCLUSION Intra-articular injection of sodium hyaluronate is an effective supplementary treatment for comminuted fracture of ankle.
ObjectiveTo compare the clinical effects of urokinase thrombolytic therapy for optic artery occlusion (OAO) and retinal artery occlusion (RAO) caused by facial microinjection with hyaluronic acid and spontaneous RAO.MethodsFrom January 2014 to February 2018, 22 eyes of 22 patients with OAO and RAO caused by facial microinjection of hyaluronic acid who received treatment in Xi'an Fourth Hospital were enrolled in this retrospective study (hyaluronic acid group). Twenty-two eyes of 22 patients with spontaneous RAO were selected as the control group. The BCVA examination was performed using the international standard visual acuity chart, which was converted into logMAR visual acuity. FFA was used to measure arm-retinal circulation time (A-Rct) and filling time of retinal artery and its branches (FT). Meanwhile, MRI examination was performed. There were significant differences in age and FT between the two groups (t=14.840, 3.263; P=0.000, 0.003). The differecens of logMAR visual acuity, onset time and A-Rct were not statistically significant between the two groups (t=0.461, 0.107, 1.101; P=0.647, 0.915, 0.277). All patients underwent urokinase thrombolysis after exclusion of thrombolytic therapy. Among the patients in the hyaluronic acid group and control group, there were 6 patients of retrograde ophthalmic thrombolysis via the superior pulchlear artery, 6 patients of retrograde ophthalmic thrombolysis via the internal carotid artery, and 10 patients of intravenous thrombolysis. FFA was reviewed 24 h after treatment, and A-Rct and FT were recorded. Visual acuity was reviewed 30 days after treatment. The occurrence of adverse reactions during and after treatment were observed. The changes of logMAR visual acuity, A-Rct and FT before and after treatment were compared between the two groups using t-test.ResultsAt 24 h after treatment, the A-Rct and FT of the hyaluronic acid group were 21.05±3.42 s and 5.05±2.52 s, which were significantly shorter than before treatment (t=4.569, 2.730; P=0.000, 0.000); the A-Rct and FT in the control group were 19.55±4.14 s and 2.55±0.91 s, which were significantly shorter than before treatment (t=4.114, 7.601; P=0.000, 0.000). There was no significant difference in A-Rct between the two groups at 24 h after treatment (t=1.311, P=0.197). The FT difference was statistically significant between the two groups at 24 h after treatment (t=4.382, P=0.000). There was no significant difference in the shortening time of A-Rct and FT between the two groups (t=0.330, 0.510; P=0.743, 0.613). At 30 days after treatment, the logMAR visual acuity in the hyaluronic acid group and the control group were 0.62±0.32 and 0.43±0.17, which were significantly higher than those before treatment (t=2.289, 5.169; P=0.029, 0.000). The difference of logMAR visual acuity between the two groups after treatment was statistically significant (t=2.872, P=0.008). The difference in logMAR visual acuity before and after treatment between the two groups was statistically significant (t=2.239, P=0.025). No ocular or systemic adverse reactions occurred during or after treatment in all patients. ConclusionsUrokinase thrombolytic therapy for OAO and RAO caused by facial microinjection with hyaluronic acid and spontaneous RAO is safe and effective, with shortening A-Rct, FT and improving visual acuity. However, the improvement of visual acuity after treatment of OAO and RAO caused by facial microinjection with hyaluronic acid is worse than that of spontaneous RAO.
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 develop a scaffold material containing collagen Ⅰ and sodium hyaluronate for the cartilage tissue engineering and to evaluate its biocompatibility by using the rabbit chondrocytes derived from amandibular condylar process. Methods The porous matrices containing collagen Ⅰ and sodium hyaluronate were fabricated by the freezedrying technique and were crosslinked by using 1-ethyl-3(3-dimethyl aminopropyl) carbodiimide (EDC). The microstructure of the scaffold was observed under thescanning electron microscope (SEM), and the enzymatic degradation test was performed to compare the ability of the scaffold resistance to collagenase before and after the crosslinking. The chondrocytes from the rabbits’ condylar process were isolated and cultured before they were seeded into the scaffold, and cell attachment and proliferation were measured by the cell count 1, 3, 5, 7 and 10 daysafter the cell being seeded; then, the biocompatibility of the scaffold was evaluated by the light microscopic examination, histological examination, and the SEM exmination. Results The porous structure of the scaffold facilitated the penetration and attachment of the seeded cells. The porosity was 83.7% and the pore size was 100-120 μm. The cell number increased from 3.7×104 per scaffold 1 day after the cell being seeded to 8.2×104 per scaffold 10 days after the cell being seeded. The crosslinking treatment could significantly enhance the scaffold resistance to the collagenase activity. The examinations under the light microscope and SEM indicated that the chondrocyte adhered and spread well on the scaffold, and the extracellular matrices were also observed around the chondrocytes. Conclusion The porous scaffold composed of collagen Ⅰ and hyaluronan has anappropriate structure and a good biocompatibility for the attachment and proliferation of the chondrocytes, which can facilitate it to become a useful scaffold in the cartilage tissue engineering.