OBJECTIVE: To observe the effects of hyaluronic acid (HA) and basic fibroblast growth factor (bFGF) on the proliferation of the cells from medial collateral ligament (MCL) and anterior cruciate ligament (ACL) cells. METHODS: The MCL cells and ACL cells of mature New Zealand white rabbit were cultured, while HA, bFGF or HA and bFGF were added to the cell culture media, the cellular proliferation was assayed by MTT method. RESULTS: HA only had no effect on the preoliferation of ACL cells, but had a small stimulatory effect on the proliferation of MCL cells. The addition of 1 ng/ml bFGF enhanced the proliferation of both MCL and ACL cells significantly, and this enhancement was maximal in the concentration of 50 ng/ml. However, the enhancement of proliferation of MCL and ACL cells could be achieved when the combination of HA in concentration of 100 micrograms/ml and bFGF in concentration of 100 ng/ml. CONCLUSION: It is evident that bFGF can enhance the proliferation of the ligament cells. HA can maintain the normal growth of ACL cells with no effect on the proliferation of the cells, while HA has a small stimulatory effect on the proliferation of MCL cells. However, when bFGF is coordinated with HA, more improvement of cellular proliferation can be achieved. HA can be used as a potential carrier for bFGF to enhance the healing of ligamentous tissue injuries.
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.
Objective To evaluate the feasibility of Seprafilm anti-adhesion membrane, a hyaluronic acid (HA) derivative, on prevention of adhesion in acute injured tendon. Methods Eighteen 4-month-old Chinese white rabbits (half males and half females, weighing 2.0-2.5 kg) were made the laceration models of the bilateral second and third toes of hindpaw. According to different treatments, the rabbits were randomly divided into 4 groups (n=18). The second toe of right hindpaw was wrapped with Seprafilm anti-adhesion membrane (group A); the third toe of right hindpaw was wrapped with polylactic acid membrane (group B); the second toe of left hindpaw was coated with sodium hyaluronate gel (group C); and the third toe of left hindpaw did not treated, as control group (group D). The general condition was observed; the range of motion (ROM) of distal interphalangeal joint was measured; the gross observation and histological observation were performed at 1, 2, and 4 weeks, then the degree of adhesion was graded. Results All rabbits survived to the end of the experiment. There was no significant difference in ROM of distal interphalangeal joint between groups A and B at 1, 2, and 4 weeks (P gt; 0.05). ROM of group A was superior to that of groups C and D at 2 and 4 weeks (P lt; 0.05). The gross and histological observations showed the same result in the grading of adhesion. At 1 week, there was no significant difference in the grading of adhesion among 4 groups (P gt; 0.05); at 2 and 4 weeks, the grading of adhesion in group A was similar to that in group B (P gt; 0.05), and the grading of adhesion in group A was significantly slighter than that in groups C and D (P lt; 0.05). Conclusion Seprafilm anti-adhesion membrane composed of HA derivative can prevent tendon adhesion and improve the joint function in acute tendon injury of rabbits.
OBJECTIVE: To study the effect of basic fibroblast growth factor (bFGF) and hyaluronic acid gel (HAG) combined with freeze-dried bone allograft in repairing segmental bone defect and to explore their mechanism. METHODS: The 15 mm segmental bone/periosteum defects were created on bilateral radius in 50 New Zealand rabbits and were treated with four different kinds of implants on 25 radius respectively (group A: bFGF and HAG combined with freeze-dried bone; group B: bFGF combined with freeze-dried bone; group C: HAG combined with freeze-dried bone; group D: simple freeze-dried bone as a control). The repair of defect was observed radiologically and histologically and were analyzed by radionuclide bone imaging and measurement of calcium contents at different periods. RESULTS: The new bone formation, bone metabolic activity and calcium contents of defects were higher in group A than in group B (P lt; 0.05), and were higher in group B than in groups C and D (P lt; 0.05). There were no significant difference between groups C and D. The bone defects healed in the 8th week in group A, in the 10th week in group B, but did not healed in the 10th week in groups C and D. CONCLUSION: As an osteogenetic factor, bFGF promotes the new bone formation; as a slow-release carrier, HAG enhances the effectiveness of bFGF. The combination of bFGF, HAG and freeze-dried bone allograft can repair the segmental bone defect more effectively.
Objective To compare the effect of decimeter wave with sodium hyaluronate product (SHP) on preventing and treating peritendinous adhesion and promoting tendon heal ing. Methods Totally 96 healthy male white 6-month-old Leghorn chickens weighing (2.24 ± 0.07) kg were randomized into group A ( decimeter wave therapy group, n=32),in which decimeter wave therapy was appl ied 1 to 21 days after operation at a frequency of 915 MHz, a power of 8 W, radiation distance of 10 cm, for 10 minutes once per day; group B (SHP group, n=32), in which 5 mL and 1.2% SHP was appl ied; and group C (control group, n=32), in which injury received no treatment. The III and IV toes of left feet of all chickens were made into tendon injury model. The general condition of animal was observed after operation; gross and histological observations were made 7, 10, 14, 18, 21 and 28 days after operation, and the biomechanical analysis was done 14 and 28 days after operation. Results Operative incision healed well, no infection and death occurred. Peritendinous adhesions in groups A, B were looser, and tendon heal ing was better than that of group C 14 and 28 days after operation. More fibroblasts with active metabol ism and more collagen formation in groups A, B than that in group C. The Pmax of group A was better than that of group B 14 and 28 days after operation (P lt; 0.05); the δmax of group A was better than that of group B 18 and 21 days after operation (P lt; 0.05), and the W0 of group A was better than that of group B 18, 21 and 28 days after operation (P lt; 0.05). There was no significant difference between group A and group B at the other time points. Conclusion Topical decimeter wave therapy and appl ication of SHP after flexor tendon repair can promote intrinsic heal ing, meanwhile they can prevent the adhesion of tendon and reduce extrinsic heal ing. Decimeter wave therapy can improve the qual ities of tendon’s wound heal ing.
OBJECTIVE: To review the recent advances of hyaluronic acid and its derivatives in medical application. METHODS: Recent original articles related to hyaluronan derivatives and their medical applications were retrieved extensively. RESULTS: Hyaluronic acid and its derivatives play important roles in visosurgery, arthritis therapy, prevention of adhesion, drug delivery, soft-tissue dilation, and percutaneous embolization. CONCLUSION: Development of hyaluronan derivatives may widen their medical application.
Objective To explore the possibility of constructing tissue engineered cartilage complex three-dimensional nano-scaffold with collagen type II and hyaluronic acid (HA) by electrospinning. Methods The three-dimensional porous nano-scaffolds were prepared by electrospinning techniques with collagen type II and HA (8 ∶ 1, W ∶ W), which was dissolved in mixed solvent of 3-trifluoroethanol and water (1 ∶ 1, V ∶ V). The morphology were observed by light microscope and scanning electron microscope (SEM). And the porosity, water absorption rate, contact angle, and degradation rate were detected. Chondrocytes were harvested from 1-week-old Japanese white rabbit, which was disgested by 0.25% trypsin 30 minutes and 1% collagenase overlight. The passage 2 chondrocytes were seeded on the nano-scaffold. The cell adhesion and proliferation were evaluated by cell counting kit 8 (CCK-8). The cell-scaffold composites were cultured for 2 weeks in vitro, and the biological morphology and extracelluar matrix (ECM) secretion were observed by histological analysis. Results The optimal electrospinning condition of nano-scaffold was 10% electrospinning solution concentration, 10 cm receiver distance, 5 mL/ h spinning injection speed. The scaffold had uniform diameter and good porosity through the light microscope and SEM. The diameter was 300-600 nm, and the porosity was 89.5% ± 25.0%. The contact angle was (35.6 ± 3.4)°, and the water absorption was 1 120% ± 34% at 24 hours, which indicated excellent hydrophilicity. The degradation rate was 42.24% ± 1.51% at 48 days. CCK-8 results showed that the adhesive rate of cells with scaffold was 169.14% ± 11.26% at 12 hours, and the cell survival rate was 126.03% ± 4.54% at 7 days. The histological and immunohistochemical staining results showed that the chondrocytes could grow well on the scaffold and secreted ECM. And the similar cartilage lacuma structure could be found at 2 weeks after co-culture, which suggested that hyaline cartilage formed. Conclusion The collage type II and HA complex three-dimensional nano-scaffold has good physicochemical properties and excellent biocompatibility, so it can be used as a tissue engineered cartilage scaffold.
Objective To observe and explore the effects of adipose-derived stem cells (ADSCs)-hyaluronic acid (HA) composite on heal ing of wound combined with radiation injury. Methods The ADSCs were harvested from the fat tissue in groin of 10 inbred Sprague Dawley (SD) rats and were isolated and cultured by enzyme digestion. The ADSCs-HA composite wasprepared with ADSCs (5 × 106 cells/mL) at passage 6 and HA (10 mg/mL). Thirty inbred SD rats, 15 males and 15 females, were randomly divided into groups A (n=10), B (n=10), and C (n=10). A 2 cm × 2 cm full-thickness skin defect was made on the rat back before 20 Gy 60Co radiation exposure. One week after debridement, wounds were treated by petrolatum gauze in group A as the control group, by HA (0.4 mL) and petrolatum gauze in group B, and by ADSCs-HA composite (0.4 mL) and petrolatum gauze in group C. The microvessel density (MVD) and the distribution of CD90 positive cells were observed at 1st, 2nd, 3rd, and 4th weeks. Results The wound heal ing was slower, and wound did not heal at 4th week and still filled with granulation tissue in group A; the wound heal ing of group B was faster than that of group A, and the wound did not heal completely with depression in the center at 4th week; the wound healed completely with epidermil izated surface and no obvious depression at 4th week in group C. The histological observation showed that MVD was significantly higher in group C than in groups A and B at the 1st, 2nd, and 3rd weeks (P lt; 0.05), and in group B than in group A at the 3rd week (P lt; 0.05); MVD was significantly higher in groups B and C than in group A (P lt; 0.05), but no significant difference was found between groups B and C (P gt; 0.05) at 4th week. No CD90 positive cell was found in groups A and B; CD 90 positive cells were observed in group C and gradually decreased with time. Conclusion ADSCs-HA composite can accelerate heal ing of wound combined with radiation injury by promoting and controll ing wound angiogenesis.
Objective To fabricate in situ crosslinking hyaluronic acid hydrogel and evaluate its biocompatibility in vitro. Methods The acrylic acid chloride and polyethylene glycol were added to prepare crosslinking agent polyethylene glycol acrylate (PEGDA), and the molecular structure of PEGDA was analyzed by Flourier transformation infrared spectroscopy and 1H nuclear magnetic resonance spectroscopy. Hyaluronic acid hydrogel was chemically modified to prepare hyaluronic acid thiolation (HA-SH). And the degree of HA-SH was analyzed qualitatively and quantitatively by Ellman method. HA-SH solution in concentrations (W/V) of 0.5%, 1.0%, and 1.5% and PEGDA solution in concentrations (W/V) of 2%, 4%, and 6% were prepared with PBS. The two solutions were mixed in different ratios, and in situ crosslinking hyaluronic acid hydrogel was obtained; the crosslinking time was recorded. The cellular toxicity of in situ crosslinking hyaluronic acid hydrogel (1.5% HA-SH and 4% PEGDA mixed) was tested by L929 cells. Meanwhile, the biocompatibility of hydrogel was tested by co-cultured with human bone mesenchymal stem cells (hBMSCs). Results Flourier transformation infrared spectroscopy showed that most hydroxyl groups were replaced by acrylate groups; 1H nuclear magnetic resonance spectroscopy showed 3 characteristic peaks of hydrogen representing acrylate and olefinic bond at 5-7 ppm. The thiolation yield of HA-SH was 65.4%. In situ crosslinking time of hyaluronic acid hydrogel was 2 to 70 minutes in the PEGDA concentrations of 2%-6% and HA-SH concentrations of 0.5%-1.5%. The hyaluronic acid hydrogel appeared to be transparent. The toxicity grade of leaching solution of hydrogel was grade 1. hBMSCs grew well and distributed evenly in hydrogel with a very high viability. Conclusion In situ crosslinking hyaluronic acid hydrogel has low cytotoxicity, good biocompatibility, and controllable crosslinking time, so it could be used as a potential tissue engineered scaffold or repairing material for tissue regeneration.
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.