Objective To const ruct art ificial derm is on co llagen2chondront in sulfate (CS) scaffo ld. Methods Co llagen w as compounded from CS and 1-ethyl-3-(13-dimethyl am inop ropyl) carbodiim ide (EDC) used as a cro sslinker. Physical and chem ical p ropert ies of the scaffo ld w ere characterized by elect ron spect ro scopy fo r chem ical analysis (ESCA ) , scanning elect ron m icrograph (SEM ) , HE staining, and mechanical p roperty test. Derm is fibroblasts w ere iso lated from human embryo and w ere cultured on the scaffo lds. Th rough h isto logical test ing, immunoh istochem ical test ing and biochem ical p roperty test ing, the p roperty of co llagen-CS art ificial derm is w as compared w ith that of colla gen spongy art ificial derm is. Results Co llagen-CS had th ree2dimension st ructure w ith po rous. Compared w ith co llagen scaffo ld, themechanical p roperty of co llagen2CS scaffo ld imp roved. There w eremo re po lar group s on the surface of co llagen-CS scaffo ld. The fibroblasts on the co llagen-CS scaffo ld grew w ell, and art ificial derm is w as const ructed. Conclus ion Co llagen-CS art ificial derm is has mo re excellent bio logical and mechanical p ropert ies. F ibroblasts at tach and p ro liferate bet ter on co llagen2CS scaffo ld than on co llagen scaffo lds.
Objective To investigate the synergetic effect and possibil ity of repairing spinal cord injury (SCI) by transplantation of olfactory ensheathing cells (OECs) and chondroitinase ABC (ChABC) in adult rats. Methods Three adult male SD rats were used to isolated olfactory bulb and primarily cultured OECs. In the 8th or 9th day, OECs were transplanted, the concentration of cells was modulated to 1 × 105/μL. Fifty-four SD rats were made the models of T8 spinal cord crush injury and divided into 4 groups. In group A (control, n=36), injured site was not treated; in groups B, C and D (n=6), OECs, ChABC and OECs+ChABC were injected into injured site, respectively. At 1, 2, 3, 7 and 14 days after injury, the BBB score system was used to evaluate the motion function. At 0, 1, 2, 3, 7, 14 days in group A and at 14 days in groups B, C, D after injury, the maximal transverse diameter and gross area of necrosis were evaluated on HE stained sections. The immunofluorescence double label ing staining for gl ial-fibrillary acidic protein (GFAP)/CS56, GFAP/growth associated protein 43(GAP-43) and GFAP/neurofilament 160(NF160) was carried out to evaluate the regeneration of nerve fiber. Results At 14 days after injury, there were significant difference in the BBB scores between group A and groups B, C, D (P lt; 0.05), and between groups B, C and group D (P lt; 0.05), HE staining showed that the formation of cavity was observed in each group at 14 days after injury. There were significant difference in the maximal transverse diameter and gross area of necrosis between groups B, C, D and group A (P lt; 0.01), and between groups B, C and group D (P lt; 0.01). The immunofluorescence staining indicated that expression of GFAP were more intense in group A than in other groups, and the cavity of the lesion site was apparent, but it was moderate in groups B and C. The expression of GAP-43 was more intense in group D than in groups B and C. The expression of NF160 was more intense in group D. Conclusion Transplantation strategy of OECs combined with ChABC was effective in the repair of SCI in some extent.
Objective To investigate the effect of chondroitinase ABC (ChABC) on the expression of growth associated protein 43 (GAP-43) and gl ial fibrillary acidic protein (GFAP) after spinal cord injury (SCI) in rats. Methods A total of 150 adult female SD rats, weighing 250-300 g, were randomly divided into ChABC treatment group (group A), sal ine treatment group (group B), and sham operation group (group C) with 50 rats in each group. In groups A and B, the rats were made the SCI models and were treated by subarachnoid injection of ChABC and sal ine; in group C, the rats were not treated as a control. At 1, 3, 7, 14, and 21 days after operation, the Basso, Beattie, and Bresnahan (BBB) score system was used toevaluate the motion function, and immunofluorescent histochemical staining was used to observe the expressions of GAP-43 and GFAP. Results At different time points, the BBB scores of groups A and B were significantly lower than those of group C (P lt; 0.05); there was no significant difference in BBB score between groups A and B after 1, 3, and 7 days of operation (P gt; 0.05), but the BBB score of group A was significantly higher than that of group B after 14 and 21 days of operation (P lt; 0.01). At different time points, the GAP-43 and GFAP positive neurons of groups A and B were significantly higher than those of group C (P lt; 0.05). After 14 and 21 days of operation, the GAP-43 positive neurons of group A were more than those of group B (P lt; 0.01). After 7, 14, and 21 days of operation, the GFAP positive neurons of group A were significantly less than those of group B (P lt; 0.01). Conclusion ChABC can degrade gl ial scar, improve the microenvironment of the injured region and enhance the expression of GAP-43, which promotes axonal growth and extension.
Objective To assess the efficacy and safety of Hirudoid for microcirculation disorder. Methods We searched The Cochrane Library (Issue 4, 2009), PubMed, EMbase, CNKI, CBM, and VIP databases up to December 2009. Randomized controlled trials (RCTs) or quasi-RCTs concerning Hirudoid for microcirculation disorder were included. The methodological quality of the included studies was assessed according to the Cochrane Reviewer’s Handbook 5.0.1, and meta-analyses were conducted using RevMan software 5.0. Results Twenty-five RCTs were included, of which only one was graded as high quality and others were of low quality. The results of meta-analyses showed: Hirudoid could be effective in preventing the occurrence of phlebitis (OR=0.18, 95%CI 0.13 to 0.25). Hirudoid for treating phlebitis was also significantly better than magnesium sulfate or placebo (OR=7.18, 95%CI 4.59 to 11.22) and the time to symptom relief of Hirudoid was significantly shorter than placebo (MD= – 29, 95%CI – 37.30 to – 20.70). Hirudoid for internal fistula in hemodialysis patients was better than the simple hot compress (OR=8.89, 95%CI 4.25 to 18.58), and also better than the magnesium sulfate plus hot compress (OR=7.62, 95%CI 2.84 to 20.44). Hirudoid could also prevent the formation of hematoma and eliminate hematoma quickly. Hirudoid for tissue injury caused by irritating fluid extravasation was significantly better than magnesium sulfate (OR=4.25, 95%CI 2.06 to 8.78). Conclusion Hirudoid can significantly improve the microcirculation disorder, especially to the phlebitis. Due to the low quality of the included studies, further, more high quality trials are required.
With silk fibroin and vancomycin (VCM) as carrier and drug model, respectively, we prepared silk fibroin microspheres (SFM) with different concentration using the water-in-oil emulsion solvent diffusion method. We further developed VCM loaded calcium sulfate hemihydrates (CSH)/SFM artificial bone composites. In this study, surface morphology of the materials was observed using scanning electron microscope (SEM). Structure of the materials was studied with Fourier transform infrared spectroscopy (FTIR). Antibacterial activity of the materials was validated with the inhibition zone test. Drug release property of materials was evaluated using ultraviolet/visible spectrophotometry. Mechanical property of the materials was tested using computer-controlled electronic universal testing machine. The results showed that silk fibroin concentration had no significant effect on molecular conformation and antibacterial property of the SFM. The average diameter of SFM increased and the release rate decreased gradually as the silk fibroin concentration increased. The release rate decreased and the compressive fracture work increased as the silk fibroin concentration increased when adding SFM to CSH. This composite had partly corrected the disadvantages of CSH including the high brittleness and initial burst release. The research would have a good application foreground in the clinical treatment of infectious bone defect.
ObjectiveTo assess the safety, feasibility, and effectiveness of medical calcium sulfate (OsteoSet) mixed with powder for injection of rifampicin after radical debridement in the treatment of sacroiliac joint tuberculosis. MethodsA retrospective analysis was made on the clinical data from 27 patients with sacroiliac joint tuberculosis who underwent debridement surgery and local bone graft of OsteoSet artificial bone impregnated with powder for injection of rifampicin between August 2006 and August 2010. There were 10 males and 17 females with an average age of 35.2 years (range, 16-64 years). The mean disease duration was 7.5 months (range, 1.5-16 months). The left sacroiliac joint was involved in 16 cases, the right side in 10 cases, and both sides in 1 case; Of them, 18 cases had iliac fossa abscess and 6 cases of buttocks abscess. According to the classification system by Kim, there were 9 cases of type Ⅲ and 18 cases of type ⅠV. The preoperative Majeed score was 61.23±6.49, including good in 4 patients, general in 19 patients, and poor in 4 patients. ResultsAll patients achieved wound healing by first intention; no complications of pelvic infection, peripheral nerve injury, and lower extremity deep vein thrombosis occurred. Twenty-six patients were followed up 16 months on average (range, 12-24 months). All the patients had normal sense and movement of both lower limbs; no perineum sensory disturbance was found. One case of multi-drug resistant had local recurrence at 3 months after operation, which was cured after adjusting anti-tuberculosis drugs, nutritional support, enhancing immunity with thymopentin, and second operation. Bone union was observed at 10.5 months on average (range, 9-12 months) in 25 patients, and clinical symptoms disappeared with no recurrence or complication, and they returned to previous work. The Majeed score at last follow-up was 92.31±3.36, showing significant difference when compared with preoperative score (t=-32.76, P=0.00). The results were excellent in 22 patients, good in 4 patients, and the excellent and good rate was 100%, showing significant follow-up was 92.31±3.36, showing significant difference when compared with preoperative score (t=-32.76, P=0.00). The results were excellent in 22 patients, good in 4 patients, and the excellent and good rate was 100%, showing significant difference compared with the preoperative one (χ2=31.93, P=0.00). ConclusionBone graft interbody fusion surgery with rifampicin loaded OsteoSet is one of the effective methods to treat sacroiliac joint tuberculosis.
ObjectiveTo research the effect of chondroitin sulfate (CS) on the proliferation of myoblasts and the formation of myotube. MethodsThe myoblasts at passage 5 were used to prepare the cells suspension (1×108 cells/mL), and the experiment was divided into 4 groups based on CS concentration in the medium:group A (0 μg/mL), group B (50 μg/mL), group C (100 μg/mL), and group D (200 μg/mL). The cell morphology and myotube formation were observed by inverted microscope at 4, 5, and 8 days after treatment; MTT was used to detect the cell proliferation at 6 days, and the number of myotube was calculated by HE staining at 8 days. ResultsCells showed spindle shape after adherent, with ovoid nuclei and dense cytoplasm under inverted microscope. When the cell adherent rate was 90%, cells arranged in whorls swirled and showed long fusiform adherent growth; and then nuclei fusion resulted in formation of multincleated myotubes. At 8 days, most myoblasts fused to form myotube in group A, but less myotube was observed in groups B and C, and the least myotube in group D. The absorbance (A) values of groups A, B, C, and D were 0.045 2±0.004 4, 0.540 4±0.096 7, 0.660 9±0.143 4, and 1.069 0±0.039 0 respectively, showing significant difference between other groups (P<0.05) except between groups B and C P>0.05). HE staining observation showed that most myoblasts fused to form myotube in group A, but less myotube in groups B and C, and the least myotube in group D. The number of myotube of groups A, B, C, and D were 222.01±30.02, 193.13±42.46, 170.26±11.96, and 136.88±16.78 respectively, showing no significant difference among groups (F=1.658, P=0.252). ConclusionCS can significantly promote the proliferation of myoblast, the promotion is the biggest when CS concentration is 200 μg/mL.
Objective To formulate a rational adjuvant therapeutic evidence-based nursing plan for a patient with grade II red and swelling type phlebitis. Methods According to the condition of the patient and using the PICO principle, we put forward clinical problems. Then we comprehensively searched the National Guideline Clearinghouse (NGC), ACP Journal Club, The Cochrane Library, DARE, PubMed, MEDLINE, CNKI and Google Scholar from 2000 to 2012. Relevant clinical guidelines, evidence summaries, systematic reviews/ meta-analyses, randomized controlled trials (RCTs), and high quality reviews on adjuvant therapy of grade II red and swelling type phlebitis were collected and their authenticity, importance and applicability were evaluated. Results One systematic review, four meta-analyses, five RCTs, and one review were totally included. According to current evidence as well as the patient’s clinical conditions and preference, a comprehensive and effective adjuvant therapeutic and nursing programme was given to the patient. For grade II red and swelling type phlebitis with blisters and severe pain, paretic infusion should be immediately stopped on the lesion-side limb, and venous indwelling needle should be extracted. Then, mucopolysaccharide polysulfate cream should be applied on the skin impaired by vein inflammation, and the local area should be gently massaged for 3 min, twice daily (once in the morning and evening, respectively). After four-day treatment and nursing care, the patient with phlebitis had already recovered. Conclusion Evidence-based medicine approaches could help us develop comprehensive therapeutic plans for patients which promote recovery of patients with phlebitis, alleviate pain, improve health, and increasepatients’ quality of life.
ObjectiveTo investigate the mechanism of magnesium sulfate in protecting rabbit cartilage by initiating autophagy.MethodsTwenty-four adult female New Zealand rabbits were used to prepare post-traumatic osteoarthritis (PTOA) models by anterior cruciate ligament transection. Then, the PTOA models were randomly divided into PTOA group, distilled water group, and magnesium sulfate group, with 8 rabbits in each group. Immediately after operation, the distilled water group and the magnesium sulfate group were injected with 0.5 mL distilled water and 20 mmol/L magnesium sulfate solution in the joint cavity 3 times a week for 4 weeks, respectively. The PTOA group was not treated. The general condition of the animals was observed after operation. After 4 weeks, the expressions of tumor necrosis factor α (TNF-α) and collagen typeⅡ in the joint fluid and the expression of collagen type Ⅱ in venous blood were detected by ELISA assay. The protein expressions of transient receptor potential channel vanilloid 5 (TRPV5) and microtubule associated protein 1 light chain 3 (LC3; LC3-Ⅱ/LC3-Ⅰ) in femoral cartilage were detected by Western blot. The mRNA expressions of interleukin 1β (IL-1β), TNF-α, matrix metalloproteinases 3 (MMP-3) in synovial tissue and collagen type Ⅱ, Aggrecan (AGN), SOX9 in cartilage tissue were detected by real-time fluorescence quantitative PCR. Cartilage tissue sections were stained with HE staining, Masson staining, and Alcian blue staining and scored according to the modified histological osteoarthritis (OA) score.ResultsAll animals survived until the experiment was completed. Compared with the other two groups, the expression of TNF-α in joint effusion and collagen type Ⅱ in joint effusion and venous blood were decreased in magnesium sulfate group; the protein expression of TRPV5 decreased, and the ratio of LC3-Ⅱ/LC3-Ⅰ increased significantly; the mRNA expressions of IL-1β, TNF-α, and MMP-3 in synovial tissue were decreased, and the mRNA expressions of collagen type Ⅱ, AGN, and SOX9 in cartilage tissue were increased; OA scores also decreased significantly. All differences were statistically significant (P<0.05). There was no significant difference in the above indicators between the PTOA group and the distilled water group (P>0.05).ConclusionIntra-articular injection of magnesium sulfate can reduce intra-articular inflammation, reduce the loss of collagen type Ⅱ and AGN, and is beneficial to cartilage regeneration in rabbits. The mechanism may be related to the initiation of chondroautophagy by inhibiting the calcium channel TRPV5.
Objective To discuss the role of heparan sulfate (HS) in bone formation and bone remodeling and summarize the research progress in the osteogenic mechanism of HS. Methods The domestic and abroad related literature about HS acting on osteoblast cell line in vitro, HS and HS composite scaffold materials acting on the ani-mal bone defect models, and the effect of HS proteoglycans on bone development were summarized and analyzed. Results Many growth factors involved in fracture healing especially heparin-binding growth factors, such as fibroblast growth factors, bone morphogenetic protein, and transforming growth factor β, are connected noncovalently with long HS chains. HS proteoglycans protect these proteins from protease degradation and are directly involved in the regulation of growth factors signaling and bone cell function. HS can promote the differentiation of stem cells into osteoblasts and enhance the differentiation of osteoblasts. In bone matrix, HS plays a significant role in promoting the formation, maintaining the stability, and accelerating the mineralization. Conclusion The osteogenesis of HS is pronounced. HS is likely to become the clinical treatment measures of fracture nonunion or delayed union, and is expected to provide more choices for bone tissue engineering with identification of its long-term safety.