ObjectiveTo evaluate the efficacy of XiaochengqiMixture (XM) on promoting healing of colonic stoma. MethodsForty Wistar rats were divided into two groups randomly after colonectomy: experimental group (n=20) and control group (n=20). In early postoperatively stage rats were given gastric administration of XM in the experimental group and pure water in the control group. On day 3, 7, and 14 after establishment of animal models, laparotomy was performed in two groups of rats, respectively. Anastomotic stoma and surrounding tissues were harvested to detect the context of hydroxyproline and collagen fiber proportion by Masson dying. ResultsOn day 3 after establishment of animal models, hyperplastic collagen with small fiber was observed while no fasciculus was found. Hydroxyproline context and collagen fiber proportion of rats were higher in experimental group than those in control group (Plt;0.05). On day 7 after operation, many fasciculuses were found in two groups of rats, hydroxyproline context and collagen fiber proportion of rats were higher in experimental group than those in control group (Plt;0.01). On day 14 after operation, fasciculuses became bigger and more regular in arrangement, but there was no significant difference between the two groups (Pgt;0.05). ConclusionXM is capable of promoting healing of colonic stoma and might prevent the occurrence of anastomotic fistula.
OBJECTIVE: To build the trestle of tissue engineering for skin with the collagen. METHODS: The collagen was obtained from the baby cattle hide pretreated by Na2S and elastinase and Protease M, then the collagen was dissolved in 0.5 mol/L acetic acid solution. The collagen was treated with Protease N to minimize its immunogenicity. The resulting collagen could be used to build the trestle of tissue engineering for skin because of good biocompatibility. The collagen molecular weight and structure were analyzed by SDS-PAGE. The bioactivity of trestle was tested in the experiment of the mice wound healing and the cell implantation. RESULTS: The SDS-PAGE result of the collagen treated by Protease M showed the typical spectrum of type I collagen. The built trestle was a collagen sponge matrix in which micropore size was 50-200 microns. It could accelerate wound healing and the implanted fibroblasts could proliferate well. CONCLUSION: The collagen treated by Protease N can get good biocompatibilily and is suitable for building the trestles of tissue engineering for skin with good bioactivity.
In order to explore the effects of clenbuterol on intramuscular collagen metabolism in denervated skeletal muscles, a randomized, double-masked and placebo-controlled group were studied. Seventy-one patients with complete function loss in muscularcutaneous nerve resulted from brachial plexus injury were administered clenbuterol or placebo 60 micrograms Bid for more than 3 months. Biopsies of the biceps brachia muscle were performed at the beginning and end of this study. The biopsied muscles were processed with anti-collagen I and IV immunohistochemical stains and image analysis as well. The result showed that the collagen proliferation of both type I and IV was much reducible in the clenbuterol-treated group than that of the placebo-treated group (P lt; 0.05). It was concluded that clenbuterol could inhibit partially the proliferation of intramuscular collagens in denervated skeletal muscle.
OBJECTIVE: To investigate the effects of basic fibroblast growth factor (bFGF) on the promoter activities of human alpha 1(I) procollagen gene and the interaction between bFGF and transforming growth factor-beta 1 (TGF-beta 1). METHODS: Fibroblasts of the hypertrophic scar and normal skin from a 3-year-old patient were primarily cultured and subcultured in vitro. Both of the fibroblasts were transient transfected with phCOL 2.5, containing -2.5 kb of 5’f lank sequence of human alpha 1(I) procollagen gene and CAT reporter gene by FuGENE transfection reagent; and treated thereafter by 16 ng/ml bFGF, 2 ng/ml TGF-beta 1 and 16 ng/ml bFGF + 2 ng/ml TGF beta 1 for 24 hours. The relative CAT expression values were determined by CAT-ELISA. RESULTS: TGF-beta 1 bly induced the CAT expression level, however, bFGF not only inhibited the basal CAT expression but also reduced the CAT expression up-regulated by TGF-beta 1 in normal skin and hypertrophic scar fibroblasts (P lt; 0.05). CONCLUSION: bFGF can reduce the promoter activities of human alpha 1(I) procollagen gene and antagonize the role of TGF-beta 1 in up-regulating the promoter activities of human alpha 1(I) procollagen gene in normal skin and hyertrophic scar fibroblasts.
In order to study the influence of severity of tendon injury on the morphology of collagen fibers during healing process of extensor tendons, 40 female Wistal rats were used for investigation. The rats were divided into 2 groups. Transection of the tendon of extensor digitorum longus was performed in one group, while partial section of the same tendon was performed in the other group. Morphometric analysis was undertaken on the 15th, 30th, 60th and 90th day after operation. The result was that there was no significant difference between the two groups both in distribution and diameter of collagen fibers on the 15th and 30th days (P gt; 0.05). However, there was significent difference between those on the 60th and 90th days (P lt; 0.05). It was concluded that the severity of the tendon injury could influence the morphology of collagen fibers during the late stage of tendon healing.
目的:观察胶原酶治疗腰椎间盘突出的疗效。方法:对1218例不同程度的腰椎间盘突出患者进行椎旁穿刺,注入胶原酶1200u进行溶解治疗。结果:91%的患者取得较好疗效,9%的患者疗效欠佳。结论:胶原酶是治疗腰椎间盘突出的一种有效方法。
Objective To construct three kinds of collagen-chitosan porous scaffolds with enhanced biostability and to investigate the histocompatibility of the scaffolds in vivo. Methods Collagen-chitosan porous scaffolds were fabricated by freeze-drying method, cross-linked using dehydrothermal treatment and glutaraldehde, respectively. The morphology of the uncross-linked scaffold (scaffold1), dehydrothermally cross-linked scaffold (scaffold 2) and glutaraldehde crosslinked scaffold (scaffold 3) was studied by scanning electron microscopy. Threekinds of scaffolds were embedded subcutaneously on dorsal surface of 12 rabbit ears. The general and local responses were recorded daily. The biostability and histocompatibility of the scaffolds were observed by using HE staining after 3, 7, 14 and 28 days of operation. Results The scaffolds had three-dimensional porous structures with a porosity of more than 90%, and possessed pore sizes of 120±10 μm, 80±15 μm and 170±20 μm, respectively. All experimental rabbits survived with good general condition during the study. All skin incisions healed well without obvious reactive red or swelling. Histological study showed that scaffold 1 was degraded rapidly with obvious inflammation. The degradation of scaffold 2 was slower than that of scaffold 1 and the inflammation of scaffold 2 was also milder than that of scaffold 1. Scaffold 3 possessed slow degradation property with slight inflammatory reaction, and rapid tissue regeneration. Conclusion The collagenchitosan porous scaffolds have three-dimensional porous structures that are suitable for tissue regeneration. The biostability and histocompatiility of the scaffolds are enhanced after cross-linked. Glutaraldehde cross-linked is better than dehydrothermally cross-linked, which can facilitate dermal tissuereconstruction.
Objective To investigate the effect of leptin on fibroblast proliferation and collagen synthesis as to elucidate that fibroblasts play a role in leptin’s effect on wound healing. Methods Purified dermal fibroblasts were derived from sucking wistar rat skin and exposedto leptin at concentration of 0, 10, 50, 100, 200, and 400 ng/ml. The survived fibroblasts were assessed by the colorimetric thiazolyl blue (MTT) assay. Replication of fibroblast was quantified by the incorporation of 3H-thymidine. Collagen synthesis of fibroblast cell was measured by the incorporation of 3H-proline into collagenasesensitive protein. Results The absorption of fibroblast exposed to leptin at concentration of 200 and 400 ng/ml 0.082±0.013, 0.091±0.018 was higher than that of control group 0.063±0.010, P＜0.05. The incorporations of 3H-thymidine of fibroblast exposed to leptin at concentration of 200 and 400 ng/ml 379±101 cpm,326±33 cpm were significantly higher than those of control group 219±56 cpm, P＜0.05. The incorporations of 3H-proline of fibroblast exposed to leptin at concentration of 200 and 400 ng/ml 911±55 cpm, 1 072±259 cpm were significantly higher than that of control group 679±176 cpm, P＜0.05. Conclusion Leptin can promote rat cutaneous fibroblast proliferation and collagen synthesis in vitro. This suggests that cutaneous fibroblast plays a role in leptin’s promoting skin wound healing and it may be one of the main mechanisms by which leptin enhances skin wound healing.
Objective To investigate the feasibil ity of alendronate (ALN) in treating osteoarthritis (OA) by observing the effects of ALN on interleukin 1β (IL-1β) induced chondrocytes of rat in vitro. Methods The chondrocytes of knee articular surface from 15 SD rats (1-month-old, female or male, weighing 100-150 g) were cultured. The chondrocytes were observed by inverted phase contrast microscope and identified by toluidine blue staining and HE staining. The third passage chondrocytes were divided into 3 groups: the chondrocytes were cultured with DMEM for 5 days in group A, with 10 ng/mL IL-1β for 2 days and with DMEM for 3 days in group B, and with 10 ng/mL IL-1β for 2 days and with 1 × 10-6 mol/L ALN for 3 days in group C. Immunocytochemistry and real-time PCR were performed to determine the expression levels of collagen type II (Col II), matrix metalloproteinase 13 (MMP-13), and β-catenin. Results Toluidine blue staining proved that the cultured cells were chondrocytes. The integrated absorbency (IA) value of Col II in group C (10.290 7 ± 0.499 2) was lower than that in group A (15.377 0 ± 0.571 8) and higher than that in group B (5.463 2 ± 0.450 4), showing significant differences (P lt; 0.05). The IA value of MMP-13 in group C (3.068 6 ± 0.205 6) was significantly lower than that in group B (6.998 1 ± 0.329 7, P lt; 0.05), but there was no significant differenc when compared with group A (2.777 5 ± 0.199 6, P gt; 0.05). The IA value of β-catenin in group C (6.611 7 ± 0.381 8) was lower than that in group B (11.799 9 ± 0.348 7) and higher than that in group A (4.390 3 ± 0.551 9), showing significant differences (P lt; 0.05). The mRNA expression of Col II in group C was significantly higher than those in groups A and B (P lt; 0.05), the mRNA expression of MMP-13 in group C was significantly lower than that in group B (P lt; 0.05) but there was no significant difference when compared with group A (P gt; 0.05). The mRNA expression of β-catenin in group C was significantly lower than that in group B (P lt; 0.05) and higher than that in group A (P lt; 0.05). Conclusion ALN can protect rat chondrocyte from OA induced by IL-1β in vitro possibly by upregulating Col II and inhibiting the expression of MMP-13 and β-catenin in the chondrocytes.