To investigate the anatomic feature of the posterior hip joint capsule and its distributional difference of collagen fibers and to probe the optimization of the capsulotomy which can reserve the best strength part. Methods Ten adult cadaver pelvises (6 males and 4 females, aged 28-64 years) fixed with formal in were used. Ten right hips were used for anatomical experiment of hip joint capsule. The posterior hip joint capsules were divided into 3 sectors(I-III sectors ) and 9 parts (IA-C, IID-F, IIIG-I). The average thickness of each part was measured and the ischiofemorale l igaments were observed. Five capsules selected from ten left hips were used for histological experiment. The content of collagen fibers in sector I and sector II was analyzed by Masson’s staining. Two fresh frozen specimens which were voluntary contributions were contrasted with the fixed specimens. The optimal incision l ine of the posterior capsule was designed and used. Results The thickness in the posterior hip joint capsule [IA (2.30 ± 0.40), IB (4.68 ± 0.81), IC (2.83 ± 0.69), IID (2.80 ± 0.79), IIE (4.22 ± 1.33), IIF (2.50 ± 0.54), IIIG (1.57 ± 0.40), IIIH (2.60 ± 0.63), IIII (1.31 ± 0.28) mm] had no uniformity (P lt; 0.01). The IIIG part and the IIII part were thinner than the IB part and the IIE part (P lt; 0.01). Two weaker parts located at obturator externus sector (sector III), the ischiofemorale l igament trunk went through two thicker parts (IB and IIE). The distribution of the collagen fibers in sector I and sector II(IA 20.34% ± 5.14%, IB 48.79% ± 12.67%, IC 19.87% ± 5.21%, IID 17.57% ± 3.56%, IIE 46.76% ± 11.47%, IIF 28.65% ± 15.79%) had no uniformity (P lt; 0.01). The content of collagen fibers in IB part and IIE part were more than that of other parts (P lt; 0.01). There were no statistically significant difference in the distribution feature of the thickness and the ischiofemorale l igaments between the fresh frozen specimens and the fixed specimens. The optimal incision l ine C-A-B-D-E of the posterior capsule was designed and put into cl inical appl ication. The remaining capsular flap comprise the most of the ischiofemorale l igament trunk and the part of gluteus minimus. Conclusion Although enhanced posterior soft tissue repairin total hip arthroplasty was investigated deeply and obtained great development, but the postoperative dislocation rate was not el iminated. It is significant for optimizing the capsulotomy to reserve the best strength part of the posterior capsule and to bring into full play the function of the ischiofemorale l igaments.
Objective To validate the advantage of repairing bone defect by staphylococcus aureus injection carried in collagen membrane. Methods Twentyfour adult New Zealand rabbits were divided into two groups randomly. After the experimental model of standard bone defect had been made by operation, collagen membrane/staphylococcus aureus injection and staphylococcus aureus injection with the same quantity were transplanted in bone defect areas of the two groups respectively. The reconstructed tissues were observed by general method, X-ray, histology, and immunohistochemistry at 2nd、4th、6th、8th week respectively. Results The experimental group showed that new bone proliferated distinctly in bone defect areaand the proliferation lasted long, and no excessive connective tissue in defectarea. X-ray observation showed that there was continual callus growth in transplantation area in early stage and the distribution of new bones was even in the group. Histological observation showed that there were many new bone growth centers in bone defect area, trabecular bones were sequentially distributed, and mature bone replacement was complete. Immunohistochemical examination showed that bone morphogenetic protein (BMP) could be seen for a long time and BMP took up a large part in the new bone tissues. Conclusion Collagen membrane could prevent parenchyma from penetrating into bone defect area and provide room for new bone growth. As the carrier of staphylococcus, collagen membrane could reduce the overflow of staphylococcus and improve its curative effect as well.
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 analyze the molecular composition of type IV collagenous fibres in internal limiting membrane (ILM) of human retina. Methods ILM was surgically removed from retina and identified under phase-contrast and transmission electron microscopes. Monoclonal antibodies against different αchains (α1-α6) of type IV collagen were immuno-localized. Results α3, α4, and α5 chains of type IV collagen were immuno-localized in human retinal ILM, while α1, α2, and α6 chains could not be immuno-localized. Conclusion Type IV collagenous fibres in human retinal ILM are composed of α3, α4, and α5chains. (Chin J Ocul Fundus Dis，2004，20：364-368)
The structure of 39 specimens of hepatobiliary duct stones with strictures were studied histologically. The elastic and collagenous fibers were studied by quantitative analysis. The results show that the epithelium of the sttnotic bile duct are intact but with proliferation. The mitochondrions are degenerated and broken, the endoplasmic reticulum are dilated, suggesting the functional impediment of these epithelium. The mucous glands are markedly proliferated fibrosis are found near the glands which are destroyed .Some of the elastic fibers are destroyed and arranged disorderly . Hyaline degeneration was observed in collagenous fibers with remarkable increase of the volume density.
Objective To investigate the effect of hepatitis C virus (HCV) F protein on proliferation and collagen expression of hepatic stellate cells. Methods After pcDNA3.1-f plasmid containing HCV f gene or empty pcDNA3.1 plasmid was transfected hepatic stellate cells LX2 by liposome, LX-f or LX-p cells were obtained by G418 screening. The proliferation of LX-f or LX-p cells was analyzed by MTT, and the contents of collagen type Ⅰand Ⅲ secreted by LX-f or LX-p cells were detected by ELISA. Results After 24 h cultivation, the proliferation rate of LX-f cells was higher than that of LX-p cells at each time point （Plt;0.01）. After 48 h cultivation, the contents of collagen typeⅠand Ⅲ secreted by LX-f were (25.89±0.42) ng/ml and (18.21±0.49) ng/ml, which was significantly higher than those of LX-p cells 〔(22.65±0.49) ng/ml and (15.29±0.62) ng/ml〕, Plt;0.01. Conclusion HCV F protein is able to promote proliferation of hepatic stellate cells, and up-regulate the excretion of collagen type Ⅰand Ⅲ in those cells, which induces hepatic fibrosis.
Objective To analyze the contents of collagen type Ⅰ, type Ⅲ and the ratio of collagen type Ⅰ to collagen type Ⅲ in posterior rectus sheath of different person. Methods One hundred and four tissues specimen of posterior rectus sheath were obtained during patients’ abdominal operation. The contents of collagen type Ⅰand type Ⅲ were detected by using immunohistochemistry methods. The differences of collagen contents between male and female, physical work group and non-physical work group, smoking group and non-smoking group were observed. The relationships between the contents of collagen and age, body mass index (BMI), and height were analyzed, respectively. Results ① The content of collagen typeⅠand the ratio of collagen type Ⅰ／Ⅲ were both lower in male than those in female （Plt;0.01）； there were no obvious differences in the content of collagen type Ⅲ and the total amount of collagen （Pgt;0.05）. ② There were no differences between physical work group and non-physical work group with the amount and the ratio of collagens （Pgt;0.05）. ③ When compared with non-smoking group, less collagen typeⅠ(Plt;0.01) and lower ratio of collagen Ⅰ／Ⅲ （Plt;0.05） were found in smoking group； but there was no difference with content of collagen Ⅲ（Pgt;0.05）, as well as the total amount of collagen （Pgt;0.05）. ④ The total amount of collagen, the content of collagen type Ⅰand the ratio of collagen Ⅰ／Ⅲ all decreased as age increases （r=0.341, 0.392, 0.212， P＜0.001, Plt;0.05）； no obvious change was observed in the content of collagen Ⅲ （r=0.089， Pgt;0.05）. ⑤ The content and ratio of collagen had no obvious relationships with BMI and height （Pgt;0.05）. Conclusion Smoking, gender and age are all influential factors of the content and ratio of collagens in the tissue.
ObjectiveTo study the preparation method of acellular dermal matrix (ADM) for cartilage tissue engineering and analyze its biocompatibility.
MethodsThe dermal tissues of the calf back were harvested, and decelluarized with 0.5% SDS, and the ADM was reconstructed with 0.5% trypsin, cross-linked with formaldehyde, and modified with 0.5% chondroitin sulfate which can promote the proliferation of chondrocytes. And the porosity, cytotoxicity, and biocompatibility were determined. Co-cultured 2nd passage chondrocytes and bone marrow stromal cells in a proportion of 3 to 7 were used as seed cells. The cells were seeded on ADM (experimental group) for 48 hours to observe the cell adhesion. The expressions of mRNA and protein of collagen type Ⅱ were tested by RT-PCR and Western blot methods, respectively. And the expressions were compared between the cells seeded on the scaffold and cultured in monolayer (control group).
ResultsAfter modification of 0.5% trypsin, the surface of ADM was smooth and had uniform pores; the porosity (85.4%±2.8%) was significantly higher than that without modification (72.8%±5.8%) (t=-4.384, P=0.005). The cell toxicity was grade 1, which accords to the requirements for cartilage tissue engineering scaffolds. With time passing, the number of inflammatory cells decreased after implanted in the back of the rats (P<0.05). The scanning electron microscope observation showed that lots of seed cells adhered to the scaffold, the cells were well stacked, displaying surface microvilli and secretion. The expressions of mRNA and protein of collagen type Ⅱ were not significantly different between experimental and control groups (t=1.265, P=0.235;t=0.935, P=0.372).
ConclusionThe ADM prepared by acellular treatment, reconstruction, cross-linking, and modification shows perfect characters. And the seed cells maintain chondrogenic phenotype on the scaffold. So it is a proper choice for cartilage tissue engineering.
Objective To study the allograft effect of two kinds of tissue engineered oral mucosa lamina proprias on skin fullthickness wounds. Methods The cultured Wistar rat oral mucosa fibroblasts (OMF) were incorporated into collag en or chitosancollagen to construct the tissue engineered oral mucosa laminaproprias, and then the OMFs were labeled with BrdU. The fullthickness round skin defects were made with a round knife (diameter, 0.8 cm) on the backs of 36 Wistar rats (2125 weeks old), which were divided into 2 experimental groups: the fibroblastpopulated collagen lattices (FPCL) group (grafted by FPCLs) and the fibroblastpopulated chitosan collagen lattices (FPCCL) group (grafted by FPCCLs), and the control group (only covered with gauges). All the wounds were observed by the naked eyes or the light microscope, and were measured 4, 7, 14, and 21 days postoperatively. Results There were no infection during the wound healing period. At 7 days after the grafting, the wounds in the 3 groups were covered by scab and/or gauze; at 14 days, the gauze and scab on the wounds in the three groups were all replaced by the new epidermis naturally except one scab each in the FPCCL group and the control groups,which was replaced at 17 days.All the centers of the new epidermis were measurable as the pink red points. At 21 days, all the new skins were smooth without hairs, and their color was similar to the normal one. At 4, 7, and 14 days,there was an indication that the wound diameters became significantly smaller in the three groups; but after the 14th day, there was no significant indication of this kind. At 7 days, the wound diameter in the FPCL group was significantly smaller than that in the FPCCL group and the control group (Plt;0.01). Under the lightmicroscope, at 4 days postoperatively, the decayed tissue on the surfaces of the recipient wounds in the FPCL group and the FPCCL group was separated from the lower granular tissue in which there were many inflammatory cells, fibroblasts, and new vessels. There was a similar-phenomenon in the control group. Each skin wound in the three groups was only partly keratinocyted at 7 days postoperativel y. The recipient wounds were wholly keratinocyted with when rete ridges observed at 14 and 21 days, but in the control group the wounds were keratinocyted with no rete ridges. Fibers in the new dermis were thin. The OMFs with Brdu appeared in the granular tissue and new dermis at 4, 7, 14, and 21 days postoperatively, which could be illustr ated by the immunohistochemical staining. The positive OMFs and the granular tissue joined in the repair of the skin defe cts without any allergic reaction during the period of the wound healing. Conclusion The oral mucosa fibroblasts as the new seed cells can join i n the repair of the skin defects effectively and feasibly. The fibroblastpopul ated collagen lattices and the fibroblastpopulated chitosan collagen lat tices can repair skin defects effectively and feasibly, too. And the quality of the new skins was better in the two experimental groups than in the control group.
Objective To repair the defects in articular cartilage with collagen complex gradient TCP in vivo andto study the regenerated cartilage histomorphologically. Methods The models of defects in articular cartilage were madeartificially in both condylus lateral is femoris of mature rabbits, male or female, with the weight of 2.0-2.5 kg. The right defects were implanted with the material of Col/TCP as the experimental group and the left defects were untreated as the control group. The rabbits were killed at 4, 6, 8, 12 and 24 weeks after operation, respectively, with 6 ones at each time, and the macroscopic, histological, ultrastructural examinations and semi-quantity cartilage scoring employing Wakitanifa repaired cartilage value system were performed. Results Four weeks after operation, the defects in the experimental group were partly filled with hyal ine cartilage. Twelve weeks after operation, the defects in the experimental group were completely filled with mature hyal ine cartilage. Twenty-four weeks after operation, regenerated cartilage had no ataplasia. However, fibrous tissues were seen in the control group all the time. At 4, 6, 8, 12 and 24 weeks ostoperatively, the Wakitanifa cartilage scores were 7.60 ± 0.98, 5.69 ± 0.58, 4.46 ± 0.85, 4.35 ± 0.12 and 4.41 ± 0.58, respectively, in the experimental group and 10.25 ± 1.05, 9.04 ± 0.96, 8.96 ± 0.88, 8.88 ± 0.68 and 8.66 ± 0.54, respectively, in the control group. At 4, 6, 8, 12 and 24 weeks postoperatively, the collagen II contents were 0.28% ± 0.01%, 0.59% ± 0.03%, 0.68% ± 0.02%, 0.89% ± 0.02% and 0.90% ± 0.01%, respectively, in the experimental group, while 0.08% ± 0.02%, 0.09% ± 0.04%, 0.11% ± 0.03%, 0.25% ± 0.03% and 0.29% ± 0.01%, respectively, in the control group. Differences between the control group and the experimental group were significant (P lt; 0.05). By then, typical chondrocyte was observed by transmission electron microscope in the experimental group and much fiber with less fibrocyte was observed in the control group. Conclusion Three-dimensional scaffold collagen complex gradient TCP may induce cartilage regeneration to repair the defects of articular cartilage in vivo.
