Objective To investigate the effect of homograft of marrow mesenchymal stem cells (MSCs) seeded onto poly-L-lactic acid (PLLA)/gelatin on repair of articular cartilage defects. Methods The MSCs derived from36 Qingzilan rabbits, aging 4 to 6 months and weighed 2.5-3.5 kg were cultured in vitroand seeded onto PLLA/gelatin. The MSCs/ PLLA/gelatin composite was cultured and transplanted into full thickness defects on intercondylar fossa. Thirty-six healthy Qingzilan rabbits were made models of cartilage defects in the intercondylar fossa. These rabbits were divided into 3 groups according to the repair materials with 12 in each group: group A, MSCs and PLLA/gelatin complex(MSCs/ PLLA/gelatin); group B, only PLLA/gelatin; and group C, nothing. At 4,8 and 12 weeks after operation, the gross, histological and immunohistochemical observations were made, and grading scales were evaluated. Results At 12 weeks after transplantation, defect was repaired and the structures of the cartilage surface and normal cartilage was in integrity. The defects in group A were repaired by the hylinelike tissue and defects in groups B and C were repaired by the fibrous tissues. Immunohistochemical staining showed that cells in the zones of repaired tissues were larger in size, arranged columnedly, riched in collagen Ⅱ matrix and integrated satisfactorily with native adjacent cartilages and subchondral bones in group A at 12 weeks postoperatively. In gross score, group A(2.75±0.89) was significantly better than group B (4.88±1.25) and group C (7.38±1.18) 12 weeks afteroperation, showing significant differences (P＜0.05); in histological score, group A (3.88±1.36) was better than group B (8.38±1.06) and group C (13.13±1.96), and group B was better than group C, showing significant differences (P＜0.05). Conclusion Transplantation of mesenchymal stem cells seeded onto PLLA/gelatin is a promising way for the treatment of cartilage defects.
Objective To prepare a self-made compound, hemostatic jelly with polylactic acid(PLA), which has the hemostatic and absorbable effect on injured cancellous bone. Methods Two bone defects of 5 mm in diameter and 4 mm in depth were subjected on 20 health rabbits by drilling through their either outside plate of the iliac, and were filled with hemostatic jelly(group A), bone wax(group B) and blank(group C) respectively. Hemostasis were observed and recorded after 1 and 10 minutes. Five specimens were harvested at 2, 4, 8 and 12 weeks postoperatively for histological observation. Results ① Hemostatic effect: Bleeding of injured spongy bone stopped within 10 minutes after the treatment of hemostatic jelly and bone wax, but bleeding of balnk did not stop. Hemostatic jelly and bone wax adhered to bone defects firmly within 10 minutes was after the treatment. ② Absorbable effect: Hemostatic jelly and bone defects have not changed visibly in the first 2 weeks. With histological observation 4 to 8 weeks after the operation, hemastatic jelly was absorbed gradually and replaced by osteogenous tissue. It was absorbed completely after 8 to 12 weeks. Bone wax was not absorbed after 12 weeks, no new bone tissue was observed at bone wax area. The blank was replaced by connective tissue and osteogenous tissue partially after 12 weeks. Conclusion The compound hemostatic jelly manifests both hemostatic and absorbable effects on injured cancellous bone and may substitute for bone wax in clinical application.
Objective To study the result of using nerve conduit coated with chitin and filled with a guide-fiber to repair peripheral nerve defect. Methods Twenty-four female adult SD rats were made the model of 14 mm-gap on bilateral sciatic nerve under sterile condition. The rats were randomly divided into 4 groups(n=6),group A: polymer polyglycolic-lactic acid(PGLA) nerve conduit coated with chitin and filled with a guide-fiber as experimental group to repair 14 mm gap of rat sciatic nerve；group B: PGLA nerve conduit coated with chitin； group C: PGLA nerve conduit； group D: autograft (control group). The repair result was evaluated by normal observation, EMG testing and S-100 histological immunostaining analysis 4 and 12 weeks after operation.Results Four weeks after the operation，there were new regenerated immature fibers in groups A,B and C, 12 weeks after the operation, the regenerated nerve fibers were seen to have bridged the gap. There were myelinated fibers equably distributed and rarely newgenerated nerve fibers in distal parts of group D. The repair result of PGLA nerve conduit coated with a chitin and filled with guide-fiber was better than that of groups B and C（Plt;0.05）. There was significant difference of nerve fiber diameter，thickness of myelin sheath and fiber density in group D from those in groups A, B and C（Plt;0.05），but there were degenerative changes such as vacuoles insheaths and myelin separation in proximal and few new regenerated nerve fibers in distal parts of group D. Conclusion PGLA nerve conduit coated with chitin and filled with a guide-fiber offers a possible substitute for the repair of peripheral nerve defect.
Objective To study the mechanism of ectopic osteogenesis of nacre/Polylactic acid (N/P) artificial bone combined with allogenic osteoblasts, and to explore the possibility as a scaffold material of bone tissue engineering. Methods The allogenic- osteoblasts seeded onto N/P artificial bone were co-cultured in vivo 1 week.The N/P artificial bone with allogenic osteoblasts were implanted subcutaneously into the left back sites of the New Zealand white rabbits in the experimental group and the simple N/P artificial bone into the right ones in the control group. The complexes were harvested and examined by gross observation, histologic analysis and immunohistochemical investigation 2, 4 and 8 weeks after implantation respectively.Results In experimental group, the osteoid formed after 4 weeks, and the mature bone tissue withbone medullary cavities formed after 8 weeks; but in control group there was nonew bone formation instead of abundant fibrous tissue after 4 weeks, and more fibrous tissue after 8 weeks.Conclusion N/P artificial bone can be used as an optical scaffold material of bone tissue engineering.
摘要:目的:研究生物降解聚DL乳酸(PDLLA)自锁式捆绑带固定骨折的生物力学性能。方法:80只新西兰大白兔随机分为两组,建立股骨干非负重骨折动物模型,应用生物降解自锁式捆绑带固定骨折为实验组,钢丝固定骨折为对照组,分别于术后1、4、8、12周行生物力学检查进行比较。结果:捆绑带组在术后4、8、12周均比钢丝组的弯曲强度高,但4周、12周时Pgt;005,无统计学差异,8周时Plt;005,提示有统计学差异。离体同种固定物不同时间段抗拉强度自身比较:钢丝固定术后4阶段抗拉强度比较Pgt;005,任何两两比较都没有统计学差异,抗拉强度未随术后时间延长发生明显下降。捆绑带固定术后4周与术后1周比较Pgt;005,抗拉强度无明显降低,但术后8周和术后12周时Plt;005,抗拉强度明显下降。结论:生物降解自锁式捆绑带在非负重骨折治疗中可发挥良好的固定作用。生物降解自锁式捆绑带降解时,应力传导促进了骨折的愈合。Abstract: Objective: To study the biomechanics function of selflocking cerclage band made of biodegradable material polyDLlactic acid (PDLLA) in the fixation of fractures. Methods: Eighty rabbits were divided into two groups. Femur fracture models were made. Fractures were fixed using biodegradable selflocking cerclage band in experimental group and metal fixation material in control group. The biomechanics was analyzed and compared after 1, 4, 8 and 12 weeks respectively． Results: The bending strength of experimental group is more ber than that of control group after 4, 8 and 12 weeks, but it was not statistically significant at 4 and 12 weeks (Pgt;005). It was statistically significant at 8 weeks (Plt;005). The tensile strength of the same cerclage instrument was compared at different stage in vitro, and the result of the control group was not statistically significant at the four stage (〖WTBX〗P〖WTBZ〗gt;005). Regarding the changes of tensile strength of the cerclage instrument at different stage, the result of the experimental group was not statistically significant after 1 and 4 weeks (Pgt;005). However, the decrease of tensile strength was statistically significant after 8 and 12 weeks (Plt;005). Conculsion: Biodegradable selflocking cerclage band could be used in thetreatment of nonweightbearing fractures. The stress force conducting promotes healing of fracture when the selflocking biodegradable cerclage band degrades.
OBJECTIVE To confirm membrane-guided tissue regeneration in the healing course of segmental bone defects and study the mechanism. METHODS Segmental, 1 cm osteoperiosteal defects were produced in both radii of 12 rabbits. One side was covered with hydroxyapatite/polylactic acid(HA/PLA) membrane encapsulated as a tube. The contralateral side served as an untreated control. Healing courses were detected by radiographic and histologic examinations. RESULTS All control sides showed nonunion, whereas there were consistent healing pattern in test sides. CONCLUSION Membrane technique can promote bone regeneration.
Objective To investigate the feasibility of fetal liver cells for liver tissue engineering, the supporting function of poly L lactic acid (PLLA) scaffold for fetal liver cells and the effects of oncostatin M (OSM), nicotinamide (NA) and dimethyl sulfoxide(DMSO) on growth and hepatic differentiation. Methods After three dimensional PLLA scaffolds having a porous structure were prepared by using NH 4HCO 3 particle, fetal liver cells obtained from E14.5 C57BL/6CrSlc murine embryos were inoculated in the scaffolds. Cells were cultured in Williams’E medium with or without OSM, NA and DMSO for 30 days. Changes in cell number, liver-specific function, and cellular morphology were observed. Results When compared with in monolayer culture, cell number and albumin secretion increased obviously in three-dimensional PLLA. Alburmin secretion increased slightly in OSM group of monolayer culture, but increased obviously in OSM groupo of PLLA culture and in OSM/NA/DMSO group of both monlayer and PLLA cultures. Conclusion The three-dimensional PLLA scaffold is a good supporting material for the cultivation of tetal liver cells. OSM, NA and DMSO remarkaly stimulated maturation of hepatic parenchymal cells in vitro in terms of morphology and liver-specific function.
ObjectiveTo evaluate the biocompatibility of poly lactic acid/bone matrix gelatin (PLA/BMG) composite biomaterial so as to lay a foundation for bone defect repair. MethodsRats'MC3T3-E1 cells were cultured with leaching solution of PLA/BMG and PLA material respectively for 7 days. The cell proliferation rate was tested by MTT and cell toxicity grading was carried out everyday. The PLA/BMG and MC3T3-E1 cells were co-cultured, the cell shape and proliferation were observed by inverted phase contrast microscope at 1, 3, and 5 days and cell adhesion by scanning electron microscope at 5 days. The PLA and PLA/BMG were implanted subcutaneously in 15 Wistar rats. The histological observation was done, and the thickness of fibrous membrane, the number of inflammatory cells, and the vascularization area were measured at postoperative 2nd, 4th, and 8th week. ResultsThe tests for cytotoxicity in vitro showed that the cell proliferation rates were over 100% and the cell cytotoxic grades were grade 0 at 1-7 days in PLA/BMG group. While in PLA group, the cell proliferation rates were less than 100% and the cell cytotoxic grades were grade 1 at 2, 4, and 7 days. After co-culture of PLA/BMG and MC3T3-E1 cells, cells grew on the surface and in the pores of PLA/BMG, and the cellular morphology was triangle or polygon with abundant microvillus on the surface. After subcutaneous implantation, the rats survived to the end of experiment, and incision healed well. PLA was wrapped by connective tissue where there were a lot of lymphocytes and neutrophilic granulocytes. The cells and tissue grew slowly in PLA. The PLA/BMG materials were wrapped by little connective tissue where there were a few inflammatory cells. The connective tissue ingrowth was observed in the center of PLA/BMG. There was no significant difference in the thickness of fibrous membrane between 2 groups at each time point (P>0.05). The number of inflammatory cells of PLA/BMG group were significantly less than those in PLA group at 2, 4, and 8 weeks (P<0.05); the vascularization area was significantly larger than that in PLA group (P<0.05). ConclusionPLA/BMG composite biomaterials prepared by super critical-CO2 technique are good in cell and tissue biocompatibilty.
【Abstract】 Objective To investigate the manufacture and biocompatibil ity of a bioabsorbable poly-D,L-lactic acid(PDLLA) plate containing rhBMP-2. Methods rhBMP-2 was composited with PDLLA (0.05 mg/plate) through vacuum to repare PDLLA plate containing rhBMP-2. Thirty-two New Zealand rabbits (male or female) weighted (3.0 ± 0.5) kg were used in he study. A 2.5 mm middle ulna osteotomy was made bilaterally. The bones as well as periosteum were removed. The right side of all the animals was experimental side (n=32), was fixed internally by PDLLA plate containing rhBMP-2.The left side of all the animals was control side (n=32), was fixed by common PDLLA plate. After a follow-up of 2, 4, 8 and 12 weeks, the ulnas were examined visually, radiographically, histologically, and by computer graph analysis to compare the biocompatibil ity. Re sults Po rosity of PDLLA plate containing rhBMP-2 was 90%, aperture was 150 μm, tensile strength was higher than 50 MPa, three point flexural strength was higher than 90 MPa and intrinsic viscosity was 1.6 dL/g (chloroform solvent). All animals had a good heal ing 1 or 2 weeks after operation. All the animal’s diet and movement were normal. All the fractures were stable. The plates in the experimental group degraded faster than those in the control group. Relative values of callus density evaluated by X-ray at 2, 4, 8 and 12 weeks after operation in the experimental group were 39.22 ± 2.48, 48.79 ± 1.26, 63.78 ± 1.78 and 78.60 ± 1.25 respectively. Those in the control defects were 33.83 ± 1.13, 41.28 ± 1.25, 55.23 ± 0.68 and 66.54 ± 1.33. At each time point, the experimental side produced better and more trabeculae than the control side did (P ＜ 0.01). Histological examination showed that the newbone formation in the experimental side at 2, 4, 8 and 12 weeks after operation was 0.106% ± 0.015%, 0.292% ± 0.019%, 0.457% ± 0.048% and 0.601% ± 0.037%, while those in the control side was 0, 0.193% ± 0.019%, 0.339% ± 0.029% and 0.574% ± 0.047%respectively. At each time point, the experimental side produced better new bone formation than the control side did (P lt; 0.05). The experimental side showed better compatibil ity between plates and surrounding tissue, faster bone formation, more bone regeneration mass and better medullary canal structure than the control side. Conclusion PDLLA plate containing rhBMP-2 has good biocompatibil ity and osteoinducibil ity to enhance fracture heal ing.
Objective To determine the incidence of vitamin B1 deficiency in critically ill patients, to compare vitamin B1 levels between septic and non-septic patients, and to explore the relationship between vitamin B1 levels and lactate levels. Methods Using a retrospective study method, critically ill patients admitted to the Department of Intensive Care of Nanjing Drum Tower Hospital from February 2022 to November 2022 were included in the study, and the patients were divided into sepsis and non-sepsis groups according to the admission diagnosis, and the differences in the vitamin B1 levels of the patients between the two groups were analyzed, as well as the correlation between the vitamin B1 levels and the lactic acid levels. Results There was a significant difference in serum vitamin B1 levels between the sepsis patients and the non-sepsis patients [(1.6±0.3)ng/mL vs. (2.1±0.2)ng/mL, P=0. 009]. For all patients, there was no correlation between vitamin B1 levels and lactate levels. But when the patient was in a hyperlactate state (lactate level ≥2 mmol/L), vitamin B1 levels were significantly negatively correlated with lactate levels (r=–0. 229, P=0. 004). Conclusions Vitamin B1 deficiency is prevalent in critically ill patients and is strongly correlated with whether or not the patient is septic. Vitamin B1 levels are significantly and negatively correlated with lactate levels when the patient's lactate level is ≥2 mmol/L.