Objective To fabricate a novel porous bioactivecomposite biomaterial consisting of poly lactic acid (PLA)bone matrix gelatin(BMG) by using the supercritical carbon dioxide fluid technique (SC-CO2) and to evaluate its osteoinductive activity. Methods The cortical bones selected from healthy adult donors were processed into BMG by the defatting, demineralizing, and deproteinizing processes. PLA and BMG were mixed at a volume radio of 3∶1; then, the PLA-BMG mixed material and the pure PLA material were respectively placed in the supercritical carbon dioxide reaction kettles, and were respectively added by the NaCl particles 100200 μm in diameter for theporosity of the materials so that the porous PLA-BMG composite material and the porous PLA composite material could be formed. The mouse osteoblastlike MC3T3-E1 cells were cultured in the dulbecco’s modified eagle medium (DMEM) supplemented with 10% fetal bovine serum. Then, 20 μl of the MC3T3E1 cell suspensions containing 2 ×106 cells /ml were delivered into the culturing plate (24 wells/plate) made of the different materials, which were co-cultured for 2 weeks. In the PLA-BMG group, 100 μg of the crushed PLA-BMG material was contained in each well; in the PLA group, 100 μg of the crushed PLA material was containedin each well; and in the DMEM group, only DMEM was contained, which served as the control group. There were 6 wells in each group. The quantitative analysis onthe calcification area was performed by the staining of the alizarin red S. Theco-cultured cells were harvested and lysated in 1 ml of 0.2% Nonidet P-40 by the ultrasonic lysating technique. Then, the ALP activity and the Ca content were measured according to the illuminations of the reagent kits. Results The porous PLABMG composite material showed a good homological porosity with a pore diameter of 50-150 μm and a good connectivity between the pores. The ALP activity, the Ca content, and the calcification area were significantly greater in the PLABMG group than in the PLA group and the control group (325.59±70.40 U/gprot, 3.51±1.64 mmol/gprot, 42.98±4.44% vs. 63.62±30.01 U/gprot, 1.04±0.21 mmol/gprot, 9.55±1.94%, and 2.40±1.47 U/gprot, 0.70±0.24 mmol/gprot, 0.86±0.41%; Plt;0.05). Meanwhile, there was a statistically significant difference between the PLA group and the control group in the ALP activity and the calcification area (Plt;0.05). Conclusion The porous PLABMG composite material prepared by the use of SC-CO2 has a good steoinductive activity and can be used as a promising bone biomaterial and a bone tissue engineered scaffold.
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.
ObjectiveTo investigate the value of the change trend of lactic acid level 24 h after operation in predicting the complications of upper gastrointestinal ulcer perforation. MethodsA total of 167 patients with upper digestive tract ulcer perforation who underwent surgical treatment in Shougang Hospital of Peking University from March 2021 to June 2023 were selected as the study objects, and were divided into mild to moderate group (n=117) and severe group (n=50) according to the severity of the disease. General data and lactic acid levels in 24 h after surgery were compared between the two groups. In addition, according to whether the patients had complications or not, they were divided into a group without complications (n=119) and a group with complications (n=48). The general data of the two groups were compared. A combined model was constructed, and Cox regression model was used to analyze the relationship between the change of lactic acid level at 24 h after operation and the complications of infection. Logistic regression model combined with restricted cubic spline model was used to analyze the dose-response relationship between lactic acid level and infection complications in patients with upper gastrointestinal ulcer perforation 24 h after operation. Log-binomial model was used to analyze the risk effect of complication types on lactic acid levels in patients with upper gastrointestinal ulcer perforation. ResultsAt 8, 16 and 24 h after operation, lactic acid levels in both groups (mild to moderate group and severe group) were significantly lower than before operation, and lactic acid level in mild to moderate group was significantly lower than that in severe group (P<0.05). Repeated measurement ANOVA showed that the time effect, intergroup effect and interaction effect of lactic acid levels were significantly different between the two groups (P<0.05). There were significant differences in exhaust time, postoperative hospital stay, C-reactive protein, lactic acid level and red blood cell distribution width between the complicated group and the uncomplicated group (P<0.05). The combined model showed that each longitudinal increase of 1 mmol/L in lactic acid level 24 h after operation, was associated with a 4% increased risk of infection complications. The limit cubic spline map showed that lactic acid level at 24 h after operation was 4.22 mmol/L as the critical point of the dose-response relationship in the infection complications of patients with upper gastrointestinal ulcer perforation. Log-binomial model analysis results showed that when lactic acid level ≥4.22 mmol/L, patients with upper gastrointestinal ulcer perforation in the complication group had the highest risk of abdominal infection. The risk ratios (95%CI) before and after adjustment were 2.09 (1.25, 2.64), 2.16 (1.28, 2.05) and 2.20 (1.32, 2.63) times of those in the uncomplicated group, and the risk ratios of different lactic acid levels were statistically significant before and after adjustment (P<0.05). ConclusionLactic acid level increased 24 h after operation in patients with upper gastrointestinal ulcer perforation is closely related to infection complications, and has a high predictive value for infection complications in patients with upper gastrointestinal ulcer perforation.
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.
In order to find an ideal biological material to prevent peridural adhesion following laminectomy, 30 rabbits were used as animal model, in each of which 2 defects with a size of 1 cm x 0.5 cm were made following laminectomy of L3, L5 spine. One of the defects was covered extradurally with chitosan, gelatin foam or PLA membrane respectively, while the other defect was exposed as control. All of these animals were sacrificed on the 2nd, 4th, 6th, 8th and 10th week after operation, and the extradural fibrosis and adhesion of every animal were evaluated by gross observation and histological examinations. It was revealed that in the chitosan and PLA membrane groups, the extradural tissue was smooth without thickening and there was no fibrous proliferation or adhesion in the epidural cavity, and that in the chitosan group, the growth of fibroblast was restrained but the growth of the epithelial cells was promoted significantly, thus, wound healing was rapid. In the control group and gelatin foam group, obvious extradural fibrosis and adhesion were observed and the extradural space had almost disappeared. Therefore, it was concluded that the biodegradable PLA membrane and chitosan were both an ideal material in the prevention of postoperative epidural adhesion.
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.
The poly-lactic acid membrane is characterized by its non-toxicity, non-carcinogenic and the property of degradation in the body in animal experiment, this membrane was wrapped around the tenorrhaphy site. The results showed that 3 to 9 weeks after operation there was an intact space surrounding the tendon, and the membrane began to degradate at 24 weeks, might achieve the purpose to prevent the formation of adhesion. From Nov. 1985 to Sept. 1989, this membrane was applied to 20 tendons of 14 patients. The follow-up results showed that the satisfactory rate was 81.8% .the indications of using poly-lacic acid membrane were primary, delayed primary and secondary suture of flexor tendon injuries in areas of the fingers.
Objective To establish an animal model for repairing the sciatic nerve defect with a biodegradable poly D,L-lactic acid/nerve growth factor (PDLLA/NGF) that can control the release conduit in rats and to observe an effect of the conduit on the sciatic nerve regeneration. Methods The PDLLA conduit and the PDLLA/NGF-controlled release conduit (NGF 450 U per conduit) were madewith the solvent-volatilixation method. Forty male SD rats were randomly and equally divided into 4 groups. The middle segments (10 mm) of the sciatic nerves of the rats were excised and were then repaired with the sciatic nerve autograft(Group A), with the PDLLA conduit (Group B), with the PDLLA conduit and an injection of NGF (30 U) into the conduit (Group C), and with the PDLLA/NGF controlled-release conduit (Group D), respectively, with the 10-mm nerve defect left behind. Three months after operation, the morphologic parameters of the nerve regeneration were observed and evaluated under light microscope and electron microscope, and the image analysis was also made. Results Three months after operation, porous adherence between the conduit and the surrounding tissues could be observed. The conduit presented a partial biodegradation but still remainedintact in the outline and the proximal nerve regenerated through the conduit cavity. Based on the histological observation, the quantity, uniformity, and maturity of the nerve fiber regeneration in Groups A and D were better than those in Groups B and C. The image analysis indicated that there were no significant differences in the nerve fiber diameter, axon diameter or myelin thickness between Group A and Group D (P＞0.05). However, all the parameters in Groups A and D were better than those in Groups B and C (P＜0.05). Conclusion The PDLLA/NGF-controlled release conduit can effectively promote the sciatic nerve regeneration of rats. Its morphological index is similar to that of the nerve autograft.
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 explore the effect of NaOH on the surface morphology of three-dimensional (3D) printed poly-L-lactic acid (PLLA) mesh scaffolds. Methods The 3D printed PLLA mesh scaffolds were prepared by fused deposition molding technology, then the scaffold surfaces were etched with the NaOH solution. The concentrations of NaOH solution were 0.01, 0.1, 0.5, 1.0, and 3.0 mol/L, and the treatment time was 1, 3, 6, 9, and 12 hours, respectively. There were a total of 25 concentration and time combinations. After treatment, the microstructure, energy spectrum, roughness, hydrophilicity, compressive strength, as well as cell adhesion and proliferation of the scaffolds were observed. The untreated scaffolds were used as a normal control. Results 3D printed PLLA mesh scaffolds were successfully prepared by using fused deposition molding technology. After NaOH etching treatment, a rough or micro porous structure was constructed on the surface of the scaffold, and with the increase of NaOH concentration and treatment time, the size and density of the pores increased. The characterization of the scaffolds by energy dispersive spectroscopy showed that the crystal contains two elements, Na and O. The surface roughness of NaOH treated scaffolds significantly increased (P<0.05) and the contact angle significantly decreased (P<0.05) compared to untreated scaffolds. There was no significant difference in compressive strength between the untreated scaffolds and treated scaffolds under conditions of 0.1 mol/L/12 h and 1.0 mol/L/3 h (P>0.05), while the compression strength of the other treated scaffolds were significantly lower than that of the untreated scaffolds (P<0.05). After co-culturing the cells with the scaffold, NaOH treatment resulted in an increase in the number of cells on the surface of the scaffold and the spreading area of individual cells, and more synapses extending from adherent cells. Conclusion NaOH treatment is beneficial for increasing the surface hydrophilicity and cell adhesion of 3D printed PLLA mesh scaffolds.