Objective To give a prel iminary experimental evidence and to prove chitosan and allogeneic morsel ized bone as potential bone substitutions in repairing rabbit radius segmental defect. Methods Chitosan and allogeneic morsel ized bone were mixed with various ratios (1 ∶ 5, 1 ∶ 10, 1 ∶ 25, 1 ∶ 50, and 1 ∶ 100). After preparation, the physicaland chemical properties of the composites were prel iminary detected; the composites at the ratios of 1 ∶ 50 and 1 ∶ 25 had good physical and chemical properties and were used for the animal experiment. The radius segmental defects of 15 mm in length were made in 50 adult New Zealand white rabbits (weighing 2.5-3.0 kg), then the animals were divided into 2 groups. In groups A and B, chitosan/allogeneic morsel ized bone composites were implanted at the ratio of 1 ∶ 50 and 1 ∶ 25, respectively. After 1, 2, 4, 8, and 12 weeks of operation, the gross, histological, immunohistochemical observations were performed. Before the rabbits were sacrified, X-ray films were taken; the serum calcium and alkal ine phosphatase (ALP) concentration were measured; and the biomechanical measurement was carried out at 12 weeks. Results The results of gross observation were essentially consistent with those of the X-ray films. The histological observation showed that the bone formation was earl ier in group A than in group B; the amount of new bone formation in group A was more than that in group B; and the bone forming area in group A was bigger than that in group B (P lt; 0.05) at 4 and 8 weeks after operation. The immunohistochemical staining showed that vascular endothel ial growth factor and insul in-l ike growth factor receptor II proteins expressed in the cytoplasm of 2 groups after 4 and 8 weeks, and the expression in group A was higher than that in group B (P lt; 0.05). There was no significant difference in the serum calcium concentration between 2 groups at each time point (P gt; 0.05). After 4 and 8 weeks, the ALP concentration in group A was significantly higher than that in group B (P lt; 0.05). After 12 weeks, the radius maximum bending loads of groups A and B were (299.75 ± 27.69) N and (278.54 ± 17.09) N, respectively, showing significant difference (t=4.045,P=0.002). Conclusion The composite of chitosan and allogeneic morsel ized bone has good osteogeneic activity and can beused as a bone tissue engineering scaffold, and the optimum ratio of chitosan to allogeneic morsel ized bone was 1 ∶ 50.
Objective To evaluate the effect of implantation of the complex of high viscous chitosan/glycerol phosphate with demineral ized bone matrix (HV-C/GP-DBM) in repairing cartilage defects of rabbits. Methods HV-C/ GPDBM was prepared by compounding HV-C/GP and DBM by 2:1 (W/W). Twenty-four 34-week-old New Zealand white adult rabbits, weighing 3.5-4.5 kg, were included. A bit with the diameter of 3.5 mm was used to drill 3-cm-deep holes in both sides of femoral condyle to make cartilage defects. The complex of HV-C/GP-DBM was then injected into the right holes as the experimental group and the left ones serve as the control group. The rabbits were killed at 4, 8 and 16 weeks after theoperation, respectively. The obtained specimens were observed macroscopically, microscopically and histologically. According to the International Cartilage Repair Society Histological Scoring (ICRS), the effect of cartilage repair was assessed at 16 weeks postoperatively. Results At 4-8 weeks postoperatively, in the experimental group, the defects were filled with hyal ine cartilage-l ike tissues; the majority of chitosan degradated; and the DBM particles were partly absorbed. However, in the control group, there were small quantities of discontinuous fibrous tissues and maldistributed chondrocytes at the border and the bottom of the defects. At 16 weeks postoperatively, 6 joints in the experimental group had smooth surface, and the defects were basically repaired by hyal ine cartilage-l ike tissues. The newly-formed tissues integrated well with the surrounding area. Under the cartilage, the new bone formation was still active and some DBM particles could be seen. However, the defects in the control group were repaired by fibrous tissues. The result of histological scoring of the specimens at 16 weeks showed that a total of 6 aspects including formation of chondrocytes and integration with the surrounding cartilages were superior in the experimental group to those in the control group, and there were significant differences between the two groups (P lt; 0.05). Conclusion The biodegradable and injectable complex of HV-C/GP-DBM with good histocompatibil ity and non-toxic side effects can repair cartilage defects and is a promising biomaterial for cartilage defect repair.
Objective To compare the effect of the composite skin graft consisting of spl it-thickness skin grafts (STSGs) and porcine acellular dermal matrix (PADM) with STSGs only, and to histologically observe the turnover of the PADM in rats. Methods Twenty female Sprague-Dawley rats, weighing 200-225 g, were included. The size of 4.0 cm × 2.5 cm PADM was implanted into hypoderm of the left side of Sprague-Dawley rats’ back. After 10-14 days, the size of 4.0 cm × 2.5 cm full-thickness skin defects were made on the left to expose the PADM under the skin and the same size of full-thickness skin defects were made on the right of the rats’ back. The excised full-thickness skin was made to STSGs about 0.2 mm by drum dermatome. The defects were grafted with composite skin (STSGs on the PADM, experimental group) and STSGs only (control group). The survival rate, the constraction degree of grafts, and the histological change in grafts area were observed at 2, 4, 8, and 20 weeks after operation. Results At 2 weeks after STSGs (0.2 mm) placed on vascularized PADM, STSGs and PADM adhered together and the composite skin had a good survival. The control group also had a good survival. Histological observations showed that STSGs and PADM grew together, neutrophil ic granulocytes and lymphocytes infiltrated in the PADM and some macrophages around the PADM. Fibrous connective tissues were filled under the STSGs in control group. At 4-8 weeks after transplantation, the composite skin had a good survival and the composite skin was thick, soft, and elastic. STSGs survived almost totally in control group, but the grafts were thin. Histological observations showed that inflammatory reactions of PADM faded gradually in experimental group; scar tissues formed under the STSGs in control group. At 20 weeks after transplantation, composite skin was flat, thick, and elastic in experimental group, but the STSGs were thinner and less elastic in control group. Histological observations showed that histological structures of the PADM were similar to the dermal matrix of rats, and the results showed that the collagen matrix of PADM was gradually replaced by the rats’ collagen matrix. Scar tissues were filled under the STSGs in control group. Wound heal ing rates of experimental group were lower than those of control group at 4 and 8 weeks (P﹤0.05); wound contraction rates of experimental group had lower tendency than those of control group, but showing no significant differences (P gt; 0.05). Conclusion Coverage wound with composite skin which composed of STSGs and PADM could improve wound heal ing qual ity; the composite skin is thicker and better elastic than STSGs only. The collagen matrix of PADM is gradually replaced by rats’ collagen matrix.