Preparation of bio-derived bone and its histocompatibility
- VernacularTitle:生物衍生骨支架材料制备及在体内的组织相容性
- Author:
Yun NIU
;
Xu HE
;
Lihong ZHANG
;
Ting GAO
;
Hua XU
;
Yulin LI
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2008;12(7):1385-1389
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: The ideal biomaterial means absence of cytotoxic effect and immunological rejection, degradation at right moment, and a well histocompatibility. Whether bio-derived bone can be used in vivo for long time and exerts functions deserves to be studied.OBJECTIVE: To investigate the local histocompatibility after bio-derived bone implanted into mouse and the effect on immunofunctions.DESIGN: A randomized controlled animal experiment. SETTING: Key Laboratory of Pathobiology, Ministry of Education, School of Basic Medical Sciences, Jilin University. MATERIALS: This study was performed at the Key Laboratory of Pathobiology, Ministry of Science, School of Basic Medical Sciences, Jilin University from March to July in 2006. Eighteen BALB/C mice (weighing 20±2 g, half male and half female), one male Kunming mouse (weighing 20 g), and one female rabbit (weighing 2.5 kg) were included in the experiment. All the experimental animals were provided by Laboratory Animal Center, School of Basic Medical Science, Jilin University. The disposal of the experimental animals in the test process accorded with ethical guidelines for the use and care of animals. Porcine cancellous bone (iliac bone) was purchased from the market. Iscove's Modified Dulbecco's Medium (IMDM, Hycolone, USA), fetal bovine serum (FBS, Gibco Co., Ltd, USA), methyl thiazolyl tetrazolium (MTT, Sigma, USA), and concanavalin A (ConA, Sigma Co., Ltd, USA) were used.METHODS: Bio-derived bone was prepared from commercial porcine bone. ① Eighteen BALB/C mice were randomly divided into three groups with 6 mice in each group: a control group (simple local muscle injury without implantation), a bio-derived bone implantation group ( implanting bio-derived bone into the lower limb), and a xenogenic bone implantation group (femoral bone from Kunming mouse was implanted into the muscle of lower limb). ② Twenty-one days after operation, the implant material and surrounding tissue were obtained for gross observation and haematoxylin-eosin staining to investigate the histocompatibility of bio-derived bone. Mouse immunofunction was assessed by complement-mediated cytotoxicity test. Absorbance was determined with an automatic ELISA reader at 570 nm to assess the cytotoxicity.MAIN OUTCOME MEASURES: ①Histocompatibility of implant surrounded tissue. ②Lymphocyte stimulation indices after induction of concanavalin A. ③ Cytotoxicity in each group after complement-dependent cytotoxicity test.RESULTS: Eighteen BALB/C mice were included in the final analysis. ①Histocompatibility of implant surrounded tissue: In the bio-derived bone implantation group, 21 days after bio-derived bone implantation, there were no presentation of congestion, degeneration, necrosis and diapyesis around the implant in gross, plenty of fibrous connective tissue invaded into the pores of the bio-derived bone, encapsulation and forming the fibrous capsule. A great quantity of neutrophils and macrophages were not detected around the implant by haematoxylin-eosin staining. Bio-derived bone was encapsulated with fibrous tissue, and part of the biomaterial began to degrade, and being replaced with fibrous tissue. Regarding xenogenic bone implantation group, necrotic tissue was detected in the cross-section of the muscle in gross. A lot of neutrophils, macrophages and necrotic tissue were detected around the implant by haematoxylin-eosin staining. ②Lymphocyte stimulation indices: The stimulation index of xenogenic bone implantation group was significantly larger than that of control group (P ＜ 0.05). There was no statistical difference between the bio-derived bone group and the control group (P ＞ 0.05). ③Cytotoxicity: The cytotoxicity of xenogenic bone implantaion group was significantly larger than that of control group (P ＜ 0.05). There was no significant difference in cytotoxicity between the bio-derived bone implantation group and the control group (P ＞ 0.05).CONCLUSION: The obtained bio-derived bone causes little immunoreactions, has no obvious cytotoxicity or inflammatory reactions, and possesses a good histocompatibility and bio-safety.
