Constructing tissue-engineered growth plate by allograft demineralized bone matrix cocultured with rabbit iliac growth-plate cells / 中国组织工程研究

ABSTRACT

AIM:Long bone growth plate injury induced by wound and infection may cause limb reduction or angular deformity. Tissue engineering provides a promising treatment of growth plate injuries. In this study,we investigated the feasibility of establishing tissue-engineered growth-plate by allograft demineralized bone matrix(DBM) co-cultured with rabbit iliac growth-plate cells. METHODS:The experiment was performed at Department of Orthopaedics,Fourth Military Medical University of Chinese PLA from June 2005 to June 2006. ①Two 3-week-old New Zealand rabbits irrespective of gender(clean grade,2.0-2.5 kg) were selected. Growth plate cells were harvested from iliac crest epiphyseal cartilage of the rabbits by dissection and digestion with type Ⅱ collagenase. The third passage cells cultured were collected and incubated on allograft demineralized bone matrix. ② Histology,immunohistochemical staining and electronic scanning microscope(SEM) examinations were performed to observe cell growth on DBM 24 hours,7,14 and 21 days after culture. RESULTS:①Growth plate chondrocytes exhibited polygonal in monolayer culture,and immunohistochemical staining for type Ⅱ collagen was positive. ②SEM examination showed that twenty-four hours after coculture,the cells adhered to DBM scaffolds;Seven days after culture,the growth-plate chondrocytes rapidly proliferated and began to secret extracellular matrix;cells covered the whole scaffold and became overlapped on the 21st day. ③HE staining showed after 14 days of culture,growth plate cells adhered DBM scaffold in spherical shape with abundant cytoplasm. CONCLUSION:Tissue-engineered growth-plate is successfully constructed by allograft mineralized bone matrix co-cultured with rabbit iliac growth-plate cells.