Yoshikawa T, Ohgushi H, Nakajima H, Yamada E, Ichijima K, Tamai S, Ohta T
First Department of Pathology, Nara Medical University, Kashihara City, Japan.
Transplantation. 2000 Jan 15;69(1):128-34. doi: 10.1097/00007890-200001150-00022.
Bone marrow cells differentiate into bone-forming osteoblasts when cultured in medium supplemented with 15% fetal bovine serum, ascorbic acid, beta-glycerophosphate, and dexamethasone.
To investigate in vivo osteoblastic activity and bone matrix formation by cultured bone marrow cells, Fischer rat marrow cells were cultured for 2 weeks in porous hydroxyapatite (HA) and then subcutaneously implanted into 7-week-old male syngeneic rats. The implants were harvested after 8 and 52 weeks for biochemical and histological analyses.
At both times, formation of lamellar bone accompanied by regeneration of marrow were seen in many of the HA pores. When a fluorochrome (calcein) was administered at 50 weeks after implantation, it was detected in the pores of implants harvested at 52 weeks. Osteoclastic resorption followed by new bone formation was seen in some pores at 52 weeks, indicating that bone remodeling was continuing. The alkaline phosphatase activity of implants harvested at 52 weeks was comparable to that at 8 weeks, whereas the osteocalcin content of the implants harvested at 52 weeks was about twice that at 8 weeks.
These results demonstrated that there was persistent in vivo osteogenic and hematopoietic activity in the prefabricated bone/HA constructs, and indicated that normal bone tissue was regenerated after grafting of the constructs, which were brittle before implantation. Tissue engineering using HA and cultured marrow cells culture may provide an alternative method of bone transplantation for patients with skeletal disorders, although further in vivo and in vitro experiments are needed.
当在补充有15%胎牛血清、抗坏血酸、β-甘油磷酸酯和地塞米松的培养基中培养时，骨髓细胞可分化为形成骨的成骨细胞。
为了研究培养的骨髓细胞在体内的成骨活性和骨基质形成，将Fischer大鼠骨髓细胞在多孔羟基磷灰石（HA）中培养2周，然后皮下植入7周龄的同基因雄性大鼠体内。在8周和52周后取出植入物进行生化和组织学分析。
在两个时间点，许多HA孔隙中均可见板层骨形成并伴有骨髓再生。在植入后50周给予荧光染料（钙黄绿素），在52周取出的植入物孔隙中可检测到该染料。在52周时，一些孔隙中可见破骨细胞吸收后有新骨形成，表明骨重塑在持续进行。52周取出的植入物的碱性磷酸酶活性与8周时相当，而52周取出的植入物的骨钙素含量约为8周时的两倍。
这些结果表明，预制骨/HA构建体在体内具有持续的成骨和造血活性，表明构建体植入后可再生正常骨组织，而构建体在植入前是脆性的。使用HA和培养的骨髓细胞培养进行组织工程可能为骨骼疾病患者提供一种替代的骨移植方法，尽管还需要进一步的体内和体外实验。
